WO2010141294A1

WO2010141294A1 - Insecticidal target genes identified from bug whole body and digestive system transcriptomes

Info

Publication number: WO2010141294A1
Application number: PCT/US2010/036275
Authority: WO
Inventors: R. Michael Roe; Kevin V. Donohue; Jacob B. Van Kretschmar; Leonardo C. Magalhaes
Original assignee: North Carolina State University
Priority date: 2009-06-01
Filing date: 2010-05-27
Publication date: 2010-12-09

Abstract

A transgenic plant such as a cotton plant is described. The plant contains a polynucleotide construct that comprises a heterologous nucleotide sequence operably linked to a promoter that drives expression in the plant. The heterologous nucleotide sequence encodes an active agent toxic to an insect pest such as a plant bug, leaf bug, grass bug, shield bug or stink bug. The active agent may be: (i) an RNA (e.g., a dsRNA) that inhibits expression of a target gene in said insect pest, and reduces or inhibits the expression of the expression product (e.g., protein, ribosomal RNA) (ii) an antibody that binds to the expression product of said target gene and inhibits the activity thereof, or (iii) an aptamer that binds to the expression product of said target gene and inhibits the activity thereof. Insecticidal compositions and bait compositions containing such active ingredients are also described.

Description

INSECTICIDAL TARGET GENES IDENTIFIED FROM BUG WHOLE BODY AND DIGESTIVE SYSTEM TRANSCRIPTOMES

R. Michael Roe, Kevin V. Donohue, Jacob B. van Kretschmar, and Leonardo C. Magalhaes

Field of the Invention

The present invention concerns transgenic plants such as transgenic cotton, and methods of controlling insect pests such as plant bugs and stink bugs.

Background of the Invention

Since the mid 1990s, transgenic Bt cotton has been widely expanded and highly efficient in controlling lepidopteran pests. For this reason — and the success of the boll weevil (Anthonomus grandis (Boheman)) eradication program — chemical insecticide use is reduced considerably. As a result, a new group of pests has recently emerged in cotton. Piercing-sucking insects, such as plant bugs and stink bugs, which were indirectly being controlled have now become important cotton pests in the US.

Among other injuries, Lygus lineolaris impact cotton by releasing salivary toxins that causes abortion and malformation of the attacked part, usually flowers. Losses can exceed $40 million depending on the region and time of infestation. Stink bugs damage cotton mainly later in the season by feeding on developing bolls, which reduces yield and fiber quality. Total cotton losses in 2004 related to stink bug damage were estimated around $10 million. Insecticide applications and yield losses in cotton production due to plant and stink bugs have been increasing sharply, and tarnished plant bug resistance to both pyrethroid and organophosphate insecticides is now widespread.

The absence of a transgenic approach using Bt toxins to control piercing-sucking insects is problematic for the sustainability of crop production. Also, even if a protein toxin was available to control piercing-sucking insects, there are important advantages to a nucleic acid approach for insect control including the significant ease with this approach of preventing target site insect resistance not possible with protein insecticides. Hence, there is a need to identify novel insecticide modes of action for use in developing nucleic acid-based, transgenic plant technology for insect control.

Summary of the Invention

A first aspect of the invention is a transgenic plant that contains a polynucleotide construct that comprises a heterologous nucleotide sequence operably linked to a promoter that drives expression in the plant, the heterologous nucleotide sequence encoding an active agent toxic to an insect pest; the active agent selected from the group consisting of: (i) an RNA that inhibits expression of a target gene in the insect pest, and reduces or inhibits the expression of the expression product (e.g., protein, ribosomal RNA) (H) an antibody that binds to the expression product of the target gene and inhibits the activity thereof, and (Ui) an aptamer that binds to the expression product of the target gene and inhibits the activity thereof.

In some embodiments of the foreoging, the insect pest is a member of the Heteroptera suborder of the Hemiptera order.

In some embodiments of the foreoging, the insect pest is a member of the Miridae family.

In some embodiments of the foreoging, the insect pest is a plant bug, leaf bug, or grass bug.

In some embodiments of the foreoging, the insect pest is a member of the Pentatomoidea superfamily.

In some embodiments of the foreoging, the insect pest is a shield bug or stink bug.

In some embodiments of the foreoging, the plant is a monocot; In some embodiments of the foreoging, wherein the plant is a dicot.

In some embodiments of the foreoging, the plant is cotton.

In some embodiments of the foreoging, the dsRNA is a double-stranded molecule with a first strand consisting essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and a second strand consisting essentially of a ribonucleotide sequence which is complementary to the nucleotide sequence of the target gene, wherein the first and the second ribonucleotide strands are separate complementary strands (e.g., on the same molecule or on separate molecules) that hybridize to each other to form the double-stranded molecule, and the double-stranded molecule inhibits expression of the target gene. In some embodiments of the foreoging, the plant further comprises:

(a) plant cells containing nucleotide sequences encoding immunoglobulin heavy- and light-chain polypeptides, wherein each polypeptide contains a leader sequence forming a secretion signal; and

(b) immunoglobulin molecules encoded by the nucleotide sequences, wherein the leader sequence is cleaved from the immunoglobulin molecules following proteolytic processing.

In some embodiments of the foreoging, the plant further comprises:

(a) plant cells containing nucleotide sequences encoding a biologically functional multimeric protein comprising at least two different polypeptides not naturally produced by plants, wherein each nucleotide sequence encoding a polypeptide of the multimeric protein encodes a leader sequence forming a secretion signal that is cleaved from the polypeptide following proteolytic processing; and

(b) biologically functional multimeric protein encoded by the nucleotide sequences and resulting from assembly of the at least two different polypeptides wherein the multimeric protein is an abzyme or immunoglobulin.

A further aspect of the invention is a transgenic seed of the plant of described above, wherein the seed comprises the heterologous polynucleotide.

A further aspect of the invention is a method for protecting a plant from an insect pest, the method comprising: introducing into the plant at least one polynucleotide construct that comprises a heterologous nucleotide sequence operably linked to a promoter that drives expression in the plant, wherein the nucleotide sequence encodes an active agent toxic to an insect pest; and wherein the plant is a plant as described above.

A further aspect of the invention is a method for protecting a plant from an insect pest, the method comprising administering to the insect pest an active agent selected from the group consisting of: (i) an RNA that inhibits expression of a target gene in the insect pest, and reduces or inhibits the expression of the expression product {e.g., protein, ribosomal RNA), (H) an antibody that binds to the expression product of the target gene and inhibits the activity thereof, and (Hi) an aptamer that binds to the expression product of the target gene and inhibits the activity thereof.

In some embodiments of the foregoing, the administering step is carried out by applying the active agent to the plant. - A -

In some embodiments of the foregoing, the applying step is carried out by applying an insecticidal bait composition to the plant, the insecticidal bait composition comprising the active agent.

A further aspect of the invention is an insecticidal composition comprising a carrier media and an active agent, wherein the active agent comprises a compound selected from the group consisting of: (i) an RNA that inhibits expression of a target gene in the insect pest, and reduces or inhibits the expression of the expression product (e.g., protein, ribosomal RNA), (H) an antibody that binds to the expression product of the target gene and inhibits the activity thereof, and (Hi) an aptamer that binds to the expression product of the target gene and inhibits the activity thereof.

A further aspect of the invention is, in an insecticidal bait composition comprising an insecticide and one or more of insect food, insect atttractant, carrier, and an insecticidal active agent, the improvement comprising employing as the active agent a compound selected from the group consisting of: (i) an RNA that inhibits expression of a target gene in the insect pest, and reduces or inhibits the expression of the expression product (e.g., protein, ribosomal RNA), (H) an antibody that binds to the expression product of the target gene and inhibits the activity thereof, and (Hi) an aptamer that binds to the expression product of the target gene and inhibits the activity thereof.

In some embodiments of the foreoging, the target gene encodes a protein of the insect pest selected from the group consisting of: transcription factors, translation factors, microvilla proteins (e.g., mucin, pseudomucin), electron transport proteins, Bacillus thuringiensis toxin (Cry toxin) receptors, insecticide target proteins (e.g., acetylcholine esterase, GABA-gated chloride channels, sodium channels, nicotinic acetylcholine receptors, chloride channels, midgut membrane proteins, mitochondrial ATP synthase, citin biosynthesis enzymes, ecdysone receptors, mitochondrial complex I, III, and IV electron transporters, Mitochondrial complex I electron transporters, voltage-dependent sodium channels, acetyl CoA carboxylase, etc.), and ribosomal RNA.

In some embodiments of the foreoging, the target gene is given in TABLE 1 herein.

The present invention is explained in great detail in the specification set forth below. The disclosures of all United States Patent references cited herein are to be incorporated by reference herein in their entirety. Brief Description of the Drawings Fig. 1. Percentage of ESTs matching putative functions listed.

Detailed Description of the Preferred Embodiments A. DEFINITIONS.

"Plant" as used herein includes both gymnosperms and angiosperms, and includes both monocots and dicots.

Dicotyledon (dicot): A flowering plant whose embryos have two seed halves or cotyledons. Examples of dicots are: tobacco; tomato; the legumes including alfalfa; oaks; maples; roses; mints; squashes; daisies; walnuts; cacti; violets; and buttercups.

Monocotyledon (monocot): A flowering plant whose embryos have one cotyledon or seed leaf. Examples of monocots are: lilies; grasses; corn; grains, including oats, wheat and barley; orchids; irises; onions and palms.

Eukaryotic hybrid vector: A DNA by means of which a DNA coding for a polypeptide (insert) can be introduced into a eukaryotic cell.

Extrachromosomal ribosomal DNA (rDNA): A DNA found in unicellular eukaryotes outside the chromosomes, carrying one or more genes coding for ribosomal RNA and replicating autonomously (independent of the replication of the chromosomes).

Palindromic DNA: A DNA sequence with one or more centers of symmetry.

DNA: Deoxyribonecleic acid.

T-DNA: A segment of transferred DNA. rDNA: Ribosomal DNA.

RNA: Ribonucleic acid. rRNA: Ribosomal RNA.

Ti-plasmid: Tumor-inducing plasmid.

Ti-DNA: A segment of DNA from Ti-plasmid.

Insert: A DNA sequence foreign to the rDNA, consisting of a structural gene and optionally additional DNA sequences.

Structural gene: A gene coding for a polypeptide and being equipped with a suitable promoter, termination sequence and optionally other regulatory DNA sequences, and having a correct reading frame.

Signal Sequence: A DNA sequence coding for an amino acid sequence attached to the polypeptide which binds the polypeptide to the endoplasmic reticulum and is essential for protein secretion. This sequence may also be referred to herein as a secretion signal or secretion signal sequence. The term "signal sequence" may also be used to refer to the sequence of amino acids that determines whether a protein will be formed on the rough endoplasmic reticulum or on free ribosomes. And while a "leader sequence" generally means a sequence near the 5' end of a nucleic acid strand or the amino terminus of a protein that functions in targeting or regulation, the term is sometimes used herein to include a "secretion signal" or a "signal sequence".

(Selective) Genetic marker. A DNA sequence coding for a phenotypic trait by means of which transformed cells can be selected from untransformed cells.

Promoter: A recognition site on a DNA sequence or group of DNA sequences that provide an expression control element for a gene and to which RNA polymerase specifically binds and initiates RNA synthesis (transcription) of that gene.

Inducible promoter: A promoter where the rate of RNA polymerase binding and initiation is modulated by external stimuli. Such stimuli include light, heat, anaerobic stress, alteration in nutrient conditions, presence or absence of a metabolite, presence of a ligand, microbial attack, wounding and the like.

Viral promoter: A promoter with a DNA sequence substantially similar to the promoter found at the 5' end of a viral gene. A typical viral promoter is found at the 5' end of the gene coding for the p21 protein of MMTV described by Huang et al., Cell 27: 245 (1981).

Synthetic promoter: A promoter that was chemically synthesized rather than biologically derived. Usually synthetic promoters incorporate sequence changes that optimize the efficiency of RNA polymerase initiation.

Constitutive promoter: A promoter where the rate of RNA polymerase binding and initiation is approximately constant and relatively independent of external stimuli. Examples of constitutive promoters include the cauliflower mosaic virus 35S and 19S promoters described by Poszkowski et al., EMBO J. 3: 2719 (1989) and Odell et al., Nature 313: 810 (1985).

Temporally regulated promoter: A promoter where the rate of RNA polymerase binding and initiation is modulated at a specific time during development. Examples of temporally regulated promoters are given in Chua et al., Science 244: 174 181 (1989).

Spatially regulated promoter: A promoter where the rate of RNA polymerase binding and initiation is modulated in a specific structure of the organism such as the leaf, stem or root Examples of spatially regulated promoters are given in Chua et al, Science 244: 174 181 (1989).

Spatiotemporally regulated promoter: A promoter where the rate of RNA polymerase binding and initiation is modulated in a specific structure of the organism at a specific time during development. A typical spatiotemporally regulated promoter is the EPSP synthase-35S promoter described by Chua et al., Id. (1989).

Single-chain antigen-binding protein: A polypeptide composed of an immunoglobulin light-chain variable region amino acid sequence (V_L) tethered to an immunoglobulin heavy- chain variable region amino acid sequence (V_H) by a peptide that links the carboxyl terminus of the V_L sequence to the amino terminus of the V_H sequence.

Single-chain antigen-binding protein-coding gene: A recombinant gene coding for a single-chain antigen-binding protein.

Multimeric protein: A globular protein containing more than one separate polypeptide or protein chain associated with each other to form a single globular protein. Both heterodimeric and homodimeric proteins are multimeric proteins.

Polypeptide and peptide: A linear series of amino acid residues connected one to the other by peptide bonds between the alpha-amino and carboxy groups of adjacent residues.

Protein: A linear series of greater than about 50 amino acid residues connected one to the other as in a polypeptide.

Immunoglobulin product: A polypeptide, protein or multimeric protein containing at least the immunologically active portion of an immunoglobulin heavy chain and is thus capable of specifically combining with an antigen. Exemplary immunoglobulin products are an immunoglobulin heavy chain, immunoglobulin molecules, substantially intact immunoglobulin molecules, any portion of an immunoglobulin that contains the paratope, including those portions known in the art as Fab fragments, Fab' fragment, F(ab')₂ fragment and Fv fragment.

Immunoglobulin molecule: A multimeric protein containing the immunologically active portions of an immunoglobulin heavy chain and immunoglobulin light chain covalently coupled together and capable of specifically combining with antigen.

Fab fragment: A multimeric protein consisting of the portion of an immunoglobulin molecule containing the immunologically active portions of an immunoglobulin heavy chain and an immunoglobulin light chain covalently coupled together and capable of specifically combining with antigen. Fab fragments are typically prepared by proteolytic digestion of substantially intact immunoglobulin molecules with papain using methods that are well known in the art. However, a Fab fragment may also be prepared by expressing in a suitable host cell the desired portions of immunoglobulin heavy chain and immunoglobulin light chain using methods well known in the art.

F_v fragment: A multimeric protein consisting of the immunologically active portions of an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region covalently coupled together and capable of specifically combining with antigen. Fv fragments are typically prepared by expressing in suitable host cell the desired portions of immunoglobulin heavy chain variable region and immunoglobulin light chain variable region using methods well known in the art.

Asexual propagation: Producing progeny by regenerating an entire plant from leaf cuttings, stem cuttings, root cuttings, single plant cells (protoplasts) and callus.

Catalytic site: The portion of a molecule that is capable of binding a reactant and improving the rate of a reaction. Catalytic sites may be present on polypeptides or proteins, enzymes, organics, organo-metal compounds, metals and the lice. A catalytic site may be made up of separate portions present on one or more polypeptide chains or compounds. These separate catalytic portions associate together to form a larger portion of a catalytic site. A catalytic site may be formed by a polypeptide or protein that is bonded to a metal.

Enzymatic site: The portion of a protein molecule that contains a catalytic site. Most enzymatic sites exhibit a very high selective substrate specificity. An enzymatic site may be comprised of two or more enzymatic site portions present on different segments of the same polypeptide chain. These enzymatic site portions are associated together to form a greater portion of an enzymatic site. A portion of an enzymatic site may also be a metal.

Self-pollination: The transfer of pollen from male flower parts to female flower parts on the same plant. This process typically produces seed. Cross-pollination: The transfer of pollen from the male flower parts of one plant to the female flower parts of another plant. This process typically produces seed from which viable progeny can be grown.

Epitope: A portion of a molecule that is specifically recognized by an immunoglobulin product It is also referred to as the determinant or antigenic determinant.

Abzyme: An immunoglobulin molecule capable of acting as an enzyme or a catalyst.

Enzyme: A protein, polypeptide, peptide RNA molecule, or multimeric protein capable of accelerating or producing by catalytic action some change in a substrate for which it is often specific. "Aptamer" as used herein refers to nucleic acid molecules having specific binding affinity to molecules through interactions other than classic Watson-Crick base pairing. Aptamers, like peptides generated by phage display or monoclonal antibodies ("MAbs"), are capable of specifically binding to selected targets and modulating the target's activity, e.g., through binding aptamers may block their target's ability to function. Created by an in vitro selection process from pools of random sequence oligonucleotides, aptamers have been generated for over 100 proteins including growth factors, transcription factors, enzymes, immunoglobulins, and receptors. A typical aptamer is 10-15 kDa in size (30-45 nucleotides), binds its target with sub-nanomolar affinity, and discriminates against closely related targets (e.g., aptamers will typically not bind other proteins from the same gene family). A series of structural studies have shown that aptamers are capable of using the same types of binding interactions (e.g., hydrogen bonding, electrostatic complementarity, hydrophobic contacts, steric exclusion) that drive affinity and specificity in antibody-antigen complexes. See, e.g., US Patent No. 7,538,211.

"Administering" as used herein with respect to active agents can be carried out directly or indirectly by any suitable means or route, including but not limited to orally or topically administering the active agent, by orally or topically administering a microbial vector such as a virus that produces the active agent (e.g., a dsRNA), etc.

B. ANTIBODY ACTIVE AGENTS.

Plants containing antibody active agents of the present invention can be produced in accordance with known techniques, or variations thereof that will be apparent to those skilled in the art. See, e.g., US Patent No.s 7,037,722 and 6,417,429 to Hein and Hiatt; see also US Patent Nos. 7,101, 688; 7,005,560; and 5,959,177.

In general, such methods can be carried out by: 1. Inserting into the genome of a first member of a plant species a gene coding for a first polypeptide to produce a first transformant 2. Inserting into the genome of a second member of a plant species a gene coding for a second polypeptide to produce a second transformant 3. Producing a population of progeny from the first and second transformants. 4. Isolating from the population, a progeny having the multimeric protein. The multimeric protein is typically an immunoglobulin molecule comprised of an immunoglobulin heavy chain and an immunoglobulin light chain. The immunoglobulin heavy and light chains are associated with each other and assume a conformation having an antigen binding site specific for, as evidenced by its ability to be competitively inhibited, a preselected or predetermined antigen (that is, the product of the insect gene). When the multimeric protein is an antigen binding protein its affinity or avidity is generally greater than 10⁵ M^'1 or usually greater than 10⁶M^'1, and preferably greater than 10⁸M-¹.

In a further embodiment, the multimeric protein is a Fab fragment consisting of a portion of an immunoglobulin heavy chain and a portion of an immunoglobulin light chain. The immunoglobulin heavy and light chains are associated with each other and assume a conformation having an antigen binding site specific for a preselected or predetermined antigen. The antigen binding site on a Fab fragment has a binding affinity or avidity similar to the antigen binding site on an immunoglobulin molecule.

In yet another embodiment, the present transgenic plant contains a multimeric protein that is a F_v fragment comprised of at least a portion of an immunoglobulin heavy chain variable region and at least a portion of an immunoglobulin light chain variable region. The immunoglobulin heavy and light chain variable regions autogenously associate with each other within the plant cell to assume a biologically active conformation having a binding site specific for a preselected or predetermined antigen. The antigen binding site on the F_v fragment has an affinity or avidity for its antigen similar to the affinity displayed by the antigen binding site present on an immunoglobulin molecule.

In still another embodiment, the multimeric protein is an enzyme that binds a substrate and catalyzes the formation of a product from the substrate. While the topology of the substrate binding site (ligand binding site) of the catalytic multimeric protein is probably more important for its activity than affinity (association constant or pKa) for the substrate, the subject multimeric protein has an association constant for its preselected substrate greater than 10³.M⁴, more usually greater than 10⁵ M^"1 or 10⁶ M^"1 and preferably greater than 10⁷ M- i

When the multimeric protein produced in accordance with the present invention is an abzyme comprised of at least a portion of the immunoglobulin heavy chain variable region in association with another polypeptide chain, this other polypeptide chain includes at least the biologically active portion of an immunoglobulin light chain variable region. Together, these two polypeptides assume a conformation having a binding affinity or association constant for a preselected ligand that is different, preferably higher, than the affinity or association constant of either of the polypeptides alone, i.e., as monomers. Useful multimeric proteins contain one or both polypeptide chains derived from the variable region of the light and heavy chains of an immunoglobulin. Typically, polypeptides comprising the light (VL) and heavy (VH) variable regions are employed together for binding the preselected antigen.

1. Inserting Genes Coding for a First Polypeptide into a First Member of a Plant Species. Methods for isolating a gene coding for a desired first polypeptide are well known in the art. See, for example, Guide To Molecular Cloning Techniques in Methods In Enzymology. Volume 152, Berger and Kimmel, eds. (1987); and Current Protocols in Molecular Biology, Ausubel et al., eds., John Wiley and Sons, New York (1987) whose disclosures are herein incorporated by reference.

Genes useful in practicing this invention include genes coding for a polypeptide contained in immunoglobulin products, immunoglobulin molecules, Fab fragments, F_v fragments, enzymes, receptors and abzymes. Particularly preferred are genes coding for immunoglobulin heavy and light chain variable regions. Typically, the genes coding for the immunoglobulin heavy chain variable region and immunoglobulin light chain variable region of an immunoglobulin capable of binding a preselected antigen are used. These genes are isolated from cells obtained from a vertebrate, preferably a mammal which has been immunized with an antigenic ligand (antigen) against which activity is sought, i.e., a preselected antigen. The immunization can be carried out conventionally and antibody titer in the animal can be monitored to determine the stage of immunization desired, which corresponds to the affinity or avidity desired. Partially immunized animals typically receive only one immunization and cells are collected therefrom shortly after a response is detected. Fully immunized animals display a peak titer which is achieved with one or more repeated injections of the antigen into the host mammal, normally at two to three week intervals.

Usually three to five days after the last challenge, the spleen is removed and the genes coding for immunoglobulin heavy and immunoglobulin light chain are isolated from the rearranged B cells present in the spleen using standard procedures. See Current Protocols in Molecular Biology, Ausubel et al., eds., John Wiley and Sons, New York (1987) and Antibodies: A Laboratory Manual, Harlowe and Lane, eds., Cold Spring Harbor, N.Y. (1983).

Genes coding for VH and VL polypeptides can be derived from cells producing IgA, IgD, IgE, IgG or IgM, most preferably from IgM and IgG, producing cells. Methods for preparing fragments of genomic DNA from which immunoglobulin variable region genes can be cloned are well known in the art. See for example, Herrmann et al., Methods in Enzymol. 152: 180 183 (1987); Frischauf, Methods in Enzymol. 152: 183 190 (1987); Frischauf, Methods in Enzymol. 152: 199 212 (1987). (The teachings of the references cited herein are hereby incorporated by reference).

Probes useful for isolating the genes coding for immunoglobulin products include the sequences coding for the constant portion of the V_H and V_L sequences coding for the framework regions of V_H and V_L and probes for the constant region of the entire rearranged immunoglobulin gene, these sequences being obtainable from available sources. See for example, Early and Hood, Genetic Engineering, Setlow and Hollaender eds., Vol. 3: 157 138, Plenum Publishing Corporation, New York (1981); and Kabat et al., Sequences of Immunological Interests, National Institutes of Health, Bethesda, Md. (1987).

Genes coding for a polypeptide subunit of a multimeric protein can be isolated from either the genomic DNA containing the gene expressing the polypeptide or the messenger RNA (mRNA) which codes for the polypeptide. The difficulty in using genomic DNA is in juxtaposing the sequences coding for the polypeptide where the sequences are separated by introns. The DNA fragment(s) containing the proper exons must be isolated, the introns excised, and the exons spliced together in the proper order and orientation. For the most part, this will be difficult so the alternative technique employing mRNA will be the method of choice because the sequence is contiguous (free of introns) for the entire polypeptide. Methods for isolating mRNA coding for peptides or proteins are well known in the art See, for example, Current Protocols in Molecular Biology, Ausubel et al., John Wiley and Sons, New York (1987), Guide to Molecular Cloning Techniques, in Methods In Enzymology, Volume 152, Berger and Kimmel, eds. (1987), and Molecular Cloning: A Laboratory Manual, Maniatis et al., eds., Cold Spring Harbor Laboratory, Cold Spring Harbor, N. Y. (1982).

The polypeptide coding genes isolated above are typically operatively linked to an expression vector. Expression vectors compatible with the host cells, preferably those E 15 compatible with plant cells are used to express the genes of the present invention. Typical expression vectors useful for expression of genes in plants are well known in the art and include vectors derived from the tumor-inducing (Ti) plasmid of Agrobacterium tumefaciens described by Rogers et al., Meth. in Enzymol. 153: 253 277 (1987).

However, several other expression vector systems are known to function in plants. See for example, Verma et al., PCT Publication No. WO87/00551; and Cocking and Davey, Science, 236: 1259 1262 (1987). The expression vectors described above contain expression control elements including the promoter. The polypeptide coding genes are operatively linked to the expression vector to allow the promoter sequence to direct RNA polymerase binding and synthesis of the desired polypeptide coding gene. Useful in expressing the polypeptide coding gene are promoters which are inducible, viral, synthetic, constitutive, temporally regulated, spatially regulated, and spatiotemporally regulated. The choice of which expression vector and ultimately to which promoter a polypeptide coding gene is operatively linked depends directly, as is well known in the art, on the functional properties desired, e.g. the location and timing of protein expression, and the host cell to be transformed, these being limitations inherent in the art of constructing recombinant DNA molecules. However, an expression vector useful in practicing the present invention is at least capable of directing the replication, and preferably also the expression of the polypeptide coding gene included in the DNA segment to which it is operatively linked.

In preferred embodiments, the expression vector used to express the polypeptide coding gene includes a selection marker that is effective in a plant cell preferably a drug resistance selection marker. A preferred drug resistance marker is the gene whose expression results in kanamycin resistance, i.e., the chimeric gene containing the nopaline synthase promoter, Tn5 neomycin phosphotransferase II and nopaline synthase 3' nontranslated region described by Rogers et al, in Methods For Plant Molecular Biology, a Weissbach and H. Weissbach, eds., Academic Press Inc., San Diego, Calif. (1988). A useful plant expression vector is commercially available from Pharmacia, Piscataway, NJ.

A variety of methods have been developed to operatively link DNA to vectors via complementary cohesive termini. For instance, complementary homopolymer tracks can be added to the DNA segment to be inserted and to the vector DNA. The vector and DNA segment are then joined by hydrogen bonding between the complementary homopolymeric tails to form recombinant DNA molecules.

Alternatively, synthetic linkers containing one or more restriction endonuclease sites can be used to join the DNA segment to the expression vector. The synthetic linkers are attached to blunt-ended DNA segments by incubating the blunt-ended DNA segments with a large excess of synthetic linker molecules in the presence of an enzyme that is able to catalyze the ligation of blunt-ended DNA molecules, such as bacteria phage T4 DNA ligase. Thus, the products of the reaction are DNA segments carrying synthetic linker sequences at their ends. These DNA segments are then cleaved with the appropriate restriction endonuclease and ligated into an expression vector that has been cleaved with an enzyme that produces termini compatible with those of the synthetic linker. Synthetic linkers containing a variety of restriction endonuclease sites are commercially available from a number of sources including New England BioLabs, Beverly, Mass.

2. Inserting a Gene Coding for a Second Polypeptide into a Second Member of a Plant Species. Useful genes include those genes coding for a second polypeptide that can autogenously associate with the first polypeptide in such a way as to form a biologically functional multimeric protein. The methods used to introduce a gene coding for this second polypeptide into a second member of a plant species are the same as the methods used to introduce a gene into the first member of the same plant species and have been described above.

3. Producing a Population of Progeny from the First and Second Transformants. A population of progeny can be produced from the first and second transformants of a plant species by methods well known in the art including those methods known as cross fertilization described by Mendel in 1865 (an English translation of Mendel's original paper together with comments and a bibliography of Mendel by others can be found in Experiments In Plant Hybridization. Edinburgh, Scotland, Oliver Boyd, eds., 1965).

4. Isolating Progeny Containing the Multimeric Protein. Progeny containing the desired multimeric protein can be identified by assaying for the presence of the biologically multimeric protein using assay methods well known in the art. Such methods include Western blotting, immunoassays, binding assays, and any assay designed to detect a biologically functional multimeric protein. See, for example, the assays described in Immunology: The Science of Self-Nonself Discrimination, Klein, John Wiley and Sons, New York, N. Y. (1982).

Preferred screening assays are those where the biologically active site on the multimeric protein is detected in such a way as to produce a detectible signal. This signal may be produced directly or indirectly and such signals include, for example, the production of a complex, formation of a catalytic reaction product, the release or uptake of energy, and the like. For example, a progeny containing an antibody molecule produced by this method may be processed in such a way to allow that antibody to bind its antigen in a standard immunoassay such as an ELISA or a radioimmunoassay similar to the immunoassays described in Antibodies: A Laboratory Manual, Harlow and Lane, eds., Cold Spring Harbor Laboratory, Cold Spring Harbor, N. Y. (1988). C. RNA ACTIVE AGENTS.

RNA active agents for carrying out the present invention are generally RNAi agents or dsRNAs (including but not limited to siRNAs, shRNAs and miRNAs, see, e.g., US Patent Application 20090118214). Production of such RNA active agents in plants is carried out in accordance with known techniques, or variations thereof that will be apparent to those skilled in the art. See, e.g., US Patent No. 6,506,559 to Fire et al., see also US Patents Nos. 7,456,335 to Kogel et al., 7,122,719 to Hakimi et al., and 6,846,482 to Liu et al.; US Patent Application 2006/0095987 to Niblett, and K. Gordon and P. Waterhouse, RNAi for insect- proof plants, Nature Biotechnology 23, 1231-1232 (November 2007).

Inhibition of gene expression refers to the absence (or observable decrease) in the level of protein and/or mRNA product from a target gene. Specificity refers to the ability to inhibit the target gene without manifest effects on other genes of the cell. The consequences of inhibition can be confirmed by examination of the outward properties of the cell or organism (as presented below in the examples) or by biochemical techniques such as RNA solution hybridization, nuclease protection, Northern hybridization, reverse transcription, gene expression monitoring with a microarray, antibody binding, enzyme linked immunosorbent assay (ELISA), Western blotting, radioimmunoassay (RIA), other immunoassays, and fluorescence activated cell analysis (FACS). For RNA-mediated inhibition in a cell line or whole organism, gene expression is conveniently assayed by use of a reporter or drug resistance gene whose protein product is easily assayed. Such reporter genes include acetohydroxyacid synthase (AHAS), alkaline phosphatase (AP), beta galactosidase (LacZ), beta glucoronidase (GUS), chloramphenicol acetyltransferase (CAT), green fluorescent protein (GFP), horseradish peroxidase (HRP), luciferase (Luc), nopaline synthase (NOS), octopine synthase (OCS), and derivatives thereof. Multiple selectable markers are available that confer resistance to ampicillin, bleomycin, chloramphenicol, gentamycin, hygromycin, kanamycin, lincomycin, methotrexate, phosphinothricin, puromycin, and tetracyclin.

Depending on the assay, quantitation of the amount of gene expression allows one to determine a degree of inhibition which is greater than 10%, 33%, 50%, 90%, 95% or 99% as compared to a cell not treated according to the present invention. Lower doses of injected material and longer times after administration of dsRNA may result in inhibition in a smaller fraction of cells (e.g., at least 10%, 20%, 50%, 75%, 90%, or 95% of targeted cells). Quantitation of gene expression in a cell may show similar amounts of inhibition at the level of accumulation of target mRNA or translation of target protein. As an example, the efficiency of inhibition may be determined by assessing the amount of gene product in the cell: mRNA may be detected with a hybridization probe having a nucleotide sequence outside the region used for the inhibitory double-stranded RNA, or translated polypeptide may be detected with an antibody raised against the polypeptide sequence of that region.

The RNA may comprise one or more strands of polymerized ribonucleotide. It may include modifications to either the phosphate-sugar backbone or the nucleoside. For example, the phosphodiester linkages of natural RNA may be modified to include at least one of a nitrogen or sulfur heteroatom. Modifications in RNA structure may be tailored to allow specific genetic inhibition while avoiding a general panic response in some organisms which is generated by dsRNA. Likewise, bases may be modified to block the activity of adenosine deaminase. RNA may be produced enzymatically or by partial/total organic synthesis, any modified ribonucleotide can be introduced by in vitro enzymatic or organic synthesis.

The double-stranded structure may be formed by a single self-complementary RNA strand or two complementary RNA strands. RNA duplex formation may be initiated either inside or outside the cell. The RNA may be introduced in an amount which allows delivery of at least one copy per cell. Higher doses (e.g., at least 5, 10, 100, 500 or 1000 copies per cell) of double-stranded material may yield more effective inhibition; lower doses may also be useful for specific applications. Inhibition is sequence-specific in that nucleotide sequences corresponding to the duplex region of the RNA are targeted for genetic inhibition.

RNA containing a nucleotide sequences identical to a portion of the target gene are preferred for inhibition. RNA sequences with insertions, deletions, and single point mutations relative to the target sequence have also been found to be effective for inhibition. Thus, sequence identity may optimized by sequence comparison and alignment algorithms known in the art (see Gribskov and Devereux, Sequence Analysis Primer, Stockton Press, 1991, and references cited therein) and calculating the percent difference between the nucleotide sequences by, for example, the Smith- Waterman algorithm as implemented in the BESTFIT software program using default parameters (e.g., University of Wisconsin Genetic Computing Group). Greater than 90% sequence identity, or even 100% sequence identity, between the inhibitory RNA and the portion of the target gene is preferred. Alternatively, the duplex region of the RNA may be defined functionally as a nucleotide sequence that is capable of hybridizing with a portion of the target gene transcript (e.g., 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, 50 degree C. or 70 degree C. hybridization for 12-16 hours; followed by washing). The length of the identical nucleotide sequences may be at least 25, 50, 100, 200, 300 or 400 bases.

As disclosed herein, 100% sequence identity between the RNA and the target gene is not required to practice the present invention. Thus the invention has the advantage of being able to tolerate sequence variations that might be expected due to genetic mutation, strain polymorphism, or evolutionary divergence.

RNA may be synthesized either in vivo or in vitro. Endogenous RNA polymerase of the cell may mediate transcription in vivo, or cloned RNA polymerase can be used for transcription in vivo or in vitro. For transcription from a transgene in vivo or an expression construct, a regulatory region (e.g., promoter, enhancer, silencer, splice donor and acceptor, polyadenylation) may be used to transcribe the RNA strand (or strands). Inhibition may be targeted by specific transcription in an organ, tissue, or cell type; stimulation of an environmental condition (e.g., infection, stress, temperature, chemical inducers); and/or engineering transcription at a developmental stage or age. The RNA strands may or may not be polyadenylated; the RNA strands may or may not be capable of being translated into a polypeptide by a cell's translational apparatus. RNA may be chemically or enzymatically synthesized by manual or automated reactions. The RNA may be synthesized by a cellular RNA polymerase or a bacteriophage RNA polymerase (e.g., T3, T7, SP6). The use and production of an expression construct are known in the art (see also WO 97/32016; U.S. Pat. Nos. 5,593,874, 5,698,425, 5,712,135, 5,789,214, and 5,804,693; and the references cited therein). If synthesized chemically or by in vitro enzymatic synthesis, the RNA may be purified prior to introduction into the cell. For example, RNA can be purified from a mixture by extraction with a solvent or resin, precipitation, electrophoresis, chromatography, or a combination thereof. Alternatively, the RNA may be used with no or a minimum of purification to avoid losses due to sample processing. The RNA may be dried for storage or dissolved in an aqueous solution. The solution may contain buffers or salts to promote annealing, and/or stabilization of the duplex strands.

RNA may be directly introduced into the cell (i.e., intracellularly); or introduced extracellularly into a cavity, interstitial space, into the circulation of an organism, introduced orally, or may be introduced by bathing an organism in a solution containing the RNA. Methods for oral introduction include direct mixing of the RNA with food of the organism, as well as engineered approaches in which a species that is used as food is engineered to express the RNA, then fed to the organism to be affected. For example, the RNA may be sprayed onto a plant or a plant may be genetically engineered to express the RNA in an amount sufficient to kill some or all of a pathogen known to infect the plant. Physical methods of introducing nucleic acids, for example, injection directly into the cell or extracellular injection into the organism, may also be used.

Physical methods of introducing nucleic acids into plant cells, and the production of transgenic plants, are discussed in greater detail below, but generally include injection of a solution containing the RNA, bombardment by particles covered by the RNA, soaking the cell or organism in a solution of the RNA, or electroporation of cell membranes in the presence of the RNA. A viral construct packaged into a viral particle would accomplish both efficient introduction of an expression construct into the cell and transcription of RNA encoded by the expression construct. Other methods known in the art for introducing nucleic acids to cells may be used, such as lipid-mediated carrier transport, chemical-mediated transport, such as calcium phosphate, and the like. Thus the RNA may be introduced along with components that perform one or more of the following activities: enhance RNA uptake by the cell, promote annealing of the duplex strands, stabilize the annealed strands, or otherwise increase inhibition of the target gene.

D. APTAMER ACTIVE AGENTS.

Aptamers can be produced in accordance with known techniques or variations thereof that will be apparent to those skilled in the art. See, e.g., US Patent No. 7,538,211; see also US Patent No. 6,949,379; 5,840,867; and 5,582,981.

One method for generating an aptamer is with the process entitled "Systematic Evolution of Ligands by Exponential Enrichment". The process is a method for the in vitro evolution of nucleic acid molecules with highly specific binding to target molecules and is described in, e.g., U.S. Pat. No. 5,475,096 entitled "Nucleic Acid Ligands", and U.S. Pat. No. 5,270,163 (see also WO 91/19813) entitled "Nucleic Acid Ligands".

The process relies as a starting point upon a large library or pool of single stranded oligonucleotides comprising randomized sequences. The oligonucleotides can be modified or unmodified DNA, RNA, or DNA/RNA hybrids. In some examples, the pool comprises 100% random or partially random oligonucleotides. In other examples, the pool comprises random or partially random oligonucleotides containing at least one fixed and/or conserved sequence incorporated within randomized sequence. In other examples, the pool comprises random or partially random oligonucleotides containing at least one fixed and/or conserved sequence at its 5' and/or 3' end which may comprise a sequence shared by all the molecules of the oligonucleotide pool. Fixed sequences are sequences such as hybridization sites for PCR primers, promoter sequences for RNA polymerases (e.g., T3, T4, T7, and SP6), restriction sites, or homopolymeric sequences, such as poly A or poly T tracts, catalytic cores, sites for selective binding to affinity columns, and other sequences to facilitate cloning and/or sequencing of an oligonucleotide of interest. Conserved sequences are sequences, other than the previously described fixed sequences, shared by a number of aptamers that bind to the same target.

The oligonucleotides of the pool preferably include a randomized sequence portion as well as fixed sequences necessary for efficient amplification. Typically the oligonucleotides of the starting pool contain fixed 5' and 3' terminal sequences which flank an internal region of 30-50 random nucleotides. The randomized nucleotides can be produced in a number of ways including chemical synthesis and size selection from randomly cleaved cellular nucleic acids. Sequence variation in test nucleic acids can also be introduced or increased by mutagenesis before or during the selection/amplification iterations.

The random sequence portion of the oligonucleotide can be of any length and can comprise ribonucleotides and/or deoxyribonucleotides and can include modified or non- natural nucleotides or nucleotide analogs. See, e.g., U.S. Pat. Nos. 5,958,691; 5,660,985; 5,958,691; 5,698,687; 5,817,635; 5,672,695, and PCT Publication WO 92/07065. Random oligonucleotides can be synthesized from phosphodiester-linked nucleotides using solid phase oligonucleotide synthesis techniques well known in the art. See, e.g., Froehler et al., Nucl. Acid Res. 14:5399-5467 (1986) and Froehler et al., Tet. Lett. 27:5575-5578 (1986). Random oligonucleotides can also be synthesized using solution phase methods such as triester synthesis methods. See, e.g., Sood et al., Nucl. Acid Res. 4:2557 (1977) and Hirose et al., Tet. Lett., 28:2449 (1978). Typical syntheses carried out on automated DNA synthesis equipment yield 10¹⁴-10¹⁶ individual molecules, a number sufficient for most processes. Sufficiently large regions of random sequence in the sequence design increases the likelihood that each synthesized molecule is likely to represent a unique sequence.

The starting library of oligonucleotides may be generated by automated chemical synthesis on a DNA synthesizer. To synthesize randomized sequences, mixtures of all four nucleotides are added at each nucleotide addition step during the synthesis process, allowing for random incorporation of nucleotides. As stated above, in one embodiment, random oligonucleotides comprise entirely random sequences; however, in other embodiments, random oligonucleotides can comprise stretches of nonrandom or partially random sequences. Partially random sequences can be created by adding the four nucleotides in different molar ratios at each addition step.

The starting library of oligonucleotides may be either RNA or DNA. In those instances where an RNA library is to be used as the starting library it is typically generated by transcribing a DNA library in vitro using T7 RNA polymerase or modified T7 RNA polymerases and purified. The RNA or DNA library is then mixed with the target under conditions favorable for binding and subjected to step-wise iterations of binding, partitioning and amplification, using the same general selection scheme, to achieve virtually any desired criterion of binding affinity and selectivity. More specifically, starting with a mixture containing the starting pool of nucleic acids, the. method includes steps of: (a) contacting the mixture with the target under conditions favorable for binding; (b) partitioning unbound nucleic acids from those nucleic acids which have bound specifically to target molecules; (c) dissociating the nucleic acid-target complexes; (d) amplifying the nucleic acids dissociated from the nucleic acid-target complexes to yield a ligand-enriched mixture of nucleic acids; and (e) reiterating the steps of binding, partitioning, dissociating and amplifying through as many cycles as desired to yield highly specific, high affinity nucleic acid ligands to the target molecule. In those instances where RNA aptamers are being selected, the method further comprises the steps of: (i) reverse transcribing the nucleic acids dissociated from the nucleic acid-target complexes before amplification in step (d); and (ii) transcribing the amplified nucleic acids from step (d) before restarting the process.

Within a nucleic acid mixture containing a large number of possible sequences and structures, there is a wide range of binding affinities for a given target. A nucleic acid mixture comprising, for example, a 20 nucleotide randomized segment can have 4²⁰ candidate possibilities. Those which have the higher affinity constants for the target are most likely to bind to the target. After partitioning, dissociation and amplification, a second nucleic acid mixture is generated, enriched for the higher binding affinity candidates. Additional rounds of selection progressively favor the best ligands until the resulting nucleic acid mixture is predominantly composed of only one or a few sequences. These can then be cloned, sequenced and individually tested for binding affinity as pure ligands or aptamers.

Cycles of selection and amplification are repeated until a desired goal is achieved. In the most general case, selection/amplification is continued until no significant improvement in binding strength is achieved on repetition of the cycle. The method is typically used to sample approximately 10 different nucleic acid species but may be used to sample as many

1 R as about 10 different nucleic acid species. Generally, nucleic acid aptamer molecules are selected in a 5 to 20 cycle procedure. In one embodiment, heterogeneity is introduced only in the initial selection stages and does not occur throughout the replicating process.

A variety of nucleic acid primary, secondary and tertiary structures are known to exist. The structures or motifs that have been shown most commonly to be involved in non- Watson-Crick type interactions are referred to as hairpin loops, symmetric and asymmetric bulges, pseudoknots and myriad combinations of the same. Almost all known cases of such motifs suggest that they can be formed in a nucleic acid sequence of no more than 30 nucleotides. For this reason, it is often preferred that procedures with contiguous randomized segments be initiated with nucleic acid sequences containing a randomized segment of between about 20 to about 50 nucleotides and in some embodiments of about 30 to about 40 nucleotides. In one example, the 5'-fixed:random:3'-fixed sequence comprises a random sequence of about 30 to about 50 nucleotides.

Variations on the foregoing aptamer selection procedures include, but are not limited to, those described in U.S. Pat. Nos. 5,707,796; 5,763,177; 5,567,588; 5,861,254; 5,496,938; 5,705,337; and 5,580,737.

The aptamers may incorporate containing modified nucleotides conferring improved characteristics on the ligand, such as improved in vivo stability or improved delivery characteristics. Examples of such modifications include chemical substitutions at the ribose and/or phosphate and/or base positions. Aptamers containing modified nucleotides are described, e.g., in U.S. Pat. No. 5,660,985, which describes oligonucleotides containing nucleotide derivatives chemically modified at the 2' position of ribose, 5 position of pyrimidines, and 8 position of purines, U.S. Pat. No. 5,756,703 which describes oligonucleotides containing various 2'-modified pyrimidines, and U.S. Pat. No. 5,580,737 which describes highly specific nucleic acid ligands containing one or more nucleotides modified with 2'-amino (2'-NH₂), 2'-fluoro (2'-F), and/or 2'-OMe (2'-OMe) substituents. Other modifications include, but are not limited to, 2'-position sugar modifications, 5-position pyrimidine modifications, 8-position purine modifications, modifications at exocyclic amines, substitution of 4-thiouridine, substitution of 5-bromo or 5-iodo-uracil; backbone modifications, phosphorothioate or alkyl phosphate modifications, methylations, and unusual base-pairing combinations such as the isobases isocytidine and isoguanidine. Modifications can also include 3' and 5' modifications such as capping.

In one embodiment, oligonucleotides are provided in which the P(O)O group is replaced by P(O)S ("thioate"), P(S)S ("dithioate"), P(O)NR₂ ("amidate"), P(O)R, P(O)OR¹, CO or CH₂ ("formacetal") or 3 '-amine (-NH-CH₂-CH₂-), wherein each R or R' is independently H or substituted or unsubstituted alkyl. Linkage groups can be attached to adjacent nucleotides through an — 0~, --N--, or -S-- linkage. Not all linkages in the oligonucleotide are required to be identical. As used herein, the term phosphorothioate encompasses one or more non-bridging oxygen atoms in a phosphodiester bond replaced by one or more sulfur atoms.

In further embodiments, the oligonucleotides comprise modified sugar groups, for example, one or more of the hydroxyl groups is replaced with halogen, aliphatic groups, or functionalized as ethers or amines. In one embodiment, the 2'-position of the furanose residue is substituted by any of an OMe, O-alkyl, O-allyl, S-alkyl, S-allyl, or halo group. Methods of synthesis of 2'-modified sugars are described, e.g., in Sproat, et al., Nucl. Acid Res. 19:733- 738 (1991); Cotten, et al., Nucl. Acid Res. 19:2629-2635 (1991); and Hobbs, et al., Biochemistry 12:5138-5145 (1973). Other modifications are known to one of ordinary skill in the art.

E. INSECTICIDAL COMPOSITIONS AND BAIT COMPOSITIONS.

Active agents as described above can be orally administered to an insect pest by any suitable means, such as by expressing the active agents in the plant through transgenic techniques (discussed below), or by applying the active agent to a plant (e.g., on said plant or immediately adjacent said plant, such as by applying the active agent to the soil or media in which the plant is grown).

The active agent can be applied to a plant as an insecticidal composition (e.g., a composition comprising the active agent plus insecticidal carriers and/or adjuvants), or as an insecticidal bait composition.

Insecticidal compositions including bait compositions are known and can be produced in accordance with known techniques. In general, an insecticidal composition comprises the active agent, a carrier (which may be simply water), and optionally a sticker. An insecticidal bait composition generally comprises the active agent and one or more of insect food and/or insect atttractant (e.g., sugar or carbohydrate), carrier, etc. Suitable carriers and adjuvants can be solid or liquid and are substances useful in formulation technology, e.g. natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, thickeners, binders or fertilizers.

A preferred method of applying an active agent of the invention, or an agrochemical composition which contains at least one of said compounds, is foliar application. The frequency of application and the rate of application will depend on the risk of infestation by the corresponding pathogen. However, the active agents can also penetrate the plant through the roots via the soil (systemic action) by drenching the locus of the plant with a liquid formulation, or by applying the compounds in solid form to the soil, e.g. in granular form (soil application). In crops of water such as rice, such granulates can be applied to the flooded rice field. The active agents may also be applied to seeds (coating) by impregnating the seeds or tubers either with a liquid formulation of the fungicide or coating them with a solid formulation.

The term locus as used herein is intended to embrace the fields on which the treated crop plants are growing, or where the seeds of cultivated plants are sown, or the place where the seed will be placed into the soil. The term seed is intended to embrace plant propagating material such as cuttings, seedlings, seeds, and germinated or soaked seeds.

The active agents are used in unmodified form or, preferably, together with the adjuvants conventionally employed in the art of formulation. To this end they are conveniently formulated in known manner to emulsifiable concentrates, coatable pastes, directly sprayable or dilutable solutions, dilute emulsions, wettable powders, soluble powders, dusts, granulates, and also encapsulations e.g. in polymeric substances. As with the type of the compositions, the methods of application, such as spraying, atomizing, dusting, scattering, coating or pouring, are chosen in accordance with the intended objectives and the prevailing circumstances.

Typical rates of application are normally from 5 g to 2 kg of active ingredient (a.i.) per hectare (ha), preferably from 10 g to 1 kg a.i./ha, most preferably from 20 g to 600 g a.i./ha. When used as seed drenching agent, convenient dosages are from 10 mg to 1 g of active substance per kg of seeds.

The formulation, i.e. the compositions containing the active agent and, if desired, a solid or liquid adjuvant, are prepared in known manner, typically by intimately mixing and/or grinding the compound with extenders, e.g. solvents, solid carriers and, optionally, surface active agents (surfactants). Suitable carriers and adjuvants may be solid or liquid and correspond to the substances ordinarily employed in formulation technology, such as, e.g. natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, thickeners binding agents or fertilizers. Such carriers are for example described in WO 97/33890.

Further surfactants customarily employed in the art of formulation are known to the expert or can be found in the relevant literature.

The agrochemical formulations will usually contain from 0.1 to 99% by weight, preferably from 0.1 to 95% by weight, of the active agent, 99.9 to 1% by weight, preferably 99.8 to 5% by weight, of a solid or liquid adjuvant, and from 0 to 25% by weight, preferably from 0.1 to 25% by weight, of a surfactant.

Whereas it is preferred to formulate commercial products as concentrates, the end user will normally use dilute formulations.

The compositions may also contain further adjuvants such as stabilizers, antifoams, viscosity regulators, binders or tackifiers as well as fertilizers, micronutrient donors or other formulations for obtaining special effects.

F. TRANSGENIC PLANTS.

Transgenic plants are produced in accordance with known techniques, or variations thereof that will be apparent to those skilled in the art. See, e.g., US Patent No. 5,750,386 to Conkling et al; see also US Patent No. 7,476,781 to Abad et al.

Vectors which may be used to transform plant tissue with DNA constructs of the present invention include, but are not limited to, both Agrobacterium vectors and ballistic vectors, as well as vectors suitable for DNA-mediated transformation.

Agrobacterium tumefaciens cells containing a DNA construct of the present invention, wherein the DNA construct comprises a Ti plasmid, are useful in methods of making transformed plants. Plant cells are infected with an Agrobacterium tumefaciens as described above to produce a transformed plant cell, and then a plant is regenerated from the transformed plant cell. Numerous Agrobacterium vector systems useful in carrying out the present invention are known. For example, U.S. Pat. No. 4,459,355 discloses a method for transforming susceptible plants, including dicots, with an Agrobacterium strain containing the Ti plasmid. The transformation of woody plants with an Agrobacterium vector is disclosed in U.S. Pat. No. 4,795,855. Further, U.S. Pat. No. 4,940,838 to Schilperoort et al. discloses a binary Agrobacterium vector (i.e., one in which the Agrobacterium contains one plasmid having the vir region of a Ti plasmid but no T region, and a second plasmid having a T region but no vir region) useful in carrying out the present invention.

Microparticles carrying a DNA construct of the present invention, which microparticle is suitable for the ballistic transformation of a plant cell, are also useful for making transformed plants of the present invention. The microparticle is propelled into a plant cell to produce a transformed plant cell, and a plant is regenerated from the transformed plant cell. Any suitable ballistic cell transformation methodology and apparatus can be used in practicing the present invention. Exemplary apparatus and procedures are disclosed in Sanford and Wolf, U.S. Pat. No. 4,945,050, and in Agracetus European Patent Application Publication No. 0,270,356, titled Pollen-mediated Plant Transformation. When using ballistic transformation procedures, the expression cassette may be incorporated into a plasmid capable of replicating in the cell to be transformed. Examples of microparticles suitable for use in such systems include 1 to 5 um gold spheres. The DNA construct may be deposited on the microparticle by any suitable technique, such as by precipitation.

Plant species may be transformed with the DNA construct of the present invention by the DNA-mediated transformation of plant cell protoplasts and subsequent regeneration of the plant from the transformed protoplasts in accordance with procedures well known in the art.

Any plant tissue capable of subsequent clonal propagation, whether by organogenesis or embryogenesis, may be transformed with a vector of the present invention. The term "organogenesis," as used herein, means a process by which shoots and roots are developed sequentially from meristematic centers; the term "embryogenesis," as used herein, means a process by which shoots and roots develop together in a concerted fashion (not sequentially), whether from somatic cells or gametes. The particular tissue chosen will vary depending on the clonal propagation systems available for, and best suited to, the particular species being transformed. Exemplary tissue targets include leaf disks, pollen, embryos, cotyledons, hypocotyls, megagametophytes, callus tissue, existing meristematic tissue (e.g., apical meristems, axillary buds, and root meristems), and induced meristem tissue (e.g., cotyledon meristem and hypocotyl meristem).

Plants of the present invention may take a variety of forms. The plants may be chimeras of transformed cells and non-transformed cells; the plants may be clonal transformants (e.g., all cells transformed to contain the expression cassette); the plants may comprise grafts of transformed and untransformed tissues (e.g., a transformed root stock grafted to an untransformed scion in citrus species). The transformed plants may be propagated by a variety of means, such as by clonal propagation or classical breeding techniques. For example, first generation (or Tl) transformed plants may be selfed to give homozygous second generation (or T2) transformed plants, and the T2 plants further propagated through classical breeding techniques. A dominant selectable marker (such as npt II) can be associated with the expression cassette to assist in breeding.

Plants which may be employed in practicing the present invention include (but are not limited to) cotton (Gossypium hirsutum), tobacco {Nicotiana tabacum), potato {Solarium tuberosum), soybean {Glycine max), peanuts {Arachis hypogaea), sweet potato {Ipomoea batatus), cassava {Manihot esculenta), coffee {Cofea spp.), coconut {Cocos nucifera), pineapple {Ananas comosus), citrus trees {Citrus spp.), cocoa {Theobroma cacao), tea {Camellia sinensis), banana {Musa spp.), Avocado {Persea americana), Fig {Ficus casica), Guava {Psidium guajava), Mango {Mangifera indica), Olive {Olea europaea), papaya {Carica papaya), Cashew {Anacardium occidentale), Macadamia {Macadamia integrifolia), Almond {Prunus amygdalus), sugar beets {Beta vulgaris), corn {Zea mays), wheat, oats, rye, barley, rice, vegetables, ornamentals, and conifers. Vegetables include tomatoes {Lycopersicon esculentum), lettuce (e.g., Lactuea sativa), green beans {Phaseolus vulgaris), lima beans {Phaseolus limensis), peas {Lathyrus spp.) and members of the genus Cucumis such as cucumber (C sativus), cantaloupe (C cantalupensis), and musk melon (C melo). Ornamentals include azalea {Rhododendron spp.), hydrangea {Macrophylla hydrangea), hibiscus {Hibiscus rosasanensis), roses {Rosa spp.), tulips {Tulipa spp.), daffodils {Narcissus spp.), petnunias {Petunia hybrida), carnation {Dianthus caryophyllus), poinsettia {Euphorbia pulcherima), and chrysanthemum. Conifers which may be employed in practicing the present invention include, for example, pines such as loblolly pine {Pinus taeda), slash pine {Pinus elliotiϊ), ponderosa pine {Pinus ponderosa), lodgepole pine {Pinus contorta), and Monterey pine {Pinus radiata); Douglas-fir {Pseudotsuga menziesii); Western hemlock {Tsuga canadensis); Sitka spruce (Picea glauca); redwood {Sequoia sempervirens); true firs such as silver fir {Abies amabilis) and balsam fir {Abies balsamea); and cedars such as Western red cedar {Thuja plicata) and Alaska yellow-cedar {Chamaecyparis nootkatensis).

The present invention is explained in greater detail in the following non-limiting Examples. EXAMPLES

Objective 1: Sequence and characterize the expressed genes from the head, salivary gland and the digestive system of the tarnished plant bug and the digestive system of the green sink bug from different developmental stages. Transcriptomes are built in accordance with known techniques. We published the ontogeny for an EST library for the digestive system of the cabbage looper in 2003, where plasmid isolation was performed using a Qiagen BioRobot 9600 followed by sequencing on an ABI PRISM 370 DNA analyzer in our Genome Research Laboratory (GRL). In this project, 957 ESTs were sequenced where 12% were digestive proteases, 9% ribosomal proteins, 7% matched previously sequenced genes of unknown function and 29% were novel proteins (Figure 1).

Over 29% of the ESTs matched lepidopteran sequences and 21% noctuid genera. More recently, using Roche/454 GS FLX sequencing, we were able to sequence in a week or less starting with mRNA, 114 million bp from the tick brain and associated endocrine tissues which represented 530,000 reads assembled into over 21,000 contigs (Table A).

Table A. Roche/454 GS FLX sequencing metrics from Dermacentor variabilis synganglion cDNA library.

Total reads 532,136

Bp sequenced 113,636,431

Bp after primer trimming 98,567,133

Assembled contigs 21,112

Putatively identified transcripts 10,674

Novel transcripts 10,445

Mean contig length (bp) 347

Mean number reads/contig 11.8

Large contigs (>500 bp) 3216

Mean large contig length (bp) 989

Largest Contig (bp) 7731

The goal of Objective 1 is to apply the same effort to build transcriptomes for the plant bug head, salivary gland and digestive system, with a comparative analysis of the digestive system of the stink bug.

Methods. Objective 1 is achieved by separate 454 sequencing of at least 500,000 cDNAs synthesized from expressed genes in the head, salivary gland and digestive system of the tarnished plant bug, Lygus lineolaris, and the digestive system of the green stink bug, Acrosternum hilare, where the cDNA will be synthesized from feeding male and female last instars and adults. All insects for these studies are collected directly from the field in North Carolina, so gene expression is most relevant to natural conditions. Cotton is scouted in June for plant bugs. Once they appear in good numbers, the grassy areas closest to the field edge are swept for feeding stage, last instars and adults of both sexes. For green stink bugs, adults are collected in July by sweeping cotton or by placing white cloths on the ground and shaking the plants. Feeding will be determined for both species by the presence of food in the digestive system. The insect collections are made in the early morning, maintained with water available ad libitum for transport to the laboratory, and dissected in the afternoon. Any residual food in the digestive tract is washed from the opened gut with physiological saline to minimize plant contamination. The cDNA library is constructed from the entire digestive system including the malpighian tubules and rectum but not that part of the digestive tract found in the head. The head does not include the salivary glands but contains part of the salivary ducts and the most anterior portion of the digestive tract found in the head. cDNA library construction is conducted from equal amounts of tissue from both feeding stage immature and adult insects of both sexes.

Ultra high throughput pyrosequencing. As a sequencing platform, the Roche/454 GS FLX has no requirement for reference genomes for transcriptomics and is therefore well suited for Objective 1. A specific protocol for the preparation of cDNA suitable for sequencing on the 454 GS FLX sequencer (posted by the NCSU Genomic Sciences Laboratory, http://gsl.cals.ncsu.edu/media/cDNA.pdf) has recently been developed, has been successfully used to construct two transcriptomes, and is used in the carrying out Objective 1.

In the current project total RNA is purified separately from each tissue using TriReagent (Sigma), and messenger RNA purified using an Oligotex mRNA isolation kit (Qiagen). Modified primers (Beldade et al. 2006) are used for first-strand synthesis and second strand synthesis/PCR amplification to limit homopolymer stretches from the polyA tail and to produce sufficient quantities of cDNA for sequencing (3-5 μg). Aliquots of the purified mRNA and resulting cDNA are stored at -80°C for further use in Objective 3. An alternative to this PCR based amplification is the Eberwine method of linear mRNA amplification followed by RNA amplification to produce cDNA. However, in our previous 454 sequencing efforts, the PCR approach produced excellent results where rare hormonal and other transcripts were identified and large full length contigs (>5 kb) could be constructed as determined by comparison to previously sequenced tick genes.

The cDNA synthesized by PCR is purified and adjusted to 50 ng/μl which is a suitable starting concentration for library preparation using the Roche/454 GS FLX library preparation protocols. Sequencing is conducted in the NCSU GRL. Contiguous sequences are assembled using GS Assembler ver. 1.1.03 (Roche) initially with default parameters. Custom PHP and PERL scripts are written to manipulate the resulting sequences, which are compared to the non-redundant and EST GenBank databases using DeCypher accelerated blast algorithms (Time Logic). To further aid in the characterization of the resulting genes, a PHP script is written that translates the resulting contigs into all six reading frames which can be searched for conserved domain architectures and signal sequences.

Data analysis and broad impact of proposed work. Objective 1 provides a comparative transcriptome for the digestive system of two agriculturally-important, emerging pest species and that for the head and salivary gland of the plant bug. Data is given in TABLE A herein. These data provide a number of new opportunities for future studies of digestion, the regulation of salivation and digestion, novel insecticide targets, and neuroendocrinology for an insect group where minimal sequence data are available.

As noted above, an aspect of the invention is a transgenic plant that contains a polynucleotide construct that comprises a heterologous nucleotide sequence operably linked to a promoter that drives expression in the plant, the heterologous nucleotide sequence encoding an active agent toxic to an insect pest. In some embodiments, the encoded active agent is an RNA {e.g., a dsRNA) that inhibits expression of a target gene in said insect pest, and reduces or inhibits the expression of the expression product {e.g., protein, ribosomal RNA) of that gene. Such plants are produced by providing a construct in accordance with known techniques, and producing transgenic plants with such a construct in accordance with known techniques. The promoter can be any suitable promoter as described above (including inducible promoters, viral promoters, synthetic promoters, constitutive promoters, temporally regulated promoters, spatially regulated promoters, and/or spatiotemporally regulated promoter, etc.). Since, for insects, a dsRNA is in some embodiments from 200 to 300 nucleotides in length (though shorter RNAs of at least 21 or 24 nucleotides in length; or longer RNAs up to 500 or 1000 nucleotides in length, or more, may also be used), the heterologous nucleotide sequence in the consruct may be any of the nucleotide sequences present in Table 1 (SEQ ID NO: 1 through SEQ ID NO: 133 herein, where sequences are given 5' to 3'), or an active fragment thereof. Examples of active fragments thereof include continuous fragments of the disclosed sequences of at least 21 or 24 contiguous nucleotides in length. Such fragments can be selected in accordance with design guidelines known in the art. For example: In some embodiments the fragments are those that begin with two contiguous "A" residues; in some embodiments the fragments are those that do not contain 4 or more contiguous "T" or "A" residues; in some embodiments the fragments are those that contain 30 to 50 percent GC content (as opposed to higher G/C content); and/or in some embodiments the fragments are those that do not contain more than 16-17 contiguous base pairs of homology to other coding sequences, particularly other coding sequences in the host plant, as determined by homology searching such as by BLAST searching). See, e.g.,Elbashir et al., EMBO J. 20, 6877-6888; see also Applied Biosystems Technical Bulletin # 506, siRNA Design Guidelines. Minor changes can be included in the sequences or fragments thereof (e.g., I₅ 2 or 3 nucleotide substitutions) in accordance with known techniques. Where a dsRNA is desired, the complementary strand can be provided in accordance with known techniques, such as by an additional self-hybridizing segment on the same nucleic acid, by transformation of the plant with an additional construct containing a nucleic acid encoding the the complementary strand driven by a promoter, etc.

A further aspect of the invention is a dsRNA compound or product encoded by a nucleotide sequence of Table 1 (SEQ ID NO: 1 through SEQ ID NO: 133) as described above, or by an active fragment thereof as described above. Such products can be produced in accordance with known techniques, including but not limited to those described above, and are useful as an active ingredient in an insecticidal bait composition as described above.

TABLE 1: NUCLEIC ACIDS ENCODING TARGET INSECT PROTEINS

Transcription factor: contie00163 lensth=1989 numreads=341

CGCACAACCGTGTGCACAGAGTTCTTTATAAAAATGGGTGCCATCATGGCGTATT

TACTTACTTCAGTCGCACAGTCGACACAAAATATAATAAAGAAGAATGGATATG

GAGTCGTCGGTCAGCCTCGATTTGAAGGTGGAGCCGGCGAGCCCCTTATGGGAG

CTTGAGCTGCTTGATGAGGTCGGGGAGATCTGGGAAGGCGATGTCACGGTCATC

GAGGGGGAACTGGATGTGGACCTGGTCGAGGATGTGGACACCGACGCCTTCTGG

GGTCAGCAGGACATCTCCGAGCTCATTGGCAAGGAAGAGCAGATACTTGGACTG

GAACTTGACGAAATCACCTTGGAGGATCAGGACGTGAAGGAAGAACCTTTCACT

AGCTGGTTGGACAATGGCGCAATCTACGATGGACCGAATTCCACCGTTTCAGGTC

TTCCAACTTTCAACACGCCATCTTTGTGGTCAGCACCAGAGTCGCCCTCCGAAGA

AAAGGCCGCAGCGTTCACGGTGCCTGCGGGTGGGCCTGCAGCGCCGGACCCGCT

GCAAGGAGGGACTGCGGACCTGTTCACGGAGTTCGAGGACGTGCTGAGCCAAGC

AGCGGGTTCCCTCACGCCGCCCGACTCTCCCCGCTCTGCATCAGCATTCGGGGCG

CAGGCGCTGCTTCAGCTCGCCCCCAACTTGCAGCCGGTGCAGCTCTCGCCCGCTT

CTGGCCAGAGATTCATCGTGTTGCAGGAAATGGCGAACTCGCCTGGCGTCAACTA

TGAGTTGGATGAATTGATGGCGGAGACCTACCAGAAGAACGGAGGTATCTCCGT

CAGCAGTAGCGGCAACAGCAGTCCCATCAGCAGCAGCAGCAGCGGTACCAAAAC

CAAAGGACGTCCTTGGGACTCACCATCTGCCTCAAGTTCTGGCTTCAGTGATAGC

GGCTACGACGATCCTGAGTGGAGCTCCACAAAATCTACGGAATCCCGTGACTCTT CTGATCCGTCGCCAGCGAAAAAAAGGTGTTTGAACTCGACTGAAGAGCGTCGAT

TGAGGAAGAAGGAGCAGAACAAGAACGCTGCTACCCGCTACCGCATGAAGAAG

AAGATGGAAATGGAAGAAATCATCGGCGAAGAGAAGGAGCTGATGGACCGCCA

CGACACCCTCAAGTCCGAAGTCGTCGAGATCACCCGAGAAATCAGATACCTCAA

GGGCCTCATGGGAGACTTATTCAGAGCCAAGGGCCTCATTCGCGCCAAGTAATTT

CCAAATCAACTCGCTCTCCCCAACCACCAAAAATCAACCTGTTCTTCTAGTAGTG

AAAATTCAATTAGTGGATGTACTAGTAGACCATAATTTCATGATATCAATAACCT

AGTTATTATAACCTAGATTCGTGTATTTTTCGGGGTTGTTTTTTCTTCAATGAACT

CCTCATTTCCTCGTTAAGTATCGTGAAATGTTTTCAATCGCTTGAAAAGTGAATTT

TTGCAGTGACATCTTCCTTGTGTGTTTTGAGGCCCTTGATCTCTTCTTCTCAAGTT

AAATCCCTTTTTTGTTACTCGTCCAAGCTTGAATTTCATTCCCCTCGCCTTTCCATG

GCCTCCCCTACTTCTCGCCATCTTGTGTACGGGTTTTTCAACCCTCTATGACGCAT

TGTATTCCAAATATGCTTTTTTGGGTCGGTTCGCCCCCGAAAAAGTTGGAATCAG

CCGATGTTTTTGGTTGATAATCCTTACATTTAGAAGGCTCAAGCATTTCAAGTTTC

CATGATCTATTTTATTTTTTAATATAACCGGTCAATCCCAATCAAAATTTTTATTG

ATCAACTTTTCTTTTTATGAACGAGGGTCATCCTCGTTTTCCATCTGTGTCAAATT

TGCCTAAATTTTCACGTTGCACTTTGGTAATCAGGATTGTACCGGAACCTCACTTT

TATTGGTTATACAATATTTAGTTATGTAATAATGTAAACTTTTTTTAAAATAAATG

TGGATCTAACTCCC (SEQ ID NO: 1) contis06596 ϊenεth=697 numreads=52

AGAGAGAGCTCGGCTTCTTCAAATTCGTCTGACTCTTCAGAAACTTCAAGAAGCA

GTTCGGTCTCCAGCCCAGGATCAAGATCTAGAGCTGAGCAACCTCCTCGTAAAAA

TGGAGGTCCTGCGAAAAAAGAGAAAAAGTCAGCCCCTCCTCAACTTGCGTCGGA

CAGAGACTCGGATATTGCTCCTACCAGGAAGAGCACAAGGTCCTCGAGTACTAG

ACGCAGCAAACACGTTGTAGCCAGGCTTGCAAGCGACTCAGACTCTGATACAGG

TAGTAAATTGAGCTCTACAAGCAAGTCCCCTGCCAAAAAAGTTACTCCCAAAGG

ACCCAACGTGAGAAAACCAAAGAACTCTCACATCTCAGAGACAAGCCATGTAAT

TGAGAATGAAGAAAGAACATGTCCGTTTGACGGTTGTGACTCTTCTGGTCATTTA

GGTGGTAGGCTTGAAAAGCATTTCACTTTGGAGGCCTGCCCTGCTTACCACAATA

CAACACCTCAACAATGCAAGGAAGCATACATTGAACATTTAAAGAGAGATGAGG

AACGGAAAAAAGCCATTGGGAACCTCAAACCGGGACAGGCAACTACACCAACTC

CTGAACAAAAGGAGTATCGGGATAAAGTTAAGGAAATCAGGATCAAATTTGTGC

CTAAGGATGATGCTCAATCGGAACCAGACAATGGAAAAGAACGAC (SEQ ID NO:

2) contJ205742 length=783 numreads=38

CGGCCGGGAGGAGTTCGAAAAGGCTGCCTGTGGGCGATGAACCCTACGAAAATA

TCCAAGATGGACGACGAAGTCGCCAAATGGTCGAGGAAAGACCCTTTGGCTATT

CGGAGAGCAATGAGATATCCAGATGATCTCGAAAAGCTGGAAAGGGGAGAAAT

GAAATTCGGCTCCATATCGCACGCTGTTGAAGAGGAGAGCGGAGATGATGAAGT

TCCCGAAGAAGAAGAAAGCGACGAGAACGGTGAAGCTGCTAAACGGATTAACA

TGTTCGAACTTGCTCACATACTTCGGGACGAAGAGAGTACCAGAGCGTTCCTGTA

CGACTATAAACTGCTGCCGCAGCAGAAGATATGCCCCAGGTGCAATTCCGAAAT

GAAGCTGACTTTCGTGAATAATTTTTACGGATGGTTCAAGTGCAATAATGGCCAC

AGGTATAACGCTGTCAAAGGAACGTGGTTCGAAGGTACGCATTTGGGTTGTCGA

GAGATCTTATTGTCGACGTACTGCTTCCTTCAGAAGTTCTCGATGGATCAAACGA

TGGCAGAAGTGACGGCGGGGAAGGACGTGTTGGAGAGGGAGGTTATAGTGAATT

GGTGGTATAACACGAGAGAGGTCTGCATAGACGCCATCGAGCGGGAGTACGACG

AACTAGCCAGCCTAGGGCCGATAGGAGGCGTCGGTTGTACGGTCATCGTCGACA AAATGAAGATCGGCGAGTTGAACAATTCCCACAGGAGGATCAAAGACAACGTCT TCATTTTGGTGATGCTCGAAAAAGGG(SEQ IDNO: 3) contis01506 Iength=333 numreads=4

GGAGACGACATCGAGCGGAAACAGCAGGAAAACAGGAGGAGCTCGCACCAGGC

GTTGATGAGGAGGGAGGTTTGGTCGTCGATCGAAATGTTTTTGAAGAGTGGGAG

CGACTTACACCACTTGACTATCTTGTAAAGTCTATGGTCGGCAATGTTACATAAA

TTGGCCATGAGATCCGTAGGCTGGGGTGCAGAGGGTCTTGGTATCCTGCCTACGT

ACTTGGGGTCGGCTTCGTTGTACTGCCAGAGGTGTTCAACCTCCATAATTTCCTGC

AGAAGTTGAGGGACGAGGTTGGTTTGTGGAGCGGATTGCTGCTGCTGTTGAAGCT

CCGCG (SEQ ID NO: 4)

contiε01035 length=736 numreads=15

ACTGCCGAACATGTTGGTGCCACCACCAGGCGACCCCATGGACGACGCTGGGGG

TGGGTTGATGCGCTTTTCCTTCTGCCTCCTATTACAGAACCAGACTCGAACGACTT

CCTTCTCCATCGTCAGACTGTCCGCTAAAATTGAGATCTCTTCAGACGTGGGTTTC

GGGTTCTGCAGGAACGCCTTCTCCAGGCTATCCGGACGCTCGTTTCTATGGACGT

CCGCTTCTTCCTCCGCCTCCCGATGGCCTCCGGGGTCGTCATCGGGTTCGTCAGA

GAGTTCGGGTTGGTCAGCGAGTTGTCCGCGTCCAACAGCCATTTTTGGAGGAGCG

GCTTCAGCTTGCACATGTTCTTGAAAGAGAGGTTCAGCGCTTCGAACCTCGAGAT

GGTGGTTTGAGAGAAGTCGTTTCCGTACAGTTTGCCCATCGCTAGGCCGACGTCA

CCCTGCGTGAAACCTAATTTGATTCGTCGCTGCTTGAAAGTTTTGGCGAACTGTTC

AAGCTCTTCGAGATCTGTCGTTTCATCGGGCTCCATAGAGTGAGGTTGGGGCATT

GGGGTTCTATGTATTTTGGCAACGGGGCTACCCTTGTTATCGCCTAAAATACCTC

CCTGTACTGAATGAGAAAGTCTAGGTGGGGGAACGTTCTGTCGTAGGCTAGGTGT

GAGGAGACCCTGATTGTGGAGGAAATACTGTTGAGGGTGATGGTGTTGGGGTGT

GGATGTGGGAGGTGGAGGAG (SEQ ID NO: 5)

contig05994 lensth=1316 numreads=44

GTGTAAGAAATGCAGAATGTATACACAGAATGGGGAAAAGGGAGGAAATTATG

GTAAGTTTCGTTCATGTTACCACCACTTCAAGTGCTAAAGTTATTGCGGTCGGAA

TGTTCACGAATATTATTCAACTGAAGTAGATTGCTACAAAATGGGGGACGAATTG

ATGTATTTTTACTTTCAACACATTCATAAGAAGAATAATGACCAGATGAGTACCT

ATAATAAAATGAAAATAGATAGGTAATAAGTATGTGCGGTTTAAACAATAGCAC

ATACCTACCTACTACCTAAGTATTATCCATAAGTACTCCCATCAGAAATTATCAA

TGATACGATGATTTTTCACAAAATATTGCAACCATGTATCCAATTACCAATCCAC

TTTCAATTCCTTACCATGTCAATACAGCATTGTGGGAAACTTGGAATCTGACTCC

ATATTATGCTCTATTTAAGAGCATTCCTTGTATTTACACCTCCCAAGCGAGCTATG

ACGTTTTGTAATCTATTCACTTGGGATGTTAGACTACAATTAGCATCTTCAAGTGA

GCAAATTTGCTGTTTGAAATCTGCATTCTCGGTAGAAAGATTTCGACTTTTCTCTC

AAGCGCATCCATGTATTCCTTCTTTTTCCTCCGTGATTCTTGAGCAGAGATCTTAT

TCTTGATTTTCCTGCGAATTTTCTTCAGACATTTTTCTTCGGCTTTTGTCAGAGGTA

GTCTCACTGGTATAGGGTACCCTTCTGCCACCAACGTTCTCTTTTCCTCTTCGGTG

AGCAATAACATCCCCGTGGATCCTTTTGGCTGAGAACTAATGAGAGGAGTTTGTA

TGGGCTGCCGACTTGTGGTGGAACCAGAGAGGAACAGACTCCGATCCGATGATG

ATGGATGAAGTGGTGACACATTTCCTTCATTATCAGAAGATGCGCAAGAAGGAG

GTGTTGGTGGTAACGGAAAGCGTGTGCTGGAAGTTGATGATGGTTCAATTTTTTA

TAATATTAGCTGAGATCATCTGGTGCTTCCTCTTCATGCCAAGACTTTTATTGCTC

AACATAGTGGTAGTTGGGAGTTGCTTGACCGGAGTTTTGCTCATAGGCTTGGAAT

TTAGAAACATATCGGTGTAGATTATCGAGGCTTTGACAGATCTAGGAGGCGGTG GGGATGGAGAAGATGGACATGATGATGCTGGAGAGCTGGCGCCGCTCATTGGCT CGGACTTGATGACCACGAATGGGATAGGAGAAGACACACGACCACTGGTGGTTT TAGGAGGTACTGCTGGAAGTAGACAATCGTCTTCTACGTCATCTTTGATGA (SEQ ID NO: 6) contisOS201 lenεth=274 numreads=2

TGGTGTCGTTTGTAACGACACCATTCTAGCGACTGGAGTATTTTCCTCTAAGGAG

CTCTGGAGACTGTCTAAAGCATCTATTGTACAAAACTGCAACGGTATGCAGCCCC

CTATAAATGTAATTAAAGTAGAAAACCCCGGCTACTATTGCAGCAGCTATACACC

CCCAGGTCAAGACCACACCACCATGATTCCTTCGGCTCAATATAGCCCTTTAGGA

CTCCCCACGAAGTGGTCAAAGCCCTGGCGAACCGCGCTCCAAGAGGGTCCGTCG

(SEQ ID NO: 7) contis03255 length=595 numreads=12

CCCGGTTACGCACTATAGTGGGCCAAAAGCCAGGGGCGGTGGCACAGGTTGTAA

CCTCTTCTGGAAGTGTGATTCAAGGGACCTCGATGTTGCGAACTGTTTCTCTGAC

AGGAAAACCAGCACGCCAGTACGTCACCATTCCTCCGAATCAGATGCCCAAGAT

TCAAGAAAGGTCCGCTACTGTTATTGCATCTCCATCTCCTAGGAATGTGGCTACT

AGCTCCCGGCCGATGACATCAGCGGGTGACAGATCACTGAAAAGAGAGATGAGC

ATTGATTTCGTGCCCGATGGGCCTAGCTCAAACAAGCGGAGGAAAACTGAAAAA

GGAGGAAAAGGCTTGAGGCACTTCTCCATGAAAGTTTGTGAAAAGGTCAAGAAG

AAGGGACTCACCTCTTACAATGAAGTGGCGGATGAACTCGTGGCGGGAGTTCAC

TGATCCCAAACACATTCCTAACAGCGTTGATCAACAATATGACCAGAAGAACAT

CAGACGTCGAGTCTACGATGCGCTGAACGTGCTCATGGCGATGAATATTATATCT

AAAGAGAAGAAAGAAATCCGATGGTTGGGGCTTCCCATGACCACTGCTCAGG

(SEQ ID NO: 8) contis02985 lensth=278 numreads=8

CGCTTGGGTGGGCACATCCGAGGATTCGAAAATAAGCGAGGTGTTCATGTTGCTG

CCATTCACAAGCAAGGACTTTTCCATCAGGAGTCTTGCTGATCGGATATCTGACT

TACCCCACCTCCAGTACCTCTACCCCGACATCTGTAAGACTCAAGCTTTCAGCAA

GTACTACACCCCCTTTAGTGATAATCAGACGACCACGAATAACGGTTACGTGAAA

CCAATGCTGGTGACTCAGATACCACGATTAGGAGGAGGCAACAGCCTCCCTCCA

ACGC (SEQ ID NO: 9)

contis06813 length=444 numreads=268

GTATCATACCTGGTGAAGTGACAGACGCCGGCAGTCATGATGGCAGGATTATAC

TCGGCCTACGACCACGACGCCAACTACGACGCGGCCTGGCTCCTTCCTCAAGACT

ATCACACGGACGAGGGATATGGGTTGTTGGACTCGATCAAGATGCCACCTCAGG

GAAGGTCCGGGTTCCACATCGTCGACATACTTGATCTCAACGACGCCGCCAAAAC

ACAACACACGGATGGCGCCGCTCTCAACCCCACCCAAGCTCACGACGATATTTCA

GGAGGTGAAATGACAAACCTCCAGCAGCTTGGGAACCACCCCGGCCTGCTGTTC

GCCGGTGGTCAAGTCACCACTCCCATGGAAGCCCTTCACCATCATTGGGGCAACT

CCATCACCGAACTTAGAGGTCACCAGCAAGCCAGCCCCGACAGCACATCACCCG

GACCGTAA (SEQ ID NO: 10)

contig05957 lenzth=899 numreads=24

GCATTTCTGCTTGCACTTCTGACCGCAGCAATGAGGTGTCACTCCAGGAGTGGGT TTAGGTTTTTGTAGACGTCCGCAGAGACACTTATATCGCAGTTTATTGGGAGTGC AGAATTTTGGTCTTCTGCAGACTGGACAAATCCACGCTCCTCCCATTTCAGTACTT CGATTTGTTTTAGCCCATCGCCTGATGCAGTGCAGATGGAAAATGTTGAAGCAGT

TGTTGCAAGGCCAAACTCTCTCTTCGGCGAGGATCAGGTCAGTGCAGATGACGCA

CTCGGCAGTGTTTGAGTTGAGTTCATCGATTAACTTAGCTCTTTGACAATCGACTG

ATTCAGGAACTTGGCGGCGAGCTGGTTCCATATTGAATCTCGGTGGAGTAAATCC

TCTGAAACCATCGCCTTGCCCTGCAGTTGGATTTGCCCGGAGTCTGGTCCGGAAG

TTTTGGTGCATGTTGGAGAACGCATCGGCCAGCTCTGCCATGCTGCTGAGCAGCT

GCACAGAGATGTTGTTGTGGAGGAATATGATGGGGTTGCCGACGTTTCTCATACT

CGTGATGATGGCTCTCTCTAAAGTAGGTTGGTCGGAATCGTCGTCATCATCAGCC

GCTTCTTCTTCTTCCTCTTCGTGGGCATTCGGTTTCTGCGGATGACAGAGATGGTG

TTGGTGAAGGATGGAGGGCATTAAGTGATAACGACACAAGCGAATGAGGTGAAG

CAGCTGTGGAGGCAGATTCGATGCTCTCCCCTGAGTCAGACTCAACGCGGTCAGT

GTCCGAAAAGAACACGTCAGACATTTCAGCGTCAGAAATGCTGCTGACGAACGT

CGTGGCGTCGCTGTCACCTGAAAGAGAGAACGTGGGGTCGTTGGATTCACTGTCT

TCATCTTCGTCCGTTCCA (SEQ ID NO: 11)

contis02526 length=277 numreads=2

ACAACAACTGGGCCACCAAGGTCCCATCGCCGTTGGGCTCCAAGGCCTTCCCTTG

GACCGTCAACCCTCTGACGACGTCGCTCAACCACCACCAGGGCGGATGCTTCAAC

CCGACGGCCGGCCAGACGATGCCGGTGTCGTCTCCGGTGACGTCCCCCGTCACCT

CGCCCTGCTACGGTGGGGCCCCGGTACATGTACCACCCCCGCCCTCCCGACGCCG

TCCTGCATGAGCTCATCCCTCGCGTCTTTGAGGCTCAAGGCGAAGCAGCACTGTC

CG (SEQ ID NO: 12)

contig02094 length=249 numreads=5

CCGTGTCCACGTCACAGTCATTTGACGAGGCGGCCGGCTTTGTTTTTGCCATTTTT TCAATATATTTTTCCATACATTTTGGGACCAGCTGTTTCACTTTCTCCAGATTTGC TTTTGTGATACTGCCAGTGTCAAGACCGTGAGTCATGCCGATTTGCTTGAGTACTT CAAAATCGTCTTTGATCTCAAATTTATCATTGAATACAAACAGGTACTCTGTCTTG TCATTGGTGATAGAGCAATCAATCA (SEQ ID NO: 13)

contig00531 length=903 numreads=54

ACAGCAATCTGGTTTTCCAAAGAAGCAATCTGAATTTGTTCCCTGGTCGATTGTT

CCACAAACTGTTTTTGTGATGCGTCTGTGTTTGTTTCGATTTTCGATAAATAGTTT

TCAACGCTTAATGCTGGCAGTGGGGTGCTGGAGACAAACTGCCTTATGGTGGCCA

TTGAAGGGGTTTGGTTGATGGATAGATCAGGAGTCACTGTTTCAGTAGGAGGACT

TGGTACTTCCCTCTTCAATTCTATGTGTTCTTCCAAGATCGTGCCCATTGTCTGGG

ATGGTCGCCTCACAGGAGAGAGTACAGCTGGTGGTGGAGGCATCAACATTACAG

GGTGCACATTGGAGTCATCAGTACATTTGCTGTTGTCTGGCTGAATGTATGACGC

AGTCGGAAGGGAAGAGTGACTCCCCAGCTGGATGAGAGGTATACCAAGAAGAA

GATTCCTCGGTGGGAAGGATACAGGGGGTTGGTTCACGCATTGAGCAAGAGCTG

CTCTGGCTGCTACTAAGGGTTCTCGTGGGAGAGCTTGGAGAGGTGGTGTACCCAG

AGGGGTATAGAGAAAATTGTGAGCTTCACTACTTCGCCCACTAGACATTACATTC

AACTTGACTACTACCGGCTCTTTGATATCCTCAACCTTGTAGGGAGGAATCGTAC

ACACTAAGTGTGACTGTTGTAAAAACTCCTTATCAGGCTGTACAACTTCTTCCCA

ATAATTCTCTGCATTCATATCCCCTTCAGAGAATATCACTCTGGTATCTTTCAAAA

AGTTTTTACCGAGTATGAACAGTTCAAGGCCTCCAGCGGAGTGACATGAAACTAC

AGACTTCTTACATATTTCAGGAACTCCAGGAGGTTGTGTGCAAATTATAGGTTGA

GAAGTCACTTGTAATGTTTCCAGG (SEQ ID NO: 14) contis04146 length=312 numreads=4

GCGTTCTCATCTTCAGCTTGGCTTCTTGGAGTGCGAAGTCTCGAACTTGCTCCAGA

TGGTTACGCAAATTGAGAACATACTCGGTCGATGGTCTTAGGCGTTGTTGAGGCG

GTCTTACTAGGGCTTGAGGCAGTCTCATATCCCGGCCATAGATTAGCTGTGCTGG

GGATAATCCTGTTGTGGCGTTTGGAGAAGATCTATAGGCCAACAGGAAAAGAGG

AATTTTCTCGTCCCAATCAGTTTGGCTCTTGTTGACAAACATTGATATGTGTTTAG

TCAAGGTTCGGTTCAGCCTTTCAACAAGCCCAGCAG (SEQ ID NO: 15) contis03476 length=652 numreads=8

AGTGGCGACTAGGTTCTTCCTGGCATCCTCATGAATAATGACACTGTGCTGGCCA

CAGTGGGCAGTAAACAGCAGGCAATCATGAACACAAAAAGGGATACCGAGCCA

GCTGTCGAACTGGAGTGCCAGTTTATTCTTCGTCTACCAGAAGAACCTGGCAAAG

CCCTCCGAGACATGTTAAGAAGTGGGATGTCCAACTTGAAAGACCGTTTCCTTGT

GAGATTGGAGAATGATGTTCGATATGGCGAAGTCCGTGTCGACAACTGGCTCCTA

CATGCCAAAGTTGTTGATCTCCTTACAATTGTTGAATCTCTGAAGACCATTGATA

ATAAAAGCTTTTACAAAACTGCTGACATCTGCCAGATGATGATATGCAAAGTGG

AAGATGATGCCCCCACAGACGAAGAATCCCCCTCCAAACATAAGAAGAAGGACC

CGAATAAAGTCGACAAAAAGTTTCTCCATCCACACGGCATCACGCCTCCTTTGAA

AAACGTCAGGAAAAGACGCTTTCGGAAGACTTTGAAAAAGAAATATGTCGAGGC

CCCAGAAATTGAAAAAGAGGTTAAACGACTGTTACGTGGCGACAATGAAGACCG

TTTACTGTCAGGTGGGAGGTGATCAACGAAGAGGACGACAAAAAAGGGAGTAA

(SEQ ID NO: 16) contis02585 length=824 numreads=46

AAAAATAATTTTTATTCATCGCTTACAGATACCTAATAGTATTATTTACAGATAAT

GTAAATAATTATGTACAGTCTTAATGTTAATAGGAGAGGTTATGATTTACAAAAA

TAATATTTTCTTTGGTGTTTTCAATTATACAGTTTTCAGCGTGACTCTGGTTTTGTC

AATTTTTTAGTGAGAAGTCGAAAGGGTAGTGGTGAGGAGGGAAGGAGGGGTGGT

GGGAGGGGATTGAAGGGAGGGAAGGGAGGTGGAAGGGTGGAGGGTAGTAAGCG

AGGGAGGGGTGGTAGAAGGCAAAGGGCTGGTAAGGCCAAAGGGCGGTAGGGTA

GTAGGGCGGAGGCGGCAACTCCCCCAGGACGGGATAAGGGCCGGGGGCGGCGA

AGGGGTGAGGAAACATCGGAGGCGGCATCGGCGACGTCGAAGGCGAAGTCAGC

GGGGCGGGAGAGGCTGAGCCGACGGACACGCCGGAGTCATCCGTCGGAGGCGG

CGGCGGGCGGGGCTTTGACAGTCGGAAGCGGGGATCCCCTTTCGCGGCGGGCCG

GCTCCTCGCGACACACGGGAGATTTTCTCTCATCGGCTGTTTCGTCGTCCGTCGCT

TCCTTCTTGGGCGTCTCATCGTCCGCTGTCGACAGCTTCGACTTCTTCTTGGTATG

CGCCATCCCCGTCTCCCGAAATCCTTTGGCGAAAGGGTTGTTATCGATCTTCAAC

TTCGTAATTTTCTCATTTTGGTAAGCTGTAACGGCGATGAATTCAGTTTCTGGGAA

GGTAACTGTGAGTGTGGGGCTCCAATGAAGGGCGAGGATGTCTGAGGCTTTGAT

GACGTG (SEQ ID NO: 17) contig02033 length=254 numreads=2

GAGGAGGAGAGGAGCAAGAGAGGAGAGAACAGGAGGAAAGAAGAGGAGGAGA GGAGTAAAGAGAGGAGAGGAGAGGAGAGGAGGAGAGTGATGAGATAGAGGAG GAAGAGGAGAGAGGAGTAGAGAGGGGGAGGAGTGATGTTAGGGAGGAGAAGTA GAAAACAGGAGAGTGAAGAGAGATCGGAGGAGTAGAGAGGAGGGGAGACGAA GAGGGTGACGCGAGAGAGGAGGATGAGATGCGAGGGAGGAGAGGA (SEQ ID

NO: 18) contig01290 length=942 numreads=36

TCAGCAAGGAGGTTGTCCAATTCTTCGGCGGTCTTCATTTCTCTCGAGTATTCGTA

AGCTTCAACAGGAACAGTTTGAGCGGGTTTGGGCAGAGGTGTTTCGCCGTCAGC

GAATCTGATGGCCATCGCTCTCTTCCAAAGTTCCAGAGCTCCGATCATGTCCCTCT

TCTTGTCAACCAACGTCGCACCGAGCAGCTCCAAGGCGTCGATACTCTCCTGCCT

GCTCACGATCTCAGGGAGGCCGATGAGGTACTCGACGATGTGGGAGTGTCCGGT

GACGCTGGCCGCCAGCAGAGGTGTCATACCGTAGGAATCGACGTCCATTTTAGC

ACCGTGGTCGACTAGCATCTTGAGAATGTTGAGAGATCCAGATTCGGCGCAGTCG

TGGAGAGCCGTGTTGCCTCTGACGGACTTTCTGTTAACGTTGGCGCCAAGTCCCA

GAAGGTAGTGGGCGATCTGGTGGTGTCCTTTGTAGCAGGCGATCATCAAACACGT

GTGACCGTGCCTGTTTGAGACCTCAATGTCAGCTCCATGTTCAACGAGGAATTTG

ACGATTTCGTAATGTCCATCAAAGCAGGCAGCTCTGAGCGGCGTGGAATTTGTTT

TGGTGGTGGAGTTGACGCAGGCGCCGCGCTTCACCAAGAATTTGACAACGGCGA

AGTGACCGGCAGCCGCTGCGCACCACAGAGGAGGCGCTCCTTCGATCGTCTCTCC

ATCAAACACAACTGAACCAGCTTGTTCGAGATCGGCGTTGCACCTTTCGACGAGG

TATTCGACAACATCATAGTGACCGTTCCGACAAGCCATGACCAGAGGTGTCGCAC

CGTTCGTTTTGGTGGCGATCAGGTTCAGTACTTCTTCGGGCGTCCTATGATCGAG

GAAAACCTGAGAGTTTGATGATTCCAAGGAGTTGGAGCTCCGGTTCCACCATCCA

AGCCTCCAC (SEQ ID NO: 19)

contig00731 length=1376 numreads=65

ACTGGGCCTGGGGCCAACCGTATAGTGCCGAACTCAGTTTGTGTTGTTGCCTAAC

TCAGTTTGTCGTGGTGTCGTCTTCAATTGGTAGTGCCAACCGAATTATTTTGTTTT

GATCTGTGTTTACTGTCCGTCGAGAGCAAGTACCTGCCATACACTTCACACAGTC

CTTCAAGCATTTCATCAAAAATGCATACCTAGAACTGCACGAGATGATGCCTCGG

AATCATCTCATTCCACAAACTATCCGACATACTGACAACCAGGATTTGGAACGTC

CTGAAGCAAAGTCGCCGATTGAATACTGGACATCTAACGATATTGCCAGCTGGCT

GATTCGGGTCTCGGATTGGTTGAAGGTGGATTTCACCGACCTACATGGAGAAAA

ATTCGTACATTTGTCCGGCACGACGCTGCTCAATATGACGGAGTCAGAGTTCACG

GATGTAACGCCGAGTTTGGGAAAAGTCTATACGAGCACATCAAGATGGAATCAA

GTCTTACCCCAGTTACATCAGGTGATTACCTCGTTCAATCGGAGACCATCAAACC

TGTTGAGCAGTGGAACGATTCGGACATCGCAGATTGGCTTGTTAGAACGTCCATT

GACATTGGAGTTGACTACACAGAACTGTGCGGTGAGAAATTCGTCCATCTAACTG

GCCCTGAGATTCTGGACATGACGGAACGAGATTTCTTGACGATCTGCCAGGAAC

ATGGAAAGCAGCTCTTCAACAGCATCCAACGTGGATTATCCTCGTGGCAAAATG

ACGTCCACGTGCCACCAATGCAGACATATGAAGATGAAAATGGCTCTGACAGGG

AAAACACTCCGTCTTCAGGTTTGTCAAAACCTGTGGTCCCCGAACATTCAAACCC

ACCAGGAATCGTGCAGAAACGAAGACGGGGAAGACCGTCCAACCCTAATAAGA

AAGGGAAGAAGAAACCAGACAATTCTTACGGACGTCTGTGGGAATTTTTGAGAG

ACCTCCTCAAGAACCCGGACATGTGCCCCAGAATTATTAAATGGGAAAACCACG

ATGATGGCTTGTTTCGATTTGTACAGAGCAATGAAGTTGCCAGAATTTGGGGTGA

GAGGAAAGGAAACAAAGATATGACCTACGAGAAACTCAGCCGTGCCATGAGGTT

TTACTACGTGAAAAAACCCTATCCTATCTACCTCAGAGGGCACTCCACGCGTCTA

GTGTACCAGTTCGGACCAGGCGTACCAAATTGGAGAGTTGAGGACCCAAACTTC

GAAAAGAGGAATAAGGACCTCGAGCAGACGTTAGACTCTATCCAGGGACTCCTG

GACAGTCTCGCTTTTGAATCCACCAACATTGGAAAAACTAATTTATAAATACATG

ATGTACGATAGT (SEQ ID NO: 20) contiε01861 Ieti2th=237 numreads=9

ATATTCAGGTGTCGGCCCCTGTTTCACCGGTTCAGGCCCCATTCAGCTCTGGCAA TTCCTGCTGGAGCTTTTGACTGACAAATCTTGCCAAGGATTCATTTCTTGGACTGG AGACGGGTGGGAGTTCAAATTAACCGACCCTGATGAAGTTGCCAGGAGGTGGGG AGTCCGAAAAAACAAGCCTAAGATGAACTATGAAAAACTGAGCCGAGGGCTTAG GTACTACTACGACAAGAA (SEQ ID NO: 21)

contis01108 lenεth=1392 numreads=57

GTGCTTCACATTCTTCTCCACATTCCATACACTCCTTTCCTGCGCTCTCTTTGTCCG

GAGCTTCAGAGACTAGATGGCAAGGTAAAGCACACTCGTGGTTAGAGCAGGCAA

GGATTCGGCCACAGTTTCTGCCACACTGCTGCGAAACAGCCCGATGACAGGGTA

AGGAGATCTCTTCGTGGCCACCGGGACACCATACTCCGACAGGGACCACACATG

GAGGACAAGGGAGTTCTTTGATTTCAACTGGAGCAGTCACTCTTTCCCAGGGTGT

CGCTGGCTTGTAGTTTGGAACTGAATTGACCAGGACAGAAGAGTGACAGGGCAG

AGGGCAACGGTGACCGCACTCCTGAGGTTTGTCACAGATTTGTCGGCACGAAGG

GCAGGGGCCCATATGACATTTATGCTCTTCCCTTTTAGGATGGTGACATTCAGGA

GGGATCATGCACAACTTGTTACATTTCAGGTTCTTCTTTTTTCGATGAGTTCCACA

GGGGACTGTGACGACAGTAGCGCCACACCGACAGGAAATTTGCTTGGTCTGTTCC

AGGGGTAGCAGGGTCCACGATGGCAGACTGAGGCACACTTGTGTTGACCACACC

GGAGTGTTTTGCCGCAAAGTTTCTCACATGGAGGACACCGGCCATCGCAACATTT

TCTATTGCAAGGATGGATGCCACAATCTCGAGTTCCTTTACACTTGGTGTCGCAC

AGGTATTCTTTGATGCAGGCAATTTCTTTCTTGTGGAGACCGCAACGGCACGACT

TCTCCAGGAACTCCATGCAGGCGCCGCAGGACCCTCGGTGGCACCGCTGGTTGCA

CGAATGAAGGCCACAATCCAACAATTTGCCGCAAGTGTTGCCACACGTGGCAAC

TTCCTGAGTACAGGGGATGACCGTTTCCTCAGACCCGCAGGGACAAGTCCTGGGC

AGGGTGCGTGGACAAGGTCCACACTCTTCTTTGTGGCAGACTAGATCACAACTGT

GTTTGTTACAGTCCAATAATTTATTGCAAACCTTGTCGCATTTCCATGAAAGCTCA

GCGCAGGGAGTGAGGCGAGATTGACTTTTACAATCGCAGGTCATCATTGATGTTT

TTGGACAAGCTGGACACTCTCCAGGATGGCAAACCGTTTCGCACTTGTGCTTTTC

ACAACTGAGGAGCTTACTGCATTTCTGACCGCACGACCATTCTTTGTAGCAGCAC

CTCACCATTTTGTCTCCTCCATTAGAGCAGTAGCATTTGGCTCTGATCATTTTGGG

GCACGGCGGACAAGGACCTGGGTGGCACAGCAACAGACACGTATGCCCACAGCT

GGGTACCAAGTCCTTGCCACATACTTCACCGCAGGAATGTGGAACGAGCCAAGG

CTGATAGGGAGGATCCAAGA (SEQ ID NO: 22) contJ202251 lenεth=256 numreads=4

GCACTGCCGGAGGAAGAATGCGTTCACTTTGGCTTCGGACATCCTCTGTCGAGGG TAGATTGGAATGTACACGTCGTTTTCTACAGCAACCTTCATCTTGTCGACGACGG ATTTCAACAGTGCGTTGAGAAATTTTGAAGAGTGCGTCACTGTAGGGTAATCCTG GATGAATTTGGTGACCAGACCAACGAGGGAGACCGTCTGAGTGCTGGACAATGG GTCCCAACACTGCGTCACTAGTTGAGTTAATTTCGGT (SEQ ID NO: 23) contis04665 Iength=272 numreads=3

GGTGGCAGGTGCCGACGGCAGGGTCCGCCGACGCCCGGGCGTCACCGCCGTCGA

GTATTTGTACTCTTGAGCTGACGACGCGGGGTTCCAGAACAGCTTTCGGTCGCCT

CTCGAGTCCGGGAGGTTGTAGTACGTGGGTTGCGAGTACTGGTCGAACGGTGCTG

TCGGGTGGTGACCGTCAACAGTACTCGAATAGTTCCAAGGCTGAGACCCCGGAT

GGTGCCGGATGACACCAGAAGATGGTTCCATGCACTCATACCCGAGCCGTTTCG

(SEQ ID NO: 24) contig00974 length=302 numreads=5

CATCACCCGGCGTATCGGAGCTGACGGACCCGGGTCCGATGATCCACGGGATCC

CGTCTCCCAGTCTCAAAGACGATCGGATAGAGGACGACGAAGAGGACCAATCCA

GGGAGCAGATGGCGTCCGGACCCGGACACAAGAAACGGAAGAGGAGGGTCCTC

TTCAGCAAACAGCAGACCTACGAGTTGGAGAGGCGGTTTCGACAGCAGAGGTAC

CTTTCGGCCCCCGAACGGGAACACCTCGCGTCGATTATCCACCTTACGCCGACCC

AAGTGAAAATCTGGTTCCAAAATCACCGATAC (SEQ ID NO: 25) contis02631 lensth=259 numreads=2

TCAAGGCGTCGTTCGAAATCAGCCCGAAGCCTTGTCGGAAAGTTAGGGAAGGTC

TGGCTAAAGATACAGGGCTCTCCGTGAGAATTGTTCAGGTTTGGTTCCAAAACCA

AAGAGCTAAAATGAAAAAAATACAGAAGAAAGCTAGGATGGATGTAGAAAAAA

CAAACAAACTACTAGGAAGCAACGGAGGTAGGACTGATGGCGACACTACAGAC

GTCCACAAGAGTGAAAGGATAAAGGACGAAGAACACAGCGGTGG (SEQ ID NO:

26) contis02098 lensth=428 numreads=17

GTAAATACCATGAAACTGGGTACCTATATAAATCATTATGTCATTTTTTCATGTTT

CGAATTTTTATACAAAAAGGAAGAAAAAGAAATACCATCAGATTGTCACTGGTG

TAGGTTCCATACTCCACTGAATACGAAAAGGTTTTTATTTGTTTTTATATTTTCGA

TAACAAAGCGGATGGTAGGGGTTTCACAACTAAGTTACGCAGAGACGTACAACC

GCATCGATTGAGAAGGTGGCAAAAAAAACTGTGGTATTCGGTCAACACTGCTGA

TAACGGTTACTTGCTGAAGGAGAGAGGTGTTGTGGAGTGGGCCATTGACTCTGAC

TACAGGTGGTGCCCATAGTATTGGTGAGGGTAGTGATACTCGGAGTGGAAGGCG

GGGTCTCCGCTCCTGTGCATCATCTGCTGGCCGTCCATCATGTAG (SEQ ID NO:

27)

contig04401 Iength=249 numreads^

TGACTCTGCTATATTCGTTCAGAGCCCAAACTGCAACCAGAGATACGGCTGGCAT CCTGCAACCGTTTGCAAGATCCCTCCTGGTTGTAATTTGAAAATTTTCAACAACC AAGAGTTTGCAGCATTGTTATCACAATCCGTTGGACAAGGCTTCGAGGCTGTTTA CCAATTAACCAGAATGTGCACCATACGAATGTCTTTTGTGAAAGGATGGGGTGCT GAATACAGGCGCCAAACTGTGACATCCAC (SEQ ID NO: 28)

contig02932 length=398 numreads=7

TTGTTGTGGGCGACGGCAGGAACGAGAAGATGGTCACTTGAGAAGGAGAAGACG

TAAAATGTGTCGTCACGACGTGGAAGCGCCCCTGGGAAAGGCGGTCTGTAGAAG

TCGAGGGCGCTGACAGTCGCCCGCAATTTTTGAGCCTTGATTGGTTTCTTTTTGGG

AATGTTGTTGTGGAGTATGAGCTGCCGCAAGGAAGGTTTAGGATGGTAGGCAGC

AGACATGTTGTGGTACCTCACCCTCTTCTCGTCCACTGCTATCCACCTTCGGAGTT

CTTTCTCTAATCTGAGAGATTCGGTTTGATTTATCCGCAAGGGACAAGTCGGGCC

TTGCTGCGAAGAGCTGGTGATGTTGAGTGGATCTTCGGCCACCCACAGTAATGTC

CGGCCTATCCGCGT (SEQ ID NO: 29)

contig06264 length=792 numreads=99

CAACGCAGAGTGAAATCAGATCACACCCTTGCTCAAAGTGCGATTTTGTAGCTCG ATGTCCTGCTAGCTTGAGAAATCATCTGACGTTGCACACGAACGAGAGGCGCTAC TCGTGCAAGAAATGCAATTACAAAGCTAAACAATATTCTCATCTGAGGAACCAC CTGTTGATACACAATTCCATCAAACCACACGCTTGTCCGCACTGTGATTACCGAG GAAAGCGCATAGATGCCCTCAGGAATCATCTTTTGAGTCATTCCAACAATGCTGC

CGCGGAAACCTACTTATGTGAAGACTGCGATTATAAGACATCTAAAATGGCCAA

TTTAAGAAGCCACAGGTTGACCCATTCCAGAATCAAGTCATTCTCATGTTCCATT

TGTGACTATAAATCTTCTTATCAGAGCGACATTTCAAGGCATACGAACAACGTTC

ATAACAAACCTGATAAACGATTTTTTTGCACGGTTTGCAACTACAAAACGTCTAG

GGCGGATCACTTGAAAGCTCATGTCAAAACTCATTCTAGTCTTAAGAAAAAGTGT

CCTCATTGTGAATACAAAGCAGTAAAAAGTATATCTGTTAGAACTCATATCAAGA

AAATCCATCCGGACAAATCTTGAGATTGTTATTGCTTCTGAAACCAAGTTTTACA

TGTTACCATAGAAAAGCTTCGAGCTTCGACTGATTTTGAGTATTGGAAAGACTAC

CTACGCTTTCCTTTAGCTTTACAAAATCTAAAAATTGTATAATTCTTTGAAAAAGA

TAGCAATAAAATGATATTTTTTT (SEQ ID NO: 30)

contis02728 lensth=287 numreads=9

TTCTCGAAAGTGAAAGAACACTAGCAACAAATGAGTGGCGGATAGACCAAGATC

CCTGGACAAATAATTTTGGAGTGAGCGAAACCGCATACAAATTTGGAGGAGATA

CATACCAGCTCTGTTGGATTGGTCTATCATAGGTTGTACGATTCTGCGTCTCGCGT

TGATGAACCAATTGTTTACTTGGAGTATGGTCAGCCCAGTGTCCTGCGCTAATTG

CTTTTTCTGATCTTCAGAAGGGTAAGGGTGAGTGAGATGCTGAAAGAGCCAAGCT

CGGAGAATGTTGG (SEQ ID NO: 31)

contis06226 length— 511 numreads=55

GAATGGAGGTCGTCAGACTACAGGGAACCGCACGACTCGTGGAGCGGTCCTTCC

ACTTCCAGGGCGAGAAGCAACAGCTCCGTCAGAAGACGCAGGAAAGAAGGCGC

TGGAGATGCTGATCGCGATGAATGGTGCAGCCAGAAGATGACGCTGACGAACGA

GCTCTGCGCTGGTGTAGCGGAATGCAGCATTTGCGTGGAACTTATAGATGCGCAG

GAGCAGGTGTACCCGTGTAAAAAATGTTTCAACATTTTCCATCTGCCCTGCATTC

GAAAATGGGCAGATCAATGCATATCAAACAATGGCACGCATTGGACATGTCCAA

ATTGCAAATATCAAACCACGCAGACCCCAAGACCATCCTGCTTGTGTGGTCAAAC

CGCCAAACCCATATCTAGTCCCGGTCGTACGCCCCATTGTTGCTCAAAGAACTGC

AGTAGATGCAACAAGCCCTGCCATCCTGGAGCGTGTGATTGATGAGCAGGATGA

CCTACTACGTAATATACTTGCT (SEQ ID NO: 32)

contig04962 length=247 numreads—2

AAGAACTCAAGCAGCTCCCTAACATGCGTATGCACAAAACATATAGAAATGGGA AAGTTTGAAATGGAGGTCTGGTACCAGAGCCCCTATCCAGATGACTACGCGCGC CTACCAAAGCTGTACTTATGTGAATATTGCCTTCGATACATGAAATCGAGAAGTA CTTTAGACCGACACTGTGTGAAGTGTGTTTGGCGCCACCCGCCTGGTGAAGAAGT TTACAGGTGCATATTGGAATATTACGGTC (SEQ ID NO: 33)

contis04941 length=264 numreads—2

ATCGGGTTCCCTTTGATGCTCCATGAAAGCAGAGAATTCAACTAGCCGCAGCTTG

TGAGTTGCTATCGCTCGTCCTTCCCATGGTGGTGGGGCTGGTACTCCAAGCTCAG

CAGAACTCCCAGATACTGAACCTGTGGGTTTCCCACTTGGGTAGGCAGGTGCACC

AGAAAAGGGCTTGACGTCTTGAGAGGTTCCTGGCTGAAGACCTGGTTGCCAAAA

ACACTTTTAATTTGGCCTGAATCTCCCGGAGTTTTCTCCGCGCAA (SEQ ID NO:

34) contig04132 length=246 numreads^

GCAAGGATGCTTCCCGCAGGAGAGGTTGTTGGGGCAGCGGGAGCCGCAGGAGTA CGACGTGGGACCTCCCATCTCCGGTGGGACGGTGTGTCCGTCGCAAGTCAGATCT CGGTCGGTTCGACCGCAGTGGCACGTTTGGTGAATGGATTTGTCGCAGGGAGGA CAAGGGCCTTGATGGCACACGGAGGTACAGGGATGCAGCCCGCAGTTGAGCGCT CGCCCACAGACGTCCTCGCAGTCGAACGT (SEQ ID NO: 35)

contig00668 Iength=668 numreads=78

CGGACGGTTCTGAACGAAAAGCAGTTGCACACGTTGCGGACATGCTACTCGGCC

AACCCGCGACCGGACGCCCTCATGAAAGAGCAACTCGTCGAGATGACCGGCCTC

AGTCCGCGAGTCATCAGGGTCTGGTTCCAGAACAAGCGGTGTAAAGACAAGAAG

AGAGCCATCGCCATGAAACAGCAGATGCAGCAGGAAAAGGACGGGAGGAAGCT

GGGCTACGGGAGCATGCAGGGTATCCCCATGGTGGCGTCTAGTCCTGTTCGGCAC

GAGAGCCCACTGGGGATGAACCCGATCGAAGTGACGTCCTACCAGCCGCCTTGG

AAAGCCCTCAGCGACTTCGCCCTGCACACCGACCTCGACCGCCTCGACCCCTCCG

CACCGCCCTTTCAACACCTTGTTAACCAACTCCATGGCCACGGATACGACATCCA

CGGACCCGCGGGACCCGCGCCACACGAAGTCCTGGGCCCCAACACCGACATGGT

CCACCCGGACTCAACGGATAGCTACGTTACCTACCTCGAAAGCGACGACAGTTTG

CAGCATGATACATCCAGCCCATAACGGCCACAAGATTTGTCGCAGAAGTGTAAA

TAATACGATTTCACTAGAAATTTCCTCACGTTCGTCAAGGAACTCTTCCTGTTTTC

CAACAAGAACAAGCA (SEQ ID NO: 36)

contiε02090 length=290 numreads—5

TCCGCACCGAGGGAGATGCCAGAGTCACGCTCGTCCGAGGAGCCTTCGCCGTCC

CAGGGAGGGCTCGCCCGAGCGCCGGGTTCTTGCTTGATGTTGTGGAGGCTGAGG

AGCGCCGTCGCTGCCACGTCCTTGTCGCCGAGGTGGCTCTTGTGTCGGAGCTTGT

GAGGGAGGCAGGAGGCAGCTGGCGGCTCGAGCGGCAGAGGGAGCTCGTAGGGG

CTCGGCGACGACCGGCTTCGACTCAGGTTCAGCACTGAGTTGCCGGCGGCTTCGA

GGACCGGAGGGTTGAGCGG (SEQ ID NO: 37)

contis05463 lenεth=611 numreads=118

TAATTTAGCTTTGCAAAACACTTTTTAAGACCGTTGCCGTTTATATCGGAAGGCA

AACGTTTTTCTAGTCAAAACGTGTCGGGTAGCAAAGACCTTCACACAAAAATTGT

TCTAGCGCACACTTTTTTTAAACATTTTTTAAATTAATTTTTTTATGTTTTTTGCTT

TTTATGAACATTTAAACCGCCTGGAGGGATTTGAAGTCCTATGTTTTGTTGCTGA

AAATTGTGAGTTCAATTTTTTAAAAATTAAAAAACAATTTTTTTTCCTTCCTAAAA

CTTTTGTTACTCCTCTCATTTGGGACCCTGTGGGCTCTCGGGACCGCTGACTTTGT

CCGAACCGTCCTTCATTTTGTTCGCAATAGTCAGCGAATATGTCGCGGAAGCGTC

CCCATGACTTCTCAGGCACTGTTATCGCTGTTCTAAAATTGGTTTTAACCTCTGAA

ATGCGCATGTAGATACCGCGATTATTTTGGCCGATGTCGAAATAAAAGTTTTTAT

TATCTACTCGCATGTGCTTCCCCTCAGGTAGCTCCCCCTTGAACCCTCCGTCATCC

GTTCCAAATTCGACGAGAAGATCGGTGAGAGCGTCTCTAAATTCTATCATCCCCC

(SEQ ID NO: (SEQ ID NO: 38)

Translation factor: contiε00310 Iensth=243 numreads=4

AGGAATGGTCGTAAAACGCTAACCACTGTGCAGGGGCTCTCATCGGAGTATGAC TTGAAGAAAATCGTCCGTGCGTGTAAAAAGGAGTTTGCATGCAATGGAACAGTG ATTGATCATCAAGAGTACGGTGAAGTTCTGCAGCTGCAAGGTGACCAGCGGGAG AACATATGCCAGTGGCTCACCAAAACAGGCCTCGCCAAGCCCGACCAGCTCAAA GTTCACGGTTTACTAAAAAAAATACCT (SEQ ID NO: 39) contig05381 length=705 numreads=34

CCGGGTATTACAGAAAATTACGGCTTGTGTGATACTTAGCGTATCGTACAGATCG

CAAAGTGTTTCTAATTTCCAATCTTCTCTCTCCACAGAGACGTAAAACTGTTTGAT

ACCTTCAAGGGTCAACTCTTCCTTCTTGACCAAGATACGGACGGGCTGCCTCATG

AAGCACGAAGTAACTTCGAGCACGTCGCTCGGCATAGTAGCAGAAAGCAGGATA

ACCTGCACTTCCTGCGGCAGATTTTTGAAGACATCGTGGATCTGCTCCTTGAAAC

CTCGAGACAACATTTCATCGGCTTCATCCAACACGAAAATCCTGATAGTGTTGGT

TTTCAGAGCTCGTCTGGTTATCATGTCCAAAACTCTTCCAGGAGTTCCGACCAGA

ACATGGACACCCAATTCCAGCTTGCGCATGTCCTCCCTGACATTGGTGCCTCCAA

TGCAAGCGTGACACATAGCCCCCATGAAATCTCCGAGGGTCAACACAACTTTCTG

GATCTGCTGGGCCAACTCTCGAGTCGGAGCGAGAATGAGTGCCTGACATTCTCGC

AAGTTGATATCAATTTGCTGGAGAATAGAAATTGAGAACGTAGCTGTCTTTCCAG

TACCAGACTGAGCCTGGGCAATGACGTCATGACCCTTGATGCAGGGGATGATAG

CGCGCTGCTGGATAGCGGAAGGCCTTTCAAAACCATAGGCGTAGAT (SEQ ID

NO: 40) contis02633 length=462 numreads=8

AGTATCGAGGAATGTTAACTTATCCTTCCCAATCACAACTGAAAATGCACCAATG

TGCTGAGTTATACCGCCGAATTCCTGTTCAACTATAGAAGACTGTCTCAGAGTAT

CCAGAAGAGTGGTTTTCCCGTGGTCCACATGACCCATGATAGTCACCACAGGAG

GACGAGGCCTGGGATCAACGTAATCCGAAGGATTTCTTCGTCCAACATCTTTAAA

ACTTTTTTCTTCAGTTTCTGTAGGCTTACTAGCCATCCTACATCGATGACCCGACT

TTTTAACTATTTCTTCAATAACTTTGAAGTTGTCGATGACATCACGGTCTTTTCTG

TACTGTTCCGTGTTGTCAATGAACGACATAACTTCTAAAACGTGTCCTACATCTTT

ACCCAGTGTATTTGCTACTTCCGCCACTGTCATACGCTTCCACACGTCAACGGGC

TCGCCTTGTTTCCATTTTGA (SEQ ID NO: 41)

contig02589 length=242 numreads=7

CTCCTGAAGGCCCTCACTAATTTACCCCATACCGACTTCATTTTGTGCAAATGTCT TCTGAATCATGAGAAGTGCAGCGAATCTCCAATCAAGGATATCATCTATTTGGCG GACTTGCTGGAATCCTGTGAATTTCAAATATTCTGGCAGCAACTGCAGGACCGCA AAATGCAGGAATTGTGTTCTCGAATCTTAGGATTCCAGGATTCAATAAGGAAGTT TGTCTGTCACGTTGTGGGTAT (SEQ ID NO: 42)

contig02032 ϊength=252 numreads=3

CGGCGGCGGCGACCGGTTTGTTGGTGGGTTTGTGTTCCTTGAGTGGAGATGCTTT GGCTCTGTCGTCATATTCAATCTCCAAGTCATCATCATCGCTTTCGTCACTGGACC CTTCTTCCTCTTCAGCCTCTTTCAGCCATGTGAGAAACGGTTCAGCCTTGTTGTGT ATTTCCTGACTAAGCTCTTTAGACACATATTTCTTGCTCACTTTCCCTCCCCATTCC AACAGGACTCCTTCATCCAGTATATCCT (SEQ ID NO: 43)

contig01435 length=212 numreads=9

TGATGTCTCTTACATCCTTCCTGTTCATTGTCTGCCAATCATACAACCTGTCTATTT TGTTCAAAGCAACGACAAAAGGAGTTTTCCTGTCCTTCAACAAGTTCAAGGACTC GATGGTTTGGGGTTCAAGCCCATGCATGATGTCCACGACAAGGATGGCGATGTC ACAAAGGGAAGACCCTCTGGATCTCAGGTTGCTGAAAGATTCGTGA (SEQ ID NO: 44) conti201089 leneth=945 numreads=27

GGGAAATCTGGACGTCGCTCAGGGTGCGCTGCTCGGTCTCGTAGTGGAAAACAG

GTTGGAAATCACCAATTGCTTCCCTTTCCCAAAAACTTACGATGAAACTGTCGAC

GAAGAGGAGTACCAATTGGATATGATGAGGAAATTGAGGAGAGTCAATGTGGAT

CACTATCACGTAGGATGGTATCAAAGCTCGGACGTCGGTTCCTTCTTGAGTTTGC

CTTTGTTGGAGTCGCAGTACCATTACCAAACGTCGATAGAAGAATCCGTTGTTCT

TATTTACGACACACAGAAATCCGAAAGAGGGTTCCTTACCCTAAAAGCCTACAG

GCTCACCCCTCAAGCCATAACCATGTACAAGGAAGGAGAATTTTCGCCTGATGCG

ATGAGAAACCTCAAAGTATCTTATGAGAGCCTCTTGATCGAAGTGCCTGTGGTCA

TCAGGAATTCCAGCTTGACTAATATTTTGATGTGCGAGCTAGAAGAAATGATTCC

TTCTCGTAAAGGTTTCAACTTCCTTGATCTTTCAACCTCGTCTGTCCTCGAAGGAC

AGCTGAGGAGCTTGATGGAGCACGTAGATGATTTGAACCAAGAAGCGATCAAAT

TCAACAGGTACCAACACGCTGTAGTTAGGCAAACCCAAGACAAACATCGCTACC

AACAAAAGAGGAATCAGGAAAATGCTGCTCGTGCTGCCAAGGACGAGCCTCTTT

TGCCCGAAGAGGACATCAACAAACTCTTCAGGCCGATCATGGTCCCTCCTCGCCT

GGTGCCCATGATCATGTCTGCCCAAGTAGACGCCTATTCCCAACACATTTCACAG

TTCTGTTCTCAAGCTTTGGCTAAATTGTACATCATGCAGGGACTTCAGACACCCG

ACAAATAATAATCCAAGACCGAAAATTGGTATTGATTATACTTTACGTTTAATAT

TCCATTAAATCATAA (SEQ ID NO: 45)

contig06240 lensth=1462 numreads=126

TCCTTCCGAGAAACCAAAGATGTTTGCCAAGGATGCTGAAATAGATATGAACCT

AGTGCTTAAAAAGTTGGGAGAAATCGTTGCAGCCAGAGGTAAAAAGAGGACCGA

CCGGAGAGAGCAGATCGAATTGCTCCATGAGCTTGCAGCCGTTGCTGACGCTCAC

CATTTAGGTCCTGCTATCAACATCAAAATCAAACTGGCCATAATATCTTCCATTTT

CGACTACAACCCGAAAGTTTCAGACGCTATGAAGCCAGAATATTGGGCCAAACT

TCTGGAAAGGATATCTGAAACATTGGACCTGCTAATCTCGACTGAAGATATTACG

ATTGGAGAAAATATCACTGAGGAGAACGAAGAGTTCGATAAAGCTCCATACAAA

GTGAGAGGTTGTGTATTGACTGTTGTGGAGCGCCTTGATGAAGAGTTTACGAAGC

TGCTGAAAGAATGTGACCCTCATTCCAATGAATACGTTCAACGCTTGACGGACGA

AAAGAGAGTTTGTGTCATTATCGATAAGGTTCAGAAGTTTTTGGAAAGAACGAGT

GTTCCTTCTGATCTGTGTAGGATTTACCTTCGTAAAGTAGAACACTTATACTACAA

GTTCGATCCGAATGTTATCAGACAAAAGAAGGGAGAACTTGAGATCGGTTCGCC

CACTAGTATCCAAATCATGGATAAGTTGTGCAAATACATCTATGACAAGGATGTG

ACTGACAGGCTGCGTACGCGTGCCATCCTCGCCCATGTTTACCACCACGCTCTTC

ATGACAACTGGTTTCAAGCTAGGGATCTCATTCTCATGTCCCACTTACAAGAAGC

TATCCAGCATTCTGATCCATCCACACAGATTTTGTACAACCGTACTATGGCACAC

TTGGGTCTTTGTGCTTTCCGGCATGCGAATATCAAGGATGCCCATAACTGCCTTGT

GGACCTAATGATGACCGGAAAAACCAAGGAGTTGCTTGCACAGGGTCTTATGCC

ACAAAGACAACATGAAAGAAGCAAGGAACAAGAAAAAGTGGAAAAGCAAAGA

CAAATGCCATTCCATATGCACATCAACCTTGAACTTATCGAATGTGCCTACTTGG

TGTCAGCTATGTTGATTGAGATCCCTTACATGGCAGCTCACGAGTTTGACGCTCG

GAGAAGAATGATCTCCAAAACATTCTACCAACAACTAAGAAGTAGCGAGCGCCA

GTCCCTCGTTGGTCCTCCAGAAAGCATGAGGGAGCACGTAGTAGCTGCTTCCAAA

GCAATGCGAAATGGAAATTGGCAAGCTTGTGCGGAGTTCATCATCAACGACAAG

ATGAATGCTAAAGTGTGGGATTTGTTCTATGAAGCGGACAAAGTCAGAGAAATG

CTCAAGAAACTCATCAAAGAGGAGTCGTTGAGAACCTACCTCTTCACGTACTCGC

ACGTCTACGCCTCCATCTCAATGGAAACGTTGTCAGAAATG (SEQ ID NO: 46) contig00950 length=726 numreads=27

TTAAGGGCCAACATTTTTATTTACATCCCATAAGATCTTGCAATTCGTATTTTCAT

CATTTTTTCAACAAAATTATAAGCACTCCAAATTCTACTAAAAATTGCAATACAT

AAAATAAAAATGTATACAAGTAATATAAACAAAAATTAACCTTCCATCCCTAAT

GTATAGCCACAATACTTCAACTTAATGACTGTTGATTCACCATACATGTATTATA

CTTACTATTAAAAAAGAGTACGAGAAACCACCATATATATATTTTTAATAAATAC

GATCTAACGTCACACATTCCGGGAATAAATCAATTTTTCAAAAGTTTCCGTATGA

TGCGTTTAGAGATAAACTGAAATCGCCGATTTTAAATCATTTCAACTTGTAATTT

ATTAAATTTTTGTATATAATGTGTGTATGTGTTCAGAAATCTTGAGGAATTCATTG

AAGTCGACCCAAATTATCCACATCAGATAGCATCTACAGCATCCCCTTCACCTTC

GTCACTAACTTCGTAACCAAATTCGATATCTTCGTCGTATCCATCCTCCAAAAAC

GTGACGGTGTCGTTGATACGAACTGTTTCAGGGAACTCTCCATAAGTTTTGAGAT

TACGCGCCTCATCTGGAGTGTATTTCTGGATAACATCGGCTTTAGTATCCTGGTA

GTCCCTGAGGCCGATAAGAATAATGTCGCCTTGATTTATCCACACCTTTTTCCTAA

GTTTCCCCC (SEQ ID NO: 47)

contig00844 length=376 numreads=6

TATGGTTTTTATTCCTTGGTACGAAGTAGGGGCTTTCAAATGTTTTGGGTTCTTGA

GGCTTCCATCCTGAAGGATAAACTTCTTTGGGTTCAGGAGTCGGAACACATTTGG

GTGGTAGAAGTCTCTCTACGAATTTCCATTCCTCAGGATTCTTGCTGATCTCAAAT

TCAGTTTTATAGGTGGGCTCGAAAGGTTTGAGGAATTTGTAATTGGTACTGGTTC

TGCACATCATTGTACCTCTCAGCGAATTCGATGAAAGAGACTTCACAAGGGTCCT

AGCTAGTTTGGATCCCACTTCAATCAAAGCCATCGTAATTCAATAAATTAGAGAC

TTCCAGTAAAACGTGAAATCAAGCACTGGAAGCCCGTTGAGACT (SEQ ID NO:

48) contig00169 Iength=373 numreads=4

CATCATAAAATGGGGTAGTTTAATACGTAATATTAACAAAAGGCGTAACAAAAG

CGTGGATGAAGGATTTCAATCTCCCTCTTCAGCGCTCTCCGCGGCGTCCTCGTCAT

CGTGATCGCCGCCGCTTCCTCTTCCTCCTCATCATCGTCGTCGCCGGCGACTTCGG

CGTTCTCCGCTTCCGCTCTCTCCATCTGTTTGGCGAGTTCCGCTTCGTCCGACGCG

TCCACCACTTTGGGTGGTTTTTCAACGCGGAACACTCCACCTAGATTGTTAATTTC

GGCTTCTATTACATCTATGGCATCCTGTAACGCTTTCAGTCCGTCAGACTTCTCAG

GTGTCGCAGTGGTCATAACATACAAAGGAGGAGCTATCA (SEQ ID NO: 49) contig05794 Iength=521 numreads=30

ATTTACAAACTTTGTATAATGTGTACCTGAATGGATATAAAGGCTTTCCAAATGT

ATTTGCACTCCAATCTGTTCTTCATCTAATAAGTATTTAACAGTAGTAGGTAGACC

TTCAGAGTACGCCTTGAACAGTTTTGAATGTAAGAAACATTATGATTGAAGATTT

TAATAAGCCCTCTGCTGATCGCCTCTTTCTCTTCGTCCCCGTCCCCAATCTTGACC

AGGGCGGTTGAAATTCTGCCTGTAAGGTCCTTGTATCCCACCACGTTGGAAGAAG

TTCCCTTGCTTCATCTCGAATATTCTTTCGTTCGAGTCCACCAAATTGTTGACCTT

GTCAGCCAATTGGAGAGCCAAGGACTGCAGACGACTGGGCTCACTTCTGTGCAT

CACTACAGTTTGAGAAGGATCGTCCAAAGAAGCCATCAGTTCCTCATTGATTATC

ATTTTGCTGACAATAGAGTGAATAACAAATCGGTCCAACTGAAACATTTCTGACA

ACGTTTTCCATTGAGATGGAGGCG (SEQ ID NO: 50) contiε02102 length=280 numreads=2

AGGAATACGCGGACTGTGTCAAGAGGGGGAAGGAGAATATTTGGATCCTTAAGC

CTACGTCGTCCCTCCTCCTAGAAGAGGACTTAAGGAAGTTGGTGACCCCGGAGA

ATATCTGCCAGTACGAGTCGATGATAGCCGGGGAGAGACGCCTGGAGGACTGTG

GGTATAAAAGTAGTAAGGGGGAGGAGAGTGAGGAAGAGGATGACGTCTACTCC

GTCCCATGGCAGTTGAGTAGGAATTTCATAAATGCGGCTAATGGCAAGGGGCTTC

TTGAACTAAG (SEQ ID NO: 51)

contig00970 lensth=579 numreads=10

CGGCTTCCACGGTCTTCTGCAATTCAATCACAAACTCAACACTTACATCATCAAG

AACCCGCCCAACTTGAACCCAGGAACTCAGGGCGCTTCTTTGACTGAAGGCAAG

CGCTCAAAGCGTAAGGGCAACAAGCAGAATGGTGACGCCACCAACGGAGACCG

CTCTGGATCTCCCAAATCAGAAGAAGAACCTGAAATCATTGTGGCACTCGGCAA

GAAGTCATCTAAGAAGGACGACGACGATGATGAAGTTGACTGGGCGGTCGATAC

ATCCGAGGAAGCCGTCCGAGCCCGACTGCAGGACCTCACTGACGGAGCCAAGCA

TCTCACTCTCACTGACGATTTGGAGAAAACTGAAAAGGAGCGCATGGACATCTTC

TACGGTTTCGTGAAGCATCGCAAGGACACTGGAGATTTAAGCAACATTGCGACT

GCCAAGGAAATTCTGTCAGAGGCCGAGCGTTTGGAGATCAAGAGCAAAGCTCCT

TTGGTCTTGGCCGAACTTTTGTTCGATGACAAAATTCACGTTCAAACCAAGCGTT

TCCGAGGACTACTTCTTCGTTTTACTCATGAGGATGC (SEQ ID NO: 52) contJ206228 length=1144 numreads=48

AGGAGGACTGGATCCACACCTCAAATATTAATTTTATTCAATTAGAAAATGGTTA

CATCTTCCGGATGGATTATATTTATTTACTGGGTAGGTATGATACTGCACGAAAA

TGTTTCTCCAGACCACATATTACTGGAAGAACGTTCCTGCATCTTCTTCTTCGGCA

TCTTCGTTCTCAGATTCGAAGGTGTTGTCAGGAATGTCATAGAGACGAATGAGGG

GCTTGTTAGGGTCCTTCATGATGAGGTATTTCCCTCCCCTTGCTTCATGCACAAGT

CGATGATGCAACGGAGGATCCCCCAGGCGTTGTCCATGTTCAACTTGATCTGAGT

TGCGAACTCCGCTGGTTTGTACTGCTGAGTCCCGAGGATGACGTGCTTCGAGGAG

TCGCGAACGTGAGCGCGGGAGACGTAGCCAAACTTGACTTGGTCAGAGCCGGCC

AGGACAGCCTGCACAGTCCACTTGGCCAGCTTACAGGCGTTGTTTCGGAGCTCGT

TGGCTAGAACTGCACCTCGCTGGATATCCAGTTTTTGCCTCCATTCGACGCCACC

AGAGAGCTTGGAGTCCCATTCATTGAGCGCCTTAATTGTCAGGAATTGAGTGTCA

CCAGCAGGTGTAGCAAGGACAGCATCGTGCTCGCACCTCGCCACGAGGATAATA

CCATTGTCAAGATCCCATTTCCTGTAACGGTATGCGACTGACGCCTCCTCACCTTC

TTCTGGGGAAGTAATGAAGGGATTGGGATTATCGAACTTGTAACGGGGCTCCCC

AGTTTTCAAAACTTGCTGAGAGAAATTGTGGTTGATGAAAGTAGCTTCGAGCGCC

AGGTTCCTTGGGCTGTTGAGACTGTTGGAGTCGTCCATAGGCGGATCCACAGAGG

TCTCGTTGACAGTCAAAAGATCGAATTCAGTGTTGTCACGCTTATCAAAGAACAG

CTTGCCTCCGATTTTTTCAATGACGATGTCCCAGGAGTAATTGGACCGAGTGCAG

CACATCAGAGTAGCCAATATAGCATCAGTAGCATAAACGTTGCCTTCAGTTTTAG

AGAGCTTACGAATGATAGGGTCGTCAGTAGTAGTGACAGTGTGGAAAATCCTAT

CGATACGCTGCAGCGGTTTCTCATTCTTGACGTTGACCCGGCCGT (SEQ ID NO:

53) contiεOS347 length=377 numreads=143

CCGATTTGTATCTCCTAAAAAATTGACTTAGCAGATCAGATCTGCAACGTTCATC GGCATTTCTTCTATGATAGTGTTGTAAAACTGTTGAATGTCCGTGAGAGTACGCT TGTCATCTTCGGTGACGAAATTGATAGCAACCCCTTTACGACCAAATCGTCCTCC TCGTCCGATCCTGTGAATGTAGTTTTCGCGGTTCGAAGGTAAGTCATAGTTTATA ACTAAGGAAACTTGCTGAACGTCAATGCCTCGAGCGAGAAGGTCAGTCGTAATC AGGACACGGCTGGATCCTGTTCTGAAAGCCTTCATAATCAAGTCTCTTTCCTTTTG TCCCATGTCTCCGTGCATAGCAGACACTGTGAAATCCCGGCCGTAAT (SEQ ID NO: 54)

contisOllOO lensth=1317 numreads=45

CTTCAACTCAACCATGAGCTGCCAGTCCGTTTTCACTAAGTCGTCTCGGAAGTAG

TCTTTGCAGGCGTCGATGGATTGCCGGGAGATTTTGGAAATTAGGAAGTCTTTTT

CATCAAAGTGGCGCCCGAACATTTTAGGGGATTCGCCAGGAATGGGCTCAATCTT

TATGCACACTTCCATCCCTTTCCGCGCACTCTCCACCGGCTTGTGGTTATTTTCTA

TGCTCGTAACTATTCCAAGATCCACGAATTCTTTGCTAGGGACACATATCGGAGT

CCCTTCTTTGACTATTCCAGCTTCGATAATAACACCCATGACGATCGGGTCACGT

GAGTTGAAAACGAACTGTGGAAGGATTTTAAGTTTGCAAGGGAAGACAGCAATC

GACTTGAACTCCTCCCTCTTACGGGCCTTCAACTCTTCGCGGAAGGCCATGAACT

TGTCGAAGAGGTGGTAGATGATGTCCGCTTGGAAGATCTTCACGCCCAGCGTGTC

AGCCAGGTCTTGCGCGTCTCTCTCGATTTTTACGTCAAAAGCGAGAATCACGGCG

TATTGGCTGTCATGCTCCAACATGATGGAAGCCTTCATAACATCTTTTTTGACAAC

TGGACCTATCCTGATGTTGGAGTAAGGTATCGAAGAGGTCCTAAGGAATTCGAG

AAGAGCTTCGAGGGAGCCGAGAGTCGATGCCTGGACGTAGACACCACGTTCGGC

CATTTTGATGGAGGCTCAGAGCGTGTTTTAGTTCCCGCGCTACCTCCTCTTTTAGA

ACCTCAACTTCATCGGGAAATTTCGCAACCCGGAGATTCAGTCCTGCTATAACTT

TTTCTAGATCTTTAGCAGCAATTTTTACACCCTGTGCAGCCCTTATTTCTTTGTATT

CGATATACGCGTTCTTAACTCTAAGTTCCTTCATGGGTTGAGGCATGAGGAGGGA

TCGGATCTGTGTGACTATGGGGGCTTCCGTGCCCGCCAGGATGGCTGTGTCTCCT

TCTCTGAGCGTACCATTTACCAGAATGACGTCGATAGTCGTTCCCAGCCCAGAAA

TGGCTTTCACCTCGAGAACGGTAGCCTGCAGTTCTTCGGAGAACATCAGACGTTT

GGCAAGCATCGTCTGACATGATTCAACGATAAGAGCCAGGAGGTTCCCCATACC

TTCCCCAGTGACGGCGGACGTGGGAACGAGGGAAACGTAACTCCTTACGTCTGG

ATTATTATAGAAGAGAGCAGCATTCATACCCTGTTCGGCAAACTGGAGGATAACT

TCGTTAGTCCTCTGTTCAAACTCCATTTGAGTGTTGGGGGCTTGTGCTTTGA (SEQ

ID NO: 55)

MicroviUa proteins: contis02056 lensth=449 numreads=19

TTCAAGGTCGTGGTTGCTTCTGTGGATTTTCCTCCTTCCGTCATTTCCTTGTCTGGA

AGCGTCAGCAGATCTCCCAGAGAATCTTCCTGGGCAGCCCGTTTCTGACGAGCAC

CGTCAGTATTGATGGTACCAGTCGTATTATTTTCTTCCGCCGTTGAATTTTCTTGA

ACTCCACTGCCTTCCGATGAGTCAGGTTTAGAAACTGCGGCAGCATCATCTGGGA

CGCCACGAACTGGGCGAGGTCGAATTTGCATAATGTGAGACTCCGATAATACAG

TTTCCATTACATATGGTGAGGATTCAGCTGTCGCAGGCGTTTCTTCATCAATTCCA

GGCTGCGTTGATGACGCTGGGTCACTGCTAGCATTGATAGAGATTGCAGCAGTAC

TGGAGGTTACTGTAGCATCGCTAGTATTGATGGACAATGTGGTAGTTGTTCACTA

GCATCG (SEQ ID NO: 56) contig01526 length=314 numreads=12

GCACCAACGGATGTTACCACTGTAACTGAAATAACGGAGTCTTCCTCTGTTACTG GATCAGGAAGTGAAACTGTAGGTACCAGCTTGATGCCACCTCTACGATTCCTCCA ATGCTACAGAAGCAAAGAGCCATCCCCCCATCAAACGGACTGTTCTCTATTTTAC TCCTGTTCAAATAACGAACCAATTCTCATGAAGTGTAGTCCTGGGATGCATTTCA ACAAATTATTAAAAATCTGTGATGTTCCTCAAAGAGCTAATTGTGATTTGCCATA TTAGTATTACTTATATTAAATTATAATATGTATAATATG (SEQ ID NO: 57) contig01939 Iength=333 numreads=7

CAATATGGACATCAGCCCAATGGACAGGACCAATATGGACAGGACCAATATGGA

CAGCCCAATGGACAGGACCAATATGGACATCAGCCCAATGGACAGGACCAATAT

GGACAGCCCAATGGACAGGATCAATATGGACAGCCCCGATATGGACAGCCCAAT

ATGGACAGAACCGATATGGACAGCCCCAATATGGACAGAACCGATATGGACAGC

AGCCCATGGACAACAGCCCCAAAACCGACCCCGTTCCCAACCAACATCCAACCA

AACCCCCAACTATTACAACAACAACAACAACAGTGACTACCAACCTCAATCACC

ATCCCATTC (SEQ ID NO: 58)

contig03364 length=521 numreads=10

CTCCCGACGTCGTTACGCGTTTGAAGATCTCTGATGTAGTCCCGACATATTTTTTC

CATCTTACTATCACATGGGTTTGATTTACACTTGTGACCACCAACTCTGCCTCCGC

AATTTGGATTTTGGCAATTACAGCGTTTCTTCCGCCGGAACAAGCAACACCAGCA

AGGACAGCAACCTCTCTTGTCATGGACGCTAACGGTGGAATCTTCTATAATAACC

TGACGTTCAGGACTACAAACTGTTTTATGGGCTTTAGTGTCTTCCCACAGCAACA

ACAGATACCCCTCCCCCTTACAGACCGGAGACACTTATACACAGAAGTTCCACAT

GGTTTTTTCGGTTTCTTGATTTTAATACGGATTCTGCTTTTCCCAATTAGCCGTTTT

TTCATCAGTTTCCCTCCCTTTCCTTTTATCGTCAGACTTGGTTCCCGATAGAATAA

TTCTTCTTCATCATCACTTGTCTCTGACATACCGCTGCTTGAACAGAAACGACCGA

AAGCTCTCTTGAAAATACAC (SEQ ID NO: 59)

contig04612 length=275 numreads=2

TGGGCTGGTGGCGTCGGCGCAGGAGCTGGCGGCACCAGCCCAAGTAATGTCGGT

GGAAGGTTCGCTGACGCCAGCAGCAGCCCCTCCAGCCTTTGAATCGCCACCACCT

CCTCCCATTGCGTCGCCCCCCAAACCAGTCAGTCCTCCTGCTGTAGTCGCGGCAC

CCCCGTCCCCTCCACCGCCGGCTCCCCCGTCGCCACCTCCGGTCGTGGAAGAGCC

GGCACCGACGAGACCTGCCGAGGAGGCGACACCTAGCGAACCAGTTGCCGTCCC

CG (SEQ ID NO: 60)

contig05924 length=388 numreads=8

CTATCACTGCTGCCACCATCCACAATATTCTGAGCACATTGTGCCAACTTAAACT

ATTTCAGTCTGTGCGCTATACATCCCCTAAGAAGAATTTAATGCTTGGTGCACTA

GCCACCCTATTTGTAATAAGTAATGTAGGTATTGCAATCGTTATTTATCATGTATT

TTTGAATTGATTTTAGAATCAATTTTCACTTCCTATGTTCATATCTAATAACATTC

CCTTGAGGGTTGTGTTACAAAATACTGTTTAATTTGTTAGTTCATGAATGGTGTTG

TTATCGTTATTTTCAAACTTTTATGTGTAAATTGTTATGGTTCCACGTTTGTCTTGT

TAACACCTCTCTTGGGGTACCTTTTTGAACTATATTGTGCTGGAGTTTTGTCA

(SEQ ID NO: 61)

contig06813 length=444 numreads=268

GTATCATACCTGGTGAAGTGACAGACGCCGGCAGTCATGATGGCAGGATTATAC

TCGGCCTACGACCACGACGCCAACTACGACGCGGCCTGGCTCCTTCCTCAAGACT

ATCACACGGACGAGGGATATGGGTTGTTGGACTCGATCAAGATGCCACCTCAGG

GAAGGTCCGGGTTCCACATCGTCGACATACTTGATCTCAACGACGCCGCCAAAAC

ACAACACACGGATGGCGCCGCTCTCAACCCCACCCAAGCTCACGACGATATTTCA

GGAGGTGAAATGACAAACCTCCAGCAGCTTGGGAACCACCCCGGCCTGCTGTTC

GCCGGTGGTCAAGTCACCACTCCCATGGAAGCCCTTCACCATCATTGGGGCAACT CCATCACCGAACTTAGAGGTCACCAGCAAGCCAGCCCCGACAGCACATCACCCG GACCGTAA (SEQ ID NO: 62)

contig06941 lensth=155 numreads=40

AGAGTCGCTCGGGCAAATCATCGTGCTGCCGCAGCCGTAACTGTCACAGCAGGTT CCTGGCTCTCTGACTGTCGGAGCCACTGTCTCGAGCTTCATGGTGCCACAGTTAG GCGGCAGAGGACACATCGAGGGCGAGCAAACGCAAACTCCGGCCG (SEQ ID NO: 63) contig06960 length=154 numreads-53

GGAGTCGCTCGGGCAAATCATCGTGCTGCCGCAGCCGTAACTGTCGCAGCAGGTT CCTGGCTCTCTGACTGTCGGAGCCACTGTCTCGAGCTTCATGGTGCCACAGTTAG GCGGCAGAGGACACATCGAGGGCGAGCAAACGCAAACTCCGGCC (SEQ ID NO: 64) contig06961 lensth=155 numreads=31

GGAGTCGCTCGGGCAAATCATCGTGCTGCCGCAACCGTAACTGTCGCAGCAGGTT CCTGGCTCTCTGACTGTCGGAGCCACTGTCTCGAGCTTCATGGTGCCACAGTTAG GCGGCAGAGGACACATCGAGGGCGAGCAAACGCAAACTCCGGCCA (SEQ ID NO: 65)

Electron transport protein: contis00337 lenεth=632 numreads=82

TTTTTTTTTTGAAATGAAAAAATATATATTTAGTATAAAAATATCAATGTTTTTCC

CCCTTACGATGCTTATCTTCCAAAGGAAATTTCCTGCTGTGGTGAATATAAATGA

AAAGAGAGAGACTGTGCAGAGCAGACCATTTTCATTCAACCCAAGAAGGGTATT

CAATAGTAACAAAACTAATAGGTAAAGTACCTATTTATGAATACTTGGTCCGATT

GATGTGTTTTAGTTTGGTGAAGGGTGATTTGTCTTTGGAACTCAAATTTTTTCGGT

GAGTTCTGGCACTGCTTTGAAGAGGTCAGCTACAAGACCATAATCAGCTACTTGA

AATATGGGCGCCTCAGGGTCCTTGTTGATAGCCACAATGGTCTTGGAATCTTTCA

TACCGGCCAAGTGCTGAATAGCACCAGATATCCCAACGGCAATATAGAGCTCAG

GCGCCACTATCTTGCCAGTTTGTCCAACCTGTAGATCATTAGCCACAAAACCTGC

ATCAACTGCTGCTCGGGAGGCACCTACAGCAGCGCCCATCTTGTCAGCAAGAGT

ATAAAGAAGTTGGAAGTTATCTCCAGATTTCATACCTCTTCCTCCACTAATTACG

ATTTTTGCGGCAGTTAGTTCCCGGCCGT (SEQ ID NO: 66)

contig06602 length=413 numreads=384

ACGGTCTGGGCGAGAGGGCCAAAATCCCCGATTGGACCATCTACAAAGTGGAAG

ACTGCCCTGAACTGGTCAGACACCGCGATAATTTGGCTTTGAAAGGCCTGAAAG

ATCCTTGGATCAGAAATGAAGCCTGGAGATTCAACCGCAAAGAGTTCGGATCTG

CTGGCTCGAGGGCCCTGCTTGTCTTCAGAGGACTGAAATGGGGCGTGTTGGCCAC

CATCATCACCGTAGGCATTGAGAAAGGCCTCAACTATGGCAAAGACGATCACCA

CGGCCACCATTGAGTTTTTTTTAAATCAATATCTATGTATTTCTGTTGATTGTAGA

AGAGTGTACATTTCGTAGAAATCTATCTACCTATACTTATTATTACCTACCTATTG

TTGTTAATAAAATACATCTAAAAGAACTAT (SEQ ID NO: 67)

contig01234 Iength=447 numreads=8

CCCCAACGAAGAGGCTGCCAGAGCTGCCAACAATGGCGCCTACCCACCTGACCT TAGTTACATCGTTCCTGGAAGACATGGAAAGGAGGATTACATCTTCTCATTGTTG ACCGGTTACTGTGATGCGCCTGCTGGTATGGTGATGCGTGAAGGACAATATTTCA ACCCTTACTTCCCTGGTGGTGCTATTTCTATGGCGCAAGCCTTGTATAATGAAATT

ATCGAGTACAGCGATGGTACTCCCGCGACAGCCTCACAACTCGCCAAAGACGTA

GCCACGTTCCTCAAATGGACCGCAGAGCCAGAGCATGATACTCGAAAACAGATG

GCTATCAAGATCATTGCTATCTTCACGTTCTTCTGCTCCGTTGCCATCTACTGGAA

AAGACACAAGTGGAGCACAATGAAATCCAGAAAAATCGTCTACCAGCCTCCGAA

AAATAATGT (SEQ ID NO: 68)

contig00750 length=493 numreads=24

TCTGTATCTAGAGAACGTTTATTAGTGCTTTAGGTAACTACCAAATTCATCATAAT

AATGCCATAAGCATCAATAATAATTATTATGCCAATTGTTCTTTGATGGGTAGTC

ACTTTTATCAACTCCAAACTTCTTGCTGGTACCGGTTTTCTCTGTCCATCTTCTCCA

ACCATTTGTCAGCTTCTGAATCGGACATCCTCCCAGCTTCAATGGCGGACTTCTTA

ACTGCGTTGCGCACGGCTTCTGGCATGTCTTTGGAGCTCCCTGCCACGAAGAAAT

GACCACTGTGAGGAGACAAGCTGGTTCACAATGAGCTCTTTTTGATCCAAAAGTA

CGTGTTGGACATAGATTTTGTGAGGTTGATCTCTGGAAAAAGCGGTGAAAACATT

GAGTTGGTGGACCATAGCCCAATCTTTCCTGAAGAAATAGTCGGCTTCTCGGTTC

CTGCTGCCATAGAACAAAGTTAGCTGTCGTTTCTCTGCACCAGACTCTT (SEQ ID

NO: 69) contig02025 lensth=357 numreads=10

AGGGTTTACGTGTATTGTGGTAAGTTGGTGGAACTCAGGTTCATTTGTGGGTGAG

CACCACTTCCTTCAATTTGTCGAAGTTGCTTCCGGAAAATGAGGTCAACTTCTTCT

TTTCTTTGATGAAAACGAAAGTCGGCATAACAGAAATCTCGTAGGCAGCAGCAA

CGTCTTCACACTCGTCGACATCGACCTCAGGAAACGATTTCAGGGTGCGCCTGGG

CGAGTTCCTCCAGTTTAGGAGCTATCATCTTGCATGGGCCACACCAGGTTGCGTA

GAAATCGATGACAACTAGGTTGGTTCCTGCTTCGGTAAGCTTAGCTTCCAAATCA

TCCCTTCGACAGACTAAAATTCGCTTT (SEQ ID NO: 70)

contig04863 length=265 numreads=3

TCACGATTTCAAAAGCCGAGGAATCGAAGTGTCTAATGTAACTTTGAACTTGGAG GCCATGATGGCTGCCAAATCCGGTTCAGTTAAGGCCCTCACTGGTGGAATCGCGC ATCTTTTCAAGGCAAACAAGGTTACCCTGGTGAAAGGACACGGAAAAATCACTA GTACCAACGAGGTCACTGCATTGAAAGAGGATGGATCGAGCGAAGTCATCAAAA CAAAGAACATCCTCATCGCTACTGGCTCAGAAGTCACACCTTTCCCG (SEQ ID NO: 71) contisOόllS length=1689 numreads=200

GGCCGGGACGTTGAGCCTGTCGCGTCTTGCGATCGTCAAGCCAAATTTGGGGTCT

GTCGTTCCCTGTTTGACGGTGCAGAGCAGAAATCAGTCGACTGAAGCTCCAGTCA

CCAAAGCGAAATATGGACCTCTTGCCGACGAGGACAGAATTTTTACCAACCTGTA

CGGACGACATGATTGGCGACTCAAAGGTGCCTTGAAAAGGGGTGATTGGTACAA

GACCAAAGAAATCATTGAGAAAGGACCTGATTTCATTCTGAATGAGGTCAAAAC

ATCTGGCTTGAGAGGACGGGGTGGAGCAGGCTTCCCATCGGGCATGAAGTGGAG

TTTCATGAACAAGCCATCTGACGGAAGGCCCAAGTACCTCGTCGTCAATGCTGAC

GAAGGTGAGCCAGGAACCTGTAAAGATAGGGAGATCATGCGTCACGACCCGCAC

AAGCTTGTTGAGGGGTGCCTGGTTGCTGGGAGGGCGATGGGAGCTCGTGCTGCTT

ACATCTACATCCGCGGAGAATTCTACAATGAGGCCTCCAACATGCAAGTGGCCAT

TGCTGAGGCCTACCAAGCTGGTCTGATAGGCAAGAATGCTTGCGGATCAGGCTA

CGATTTTGACGTGATCATGCACAGAGGAGCCGGAGCTTACATTTGCGGCGAAGA

AACTGCTCTCATCGAGTCTCTGGAAGGTAAACAAGGAAAGCCCAGGCTCAAGCC GCCATTCCCTGCTGACATCGGTGTCTTTGGATGCCCTACCACTGTTGCCAACGTCG

AAACCGTGTCAGTCGCACCTTCGATTTGCCGCAGAGGTGGAAAGTGGTTCGTTGG

TCTGGGCCGTCCCCGAAATTCCGGAACTAAATTGTTCAACATCTCGGGTCACGTC

AACAATCCTTGCACAGTGGAAGAGGAGATGTCAATACCGCTGCGCGAGCTGATA

GAACGGCACGCCGGAGGTGTGCGTGGCGGATGGGAAAACCTATTGGCTGTCATC

CCTGGAGGGTCATCTACACCTCTGATTCCACAAAGTGTCTGCAATGAGGCCATAA

TGGATTTCGATGGGTTGGTTGCGGTCCAGACGAGCTTGGGAACTGCTGCTCTGAT

TGTGATGGACAAGTCGACGGACATTGTCAAGGCTATCACTCGACTCATCATGTTC

TACAAGCACGAGTCCTGCGGCCAGTGCACACCATGCAGGGAAGGAATAAGCTG

GATGAACAAGATCATGCACAGGTTCGTAACTGGCAACGCTCAGCCCTCTGAAAT

CGACATGCTGTGGGAACTGAGTAAGCAGATTGAAGGACACACCATCTGTGCTTT

GGGAGACGGTGCTGCCTGGCCTGTACAGGGTCTGATTCGCCACTTCCGACCAGAG

CTGGAAGAGAGGATGCGCGTCTTCCACCAGAAGGACAAGAAAGCAGCAGCCTCC

AATTAAATCTCCTCCTCACTCCAAAATGTAGAAACTCCACCTCACTATTATGGTA

TTACCTCTTGAGAGGAACTCTACTAGGATAAAGCATAATAAATAAGTACCTAGCA

TTCGAACTCTGCTGTACGCGCTTAACGTTGTAATTTATTGTTGCAAAACAAGTAG

GGGATTTAAAAACAGGAATGTTGATTCTATTCAGTTCTGCTTTATCCTAAATATC

ACACATGAAATGTATAATGAAACTCGAAAATGTTAATAAAGTTATGTTTCAA

(SEQ ID NO: 72) contiε05464 length=362 numreads=6

_{TTTTTTTTTTTTTACT}^_{CAAAGTGTTTCAATGTATT}^

ATGAGACGATCAACAATGACGAAATATAGGTAGTTACTTATTTTGGCGTTGGTTG

ATGCCAGGATGAAACGGATGTTTGATAACTGACTAACGAAGCGAGTGGTGACAC

CCCAGAAGGAAACTGCCGAGACACTTCACTTCTTTTCTTTAGGGGCGTCCGCCTT

CTCGACGGGGATTTCCGAAGAGGCGTCTTTGAAGTTGGGGAACTGTTCCCACGGA

GTCGAGAGATCGAATTTACGGAACTCTTGAGCCAACTCTAGCGGCTCGCAAACA

CCCTTTTCCTTTCATCGTCGTAGCGGACCTCGA (SEQ ID NO: 73) contiε01256 lenεth=352 numreads=28

ATACTAAAAATTATCATATCAGTTTCAATAACAATTATATTATCAATCATCATAA

TAACATTATGTATATTAATCAGAAAAAAATCAATAATAGATCGTGAAAAAATAT

CACCGTTTGAATGTGGGTTTCGACCCAAAAAGATCATCACGAGTACCTTTTTCTT

CACAATTCTTCTTAATTGCAGTCCTATTCCTAATCTTTGATATCGAAATCGTAATT

ATCTTACCAATAATCATCACGATCAAAACAAGAAATATAATATATTGATTTATAA

CATCAACCATATTCTTAATCATTTTAATTATTGGATTATATCACGAATGAAAAAA

TGGGATCCTAGAATGAGCCAAC (SEQ ID NO: 74)

contis05979 Ienεth=375 numreads=12

TTCCTTTATTTTTTGTTGATTTAGATCTTGTTTGGCAGAAACAAATCGGTCACCCT

TTGATTTTGCTGAAGGTGAATCTGAGTTGGTTTCGGGATTTAATGTCGAATATAG

AAGAGGGGGATTTGCTTTTATTTTTTATCAGAATATATAAATATTATTTTTATATC

TTTGTTAACTTGTATTGTTTTTATAGGTTGTAATATTTATTCTATCACCTTTTTTTG

AAGTGGGTTTTTTTGGTTTTTTCATTTATTTGGGTTCGCGGGACTTTACCCCGCTTT

CGTTATGATAAGTTAATATATTTGACTTGAAAGATATTTTTGCCTTTATCTTTAAA

TTATTTGTTATTTTTTATTGGATTATATTTCTATATAA (SEQ ID NO: 75)

contiε02238 length— 224 numreads=7

TGAATTTTCACCCCATCATCATCTAAATCAAGCTCCCATGCTTGGAAGTGGTTTCC CTGAGCAGCCAATAGAGCTTTAGCTTTTGTACACCAGGGGCACCAAGACTTGCTG AAAATTGCAATCTGATTACCCTTAACGAACTCTTCGATTCTGTTGACAAAAATCT TGCTGTCTGTGTTTGAAGAATCAGATGACCTTCCAACGTTGATCAATTCCTTAAGT CG (SEQ ID NO: 76)

contiε06162 lensth=307 numreads=20

CGGCAAAGGTCCAGATCAGCCATGCGGTGGCATTAGCGGAAGATACCTCGGGCG

GGGCAAATGCGGCTCCGGAATGGAATGCAGCGGAGAATACAAGGGTTACTGCAA

GCACTGCTCATCGGTAACGCTGGAATGTGCCAAATGGCAGCACGCACGCAGAGA

ATTGGATGATATTGACAGCGGGTTGATGGACTACGGGTTCTGAGCAATAACAAA

ATACACCCAAAGTCTCCACCAACTGCGACTTCAAAGAGGAAGATGAAGACTATA

TCTTGTACCATTATTTACGACATTAAACATGCTCGTA (SEQ ID NO: 77)

contig01186 length=1349 numreads=17

CTTGAGAGAACCTTCCTTCACATAGTTCTCCAGCTCTTCTTCGTATAAGAAGTCTT

CGCTCTTCTTTCTGCATCCAAAATACAGTATCGTTTCTCCAACTGGTTTACTGTCT

TTTTTGGCGAGGTGGCGCTCCTGAATGAAACCCCGGAAGGGAGCAAAACCTGTT

CCTGGCCCGATCATGATAATTGGCGTTTGTGGCCTTGTAGGAAGTTTGAATTGCG

ATTTTCTGACGAAGATAGGAGCAACCACAGGGGCGACGCCATTTTCTGAAGGTA

CTTTGTTAGACAGCCAGGACGTGCACACACCCTTGTTCCTGCGACCAGTAGGAGT

ATCGTACTCTACTTTGACTGCCGTCACGTGCACTGCCGTGGGAGAAACCTTGGGG

GATGACGAAATTGAATAGTAACGACACTGGAGTCTGGGAAGCAACTCGCAAAGG

CGATCCAATTGAGGTTTGACGGAAGGAAGATCTTCAAGGATATGGACGATGTTA

CGGTTGTCCTGAACTATCCACTGTTGGTACAGAGCTTTGCCCTCCGGGGTTGTGCT

TGCCATAAGCTTCAATTTTTCAGCTTCCTTGGGATCTTTAGCGTACTCTGCCAATT

CTTTGAGGATGTGGGTTCTGGGGTTTGACGTGATGTCGATGTAGTAAGTGAGAGC

CGTCTTGTAGGAGCACGGACAAGGGAATGGGTGCTTCTTACTAGATTCCTCATCT

GTATTCGTGAGAGTGAAAATAGTTTCCAAAGCTGTATTCGACATATCGAGACATT

TTCCAATCCTGTTCACAAGCTCCTCGGAGTTCTGTGGGTATATCGCGAGATGGTC

GCCAACATCATATCTGATTTTAGAGCCCTCAATGTCGAATTCAATGTGCATGCAG

GAACGATCGCCTCCTTTGTACAACTCGCGGTTGACTTTGATCGGTGCCAGGTAAG

GGTTCTTCAGGTCGAATGGTGGTCTTTGATTGGTGAATGAGTAAAGCCTGGCGAC

CTCTCCGGTGTACACACGTTCTGGGATCTCCTGGATGTGTTCAGTGAGTTTGTATT

GACGGATGGACACATCTTCTCCTGCTCCTTCGATACCGAAGTACTCACATACAGT

GGGCCAGAACTTGTCCTTCCATGTGATGAAGTCATCTTCAATGTTAGCATCATCG

TCTCCAAGTCCAAGTTCAATGACCCTGGTGGCACCCAGTTCTTCGAGGCGTTTAT

CAACGTAGATCCCCACTTCGTTGTAATGCTCGTACGTTTTGTTCCCCAACCCAAAC

ACAGCGTAGTTCAGGCCAGTGAGATCGGCCTCTCCGTTTTGTAGCCACTCGTAAA

AGTCCATGGCGTTATCAGTAGGGTCAC (SEQ ID NO: 78)

contig01654 length=356 numreads=6

TTGAGGACCGTAAAAAACTGGATGGACTGTACGAATGTATCCTATGCGCCTGCTG

CAGCACCTCCTGTCCATCATACTGGTGGAACGGTGACAAGTACCTGGGCCCTGCA

GTGTTGATGCAGGCTTACCGCTGGGTCATCGACTCTCGCGATGAAATGACTCAAG

AGAGATTGGAAAGGTTGAAGGACCCTTTCTCAGTTTTCCGGTGTCACACCATTCT

CAACTGCACTAAAACGTGTCCTAAGGGTTTGAACCCGGCCAAAGCCATCTCCGA

GATGAAACAACTCATCTCTGGCACAAAGCACAAGCCGGCCCCTGAACTCAAAGC

TGTGTAAAATCGCCGACACCGTTCACCT (SEQ ID NO: 79) contis00571 lensth=2S5 numreads=44

CCGGGAAGAATTCGATAAGAACAAGGCTGTGAGGGACATTCGTGTGATCGATAT GCTGGTGATAAAGGGTCAAATGGAGCTTAAAGAAACAGTCAACATATGGAAGCA GAAAGGTACTGTCATGGCTTATTTCAAGGAGACTATCGAACCCCAGCCCAAAGA TTTCTTGTCCAAGTTCTTCTCGAATGTAAAGTAAAAGATTTAGTTCATTCAAATTA TGTAAAATTGTTTTAATATACTTAGTTGTTGGTTCTT (SEQ ID NO: 80)

contiε04258 hnsth=340 numreads=15

CGAAGACCATCTTGGTGCCGGGGATGTATTTCTTGGGGTTCTCGAGGTACTCGAA

GAGAGTGTCCTTGCTCCAAGTGATGCCTTTACTGACGTTGGCGTCGGAGTAGGAG

AATCCTGGGGCTTGACCAGTCTTCCTTCCAATAAGACCACTGAGGTTGGGTCCTA

CTTTATGTTTGCCTCCCTTCTCCACCGTGTGGCACTGGGAGCATTTCTGGACGAAC

ACCTTCTTTCCCTTCTCAGCATCTCCTGCGGGAACACCCATGACTTCGAGTTATGG

ATTAGGTAGGTGGCTGAACGTCCGAAAAATCGAAGAGGTCGGTCGACTGCGCGA

TCACTGGCA (SEQ ID NO: 81)

contig06671 length=469 numreads=16

AAAACCATTAAAGAAATAATATCAATTTTTAAAAAAAAGATATTTAAAGGAAAT

CAATGTAAAAATAACAAATAATATTCAACATAATAACCATAAAAACCACAAAAC

AAACCCCTATAAACAACCCCATGATTAATAGAAGAATATAGATAAATACTATAT

ATACATCCTATAAAAGAAGCAAATATCAAATAGAAAAAAGAAATATATCTCCAA

GATATAATTCCATTAATCAATAAAATTTCACCTATCAAATTCATTGAAGGAGGTC

TAGCCATATTATTAATCATCAATAAAAATCAAAATAAACATATTCTAGGTATAAA

AGTTAATAAACCCTTATTAATAAAAATTCTTCGAGATAAAGTTCGTTCATAGGCA

ATATTTGCTAAACAAAATATACCAGAAGAACATAAACCATGACCAACCATCAGA

ATCAATGAACCAATAATACCTAATATATTCACAC (SEQ ID NO: 82) contis06264 length=792 numreads=99

CAACGCAGAGTGAAATCAGATCACACCCTTGCTCAAAGTGCGATTTTGTAGCTCG

ATGTCCTGCTAGCTTGAGAAATCATCTGACGTTGCACACGAACGAGAGGCGCTAC

TCGTGCAAGAAATGCAATTACAAAGCTAAACAATATTCTCATCTGAGGAACCAC

CTGTTGATACACAATTCCATCAAACCACACGCTTGTCCGCACTGTGATTACCGAG

GAAAGCGCATAGATGCCCTCAGGAATCATCTTTTGAGTCATTCCAACAATGCTGC

CGCGGAAACCTACTTATGTGAAGACTGCGATTATAAGACATCTAAAATGGCCAA

TTTAAGAAGCCACAGGTTGACCCATTCCAGAATCAAGTCATTCTCATGTTCCATT

TGTGACTATAAATCTTCTTATCAGAGCGACATTTCAAGGCATACGAACAACGTTC

ATAACAAACCTGATAAACGATTTTTTTGCACGGTTTGCAACTACAAAACGTCTAG

GGCGGATCACTTGAAAGCTCATGTCAAAACTCATTCTAGTCTTAAGAAAAAGTGT

CCTCATTGTGAATACAAAGCAGTAAAAAGTATATCTGTTAGAACTCATATCAAGA

AAATCCATCCGGACAAATCTTGAGATTGTTATTGCTTCTGAAACCAAGTTTTACA

TGTTACCATAGAAAAGCTTCGAGCTTCGACTGATTTTGAGTATTGGAAAGACTAC

CTACGCTTTCCTTTAGCTTTACAAAATCTAAAAATTGTATAATTCTTTGAAAAAGA

TAGCAATAAAATGATATTTTTTT (SEQ ID NO: 83)

contig04729 length^όl numreads=3

TCCGAAGGAATTCAACTAATCCGAAGGGTGTTTTAGGGTCTAAAGCCACCGGCG AGCCACCGCTGTGTATGACGATGAGCCTGCAGCACGCACTTAGACAAGCATTGA GGTGTGCGAGAGTGGAATCAGGCCTAGCAGACGAATGGTTCGACTACCATCAAC CATGCACTTTTGAGAAGACATTACTAACATCAGGAACCAGCCCGAAACAGTTTA ATTTGTGACGGCACCACGATAAACTCAACAAAGTTGGAGGGTTCG (SEQ ID NO: 84)

contis01756 length=577 numreads=19

AAATTTATTAGTAATATAAATAATGGGAATATTATATTAATATATAATATATTAG

TCGTTGATATTTTGTTTGTTTTATTTAAAATAACTCATTTTTGTGAGTTTCTTATTA

TTATGTTGATAGATCTAATTATTCGTATATAATATGAAAGTGTGACTAATGATGA

CAATATTATCACTGTTAATATTATTATTTCATTTGAAATAATTATATATTCGATAG

TTATTCATTTTGGTATGAACCCAAGAAAAGGGGGCATCCCTCCTATTCTTATTATT

ATAATTATAAATGATAATTTTTCTGTTATTCTTAGGGATTT AATATTAAATTGATT

AATATATATAATATTATATTTACTTAAATTTATGCATATTGGAATGATTATTAGTG

AATATAATATTATATATATTATTCATAATTTTTTATTGATTATAGCAGCTGCTATC

ATTCAACCTACATGATTAATAGATGAGTAAGCTATGATTTTCCGGATTGAGGTAT

GATTTAATCCTAAAATTGCACCTATGATGGTAGATAATATAATTATCATTATTATT

AATTTGTTGTTTTCTATAA (SEQ ID NO: 85)

contis01582 length=718 numreads=22

TACATTTTTCGGATGTTATAGTGCAATTTACTTGATTTTTCGAATTAATTTGGTGC

GTGGGAATTTTAGAAATGGCTCTTTTGAAGCCTACCTTGTTGTTTAGCATTACCCG

TCGATCCATTGGCTCCTCCCATGTCTTGGGAGCTTACTGGAACAAGGACTGGGAG

CCTGGCCCCTATCCACGGACTGAGGAAGAGAAGGCGGCTGCAGCAAAGAAGTAT

GGTTTGTTGAAAGAAGAATATATACCCATCCCCGATGATGGGGACGGTGCTGGA

GATTACCCTGCATTCAAGCCGTATTCTGTGGAAGGTAGAGACCCCCATTACCCTT

GGGATTTCCCAGAGTACCGAAGGAACTTCAATGAAGCCATGCACGAGAACTTCA

ATATGTACACTGAAGAAAAATATAACGTCGAGAGGAAATTTCGGTATCCGATGA

GTGCCATGTTGGCCGCCTTCCTTGGAGTCATGGGAGGCACTTTCTTCCTGGTCGA

CCAATGCAAATACACCGCGTTCCTTCCTGCTATGCCCAGACACCTTCCAGAGCCT

GGAGTCTCGCACTACAAGTACTCTCGCACTTAAGTAACGTACCCCCATATTAACA

TAGTTGTACAATATTTCAACCCTTTCCTACCCTATCTACCGATTGGTCTTAGACCA

TTGTAATCATTTATGGGGGAACTGTAAAATTTCGTTATTAATAAATGTTGATATTT

TAC (SEQ ID NO: 86)

contig02061 Iensth=508 numreads=5

ACTCTCCAATATTTGATGGCTGGAGCATCGGCTGTTCAGATATGTAGTTCTGTGC

AAAATCAAGATTTCACCGTCGTGGATGATTACATCTCTGGCTTGAAAGCTCAAAT

TTACATTAAATCCTTGGAACACTCCGAAAAATGGTTGGGACAGTCACCACCTACC

CAAGTCCACCAAAAAGGAAAACCGACTTTGACAACAAAGAACCAGAATGGGAA

GACACTGCCTGTTTTCGGAAAATACAAAGAAGAACGAGAAAAAATCATCGCTCT

TTCAAAGAAATCAGGACAAAAAGCGAAAACACTCAAGCCCCGGGTGATATCTGA

GCAAGTGAGGCCAGTCATGAGGCTCGAGGAGCTGATAGGCTCCTCATTGGAGCA

CATCACGAATTACAAAAGTCTGGACAACACCGAACAAGTTGTAGCCCTAGTCAA

CGAGGATATGTGCATCAATTGTGGGAAGTGCTACATGGCATGCAATGATTCTGGT

TATCAAGCAATCACATTTG (SEQ ID NO: 87)

Known chemical insecticide targets: contiε01103 lensth=312 numreads=15

TATCTGGCGAGCGAGCAATGTGGTGTCGTCCTCTCGTTTTTCTGTCGGTCGTCGCC CTTTCGCGCGGGGCGTCGGCGCCAGTGGTTGACACCCCGGCGGCGCCAGTCGTCG TCACCCCTTTGGGGAAAATCAGCGGTTGGGAGAGCGATTGGGCGACGCCGAGCG GGCGCGCGTACCAGGGTTGGACCAAAGTGCCCTACGCTCAGCCTCCAGTTGGAA GTCTCAGATTCAAGCCACCCAAGCCTTTAGAAAAATGGGACGGCGTGTTGGAGG CCACAGGAGAGATGCCCCATTGCATACAGCGCGGTCCTG (SEQ ID NO: 88)

contis05244 lensth=510 numreads=9

CGTGATGCACGGGGACGAGATCAATTACATCTTTGGCGAACCGCTGAACCCGAC

CAAAAATTACCAGCAGAACGAAGCCGACCTGAGCAAAAGGATAATGAACTATTG

GGCGAATTTCGCCAAAACGGGGAACCCAAACCTGCGGAGTAATAATTCGTGGAC

ATCGACGTACTGGCCGCAGCACACGGCATACGGCCGCGAGTACATCACCCTCGA

TATCAACACTACCGCCTCGACGGGGAGGGGCCCGCGCGTGAAGCAGTGTGCCTT

TTGGAAAAAATATCTCCCCAGCTGATTTCTATGACAGAACAACGACAGCAAAAC

GTGCAAGAATGCCCGCCCATAGCTTCTGGAGTTTCCCTGAGGGACTCGTACACGT

ACATCCTGGCGCCCATCTTCAGCTTCTTCACGTTCCTCCTGGTCCTCCTGACATCT

GTGTAAACTGACAACAGCCGAGGAGACCACCCGAAATGAAATTTTAAGAGAGAG

GCACCAGAAAAACCTACAAAT (SEQ ID NO: 89)

contis00372 length=361 numreads-4

ATCCAAGGTTTGTCGAGGGCGCATCGACAGATCTTCAGTAGCAAACGCGAGCTC

CGAGCCTTCAGGTTCGATGACTGGCACGTAGGGGATGGTGTCAGGTGGACTACC

ATCGTTTGGGCTTGTCCAATAGCCATTAAAAGAGCCTTGGCGTCCACCTCCTGCA

AGCACTTTAGAGTTTCTTCGTTGTCCTTCCGGGAGCAGCCTGTCAATTTCACTACA

TTCATCGTGGTGGATTTGACTTTTTCGGTCGCGAAGAGAGACCAAACGTGGTTGA

TAGTCCCTGAATGTGATACAGCCCTCGTCACAAGGTCTTCGCTTAGTGGAGATTT

CAGCAAAAAGTGCGACGAGGCGCCCCCAGCAG (SEQ ID NO: 90) contie05930 lensth=890 numreads=27

TGTTTCCTTGGGTGACACAGCGTACAGACTTCGATCGTTGAAATAGTATAGCTTT

TGATCGGCGTGCGGTGCAACATTTATGCTGTTTCCTTTAGGCCGTAAAGTTCTTGA

GCTCGAGCCTCGAGATTTCGCCAACATACTCAAAATGGTAGTACCACTTTGGACC

TTGGTGATACTGAAGATTGTAAAGTGCTGGGAAGAAAAACATGGTATCAGCTCC

AACTTTTGCTAGTTCGATAGAGAAATTTTGTAAGGGCTCCATAACGGGAAAGTAT

AATTTCTTCAATATCGCAGCTCCTTCTCTCATTCTCTCTGGATCCTCATAGCCCAA

ACGAGTGGCGATTACCCGTTCCAAGTAAGCGTCCAAATTTTGCAGTGCATCGGAC

ACGTCGTTAGGGGACATCATAAAAATATTTAAGAGAAATTCGCCATTAGCGTTAC

TCGTGATCCATGGTTTATTGGAAGGAAGCTCGAACAGGTCTTCTTTGATCACAGC

ATCCTCCGCGTCCTCAGGTTCAAAGCAGGGTGCAAAACTATAAACCCCAGACAG

TCCGTCTGGCATTGCACCAATCAAGTCTTCACCGCTGGCTCCTTGAAGGCACTCG

AAAATCTCCAAGTTGCTGCTTGTGATTCGACAGCCAAGTTTCGCAGCTAACTGCA

ATGTGCCATTTCTGATGATTTCCGGTTTAATTTCGGACCATATGTAGTTAATTGTT

CCAGAATCAGAAAGTGCCCTGTGTACCATCCCTTCAGTAGCTGGTGTCTTCAACA

GGATATGACTGCTAGCACCTCCAGCTGATTCACCAACGACTGTAACGCTATTCGG

GTCACCACCGAAAGAAGCGATATTCTGCTGCACCCATTTCAACGCCATAACTTGA

TCCTTAATC (SEQ ID NO: 91)

contis03360 lensth=249 numreads=6

CGTCGCAGTGATGGTTGGGCTAACTTCTCTGTGGCGCTCAGCGCCGTTTTCGCCT GTTCGGTCGCTCAACATGAACAACAACCCAAGTAACGACGTCTCTCGGGACGTTG CAAGGATCGGTTCGGCATTCAATTTCAGGCAGAGAATTCCACGCTTTCGAGGGCA TACCCTTCGCAAAACCTCCAGTAGGAAAACATCGGTTCAAGCGAGCTGTTCCGAA TGGCCCGTGGACTGGAGTCCTCCACGCAA (SEQ ID NO: 92) contig05458 lenεth—272 numreads=7

TCCAAAACGGCCGACATCGAGAGGATGATCAGGTCCTCCGCCTCTTCGGCGACTT

CGTCCTCGAAGGGAAGCAGTTGGTCGAGCAGCCAACGAGCCGTGCCAGGAGCGT

CTGCCGAGAGCAAACGGAGGCTAACGGAATCCGCGACCCGGGCGCTGACGGACG

CCCCCGCCACGCGCCCAGTCGGCGTCTGGAAACGCAGGGAACTCATCGCATCCC

AACCAAAGAGCCCCCTCTAGTCATTTACCCGAAATCAAAAAACCCATCTTTTTAT

(SEQ ID NO: 93) contig01440 lenεth=249 numreads=3

TAGGAGGTGAGATACTCCTCTTTCTGGTTTGCATTCTCATAGGGGACGAACCCCC TCCACTGCTCCTCCCCTTCCTCTCAGCTTCTCGGGCACTGCTCGAACTCATCTTCC TTGGTGGAGAATCTTTATGGCTGCTGCTTTTCATGATGGATTTCTTCTTGTCTTCT GGGGATGACGAGGACTGACGCCTCTTCTTATCGTCGGTCTTCTTGACAACGCTGG CACTCGACCTCGACGAGGACGAGCTGG (SEQ ID NO: 94) contig02231 lensth=430 numreads=6

ACCTCCGTATCAAACACTACAAGGTAGTAACAAAATTCATCATGATGACAGGAG

TCATCGTTCTGCTGCTCACTGTATTCGCGTGCTTCGCCGCCAGCAGTGTCGTCAGC

ATTCAAGGACCACGTGTCGGCCCGATCATACCACCCCAGAGGATCGCAGTTGCA

CGTGGCAAGCTTGTGTGACGGATAAATCAGTTTGTTTCTGCATGCCATCCCTCCTC

GGCGTCCCTCAAATCAAGTGCAATCCCGTTAAGGATCCGAAACAAGACGTTTTCC

CGAAGTGCATAGATGTGATGTTGGGATGATCATCATCGACGGATTGATCAAGGC

GATTGATCCAGATGGAGAAACTGAGTCTTTTTGGACGTATTATATTATTGTCGCC

TAAATTTTGTATGTAATAAAACATTTTAATGCATAATAAAAATTCT (SEQ ID NO:

95) contig05130 lenεth=260 numreads=2

CTCGACGAAAGCCTGGTAGTAAAATTCAAGGGAGGGGCAGACCTTGAAACCCTT

GGACTACAGACGGTTTCGTTGTCTGACATCGAAGAGGATGACGAGCCCCATCCG

GGTAAAAGCTTCATGCAAACTACCCACGTCTACACGTATACCAACAGCACAGTA

ATGCATCCGGACGACTGTACAACTACAACCAACGTAGTTTCCAGCTACCGAGGA

AGCCGCATGAGCACAGCAGCCAGCAGCAGGATGAGTTCCGTGTG (SEQ ID NO:

96) contig00542 ϊenεth=270 numreads=3

ACCAATGATTTTGACGACTCTCCAGACGACGGGTCGCCAGTCGATCTCAAGTTTG

AAGTACTTAAACGGAAAATTGGCCAGCTTGAAGATGAAAACAAGAGCCTGAAAA

ATGAGACTTTCCGCCTTTCTGAGGAAACGGAGGCTTGCGAGGAAGCTGAAGCCA

AACTCGTGGAAGACCTTGTGGCACAACTCGAAGCCTGCAGGTTCGATTTGGATTC

TACGGGTATCGACATCGAAAAATCCAAACAACGAGAGGCTGAGCTGAAAGAG

(SEQ ID NO: 97) contig04701 ϊenεth=259 numreads=2

GCCAATGACGACATCTACTGACATTCTCACCTCCTTCGGTATGGACACGCCTGAG AATGACAGAAACGTCGAAAACTACCCGATCCCGAAAGAAATTAGTTCACGTTCT AAGATTAGAGAATTTGATGAGAGGAAAAGAAAGGTACCGACAGGAAGGAAATT CGGTCCTCTGGCTCCCATACCAGTTCCCAAAGAACTGGAAGATGTGGACATTACT GAACTCAATTCAATTGAAGAGGTGGCTAAGCAGATAGCAGAG (SEQ ID NO: 98) Glycolysis/sluconeoεenesis: contiε05492 length=915 numreads=475

CCTAGTTCCTCTTCGATACGAAGAATCTGGTTGTATTTGGCACAGCGTTCAGATCT

GCATGGTGCACCAGTCTTGATCTGTCCAGTTCCAAGTCCAACGACCAGATCAGCG

ATGAAGGTGTCCTCAGTTTCTCCGGACCTGTGCGACACCATGGTGCCCCAGCCGT

TCTTTTTGGCGAGGTTGTGGGCTTCAATCGATTCCGTCACCGAGCCAATTTGATTG

ACTTTGAGGAGGAGACAGTTACAAGACTTTTTGCCCACCGCCATTTCTATCCTCTT

GGGGTTCGTGACAGTAAGGTCGTCGCCAACAATCTGAATGCTGGTACTTCCAGTA

AGTCCAGACCAAGCTTCCCAATCGTCCTGGTCGAAAGGATCCTCGATGGAAACG

ATAGGGAATTCCTTGATGAAAGACTGGTAGAGATTAGTCAGCTCAGCAGGGCTG

ATGTAGGTGGAAGGGTCGGACTTTGGGTTCTTGAAGTCCAAGTCATACTTCCCAT

CCTTGAAGAATTCAGAAGCAGCGCAATCCATTCCAATCTCGATTTTACCGGTGTA

GCCGGCCTTCTCGATGGCAGTGGTGATCAAAACGAGACCTTCTTTGTTGTCGAGG

ATGTTGGGTGCGAACCCACCTTCGTCGCCCACAGCTGTGGCGTCAAGGCCAAACT

TGCCTTTGATGACCGCCTTCAAGTGGTGGTACACTTCAGAACCCATCCTCATCGC

CTCGCAGAAATTGGAAGCTCCAGTTGGAAGGATCATGAATTCCTGCATGGCGAG

TTTGTTGCCGGCATGGGAGCCACCGTTGATGACATTGAAGGCAGGGCAGGGAAG

GATGATGTTTTTGTTGCCAGCAAGTTCAGCAATCCATTTGTACAGAGGAAGGCCT

TTTTTAGCAGCTCCCGCTTTGCATACGGCAAGTGAC (SEQ ID NO: 99)

contis01170 length=499 numreads=19

GATAGTTTTACTGTATTTTTCATTACTGACAAAATGTAGAAAATGAGCATTATGC

TGCTTTAAAATATATACTTTTTACATCAGTTTGTTGTTCTGCTTATAGCATGGAAG

TTGTAACACTCCCAAGTGTTAAGTTGAGGTTCAGTACAGATAACTAGTATGGGAG

ACAGAAAGCGACTTTATGGCAGTTTACTTGTCCTTGGTCTGCATGTACTTGATGA

GGTCGACAACACGGTTGGAGTAACCAAACTCGTTATCATACCAAGACACAAGCT

TGACGAATTTGTTGTTGAGAGGGATACCAGCCTGAGCGTCAAAGATGGAGCTGT

GGGTGTCACCAATGAAATCAGCGCTGACGACCTGGTCTTCAGTGTAGCCCAAGAT

TCCCTTGAGAGGTCCTTCGGAAGCCTCCTTCACTTTCTGCTTGATTTCGTCGTAAG

TGGCTTCTTTGGTCAGCCTGACAGTGAGGTCCACGACTGAAACGTTGGGGACAGG

AACT (SEQ ID NO: 100)

contis06703 lensth=1080 numreads=132

AAAAAAAAAGAAACTATTGGTAAGAAAAGTTTACGGAGTAGGTACAATATGGTA

GCAAAGTACCTACTCAAAATCAATAGGTACATAATATATACTCATAATATCATTT

AATTGGACATTGAATGTCATCTCAGTCGATCGAATGCAACAATATCGCAGCTTTC

TCAATTCATTGCACATCCGCTTGTAGAACTCACAGTTTCACGGCAACTTTAAGGC

AAAATAAATTGTTGGAAACGACGTTTTTTCCAGGTAAGTACGAAGTTCCGAACTT

AGGGTCCTTGTAAAGCTAAGAAAAACAGTGCGATGGAACGAAAACAAATATCAT

TACAACTATATACATAATGTAATTTTACACAAGTTATTTTTTTTTGTTTTTTTAAAT

TTTTTTTCTTTTTCATTTTTCAATTTTTTGAAATAAATTAAATAAATATCTTACTCA

TTTTAACGAACTGATAGGGCAAATAATAAGGTGTCAGCTACAGAATGCGAAAGA

ATTGACTATCTCTTAATGCTCGTAATAAATTCTGTTTTTTTAATGATAAAAATTAT

AAGGTGAACTTTTAAAATAAAAATTCAAAAATCACTAGACGAGTTGAAGCACTA

ACGTGTTGATGTGGGATGAATAGGTTGTTTTAAAAACGTAGTCGGACATCTTGAA

GAAAATCCGATGAATAAAAAAAACTCCACGGAGGAGTTATCAGCGAATAATTCT

AAACTGTTTAAACCTTCTTGCGACTGTGAAAGGTAACATTTTTAAAGGTTATATTT

CTTTGTGATGCGGGGTTCGGAGATGAGGGTGAACTTGGGGAAAGTCTATTTAGA

GGGACATGGACTCTTCCAAGAGGGAAGCGAGTTCTTTGAGATGCATCATTTTGGA

GCCGGTGGCCGGAGAGGCGGCAACGGGTCTGCCAGCATAGCTGACATCCCGACT CCTGCTGAGGGTGGTTTGGAACCTGGGGCAGACGTAAGACCATGCTCCCTGGTTC TTGTGCTCTTCTTGGGCCCAGCACAGCTTGGCATTAGAGTACTTGGCGCATTCCTT CTTGATGAGGTCGTAGGGGAAAGGAGAAATCTGTT (SEQ ID NO: 101) contJ206954 length=149 numreads=90

TTGATGGCTCCTGCAGTTCTAATGAAAGATGCGAAGACCATGTCGACTCCTTGCT GAACTCCAAACTCCAAATCCTGTTTGtCCTTTTCAGACACGGCAGGAAGATCGAC ATTGATGCCTGGAAGgTTGACACCTTTCTTGCTTCCCAA (SEQ ID NO: 102) contis06212 lensth=1248 numreads=63

ATAGTATTTTATTGTAGATTTATAAGACGCATGGAAAATATGGGATAAGGGGTAA

ATTATTGATAGTTCAACGTGAGGAAATCGTAAATGATTGATAATTAGTAAGCGTG

GTTTTTGATAAACAGATCTTGGTCGGCTGCTGATCCTTTGACGGTGCCGGCGACG

TACTTCCCAAGGGCGGCCTGTCCGTTGGCACGCGCCCTGTTCAGCAGCTCCTGCT

GAGCAGCTTTCAAATTGCCATCTTTGCCTTCCAAGCGCGAGGACGGAGGCTTGGA

GAGCTCTACCAAACGAGAATGTGAGCGGCCAGGGTTTCTTTCCTTGGAATTTGTT

GATGGCATCCAAATTCAGGGTCGCTTCTTCCTCGGACTGACCGCCAGACAAGAAG

GTGATGCCCGCGACGGCTGCTGGAACTGTTCTGGAGAAGGCGAGACAAGTGGCC

CTAGCTACTTCCTCTGGCCTGGCTCTTTTGGGGCATTGCATGCCTGGAGTGACCAT

GTTGGGCTTGAGAAGCGTTCCCTCGAGGTAAACGTGGTGGTCGTTCAGTGCCTTG

TACACGGCTGCCAGTACAGTCTCCGTCACTTTTTGCGCTCTGTCCAGGTCGTGCG

AGCCATCAGGCAGTACCTCTGGCTCCACGATTGGCACGATTCTGTTGGCCTGGCA

GATCGATGCGTAGCGAGCGAGCACATTAGCGTTCTCCAGTATGGCTTGGTACGAA

GGAATGTTCTCGCCGATTTTGATGACGCAACGCCACTTGGCGAACTGGCAGCCTT

CCTTCTTGTACGAGGCGCATCTCTCAGCCAGATCGTCGAGCCCCTGAGTCGTGCA

TTCGTCCTCAGACCCAAAAAGTGGGACAACCCCCTTGTCGACTTTGATGCCGGGG

ATGATTCCCTTCTTCTTGAGGACGTCGACGAACCGCGTGCCGTCGTCCGTCTTCTG

GAAGAGGGTCTCGTGGAAGAAAATGACGCCGGAGATGTTCTCCGCCACGAGGTT

ATCCGACGTGAAGAGGACTTGCCGGTACCGTCTTCGATTCTCTTCCGTGTTCTCCA

CGTTGATATCCTTGAAACGCTTTCCAATAGTGCCGGTTGATTCATCCGCAGCCAG

AATTCCCTTACCAGGAGCTACGATAGCCAGCGCGATTTCCCTGAGCTCCTTTTGG

AGTTCTGGAGGGGGGTAATTGAAATACGTCGTCATTCTGAAACTATCAATCAGTA

GGAATACAGGCTGAGGTCCGAGGAGAGGTTGTTGGGA (SEQ ID NO: 103)

contigOόlOl length=1620 numreads-159

ATGGTACCAGCGCCCTCTATTTGACGAGAAACGACGAAACCTTCAGTATGGAAC

AACGACGGTTTAAATCTGTCATTTTCTCCCGACAGATTCTCATGAAACGAATCCT

GTTTACACTTTACAGGGTCAAAAAACCCTCAACAATTGATCACAATTTTTTCTGG

ATCATTAAAAATGATCGTACATTTACGTATTACTAGATTGAACGAGGTTGAAATT

ACATATTATGATGAAGTTGTGAACATTTTAACATGTTTTAACAAAATGTTTAAGC

CAGTCTGTACGAACCCAAGCTATGAATTGCGGGGTCATGTTGGTTGTTTCGTGTG

TTGATTACTGTTGATGAGTGAGCAGCCTGAGACCTTCTGGTTCTTCACCTTCCACC

TCATCACGACACTACATCAGGAAGGGATGGAGAGAAGCCTCACGACAGCCTCAA

GCAGTTCACCCTTCAACCTGAACTAACGTGAGGTGACTTCATTTTTTGGTAGGTA

CCTACGTTTTGTAGGTTACAGGCAGTATATTTTTAAAAATAAAACATTAATGTCA

TACGGGTGACCCAGGATGAGTCCAGTACGAGATTGTGAAGTCTTTTAGATCTTTC

GTCCAAAATATACGACGGTAGGTACCTAGTAGTAGTTAGAATAAATTAAAATTAT

AGAATTGGAAGTTATTGTAGTCAACTTGAATACGTTCACTAAAAACTACGTAGAA

TGGCTTGCTGCTAGGAGGTGCTGGTGCGCTCAGAGGGAGAGGACTGAGAAGCTG

CTGTCGAGGTTCTTGGGGACGCTGACGACGCGGATGGTGTTGGTGTAGCCAGAA CCGGACTTCCAGCCGGTGAGGACGACGACTTGGTCGCCTTCCTTGAGGAAGCCCT

TCTTGGAGACGACCTGAAGGCCGTGGATGATCCTCACGTCGACGTCTTTCAACCA

GTCGGTGAGCGGCTCCTTGTCAAACACAACAGGGAGGATACCCCGGTAGAGATG

AGCTTGCCTGGCGACCTGTGGGTGACGGGTGACAGCGATGATGGGGCACCTCGG

CCTGTATTTGGCGACGAGGTGGGCAGAGCGACCACTGGTAGTGATGACCAAGAT

GGCGGACGCGAGAGCCTTGGTGGCTGCTTCGACAGCAGCGATGGCTACGCTGTG

AGTTGCATCGATGGGTGGAAGAACCTTTCCAGCCAAGTCAGAGAACAGCTGATT

GTGCCAAATAGCGGCTTCAGCCTCTTTACAGATGTTGGCCATGGTCTGGACGCAA

TCCAAGGGATACTCGCCCTTGGCCGTCTCTCCCGACAACATGACGCAGTCCGCGC

CGTCGAGGATGGCGTTGGCGACGTCAGAGCTTTCAGCTCGCGTGGGTCGGGGCTT

CTTGATCATGGATTCCAACATTTGAGTGGCGCAAATGACTGGTTTTCCTTGTTTAT

TACACTTGGCAATCATGGCTTTCTGGGCCAAGAAGACCTTTTCAGTTGGTATTTC

AATACCAAGATCACCCCTAGCAACCATGATGCCATCGGCAGCTTCAATGATTTCA

TCCAAATTCTGCATGCCTTGATGATTTTCGATCTTGGCAATGATCAGGATGTTCCT

TCCAGCTTCGCCCAGAATTTCTCTTATTTCT (SEQ ID NO: 104) contJ204894 length=217 numreads=6

AATGTCTAGGGACGACCTTTTTACTGTCAACGCTTCGGTCGTGAGAGATCTGGTA ACCGCGTGTGCGTGTTTTTGCCCTAAAGCCCTCATCGGAATCATCTCTAATCCAAT CAACAGCACCATTCCCATCGCTGCTGAAGTCATGAAACTGTCCAACGTGTACGAC CCCAAGAGAATATTCGGGATCACGTCGCTGGATCTCGTAAGAGCCAGTACG (SEQ ID NO: 105) contis04593 lenεth=245 numreads=2

TTGGATGGGTTTGGATATTGGAAACAAATCGATTGATTGTTTCTCCCAAGTCATTT TAAGGTCCAAGACTATAGTTTGGAACGGTCCCCCAGGTGTTTTTGAGTTCGAGAA ATTCTCTCGAGGGACTCGAGGTATGTTGGACGCCGTCGTCGAAGCTACTTGCGAT GGCGCGCTGACAATCATTGGAGGGGGCGACACGGCGACTGTGGCCAAGAAGTTC AAAGCAGAAGATAAGGTATCTCACG (SEQ ID NO: 106)

contis03352 length=389 numreads=12

CTCAATGCTGGGGTTGCAGAGTTCAGGGATCTTTTCGTGCATGATGTTGTGGAAG

TGAGGGTGGAAACGGTACAGTGAACCATCGACTCCAACGGTGACAGAGGGGATG

TCCATCTTGTTGATGAGCGTGGCCACGCCCGCTGAGGCAAGGTAGGCTGCTCGCC

GGCTCACGCTTTCGCAGATGTACCTGATGTTGGAGCAGTCTTGGTCGGTGGGGTC

GAGGATGTTCAGTTTTCCCAGAACGTTCCGGCACTGCTTGAAGACTCCTCTGGGA

TCATTTTCGATTTCAGAAATGTATTCAGTGAGGAAGCTCCATCGCTCTTGAAGTTT

GGGGTTGATTTGACCTCCAAATAATACTTTGTCTTTGGTGAACTTGATGATGAGA

AGTC (SEQ ID NO: 107)

contis00761 lensth=218 numreads=49

TGGCCACAACCGGGAAACAAGACATTCTGGATATCGTTCCCACCAAAATCCACC AAAGAAGCCCCATCATTCTCGGTTCCAAGCGAGATGTTCAAGAACTGGTTGACCT GTACCAAGATCCGAAATTCAGGGATTGAAACTCAACTGTTAAAGACTGCTAATTA TTAAGTTATTCGTTGTTATGTTCTGTTAGACACAAATAAAATAATTTAAAATTA (SEQ ID NO: 108) contis00842 lensth=255 numreads=3

CCGTCAACTCCAACAGTGACGGACGGGATGTCCATCTTGTTGATGAGCGTTGCCA CACCGGCGGACGCGAGGTAGGCCGCTCGCCTGCTCACACTTTCGCAGATGTATCT GATGTTCGAACAGTCTTGATCGGTTGGGTCGAGGATGTTCAATTTCCCCAGAACG TTCCGACACTGCTTGAAAACACCTCTGGGATCACTTTCAATGTCAGAAACGTATT CCGTAAGGAATCCCCATCGCTCCTGGAGCTTAGGG (SEQ ID NO: 109) contig04919 length=243 numreads=4

GTGAGGTGATGGATCTTCAGCAAGGAGCTGCATTCACCCCGTCCATTAAAGTCTA TGGCAGCACTGACTATGCAGTGACCAAGGACTCAAAGATTTGTATAATCACTGCA GGCGCTCGGCAGAAGGAAGGCGAAACGCGCCTACAGCTGATTGAAAAGAACGTC CAGATTTTGAAGCAAGTTGTGCCTGAGCTCCTCAAGTATAGTCCCAACGCCATCC TACTGTTGGTAACCAACCCTGTTG (SEQ ID NO: 110)

contigf)1640 Iength=286 numreads=4

TGCTCGTGGTCTCCTGTTCGTTGGGTCAGGCGTGAGCGGCGGTGAAGAGGGAGCC

AGGCGAGGACCTTCTCTCATGCCTGGAGGCGAACCAAAAGCCTGGCCTCATCTTG

CCCCTATTTTCCAGTCCATCAGTGCCAAAGTGGAGGGCGAATCCTGTTGCGACTG

GGTTGGCGATGGCGGCTCTGGTCATTTCGTCAAGATGGTCCACAATGGGATCGAG

TACGGCGACATGCAGCTGATCTGTGAGGCTTACCATCTCATGAAGGACCTTCTGG

GACTCGACCAC (SEQ ID NO: 111)

contigOόlll length=1429 numreads=121

TCTCGAGTCGCCGGGGAGCCAACCACTTCAATCTTCACTTCAATCGTCACTTCAA

TCGCCACTTCCATCGCCAAAGGTCAAAAGGTTCAACCATGAGGAATAAAGACGA

AATGGCGCTGGCGATCGATGCAGCGATCTCGCAGGAACTGCAGCCTTTGGTACAT

TCCGACCCGAATGTTCAAAATAAGCTAGAAGAGATTCTCCACTGCCTGAGGCTCT

CCGGTGATGACGTCATCCGCGTCGGGGACGTTTTCGGAGAGGAAATGACTCGCG

GCATGAGAGACCAACCTTCGTCCCTGCGAATGGAAAATACGTACATTCCTGAAAT

ATCCTGTTCAGAAGAAGGCGAGTACTTGGGGCTGGACCTCGGAGGAACGAATTT

CCGAGTTCTCCTGGTGAAACTGAAGGACGGCGAGGTGGTGGAGGAGAAGTTCCT

CAGCTACGACGTCGACACGGAGACCCGTCAGGGCGAAGGCTCAAGACTCTTCGA

TTTCCTCGCTGACTGCGTCTCCGACTTCGCGCACAAAGAAGGCGTTGCAGACAAA

GGCCTCAACATGGGTTTCACGTTTTCTTTTCCCATGCACCAGAAGTCCCTCCACAA

CGGGATTCTCACCCAATGGACCAAAAGCTTTTCCGCCACGAATGTCGTCGGTAAC

GACGTCGTCAAGATGCTCCAAGAAGCGCTGGAGAGGAGGGGCGACGTCAAAGTT

AACGTCAACGCCATCTTGAACGATACTACGGGGACGCTGTTGCAAGGCTGCGCG

ATGGACAGAAGAACGAGAGTCGGTCTGATCCTCGGAACAGGAAGTAACGCTGCT

TACCTCGAGAGAGCGGACCGCGTCTGCCATTGGGAATCAGAGAGACACGGCGAA

CAAGAGATTGTGATCGACATTGAATGGGGTGCGTTCGGAGACAACGGCGTCATA

GATTTCATCAAGTCACACTTCGACCGTCAGCTGGACAACGGTTCATTGCACATCA

ACTCCTTCTCGTTTGAAAAGTACATCAGCGGTTTGTACCTCGGCGAGCTGGTGAG

GATCGTCTTGCTCCACCTCGCCAACAACAGGTTGCTCTTCAACGGCATCCTAAGC

AAGGAAATTCAAGTCAAGGACTCATTTTTGTCTTCATACGTCTCAAGAGTCGAAA

AGGACAACGTAGACACCCAATCTCATGAGACTGAAACGTTGCTGAGGAAATTCA

ATTTGGATTTCTGCGATGATGATGTAGCTGTGATCAAATATGTCTGCGAGTTGAT

TTCCCTTAGAGCTTCTCTTCTCGTGTCAGCTTGTTGCGCCCGGCTATTGAAGCAAA

TGGACAAGCCCGACGTAACAATCGCCGTAGATGGATCTGTATTCAAATTCCACCC

TCGCTACCGGTTGTGGATGGAGAAGTTCATCAGGTTATTGGCACCCAGCCATAAA

TTCCAACT (SEQ ID NO: 112) contiε04328 lensth=244 numreads=5

CACGCTTACTTTCCCTGTCATTAACAACGCTCGTAACTGCATTTTCGCGCTTACTG GTCAAGGAAAGGCAGAAATGGTTAAGAAAATACTGAAAAACAATGAAAACCTG CCCGCAGGAATGGTGCTCCCTCGTGAAGGCGAAGTCTTATGGATTTTGGATAAAC CAGCTGCATCTCTTCTGTAATCAGTAATTTGAATCATTTTACCCCCTAAACTCGAT GGCAAACCGCACAAATGTGACAAA (SEQ ID NO: 113)

contig00122 length=246 numreads=2

TCAACAAAGCCCTGGAGAATGAGAAAATAAAATTGAGCGTGAATGATTTCGTCA TTAAAGCTACAGCTCTCGCATCCCTCAAAGTCAAAGAAGCTAATTCTGCGTGGAT GGGAGGTTTCATTAGACAATATAACGGTGTAGATGTGAGTGTGGCTGTAAGCAC GCCCAAAGGCCTGATCACGCCCATAGTATTCAACGCGCACATTAAGGGGCTTATC GAAATTAGCACTGACGTCAAATCGCTCG (SEQ ID NO: 114)

contig01176 lenεth=711 numreads=8

CTACGAATCCTTTGACTGGGAGCACGGTGTGCTCGTTGGGGCGGGAATGCGAAG

CGAAGCCACAGCAGCAGCTGAGCACCGAGGAAAGGAAATCATGCACGATCCCTT

CGCGATGAGACCGTTCTTTGGGTACAATTTCGGCCACTACATGTCCCACTGGCTG

TCCCAGCAGACTCGCCATGGAGATGTCAAGCTTCCCAAGATCTTCCACGTCAACT

GGTTCAGAAAAGATGCTGACGGAAAATTCCTCTGGCCTGGATTCGGTGAGAACG

TCAGAGTTTTGGACTGGATTTTACGGCGAGTCGACAACGACTGTTCCATCATCGA

GCCAACACCAATAGGTTACGTGCCCAAAGAGGGCAGCCTAAACCTGGATGGCTC

TCTCCTCGGCCAGACATGAAGAAACTCCTCAGCGTTCCTAAAGACTTCTGGAAGA

AAGAAGCGGAGTCCATCGAGAAGTATTTGAAAGAACAAGTCGACAAAGACGTTC

CCGTGGAGGTTTTGAATCAGCTCAAAGCCCTCAAACTGCGCCTCAGCTAAACATC

ATTGACAAGCGGCCATCATCTACTGAAGTCATCCATGGCTAATCGTATCAAATCC

AACTAATTTTCATTTTTGTAGTAAACTCTGTTCACTGTCTGTCGTTATGGAACTCA

ATTCACACAATAAAAACAACCAGTTGTCAATCTGGTTACAATGGGAAAAAAAAA

A (SEQ ID NO: 115)

contig02838 length=330 numreads=4

CGCGATGTTGTTCGAGTTCCAGAGCGCGCTGAAGATGCGGTTGGCGAAGCGCGT

CGTGATGATGTTCTGCACCATCTCCTTGCCAAGGTAGTGGTCGATGCGGTAAATC

TGCGACTCGTCAAAGTAGGGCTCGATCGCCTTGCACAGCTCCGCGGAGCTCTGCG

TGTCGTGACCGAACGGCTTCTCAATAATCACACGAATCCACCCACCCTCCTGAGG

CATCGCGCCACCGCGGATGCTGCCGCACGCCCCCGCGAAGGCGCTTGGCGGCAG

GGCAAGGTAGAAGAGGCGGTCGCCGCCCTTCTCGGGCCCCTCGAAGGCGTCCTC

CAG (SEQ ID NO: 116)

contig04580 length— 248 numreads=2

GGTGAACCCAGCACCCTGGCGCCACCCGCTGACAAGCACCAACGGATCGCCAGG CTTGATGATGCCCTCCAATTTTCCGGCATTGATTCCAAATTGAAGCCTCAAGTCA ACATCCAAGCACCAGTTTGGATTAGGAGCCACAATGTAGTGAAGGGGCCAAACT GCTCTGAACATGTGGAGTCTTCTGGCGACGCTGCCATACCTGGTGATGGCGATAA CGGGGCACCTGGGACGGTACTTGGCCAAAT (SEQ ID NO: 117) contig00231 length=596 numreads=15

AAGAAATGGTTCAGAATCTTATGGTTCTAAGATTCGGCAATGGCATATTCAGTCC

GACGTGGAATCGCTTGTTTATTGCATCGATCCAGATCACTTTCAAAGAGCCTTTC

GGTACCGAAGGAAGAGGAGGTTACTTTGACGAATTTGGAATTATCAGAGACGTT ATGCAGAACCACTTGGTTCAAATACTCACTCTGGTGGCAATGGAAAAGCCTGCTT

CGATACACCCTGATGATATTCGAAATGAGAAGGTTAAAGTGTTAAAATGCATCA

AGCCTGTCCCTATCGATGATGTTGTGTTGGGGCAGTACGTCGGTAACCCGGATGG

GGAAGGAGAAGCCAAAATTGGTTACTTGGATGATCCCACCGTTCCAAAAGGATC

AACCACTCCCACGTTTTGCACTGCCATACTCAAAATTGATAACGAAAGGTGGGAA

GGGGTCCCATTCATATTACGCTGCGGGAAAGCACTCAACGAACGGAAAGCTGAG

ATCAGGATTCAATACAACGAAGTTCCCGGCGATATCTTTGGGGACAGTGCAAGA

GGAATGAACTGGTCATTCGAGTGCAGCCAGGAGAAGCAGTCTATGTGAAAA

(SEQ ID NO: 118)

contig01580 length=309 numreads=7

ATCATCGCTGTCACCCGTCACCCACAGGTCGCCAGGCAAGCTCATCTCTACCGGG GTATCCTCCCTGTTGTGTTTGACAGAGATCCTGCAAACGAGTGGATGAGCGATGT AGAAGAGCGAATCAAATACGGTCTGACTGTGGGCTTCACGAGGGGGATCCTCCA GAGTAACGACCCAGTCGTGGTCATGACCGGCTGGAAGCAGGGGGTCGGCTTCAC TAACACGCTACGCCTCGTGTACGTCAGGGACGACAAAGACAAAATGGGACTCTA ATCATCTGGAAATTCTCAATAAAATCTGGTCAATTTT (SEQ ID NO: 119)

contig05630 length=638 numreads—13

GTGAGTTGAACGATTTTATTTTCAAACTTTAAATTCAAGATATAGAATAATTTGT

GACACGAGTAAAGTAGGCCGGGTACAGTAAGGTAACCAGTACGGATACCAAATT

GTACCTAACCAATAAACAATTCGAAATTGAATCCACCTCTTATTTGCTTTTCTTGA

CGAATTCCACCCCTTTCTCAATGCTCTTCTGCAGTTCTTGTGCGGCGTTAGTAACT

AACTCTTGCTCATATTTGTTCAACTCTCCGTTTCCCAAGTTTTCAGCTACACCGTC

TTTATTTAGCAAAAGTTTACTTGAGAAATGAGACACGCAATCGACGACGTTGGAA

GCGACGTAGGCCCAATTCGCAAACGTTCGGTTCGCCTTTCAAAGCTTTGCACAGA

GAAAATGTGAATTTAGCTGCCGCATAGGCCATTGATAGAGTAGCTGAACCTGTTC

CCGCTTTAGCTTTTACCACTTCGGTTCCGGCTTCCTGGATCCTGTGGGTCAACTTG

GCCACTTCATCTTCGGAAAAGCAGAGGCCTGAGGGTTTAGTTTGTGATAAGACTG

GAATGATCGTCTTGCCCGCGTGTCCTCCGATCACCGGAACTTCAACGTCCGCAGG

GTCGAGGTTCTTCGCCTCGGCTATGAACGT (SEQ ID NO: 120) contis03919 length=253 numreads=3

CGTGCATTACAGCCTCAGGGTCGTCGGAGTTGACATGGAAAATGGGAGCATTCA CCACGCGAGCAACGTCTGTACAATACGGAGAGGAACGGGAGAACCTGGGATCGG TAGTAAATCCAATCTGGTTGTTGACAACAATGTGGATGGTTCCGTGGGTCGTGTA GTCTGGGAGGTCAGACAAGTGGAAAGTCTCAAACACGACGCCCTGACCACAGAA AGCTGCGTCTCCGTGCAGAAGGATGGACATCACCTT (SEQ ID NO: 121) contiε01317 length=250 numreads=4

AGTATATTCGGATGGACATTGAAGTCGTTGGGAACTCGAACAGAGCTCAATCCA ATAACTGTCAGCAAATCCTTGTCAACCCCTGTATCCCAAGTTGTGATAGCACTTG GGGCAGGTTTGTAACCAGTCCATAATTGCCTTTAGGATCTGGTGGAATAAAATTG CTAGCATTCTCTAAAGCTGCAGTTAACTGCGCATGGTACTCATTTTTGATATCTTC AATATCTTTTTCAGTAATTAAACCACTCCT (SEQ ID NO: 122)

contig02816 length=256 numreads=ll

AAGGTCTCATATTTGCTGATCGACCGACAGAATGCAGAGCCTTAACGAAATCGGT

TCCTTGTTGCAGTTTTTCTATAACTGGTTCTCCAACTCTGGTCATAATCCTCATAG

AGGCTGCTACATAAGGCGAGTCCGTCACCTGAACACCGATTTTGGACATCGGACC GCCAACTGGACCCATGGAGAAAGGGATGACGAACATTGTACGACCTTGCATGCA ATTGGGGAATCGTTCGTTAAGGCTCTGCATTCTGTC (SEQ ID NO: 123) contig06121 length=594 numreads—439

CCAGAACAGGGCTCAGGCTGCAGTCGCCAACCGTCTGGGTGTGAGGATCAACCA

GGTGAAGAACGTCATCATCTGGGGCAATCACTCTTCCACCCAGTTCCCTGACGCT

CTCAACGCCTCTGTCGTAGTTGAGGGTGCCACCAAGTCTGTCCCTCAAGCCCTCA

ACAACGACCCCTGGCTCAAGGAAGAATTTGTCACGACTGTACAAAACCGTGGAG

CAGCTGTGATCGCGGCCAGGAAGATGTCTTCCGCCATGTCTGCGGCCAAAGCGG

CAGCAGACCACATGAAGGACATATGGAACGGAACTGCTCCCGGGACCTACACCT

CCATGGCTGTCCTCTCTGATGGCTCTTACGGCGTCCCTGCGGATCTCATGTTCTCT

TTCCCCGTTACAATTGCCAACCGAGAGTGGAAAATCGTCCAGGGACTCCCCATCG

ACGACTTCGCCCGGGCTAAGCTGGACATCACCGCCAAGGAGCTCCTCGAAGAGA

AGGAGGATGCCGTTGCGCTCTGTCAGGATTGACTAGACGTAAAGCCCAATCAAA

GGAGTAAATTGTAACCAACTCAAACTGATTAGTGCCATCGGACAAGTGG (SEQ ID

NO: 124) contiε02214 length=253 numreads=4

AGAGAATTTTGGGAGCGACCACAATCAAAGGTTTCCTGTAGTTTCTGACCATTTG CCGCCGAAGGAGGTGGAAATACTGAGCGGGAGTAGTGGGGTTCACGATGTGGAT ATTCACATCTTCTCCATCTGGGCGTACCTCAGATGAATCAGACAGCTGTAGAAAT CTCTCCAATCTGCACGAACTGTGTTCTGGACCAGCTCCATCGTATCCATGGGGTA GCAACATGGTAAGACCACTCATCGTCATCCATTT (SEQ ID NO: 125) contig06953 length=148 numreads=26

TGATGGCTCCTGCAGTTCTAATGAAAGATGCGAAGACCATGTCGACTCCTTGCTG AACTCCAAACTCCAAATCCTGTTTGTCCTTTTCAGACACGGCAGGAAGATCGACA TGATGCCAGGAAGGTTGACACCTTTCTTGCTTCCCAG (SEQ ID NO: 126)

contigOllSO length=1037 numreads~14

TCCTTGATGAACACAGATCGGAGCTGACCGTTCATCTCGAAGAAGACTTCCCTTT

CTCCAGATGGCGTGAGGTCGTCAGCCGTCGCCAGACACTTGATGGAGAGGAGTT

TACCCTTCTCTAGTGGAACCTCGAACTCTTCACCGATCTTGGGGCCGGAGAGGAA

GGTTCTGGTGTCGAGCTTGTCGACGGGGCCGAACTCGTCCCTGAAGTTGAGGAAG

TCGTCGGTGACTGTGGGATAGAGGGCCGAGCTGATGACATCCTTATCGGTCACAA

ACGTGTGCTTCTCCTGCAGCGACTCTTTCAACTTTCCAAAGTCCAGAGGTTTGAG

AGACTCTCCAGGTCGCCCGTCTATCCTTGGCATGTCTTTCAGCACCTGGGATCTG

AAGGGTTCTGGGAATCCTAGGTAGGGTTCGCCGATCGCACCCTGCAGGAACTCA

ACGACGGATTTGGGGAAGGACAACTCCCCAGCGCGCTTGAGGACCTCCTCGGAG

CTCAACTTGTTCTGAACCATGAACTGAGCCAAATCTCCTACCACTTTG

GATGATGGAGTCACCTTGATGATGTCTCCGAGAATCAAGTTGGCTTCTCTGTAGG

CTTTCTTGACTTCCTCGAACTGTTCTCCCAGACCGAGAGAGTAGGCTTGGAACTG

CAGGTTCGTATACTGACCGCCAGGGATTTCATTCAGGTAGACGTCGGCGTTGCCA

GATTTCATCGTGGTGGTGCATTCAAAGGGCGCGTAGAGCGTTCTAGTTTGTTCCC

AGTAAGCTGAGTACTCGGACACGTTCCTCAGATCAAACCCTGTATCCAGCCTGGT

GCCAGCGAGAGAAGCGACGACGGCTCCCATGGAAGGCTGTGACGTCATCCCTGA

CATGGAGTCGACGGCTGTGTCCACTACGTCAGCGCCTGCATCGGCGCAGGCGAG

CATTGAAGCAACCCCAGCGCCGCTTGTGTCGTGCGTGTGGATGTGGAGGGGGAT

GTCTGGCACTGCGTCTTTCACAGCCTTCACCAGAAGTTTGGCCGCTTGCGGCTTC

AGGAG (SEQ ID NO: 127) contJ203691 length=251 numreads=3

AGAAACGATTACGAAGTCTGTCATGAAAACCAACCATCTCGTCTCAGTGGAACA AGGTTGGCCTCAAAGCGGCGTCGGTGCTGAGATCTGCGCGAGGATCATGGAAAG CGAAGCTTTCTTCCACCTAGACGCCCCCGTCATCCGAATCACCGGTGCTGACCTT CCAATGCCCTACAACCATCATCTGGAAGCACTCGCCCTCCCTCAAGTCAAACACG TCGTAGAAACTGTCAAGAAAGTGCTCGGCGCGT (SEQ ID NO: 128)

Bt receptor: contis04242 length=257 numreads=5

TGAACAAGTGTTTTGCTCACTGTTATTACCAACAGACACCGTGATAAGCGGATAA GATACTACTTGGATAATTTCCAAGACATGTTCAGCGTGTTGGCAAGCGGAGACAT CGTTTTGCGTAAGCACTTGGACTATGAGACGGAAGACACATACCTGTTCCAAGTG TATGCCACTGATGGGACGATGAATGCTAGTGCAGCCGTTAACATAACGGTGCTG AACGTGAACGATTGGGACCCCCGATTCAGGTTTCCTCA (SEQ ID NO: 129) contig05324 length=1261 numreads=543

TCATTAAAAATTTACGGTTTAATAAAAATTAATTCTGAATTTCGAGAATATCCAA

CTAGGTAGTAAAGTTCAGTAGAAGTAGACAAGACGTTTTCGATCGATAACGAGG

GTGACTTCGTCGTATCTACAGGACGGTAACTTTGATAGCTGCTCTGTGAGATTTG

CAGATGGCAGGCATTTCTCTCAGCAGGTTATCGACGTCGGTACTCAACAGGCAAA

TGTTGGCGAACTTGTAAAAGTGTTGTAGGAAATATTCATTCTCATGCCCGAAGCA

GAAAGATCTGAAGCGGTGAAGCAGCTTCTTGTAACAAGGGAAGATGTCCTTCCG

CAAGGCCAGCTTCTCCTCAGAGATTACAGACCTTACTTTATGGGAGTTTGCAGTA

GCAGCCATTACGAATTCATCAAACAATTGGTCACCTAATTTTCTGGCCTTGATAA

TTTCATCAACCTCGTTCTGTAACTTTTGTTTTTCAGCCACATCGTTTGACTTGCTAA

TTTTATGCCGAAGGTAGTGCAAAGCAGCCTCTCTGCTGCTGACCGAAACAACATC

ACCGGGTACCGTTTCAGAATACGTTGGGTGCTTCTTGTTAGGCGTCAGCTCTTCC

GGCCCGAGTGAAAAATCGCCGTAGACCTGTACGTGGCTTTTCGTTACGGCGGTCC

TCACGTGTTGGTACTGGTGGAAAAACGTTTCCAGAGTGCCTGTCTTTTCCGAGTC

CATGTGCTCCATCCACCTGACGCTAAATTCATCTCCTAAGCAGGTGCCGTCATCG

CAGTACCATCCCCAAGAACTCTCGTCCCTAGCTGCAGCAGTCATCACGTAAATGT

CTGTATGTTCTGGCAAGATGTTGTCGAACATAGATCCACTTTCACAAGCCTCAAC

ATAGAAGATAAACTTGTCGTACTTCTTCTTTCTACGCATCTGACTGAGAGTATTTT

CCAAATCGTCCGCCATCAGGTTGTCATTGGGGAAACAAAGCATTCCAGTACCGCC

GTGGTCCGCGAAGTACACGAATAAGTTGGTGTTTTCATCACTTTCAATAACTCGT

CCGGTTCCAATTCCTTTCATACCTGTTTTGTTTCCGCTGAGGATGTTCAGGAAATT

TTGTGGCGTGAGATCATTTCCTGTATAGTCAATTTTAACACCAGCGTAGACGTTG

GTTCCATTTGGTTTGTTGATGATGACTCCAGGCGTCGGATTATCGGTATTATAGGC

GATGTCGTTGTACATCATCACAATTATCTTTTCTGCTGGGATTCCTTC (SEQ ID

NO: 130) contig05633 length=430 numreads=8

GAAGACCTGGTGGATGATGTAATTGGCCGCCAATCCGAAATCTCCCTTCGATATC

TGATAAATCGATACTCGACGCCATCTTGAACTCAGTGCCTTCTCGATTCGTCTTCC

GGAGGACAGTTTAGTTTTCGTCGTAGAGGGGAGTCGAGTCGCCGAGGGACATGG

ATTGAGGGCTGTTGATGGTTCGAGGGGTCGACCGGGCTGAGCGAGGGGTGCCGG

AAACGCGGGGAGTGTTCGTTCCGAGGTCGAAAGGATCGTATTTCCCTTGAGTCGG

TGGCAGTCGAGGGGATCTCCCGGTTCGATCGTTCCCTTGAGATCCCATATTAGTC

GCTCTACCCCATCCTACTCTAGGAGACTGCATGCTGCGAGGACTGAAATCCACGT CGTTGTACCGCGGAGTTCTCTGTTCCTCCCGTCATCCCGAGGAGTG (SEQ ID NO: 131)

contig06107 Iensth=1094 numreads=95

CTCCGTTTAGGGCTCTTCGTCTGTTTCGTGGTGACGGTGAGCGGTTTGGAACCGG

TGGAAGACACGAGGTATTATGTTGAACTCGTTAATTCAGCAAACACTACGTGGAC

GGCCGGTTACAACTTTGAAAACTCCTCTGTTGAGTATCTGAGGCAATTCGTGGGC

ACTGAAGTCAACCAGTTCGAACGTATTTTGCCGAATTTTGAAAGTGCTGGTGGCC

ATGAAGACATCGATATCCCGACTAGTTTCGACGCAAGCAAGAAATGGCCCAAGT

GCGCTCACATCAAACACGTCAGAGACCAGAGTGCCTGCGGCTCTTGTTGGGCTGT

GTCTCTGGCGTCGGTTTTGACTGACCGCCTGTGCATCGCCTCAGGAGGAACCTTC

AGGGCGCCAATGTCCGCGGAAACCTGATGTCATGTTGCTCTGAATGCAGTAATGG

CAACGGGTGTCAGGGTGGAAATCACGCTGTCGCTTGGGTCTACGCCAAAACCCA

AGGCATCGTTACTGGTGGGGACTACCACTCTCATATTGGTTGCCAACCCTACCTA

CTTCAGCCTTGCGGACATTCTGCCGACTCCAAGCGTGTTTCCTGTGGAGAATTATT

GCCCTCAGTGAAGGCCCCAGTTTGCGAGAACACCAAGTGCTTGAACCCTTTCTAC

AAGAAAAATACATTCTCCAAAGACCATAAATTCGTAAAAGAAGCCTATGCTATT

ATGAACGACGAAAAAGCAATTCAACGCGAAATCCTGCAGAGGGGACCTGTGGCT

GCAGCTTTCGTCGTTTACGCAGATTTCCCCTCCTATAAATCAGGTGTGTACCAGA

GAATGTCGGAGGCTGTGGTTGGAGGTCACGGCGTTAAAATCATCGGATGGGGAA

CTGAAGGGGGTGTCGACTACTGGCTGGTCGTCAACTCCTGGAACACCGACTGGG

GTGAGAAAGGGCTCTTCAAAATCCGCCGAATGACCGCAGGCGTCGATTTCGAAC

AGATGATCACTTTTGGCGTCGTATAATTTGTTACCGTTCTGTTATATAGGATTAGG

TACCAATGTTATCTCACAATTAATTTAATGTGAATGATATATATTATTTTAAAC

(SEQ ID NO: 132) contJ206354 Ienεth=444 numreads=275

TGAAAGTGTAGGCATTGGTAAGTTCAACTGCCTCGATAGGTGGAATATCCGTATG

GGCTGAGCAGCAATGGCACGTGATAATGTGTGTTTCCATCTTCAATGTTGAAAAC

GACTTCAACATAAGGATAAAACGTTTTGGCTCCCTTCGACGAGAAATACTGCTCA

GTCTCGAATTTCATCTTATGGATTCCGGGAGAGATAGATCCAGGCTTGGTGAAAC

CAGGGAGGCGTCCGTCCGAATTCGTTTTGTCTTTGTTGATCTCCACCCAATGGTTC

CCAACGTCTGGTGCCATTTTGTAAAGTGTAACAGACAGTCCAGCTGCTGGAAGGC

CAGAAGATGTGTCCAGAACGTGGCAAGTGATTGGAATCACGTTAGACGCCATGG

TGTTTTTCACGGAAAGATGTTGGGAAATCACTTTGAGTCGTTCCCCGGCCGCTGA

CATA (SEQ ID NO: 133)

The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.

Claims

THAT WHICH IS CLAIMED IS:

1. A transgenic plant that contains a polynucleotide construct that comprises a heterologous nucleotide sequence operably linked to a promoter that drives expression in the plant, said heterologous nucleotide sequence encoding an active agent toxic to an insect pest; said active agent selected from the group consisting of: (i) an RNA that inhibits expression of a target gene in said insect pest, and reduces or inhibits the expression of the expression product {e.g., protein, ribosomal RNA) (U) an antibody that binds to the expression product of said target gene and inhibits the activity thereof, and (Hi) an aptamer that binds to the expression product of said target gene and inhibits the activity thereof.

2. The plant of claim 1, wherein said insect pest is a member of the Heteroptera suborder of the Hemiptera order.

3. The plant of claim 2, wherein said insect pest is a member of the Miridae family.

4. The plant of claim 3, wherein said insect pest is a plant bug, leaf bug, or grass bug.

5. The plant of claim 2, wherein said insect pest is a member of the Pentatomoidea superfamily.

6. The plant of claim 5, wherein said insect pest is a shield bug or stink bug.

7. The plant of claim 1-6, wherein said target gene encodes a protein of said insect pest selected from the group consisting of: transcription factors, translation factors, microvilla proteins (e.g., mucin, pseudomucin), electron transport proteins, Bacillus thuringiensis toxin (Cry toxin) receptors, insecticide target proteins (e.g., acetylcholine esterase, GABA-gated chloride channels, sodium channels, nicotinic acetylcholine receptors, chloride channels, midgut membrane proteins, mitochondrial ATP synthase, citin biosynthesis enzymes, ecdysone receptors, mitochondrial complex I, III, and IV electron transporters, Mitochondrial complex I electron transporters, voltage-dependent sodium channels, acetyl CoA carboxylase, etc.), and ribosomal RNA.

8. The plant of claim 1-7, wherein said target gene is given in TABLE 1 herein.

9. The plant of claim 1-8, wherein said plant is a monocot.

10. The plant of claim 9, wherein the plant is a monocot selected from the group consisting of wheat, oats, millet, barley, rye, maize, rice, buckwheat, sorghum, triticale, spelt, linseed, and sugar cane.

11. The plant of claim 1-8, wherein said plant is a dicot.

12. The plant of claim 11, wherein said dicot is potato, soybean, Brassica, sunflower, cotton, or alfalfa.

13. The plant of claim 12, wherein said plant is cotton.

14. The plant of claim 1-13, wherein said active agent is a dsRNA.

15. The plant of claim 14, wherein said dsRNA is a double-stranded molecule with a first strand consisting essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and a second strand consisting essentially of a ribonucleotide sequence which is complementary to the nucleotide sequence of the target gene, wherein the first and the second ribonucleotide strands are separate complementary strands (e.g., on the same molecule or on separate molecules) that hybridize to each other to form said double-stranded molecule, and the double- stranded molecule inhibits expression of the target gene.

16. The plant of claim 1-13, wherein said active agent is a secretory antibody.

17. The plant of claim 16, wherein said plant further comprises:

(a) plant cells containing nucleotide sequences encoding immunoglobulin heavy- and light-chain polypeptides, wherein each polypeptide contains a leader sequence forming a secretion signal; and (b) immunoglobulin molecules encoded by said nucleotide sequences, wherein said leader sequence is cleaved from said immunoglobulin molecules following proteolytic processing.

18. The plant of claim 16, wherein said plant further comprises:

(a) plant cells containing nucleotide sequences encoding a biologically functional multimeric protein comprising at least two different polypeptides not naturally produced by plants, wherein each nucleotide sequence encoding a polypeptide of the multimeric protein encodes a leader sequence forming a secretion signal that is cleaved from said polypeptide following proteolytic processing; and

(b) biologically functional multimeric protein encoded by said nucleotide sequences and resulting from assembly of said at least two different polypeptides wherein the multimeric protein is an abzyme or immunoglobulin.

19. A transgenic seed of the plant of claim 1-18, wherein the seed comprises said heterologous polynucleotide.

20. A method for protecting a plant from an insect pest, said method comprising: introducing into said plant at least one polynucleotide construct that comprises a heterologous nucleotide sequence operably linked to a promoter that drives expression in the plant, wherein said nucleotide sequence encodes an active agent toxic to an insect pest; and wherein said plant is a plant of claim 1-18.

21. A method for protecting a plant from an insect pest, said method comprising administering to said insect pest an active agent selected from the group consisting of: (i) an RNA that inhibits expression of a target gene in said insect pest, and reduces or inhibits the expression of the expression product {e.g., protein, ribosomal RNA), (H) an antibody that binds to the expression product of said target gene and inhibits the activity thereof, and (Hi) an aptamer that binds to the expression product of said target gene and inhibits the activity thereof.

22. The method of claim 21, wherein said administering step is carried out by oral or topical administering said active agent.

23. The method of claim 21, wherein said active agent is a dsRNA.

24. The method of claim 21, wherein said active agent is a secretory antibody.

25. The method of claim 21-24, wherein said target gene encodes a protein of said insect pest selected from the group consisting of: transcription factors, translation factors, micro villa proteins (e.g., mucin, pseudomucin), electron transport proteins, Bacillus thuringiensis toxin (Cry toxin) receptors, insecticide target proteins (e.g., acetylcholine esterase, GABA-gated chloride channels, sodium channels, nicotinic acetylcholine receptors, chloride channels, midgut membrane proteins, mitochondrial ATP synthase, citin biosynthesis enzymes, ecdysone receptors, mitochondrial complex I, III, and IV electron transporters, Mitochondrial complex I electron transporters, voltage-dependent sodium channels, acetyl CoA carboxylase, etc.), and ribosomal RNA.

26. The method of claim 21-25, wherein said target gene is given in TABLE 1 herein.

27. The method of claim 21-26, wherein said administering step is carried out by applying said active agent to said plant.

28. The method of claim 27, wherein said applying step is carried out by applying an insecticidal bait composition to said plant, said insecticidal bait composition comprising said active agent.

29. An insecticidal composition comprising a carrier media and an active agent, wherein said active agent comprises a compound selected from the group consisting of: (i) an RNA that inhibits expression of a target gene in said insect pest, and reduces or inhibits the expression of the expression product {e.g., protein, ribosomal RNA), (H) an antibody that binds to the expression product of said target gene and inhibits the activity thereof, and (Hi) an aptamer that binds to the expression product of said target gene and inhibits the activity thereof.

30. In an insecticidal bait composition comprising an insecticide and one or more of insect food, insect atttractant, carrier, and an insecticidal active agent, the improvement comprising employing as said active agent a compound selected from the group consisting of: (i) an RNA that inhibits expression of a target gene in said insect pest, and reduces or inhibits the expression of the expression product (e.g., protein, ribosomal RNA), (H) an antibody that binds to the expression product of said target gene and inhibits the activity thereof, and (Ui) an aptamer that binds to the expression product of said target gene and inhibits the activity thereof.

31. The composition of claim 29-30, wherein said active agent is a dsRNA.

32. The composition of claim 29-30, wherein said active agent is a secretory antibody.

33. The composition of claim 29-32, wherein said target gene encodes a protein of said insect pest selected from the group consisting of: transcription factors, translation factors, micro villa proteins (e.g., mucin, pseudomucin), electron transport proteins, Bacillus thuringiensis toxin (Cry toxin) receptors, insecticide target proteins (e.g., acetylcholine esterase, GABA-gated chloride channels, sodium channels, nicotinic acetylcholine receptors, chloride channels, midgut membrane proteins, mitochondrial ATP synthase, citin biosynthesis enzymes, ecdysone receptors, mitochondrial complex I, III, and IV electron transporters, Mitochondrial complex I electron transporters, voltage-dependent sodium channels, acetyl CoA carboxylase, etc.), and ribosomal RNA.

34. The composition of claim 29-32, wherein said target gene is given in TABLE 1 herein.