JPH0956382A - Gene coding for protein controlling morphogenesis of plant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject new gene, having a specific amino acid sequence or an amino acid sequence containing a mutation of an amino acid residues without affecting the activities and capable of manifesting activities in controlling the morphogenesis of a plant, and coding for a protein capable of performing the control, etc., over the elongation of a stem. SOLUTION: This gene is a new DNA, having activities in controlling the morphogenesis of a plant and coding for a protein containing an amino acid sequence represented by the formula or an amino acid sequence having the substitution, deficiency or insertion of one or more amino acid residues without affecting activities in controlling the morphogenesis of the plant in the amino acid sequence. This gene is useful for regulation, etc., of the stem length of the plant and an inflorescent form. The DNA is obtained by preparing an mRNA from a terrestrial tissue of Arabidopsis thaliana, preparing a cDNA library therewith and screening the cDNA library with an oligonucleotide synthesized based on a base sequence of a mutable gene isolated from a plant body having the mutation related to the control of the morphogenesis.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a gene DNA that controls the ecology of a plant, a DNA encoding an antisense RNA against the gene, and these DNAs.
The present invention provides a technique for controlling the stem length and inflorescence morphology of a plant transformed with.

[0002]

BACKGROUND OF THE INVENTION Plant morphology is believed to be influenced by genetic and environmental factors. Conventionally, as one of the methods for producing a plant with a short stem length or a plant in which the inflorescence morphology is changed, there is a method of changing the morphology of a useful variety by crossing with a plant whose morphology is genetically different, It is difficult to stably obtain individuals with better properties than the parent varieties.

[0003] It is also possible to select individuals who exhibit morphological changes due to mutations in genes affecting morphology due to natural mutation or mutagenesis, and their gene function is reduced. It is extremely difficult to arbitrarily generate an individual in which a mutation occurs only in a gene that controls a specific morphogenesis while retaining the trait.

Therefore, if a gene that controls plant morphogenesis can be isolated, by transforming a plant with a vector (antisense RNA expression vector) into which an antisense RNA for the gene has been integrated, It is believed that it will be possible to produce plants with shorter stems.

Arabidopsis thaliana (Arabidopsis thaliana) is a vegetaive
stage to transition to reproductive stage, accompanied by flower bud elongation, bud formation and subsequent internode (intern)
The expansion of ode) is strongly coordinated and exhibits a highly ordered branching pattern. Standard Arabidopsis ecotype
Known as Landsberg erecta
The Electa strain retains the endogenous erecta (dwarf) mutation and exhibits a different flower bud structure. Form compact flower buds with densely populated flowers on top. Mutations are pleiotropic with round leaves and short flat liquefions (Huang, I. et al., (199
1) Abstracts presented at the UCLA Keystone Symposium).

[0006] Further, dwarf mutants genetically known to have a mutation at the same locus as that of the Landsberg erecta strain have been obtained, and er-101 strain, er-102 strain and
It is named er-103 strain.

[0007]

As described above, the relationship between plant dwarfing and a specific gene has been empirically known to some extent, but the gene itself has not yet been obtained.

The present invention has been made from the above viewpoint, and an object of the present invention is to provide a gene that controls morphogenesis of plants. Further, by transforming a plant with a DNA encoding the gene or an antisense RNA against the gene, and promoting or suppressing the expression of a gene that controls morphogenesis, a plant having a long stem or a stem It is also an object to provide a plant whose length is shortened or a plant whose inflorescence morphology is changed.

[0009]

In order to achieve the above object, the present inventors cloned a gene controlling plant morphogenesis from chromosomal DNA of Arabidopsis to obtain an antisense expression vector incorporating the gene. After
It was found that the morphology of the plant individual transformed with the vector DNA was changed, and the present invention was completed.

That is, the present invention has one or two or more amino acid sequences having an activity of controlling plant morphogenesis and having no influence on the amino acid sequence shown in SEQ ID NO: 1 or the activity of controlling amino acid sequence of plant in this amino acid sequence. Substitution of amino acid residues,
A DNA encoding a protein containing an amino acid sequence having a deletion or insertion.

The present invention also provides a DNA encoding an antisense RNA that suppresses the expression of the above DNA. As this DNA, D having a base sequence substantially complementary to at least a part of the base sequence shown in SEQ ID NO: 1
NA is mentioned.

The present invention further provides a plant transformed with the DNA encoding the above protein, and a plant transformed with the DNA encoding the antisense RNA.

The protein encoded by the DNA of the present invention is a protein that is highly expressed in stems, flowers, etc. in which traits are significantly changed, and is involved in the control of plant morphogenesis, particularly in stem elongation. It is expected that stem elongation can be promoted by transforming a plant with the encoding gene and increasing the expression level of the gene.

On the other hand, by transforming a plant with a DNA encoding the antisense RNA, that is, a DNA that suppresses the expression of a gene relating to the control of plant morphogenesis, it is possible to suppress the stem elongation of the transformed plant. Expected to be possible.

In the present specification, "chromosomal DNA"
And "chromosomal gene" refer to DNA contained in the nuclear chromosome of plant cells and the gene existing on this DNA. Further, the gene relating to the control of plant morphogenesis provided by the present invention may be referred to as “morphological control gene” or “gene of the present invention”.

[0016]

Embodiments of the present invention will be described below. The gene of the present invention can be obtained by, for example, isolating a mutant gene involved in its phenotypic mutation from a plant having a mutation related to the control of morphogenesis. Furthermore, hybridization using an oligonucleotide prepared based on the obtained nucleotide sequence of the mutated gene as a probe, or PCR using a pair of oligonucleotides produced based on the nucleotide sequence of the mutated gene as a primer (polymerase chain reaction)
Thus, the wild-type gene can be obtained from the chromosomal DNA of the wild-type plant.

Preparation of a plant having a mutation relating to the control of morphogenesis, a method for isolating the gene of the present invention from this mutant,
Also, the method of using the gene of the present invention will be described in detail. Incidentally, D
The methods required for general gene recombination such as NA cleavage, ligation, transformation, gene nucleotide sequence determination, and hybridization are described in the instructions attached to commercially available enzymes used for each operation, and Molecular cloning. (Maniatis T. et
al . Cold Spring Harbor Laboratory Press).

<1> Isolation / Identification of Genes Controlling Morphogenesis of Plants (1) Preparation of Plants Having Mutations Related to Regulation of Morphogenesis Mutation is caused in a gene controlling morphogenesis of plants, for example, Arabidopsis thaliana. For this, a mutagenesis method (gene disruption) in which a foreign gene is introduced into plant cells and inserted into chromosomal DNA to disrupt the gene at the insertion site is used. Examples of applicable gene transfer methods include a method using Agrobacterium, an electroporation method for plant protoplast cells, a polyethylene glycol method, and a microinjection method. Among these methods, the method using Agrobacterium is effective for Arabidopsis thaliana because of high transformation efficiency and low induction of mutations other than mutations due to gene transfer.

Here, a binary vector system (a vector system containing T-DNA capable of introducing DNA into plant cells, an origin of replication functional in microorganisms such as Escherichia coli, and preferably a marker gene for selection of plant cells or microbial cells) The method for introducing a foreign gene into a plant cell by the Agrobacterium infection method using) to produce a mutant plant having a gene that controls morphogenesis is shown below.

[0020] Arabidopsis thaliana
In a wild-type strain of Ti plasmid, a binary vector derived from Ti plasmid (having, for example, a hygromycin resistance gene and an ampicillin resistance gene as a marker) was added to the in planta Agrobacterium infection method (Chang S.
S., Park SK et al. Plant J. 5, No. 4 (1994)) and the like, and after about 6 weeks of breeding, seeds are collected. The seeds obtained are sown on an agar medium containing hygromycin to grow. Transformed plants exhibiting hygromycin resistance are transplanted to rockwool and cultivated, and mutants (erecta mutants) whose stem elongation is different from that of wild-type plants are visually selected.

The mutant thus obtained is highly likely to be mutated by inserting a binary vector into a gene that controls morphogenesis of plants.

(2) Isolation of mutant gene of morphological gene From the mutant with altered morphology obtained as described above,
The mutant gene involved in the mutation is isolated using the so-called plasmid rescue method. That is, chromosomal DNA is prepared from a mutant plant by the method described in Cell, 35 (1983) p.35 or the like, cleaved with a restriction enzyme, and the ends in the molecule are ligated by self-ligation. Obtained circular DNA
However, if it contains a binary vector inserted into the chromosome, this DNA molecule functions as a plasmid capable of autonomous replication in Escherichia coli cells, and the transformant is resistant to a marker drug (for example, ampicillin).

Escherichia coli was transformed with the above-mentioned circular DNA, and the transformant resistant to the marker drug was transformed into recombinant plasmid D.
By recovering NA, a chromosomal DNA fragment containing a morphological control gene can be obtained together with the binary vector.

Alternatively, a chromosomal library is prepared by preparing chromosomal DNA from a mutant plant, cutting it with an appropriate restriction enzyme, ligating it with a plasmid or a phage vector, and transforming E. coli with this. A clone having a mutant gene fragment can also be selected by selecting a clone from this library using T-DNA or the like as a probe.

(3) Isolation of wild-type gene of morphology-controlling gene A chromosomal DNA library was prepared from wild-type Arabidopsis thaliana using a P1 phage vector and the mutant gene fragment obtained as described above was used as a probe. By the hybridization, the wild type gene of the morphological control gene can be isolated. The gene of the present invention can also be obtained by preparing a primer based on the nucleotide sequence of the mutant gene and amplifying the wild-type gene from the chromosomal DNA of the wild-type plant by PCR.

The nucleotide sequence of the DNA fragment containing the Arabidopsis thaliana morphogenetic control gene obtained in the Examples below is shown in SEQ ID NO: 2 in the Sequence Listing. This gene has 27 exons (ex
on) and 26 introns.

As mentioned above, the gene of the present invention contains a large number of introns. To obtain a DNA encoding an exon portion, that is, a protein that controls plant morphogenesis, cDNA of a morphological control gene may be isolated.
The cDNA library is prepared by extracting mRNA from the above-ground tissue of Arabidopsis thaliana, synthesizing cDNA using reverse transcriptase, inserting the double-stranded product by polymerase reaction into a vector, and transforming E. coli. Can be made. Since cDNA cloning kits are commercially available, these may be used. A morphological control gene cDNA clone is obtained from the obtained cDNA library using the chromosomal gene as a probe.

C obtained in the following examples as described above
The nucleotide sequence of DNA and the amino acid sequence deduced from this nucleotide sequence are shown in SEQ ID NO: 1 in the Sequence Listing. The translation product of this gene showed homology with RLK5 described in Nature, 345 (1990) p.743. The RLK5 gene was isolated as a receptor-like protein kinase existing on the cell membrane, but it was isolated due to the homology with the nucleotide sequence of a known protein kinase gene. Is completely unknown. The expression pattern of this RLK5 gene is different from the genes involved in the control of morphogenesis, and it is expressed not only in the aerial part but also in the root, and it is considered that the two are functionally different genes.

<2> Utilization of Morphology Control Gene The gene of the present invention is a gene involved in control of plant morphogenesis, particularly stem elongation, and a gene is transformed with this gene to increase the expression level of the gene. By doing so, it is expected that stem elongation can be promoted.

To transform plants with the gene of the present invention, DNA is introduced into protoplasts by a method such as electroporation (electroporation) or a method using Ti plasmid of Agrobacterium. Good. In that case, a chromosomal gene may be used as the gene of the present invention, or cDNA prepared from mRNA may be used.
A may be used.

On the other hand, a plant is expressed with a DNA that expresses antisense RNA that suppresses the expression of the gene of the present invention, that is, RNA that has a sequence complementary to the full length of mRNA transcribed from a morphology control gene or at least a part thereof. It is expected that the stem elongation of the transformed plant can be suppressed by the transformation.

DNA expressing antisense RNA is
It can be obtained by linking the antisense strand (the strand having a base sequence complementary to the sense strand (coding strand)) or at least a part thereof downstream of the promoter. In other words,
2 containing a sequence homologous to the coding strand or at least a part thereof
It can be obtained by ligating the double-stranded DNA in the reverse direction of the original transcription and ligating it downstream of the promoter. The antisense strand can also be obtained from chromosomal DNA or cDNA, but when chromosomal DNA is used, the intron cannot be expected to fulfill the function of suppressing the expression of the gene of the present invention, so it is preferable to use the exon portion. Further, as the at least part of the antisense strand, any of the coding region, 5 ′ untranslated region and 3 ′ untranslated region can be used. In addition, D encoding antisense RNA
NA may contain, in addition to the 3'untranslated region, a sequence encoding poly dU complementary to the poly A chain added to the 3'end of mRNA.

Examples of promoters that can be used in the present invention include CaMV 35S promoter and the like. The plant can be transformed by transforming the plant with the DNA encoding the antisense RNA in the same manner as the transformation with the gene of the present invention.

Further, a region controlling the expression of the gene of the present invention is included upstream of the coding region of the gene of the present invention. This region includes base numbers 1 to 17 of SEQ ID NO: 2.
A region containing at least a part of the sequence represented by 52, more specifically, a region of base numbers 396 to 1752 can be mentioned. It is expected that this region can be used to control gene expression in plant cells.

[0035]

EXAMPLES The present invention will be described more specifically with reference to the following examples. (1) Preparation of mutant plants for regulation of morphogenesis Arabidopsis (Arabidopsis) ecotype WS (Was
Agrobacterium EHA101 strain (Agrobacterium) having pGDW32 which is a binary vector derived from Ti plasmid (has hygromycin resistance gene and ampicillin resistance gene and can autonomously replicate in E. coli cells) in silewskija, purchased from LEHLE SEEDS. tumefacience) to the in planta Agrobacterium infection method (Cha
ng SS, Park SK et al. Plant J. 5, No. 4 (1994))
Infected.

Specifically, it was carried out as follows. pG
Agrobacterium introduced with DW32 was grown overnight in an LB liquid medium containing an antibiotic (hygromycin).
10 μl of this culture solution was diluted (1/20) to 190 μl of Gamborg's B5 medium (containing 2% sucrose, pH 5.5). 19 to 23 days after sowing, Agrobacterium treatment was performed at the growth stage just after starting the drawing.

The flower stem that had just begun to grow was cut out at the base using a No. 11 scalpel, and a 26 G × 1/2 size injection needle was used to penetrate the center of the rosette stem from the cut end. 1 μl of Agrobacterium diluted solution was injected into the wound. During this time, the illuminance was set to 3000 to 4000 lux. Three days after the inoculation, the plants were transplanted to rock fiber minipots, and thereafter, ordinary cultivation was performed. Approximately 6 weeks after inoculation, seeds were harvested from approximately half of the early formed pods.

The obtained seeds were sown on a B5 agar medium containing 10 μg / ml hygromycin, and the temperature was 22 ° C. and the humidity was 30-40.
%, Illumination: 6000-12000 lux, 12 hours light / dark
It was grown in a 12-hour photoperiod. As the nutrient, HYPONeX (manufactured by Murakami Bussan Co., Ltd.) diluted 1/1000 was used. Transformed plants having a hygromycin-resistant pGDW32 T-DNA insertion sequence were transplanted to rockwool and grown to produce T4 seeds (4th
Seeds of the generation). At this time, a visual search was carried out to obtain a mutant in which the elongation state of the stem was different from that of the wild type plant.

(2) Isolation of Mutant Genes Arabidopsis genomic DNA was cloned into Cell, 35 (1983) p.35.
It was prepared by the method described in 1.

5 g of Arabidopsis tissue (root) frozen at -80 ° C. was crushed in a liquid mortar in liquid nitrogen until it became a fine powder, and 25 ml of this buffer for DNA isolation (50
mM Tris-HCl, pH7.5, 0.2M NaCl, 20mM EDTA-Na2, 2% N-
Lauroyl sarcosine sodium salt, 3g / ml urea, 5% T
(E-saturated phenol) and stir to add 25 ml of phenol.
/ After adding chloroform, 1.5 ml of 10% SDS (sodium dodecyl sulfate) was added and gently stirred at room temperature for 10 minutes. After centrifuging this at 6000prm for 10 minutes, add 25 ml to the water layer.
6000prm again after adding phenol / chloroform
It was centrifuged at 10 minutes. Add 15 ml of ethanol to this water layer,
Immediately after stirring, it was centrifuged at 6000 prm for 10 minutes. The supernatant was discarded, 25 ml of 70% ethanol was added to the precipitate, the mixture was vortexed, centrifuged at 6000 prm for 10 minutes, the supernatant was discarded, and the precipitate was dried under reduced pressure. Add 400 μl TE, pH 8.0 (10 μl
g / μl RNase).

200 μg of chromosomal DNA prepared from the mutants involved in controlling morphogenesis as described above
Purified by CsCl ultracentrifugation. Of this, 1 μg
The chromosomal DNA is cleaved with EcoRI or XbaI, the restriction enzyme fragment is purified by phenol extraction and ethanol precipitation, the ends of the molecule are ligated by self-ligation, and the resulting circular DNA is used to transform E. coli (XL1-Blue MRF '( STRATAGEN
(Purchased from company E))) was transformed. As a result, about 1000 colonies of transformants showing ampicillin resistance could be obtained. Plasmid DNA from the obtained resistant colonies
Was extracted and analyzed.

Among the plasmids rescued as described above, the one obtained from the DNA cleaved with EcoRI is pR.
PRX obtained from DNA digested with Ea, pREb, and XbaI
They are named a and pRXb.

(3) Isolation of Mutant Gene Using the chromosomal DNA fragment contained in the plasmid rescued as described above as a probe, chromosomal DN
Mutant genes were isolated from the A library. P1
Using a phage vector (purchased from DU PONT), a nuclear DNA library of Arabidopsis ecotype Columbia strain was prepared according to the method described in The Plant Journal, 7 (1995) p.351. On the other hand, the above pR
The EcoRI-XbaI fragment prepared from Xb and pREa was labeled with ³² P, and plaque hybridization was performed using these as probes. As a result, two positive clones were obtained and named 28D7 and 61H10, respectively. A restriction map of these clones was prepared, and 28D7 was about 25 kb, 61
H10 was found to have an insert fragment of about 75 kb derived from the Arabidopsis chromosomal DNA (Fig. 1).

Further, the inserted DNA for nucleotide sequence determination
Subcloning of the mutant gene contained in the fragment was performed. At this time, a restriction enzyme map of sequences adjacent to T-DNA,
Southern analysis of the insert was performed to estimate the T-DNA insertion site on the insert (Fig. 2). The chromosomal DNA sequence near this insertion site was subcloned into Bluescript II SK +. The position on the insert of the obtained subclone is shown in FIG.
Shown in

(4) cD of a gene relating to regulation of morphogenesis
Isolation of NA clones mRNA was prepared from aerial tissues of Arabidopsis ecotype Colombia strain, and molecular cloning (Sambroo
k, J., Fritsch, EF and Maniatis, T. (1989), AL
aboraroty Manual, second edition, Cold Spring Harb
or LaboratoryPress, Cold Spring Harbor, NY) to prepare cDNA, and a cDNA library was prepared using λYES (purchased from CLONTECH) as a vector. On the other hand, a sequence adjacent to T-DNA was excised from pRXb and pREa and labeled with ³² P. Using these as probes, plaque hybridization was performed on 300,000 plaques of the phage library. The plasmid thus selected was designated as pKUT161.

(5) Analysis of genes relating to regulation of morphogenesis The nucleotide sequences of cDNA clones and chromosomal genes contained in pKUT161 were determined. The nucleotide sequence of cDNA and the amino acid sequence deduced from this sequence are shown in SEQ ID NO: 1
Shown in The nucleotide sequence of the DNA fragment containing the chromosomal gene is shown in SEQ ID NO: 2.

When the amino acid sequence deduced from the nucleotide sequence of the cDNA was analyzed, the translation product of this gene was Na
ture, 345 (1990) p.743 and showed homology with RLK5. The RLK5 gene was isolated as a receptor-like protein kinase present on the cell membrane,
It was isolated by homology to the nucleotide sequence of a known protein kinase gene, and the function of the gene and its translation product are completely unknown. The expression pattern of this RLK5 gene is different from the genes involved in the control of morphogenesis, and it is expressed not only in the aerial part but also in the root, and it is considered that the two are functionally different genes.

(6) Analysis of mutant for regulation of morphogenesis: The mutant for regulation of morphogenesis obtained in (1) above is a mutation in the same locus as the electa mutation of the Landsberg erecta strain from the elongation state of the stem. Was thought to be. Therefore, first, a genetic complementation test was performed on the Landsberg erecta strain, er-103 strain, and the mutants isolated in the present invention. As a result, it was found that all of these mutations were caused by mutations at the same gene locus, and the isolated mutant was designated as er-104 strain.

Landsberg erecta strain, er-103 strain to mRN
A was isolated and amplified by the RT-PCR (reverse transcription-PCR) method. That is, reverse transcription was carried out using mRNA as a template and an oligonucleotide having the base sequence shown in SEQ ID NO: 4 as a primer. Then, using the obtained cDNA as a template, the oligonucleotide was used as a 3'side primer and SEQ ID NO: 3. A PCR reaction was performed using an oligonucleotide having the base sequence shown as the 5'-side primer.

Using the obtained amplification product as a template, the nucleotide sequence was determined by the direct sequencing method using the above oligonucleotide primer. As a result, Landsberg
In the erecta strain, the isoleucine residue at amino acid number 750 was replaced with a lysine residue as the T at base number 2299 of SEQ ID NO: 1 was replaced with A. In the er-103 strain, the methionine residue at amino acid number 282 was replaced with an isoleucine residue by substituting A for G at base number 896 in SEQ ID NO: 1 of the Sequence Listing.

Among all these mutants, it was found that a part of the nucleotide sequence of the gene relating to the control of morphogenesis has a mutation that causes a change in the amino acid sequence, and this gene controls the morphogenesis (morphology). Could be identified as a regulatory gene).

(7) Expression analysis of morphological control gene As another method for confirming that the cDNA isolated as described above and the chromosomal gene are genes that control morphogenesis, The expression level analysis in tissues and the change in gene expression level between wild type plants and mutant plants were compared and analyzed by Northern analysis. A cDNA clone was used as a probe. From the results of Northern analysis using the Landsberg erecta mutant and the whole wild type plant, the expression of the morphological control gene (mRNA production) was strongly observed in the wild type plant, whereas it was almost observed in the mutant. I wanted to be

Further, as a result of analysis of the expression level of the morphological control gene in each tissue of the wild type plant, the expression level was high in the stems, flowers, etc. in which the traits were markedly changed in the mutants. Was confirmed to be a gene involved in stem elongation.

Furthermore, as described above, since the expression of the morphological control gene is site- and time-specific, the expression control region of the gene of the present invention can be used for controlling the expression of a foreign gene in plants. Such an expression control region includes a region containing at least a part of the sequence represented by base numbers 1 to 1752 of SEQ ID NO: 2, more specifically, base number 396.
-1752 areas.

[0055]

INDUSTRIAL APPLICABILITY The present invention provides a gene that controls plant morphogenesis. It is expected that the stem elongation can be promoted by increasing the expression level of this gene.
In addition, it is expected that stem elongation can be suppressed by transforming a plant with a DNA sequence that expresses antisense RNA for this gene.

[0056]

[Sequence list]

 SEQ ID NO: 1 Sequence length: 3176 Sequence type: Nucleic acid Number of strands: Double-stranded Topology: Linear Sequence type: cDNA to mRNA Origin Organism name: Arabidopsis thaliana Strain name: Colombia Sequence characteristics : Characteristic symbol: CDS Location: 51..2978 CTTTTAAAGT ATATCTAAAA ACGCAGTCGT TTTAAGACTG TGTGTGAGAA ATG GCT 56 Met Ala 1 CTG TTT AGA GAT ATT GTT CTT CTT GGG TTT CTC TTC TGC TTG AGC TTA 104 Leu Phe Leu As Gly Phe Leu Phe Cys Leu Ser Leu 5 10 15 GTA GCT ACT GTG ACT TCA GAG GAG GGA GCA ACG TTG CTG GAG ATT AAG 152 Val Ala Thr Val Thr Ser Glu Glu Gly Ala Thr Leu Leu Glu Ile Lys 20 25 30 AAG TCA TTC AAA GAT GTG AAC AAT GTT CTT TAT GAC TGG ACA ACT TCA 200 Lys Ser Phe Lys Asp Val Asn Asn Val Leu Tyr Asp Trp Thr Thr Ser 35 40 45 50 CCT TCT TCG GAT TAT TGT GTC TGG AGA GGT GTG TCT TGT GAA AAT GTC 248 Pro Ser Ser Asp Tyr Cys Val Trp Arg Gly Val Ser Cys Glu Asn Val 55 60 65 ACC TTC AAT GTT GTT GCT CTT AAT TTG TCA GAT TTG AAT CTT GAT GGA 296 Thr Phe Asn Val Val Ala Leu Asn Leu Ser Asp Leu Asn Leu Asp Gly 70 75 80 GAA ATC TCA CCT GCT ATT GGA GAT CTC AAG AGT CTC TTG TCA ATT GAT 344 Glu Ile Ser Pro Ala Ile Gly Asp Leu Lys Ser Leu Leu Ser Ile Asp 85 90 95 CTG CGA GGT AAT CGC TTG TCT GGA CAA ATC CCT GAT GAG ATT GGT GAC 392 Leu Arg Gly Asn Arg Leu Ser Gly Gln Ile Pro Asp Glu Ile Gly Asp 100 105 110 TGT TCT TCT TTG CAA AAC TTA GAC TTA TCC TTC AAT GAA TTA AGT GGT 440 Cys Ser Ser Leu Gln Asn Leu Asp Leu Ser Phe Asn Glu Leu Ser Gly 115 120 125 130 GAC ATA CCG TTT TCG ATT TCG AAG TTG AAG CAA CTT GAG CAG CTG ATT 488 Asp Ile Pro Phe Ser Ile Ser Lys Leu Lys Gln Leu Glu Gln Leu Ile 135 140 145 CTG AAG AAT AAC CAA TTG ATA GGA CCG ATC CCT TCA ACA CTT TCA CAG 536 Leu Lys Asn Asn Gln Leu Ile Gly Pro Ile Pro Ser Thr Leu Ser Gln 150 155 160 ATT CCA AAC CTG AAA ATT CTG GAC TTG GCA CAG AAT AAA CTC AGT GGT 584 Ile Pro Asn Leu Lys Ile Leu Asp Leu Ala Gln Asn Lys Leu Ser Gly 165 170 175 GAG ATA CCA AGA CTT ATT TAC TGG AAT GAA G TT CTT CAG TAT CTT GGG 632 Glu Ile Pro Arg Leu Ile Tyr Trp Asn Glu Val Leu Gln Tyr Leu Gly 180 185 190 TTG CGA GGA AAC AAC TTA GTC GGT AAC ATT TCT CCA GAT TTG TGT CAA 680 Leu Arg Gly Asn Asn Leu Val Gly Asn Ile Ser Pro Asp Leu Cys Gln 195 200 205 210 CTG ACT GGT CTT TGG TAT TTT GAC GTA AGA AAC AAC AGT TTG ACT GGT 728 Leu Thr Gly Leu Trp Tyr Phe Asp Val Arg Asn Asn Ser Leu Thr Gly 215 220 225 AGT ATA CCT GAG ACG ATA GGA AAT TGC ACT GCC TTC CAG GTT TTG GAC 776 Ser Ile Pro Glu Thr Ile Gly Asn Cys Thr Ala Phe Gln Val Leu Asp 230 235 240 TTG TCC TAC AAT CAG CTA ACT GGT GAG ATC CCT TTT GAC ATC GGC TTC 824 Leu Ser Tyr Asn Gln Leu Thr Gly Glu Ile Pro Phe Asp Ile Gly Phe 245 250 255 CTG CAA GTT GCA ACA TTA TCA TTG CAA GGC AAT CAA CTC TCT GGG AAG 872 Leu Gln Val Ala Thr Leu Ser Leu Gln Gly Asn Gln Leu Ser Gly Lys 260 265 270 ATT CCA TCA GTG ATT GGT CTC ATG CAA GCC CTT GCA GTC TTA GAT CTA 920 Ile Pro Ser Val Ile Gly Leu Met Gln Ala Leu Ala Val Leu Asp Leu 275 280 285 290 AGT GGC AAC TTG TTG AGTGGA TCT ATT CCT CCG ATT CTC GGA AAT CTT 968 Ser Gly Asn Leu Leu Ser Gly Ser Ile Pro Pro Ile Leu Gly Asn Leu 295 300 305 ACT TTC ACC GAG AAA TTG TAT TTG CAC AGT AAC AAG CTG ACT GGT TCA 1016 Thr Phe Thr Glu Lys Leu Tyr Leu His Ser Asn Lys Leu Thr Gly Ser 310 315 320 ATT CCA CCT GAG CTT GGA AAC ATG TCA AAA CTC CAT TAC CTG GAA CTC 1064 Ile Pro Pro Glu Leu Gly Asn Met Ser Lys Leu His Tyr Leu Glu Leu 325 330 335 AAT GAT AAT CAT CTC ACG GGT CAT ATA CCA CCA GAG CTT GGG AAG CTT 1112 Asn Asp Asn His Leu Thr Gly His Ile Pro Pro Glu Leu Gly Lys Leu 340 345 350 ACT GAC TTG TTT GAT CTG AAT GTG GCC AAC AAT GAT CTG GAA GGA CCT 1160 Thr Asp Leu Phe Asp Leu Asn Val Ala Asn Asn Asp Leu Glu Gly Pro 355 360 365 370 ATA CCT GAT CAT CTG AGC TCT TGC ACA AAT CTA AAC AGC TTA AAT GTT 1208 Ile Pro Asp His Leu Ser Ser Cys Thr Asn Leu Asn Ser Leu Asn Val 375 380 385 CAT GGG AAC AAG TTT AGT GGC ACT ATA CCC CGA GCA TTT CAA AAG CTA 1256 His Gly Asn Lys Phe Ser Gly Thr Ile Pro Arg Ala Phe Gln Lys Leu 390 395 400 GAA AG T ATG ACT TAC CTT AAT CTG TCC AGC AAC AAT ATC AAA GGT CCA 1304 Glu Ser Met Thr Tyr Leu Asn Leu Ser Ser Asn Asn Ile Lys Gly Pro 405 410 415 ATC CCG GTT GAG CTA TCT CGT ATC GGT AAC TTA GAT ACA TTG GAT CTT 1352 Ile Pro Val Glu Leu Ser Arg Ile Gly Asn Leu Asp Thr Leu Asp Leu 420 425 430 TCC AAC AAC AAG ATA AAT GGA ATC ATT CCT TCT TCC CTT GGT GAT TTG 1400 Ser Asn Asn Lys Ile Asn Gly Ile Ile Pro Ser Ser Leu Gly Asp Leu 435 440 445 450 GAG CAT CTT CTC AAG ATG AAC TTG AGT AGA AAT CAT ATA ACT GGT GTA 1448 Glu His Leu Leu Lys Met Asn Leu Ser Arg Asn His Ile Thr Gly Val 455 460 465 GTT CCA GGC GAC TTT GGA AAT CTA AGA AGC ATC ATG GAA ATA GAT CTT 1496 Val Pro Gly Asp Phe Gly Asn Leu Arg Ser Ile Met Glu Ile Asp Leu 470 475 480 TCA AAT AAT GAT ATC TCT GGC CCA ATT CCA GAA GAG CTT AAC CAA TTA 1544 Ser Asn Asn Asp Ile Ser Gly Pro Ile Pro Glu Glu Leu Asn Gln Leu 485 490 495 CAG AAC ATA ATT TTG CTG AGA CTG GAA AAT AAT AAC CTG ACT GGT AAT 1592 Gln Asn Ile Ile Leu Leu Arg Leu Glu Asn Asn Asn Leu Thr Gly Asn 500 505 510 GTT GGT TCA TTA GCC AAC TGT CTC AGT CTC ACT GTA TTG AAT GTA TCT 1640 Val Gly Ser Leu Ala Asn Cys Leu Ser Leu Thr Val Leu Asn Val Ser 515 520 525 530 CAT AAC AAC CTC GTA GGT GAT ATC CCT AAG AAC AAT AAC TTC TCA AGA 1688 His Asn Asn Leu Val Gly Asp Ile Pro Lys Asn Asn Asn Phe Ser Arg 535 540 545 TTT TCA CCA GAC AGC TTC ATT GGC AAT CCT GGT CTT TGC GGT AGT TGG 1736 Phe Ser Pro Asp Ser Phe Ile Gly Asn Pro Gly Leu Cys Gly Ser Trp 550 555 560 CTA AAC TCA CCG TGT CAT GAT TCT CGT CGA ACT GTA CGA GTG TCA ATC 1784 Leu Asn Ser Pro Cys His Asp Ser Arg Arg Thr Val Arg Val Ser Ile 565 570 575 TCT AGA GCA GCT ATT CTT GGA ATA GCT ATT GGG GGA CTT GTG ATC CTT 1832 Ser Arg Ala Ala Ile Leu Gly Ile Ala Ile Gly Gly Leu Val Ile Leu 580 585 590 CTC ATG GTC TTA ATA GCA GCT TGC CGA CCG CAT AAT CCT CCT CCT TTT 1880 Leu Met Val Leu Ile Ala Ala Cys Arg Pro His Asn Pro Pro Pro Phe 595 600 605 610 CTT GAT GGA TCA CTT GAC AAA CCA GTA ACT TAT TCG ACA CCG AAG CTC 1928 Leu Asp Gly Ser Leu Asp Lys Pro Val Thr Tyr Ser Thr Pro Lys Leu 615 620 625 GTC ATC CTT CAT ATG AAC ATG GCA CTC CAC GTT TAC GAG GAT ATC ATG 1976 Val Ile Leu His Met Asn Met Ala Leu His Val Tyr Glu Asp Ile Met 630 635 640 AGA ATG ACA GAG AAT CTA AGT GAG AAG TAT ATC ATT GGG CAC GGA GCA 2024 Arg Met Thr Glu Asn Leu Ser Glu Lys Tyr Ile Ile Gly His Gly Ala 645 650 655 TCA AGC ACT GTA TAC AAA TGT GTT TTG AAG AAT TGT AAA CCG GTT GCG 2072 Ser Ser Thr Val Tyr Lys Cys Val Leu Lys Asn Cys Lys Pro Val Ala 660 665 670 ATT AAG CGG CTT TAC TCT CAC AAC CCA CAG TCA ATG AAA CAG TTT GAA 2120 Ile Lys Arg Leu Tyr Ser His Asn Pro Gln Ser Met Lys Gln Phe Glu 675 680 685 690 ACA GAA CTC GAG ATG CTA AGT AGC ATC AAG CAC AGA AAT CTT GTG AGC 2168 Thr Glu Leu Glu Met Leu Ser Ser Ile Lys His Arg Asn Leu Val Ser 695 700 705 CTA CAA GCT TAT TCC CTC TCT CAC TTG GGG AGT CTT CTG TTC TAT GAC 2216 Leu Gln Ala Tyr Ser Leu Ser His Leu Gly Ser Leu Leu Phe Tyr Asp 710 715 720 TAT TTG GAA AAT GGT AGC CTC TGG GAT CTT CTT CAT GGC CCT ACG AAG 2264 Tyr Leu Glu Asn Gly S er Leu Trp Asp Leu Leu His Gly Pro Thr Lys 725 730 735 AAA AAG ACT CTT GAT TGG GAC ACA CGG CTT AAG ATA GCA TAT GGT GCA 2312 Lys Lys Thr Leu Asp Trp Asp Thr Arg Leu Lys Ile Ala Tyr Gly Ala 740 745 750 GCA CAA GGT TTA GCT TAT CTA CAC CAT GAC TGT AGT CCA AGG ATC ATT 2360 Ala Gln Gly Leu Ala Tyr Leu His His Asp Cys Ser Pro Arg Ile Ile 755 760 765 770 CAC AGA GAC GTG AAG TCG TCC AAC ATT CTC TTG GAC AAA GAC TTA GAG 2408 His Arg Asp Val Lys Ser Ser Asn Ile Leu Leu Asp Lys Asp Leu Glu 775 780 785 GCT CGT TTG ACA GAT TTT GGA ATA GCG AAA AGC TTG TGT GTG TCA AAG 2456 Ala Arg Leu Thr Asp Phe Gly Ile Ala Lys Ser Leu Cys Val Ser Lys 790 795 800 TCA CAT ACT TCA ACT TAC GTG ATG GGC ACG ATA GGT TAC ATA GAC CCC 2504 Ser His Thr Ser Thr Tyr Val Met Gly Thr Ile Gly Tyr Ile Asp Pro 805 810 815 GAG TAT GCT CGC ACT TCA CGG CTC ACT GAG AAA TCC GAT GTC TAC AGT 2552 Glu Tyr Ala Arg Thr Ser Arg Leu Thr Glu Lys Ser Asp Val Tyr Ser 820 825 830 TAT GGA ATA GTC CTT CTT GAG CTG TTA ACC CGA AGG AAA GCC GTT GAT 2600 T yr Gly Ile Val Leu Leu Glu Leu Leu Thr Arg Arg Lys Ala Val Asp 835 840 845 850 GAC GAA TCC AAT CTC CAC CAT CTG ATA ATG TCA AAG ACG GGG AAC AAT 2648 Asp Glu Ser Asn Leu His His Leu Ile Met Ser Lys Thr Gly Asn Asn 855 860 865 GAA GTG ATG GAA ATG GCA GAT CCA GAC ATC ACA TCG ACG TGT AAA GAT 2696 Glu Val Met Glu Met Ala Asp Pro Asp Ile Thr Ser Thr Cys Lys Asp 870 875 880 CTC GGT GTG GTG AAG AAA GTT TTC CAA CTG GCA CTC CTA TGC ACC AAA 2744 Leu Gly Val Val Lys Lys Val Phe Gln Leu Ala Leu Leu Cys Thr Lys 885 890 895 AGA CAG CCG AAT GAT CGA CCC ACA ATG CAC CAG GTG ACT CGT GTT CTC 2792 Arg Gln Pro Asn Asp Arg Pro Thr Met His Gln Val Thr Arg Val Leu 900 905 910 GGC AGT TTT ATG CTA TCG GAA CAA CCA CCT GCT GCG ACT GAC ACG TCA 2840 Gly Ser Phe Met Leu Ser Glu Gln Pro Pro Ala Ala Thr Asp Thr Ser 915 920 925 930 GCG ACG CTG GCT GGT TCG TGC TAC GTC GAT GAG TAT GCA AAT CTC AAG 2888 Ala Thr Leu Ala Gly Ser Cys Tyr Val Asp Glu Tyr Ala Asn Leu Lys 935 940 945 ACT CCT CAT TCT GTC AAT TGC TCT TCC ATG AGT GCT TCT GAT GCT CAA 2936 Thr Pro His Ser Val Asn Cys Ser Ser Met Ser Ala Ser Asp Ala Gln 950 955 960 CTG TTT CTT CGG TTT GGA CAA GTT ATT TCT CAG AAC AGT GAG 2978 Leu Phe Leu Arg Phe Gly Gln Val Ile Ser Gln Asn Ser Glu 965 970 975 TAGTTTTTCG TTAGGAGGAG AATCTTTAAA ACGGTATCTT TTCGTTGCGT TAAGCTGTTA 3038 GAAAAATTAA TGTCTCATGT AAAGTATTAT GCACTGCCTT ATTATTATTA GACAAGTGTG 3098 TGGTGTGAAT ATGTCTTCAG ACTGAAAAAAAAAAAAAAAAAAAAAAAAAATCAAAAAAAAAAAACAAAAAAAAAAAAACAAA

SEQ ID NO: 2 Sequence length: 9295 Sequence type: Nucleic acid Number of strands: Double strand Topology: Linear Sequence type: Genomic DNA Origin organism name: Arabidopsis thaliana Strain name: Colombia Sequence Features of: feature symbol: exon Location: 1803..1881 Sequence features: Feature symbol: intron Location: 1882..2227 Sequence features: Feature symbol: exon Location: 2228..2366 Sequence features: Feature symbol: intron Location: 2367..2467 Sequence features: Feature symbol: intron Location: 2540..2643 Sequence features: Feature symbol: exon Location: 2468 .. 2539 Sequence features: Feature symbol: exon Location: 2644..2715 Sequence features: Feature symbol: intron Location: 2716..2809 Sequence features: Feature symbol: exon Location: 2810. .2878 Sequence features: Feature symbol: intron Location: 2879..2968 Sequence features: Feature symbol: exon Location: 2969..3040 Sequence features: Feature symbol: intron Location: 3041..3118 Sequence features: Feature symbol: exon Location: 3119..3190 Sequence features: Feature symbol: intron Location: 3191..3266 Sequence features: Feature symbol: exon Location: 3267..3338 Sequence feature: Feature symbol : Intron Location: 3339..3421 Sequence features: Feature symbol: exon Location: 3422..3493 Sequence features: Feature symbol: intron Location: 3494..3586 Sequence features: Feature Symbol: exon Location: 3587..3655 Sequence feature: Feature symbol: intron Location: 3656..3740 Sequence feature: Feature symbol: exon Location: 3741..3812 Sequence feature: Feature Symbol: intron Location: 3813..3888 Sequence feature: Feature symbol: exon Location: 3889..3960 Sequence feature: Character that represents the feature: intron Location: 3961..4048 Sequence feature: Character that represents the feature: exon Location: 4049..4120 Sequence feature: Character that represents the feature: intron Location: 4121..4209 Sequence features: Feature symbol: exon Location: 4210..4281 Sequence features: Feature symbol: intron Location: 4282..4349 Sequence feature: Feature symbol : Exon Location: 4350..4421 Sequence features: Symbol that represents the feature: intron Location: 4422..4508 Sequence features: Feature symbol: exon Location: 4509..4580 Sequence features: Show the feature Symbol: intron Location: 4581..4706 Sequence feature: Feature symbol: exon Location: 4707..4778 Sequence feature: Feature symbol: intron Location: 4779..4860 Sequence feature: Feature Symbol: exon Location: 4861..4932 Sequence feature: Feature symbol: intron Location: 4933..5018 Sequence features: Feature symbol: exon Location: 5019..5090 Sequence features: Feature symbol: intron Location: 5091..5176 Sequence features: Feature symbol: exon Location: 5177..5248 Sequence features: Feature symbol: intron Location: 5249..5412 Sequence features: Feature symbol: exon Location: 5413..5481 Sequence feature: Feature symbol : Intron Location: 5482..5576 Sequence features: Feature symbol: exon Location: 5577..5648 Sequence features: Feature symbol: intron Location: 5649..5726 Sequence features: Feature Symbol: exon Location: 5727..5800 Sequence feature: Feature symbol: intron Location: 5801..5882 Sequence feature: Feature symbol: exon Location: 5883..6011 Sequence feature: Feature Symbol: exon Location: 6096..6443 Sequence features: Feature symbol: intron Location: 6012..6095 Sequence features: Symbol representing the feature: intron Location: 6444..6519 Sequence features: Feature symbol: exon Location: 6520..6890 Sequence features: Feature symbol: intron Location: 6891..6974 Sequence feature: Character representing the feature: exon Location: 6975..7328 Sequence GAATTCAAAG GAATAAGCAT CGGAGACGAT TTAATGTTACCTGACTATCTGAGAGACATCATGACGACATGACATCATACATATCATACATATCATACATATCATACATACATCACATCACATCACATACATACATA TGTACCGTTA GGGAGGAGGA GGAGACGGGA 240 GAGAAAAAAG GAGACGGAGA TAACTCGCCG GCTCTGTTTC CATGGCGGAG GTGATAATGT 300 AGCTGCGCAC GTTAGCTTTT TGTGGTTTGA GTTGGAGAAC AGTGGGAGGC TCACGGTAGC 360 GTGGAGTGAC GACATTGGGG ATAACACCAG AGGCGTCTTA TCTCCGTTGG ACAAATTATT 420 ATTATGGCTA TGAACATTCA ACATATAATT TAATTAGAAA TTTGCGGATG AAAAAGAGGT 480 AAACAATTGC AGAAATGGTT AAAAATATTA ACGTTGTACA GCAAATGATA ATAAAAAGTG 540 TAACGTACAG TGTGTAAGGA ATGGAAAA AT AATAATTTGG GTTAAAATAA ATATGTAGTT 600 TTCTAACTAT ATAGTACTTT TTGAGAAAAG ATAATATTAT GTGTATTTTT ATTGAAACAA 660 ATAAATGATT TAACAAAAAA AAAAAGAGAA GTTAAAATGA AAAGGAATTA TTATTTTTTA 720 AGTTCTTCCT TCTTTTGTTG GGCCTGTGAC CCTTTTAGTT TTAGTCCACT TCGTTCTCAA 780 AGCTTCAAAA TATTAATTTT GTGACAAACC GACCGGAGCC AACCAAACCG GTTAACATCC 840 TAAAACCAAT CATATTTTAT TAAGTTTTGT GTTGATGCTA AACCAAAAAT CATTGGCATG 900 CATATTTCTA AATTTAGTAA TAAACAAAAA CACTTAGAAA TCACACGTTC ACTATACTAA 960 AAAACGTTGA CAAAAACACA ACAACTATAC TAATAATTAA AGAAGAGAAA ACTGAACCAA 1020 ACTTTTTGTA AACTCCTGAA TTTAAATTAG TAATTGAAGT AAGAAGATGA AGAAGAACAT 1080 GTTAAGCAAA CAAAAAAATT ACACTAAAAT CATATAAAAA TACATAATTA CAAAAGTACC 1140 CATAAGATGG ATTTATTGAT ATGGGTCATC TGTGAAACAA GCCACAGAGA GACAAAGACT 1200 CGTAAGTATT GGGCAACGAA AGCGACCTCC TTTATTCACC ACTGCCATTA ACATGTTCTT 1260 CTTCTCCTTC TTCTTCTACA TTTTATGACC GTTTTACCCT TCAAGAGAGA GAAACAAAAT 1320 CACTCCCTCT CACTCACTCT ATCTCTCTCT TCTGCAAAGC TTCAGAACTC TGGCAGAGAG 1380 ATAAAAGATG ATGGGGTTTT TAACTTTATC CTCCCCAAAT AATTCTTCTT CCCTTCATCT 1440 CTCTCTCTTA CACAACAGGT CCCTACATTT GTACAATCTC CTCTCTTTAA AGACTCTCTC 1500 TCTTTCTCTC TCCATCTCTA TCTTACTCTG TATTTCTGTC GTCTGAGCAC TCAATGAAAC 1560 CACTGTAAAT TTCCGCCAGA ATTTGATGTG ATGGAACGAT AAAAATCATT TTTTCTCGGT 1620 TAAAGTAAAA AAACAAAAAC AAATTTCTGT AGAAATCATA ATAAAAGAAA GAAAAAAAAT 1680 CTAATGTCGG TACATAATAC GGTTCTCTTC TTCTTCTCTA TCCTCTGTTT CTTCTTCATG 1740 GAGACTTGAA AGCTTTTAAA GTATATCTAA AAACGCAGTC GTTTTAAGAC TGTGTGTGAG 1800 AAATGGCTCT GTTTAGAGAT ATTGTTCTTC TTGGGTTTCT CTTCTGCTTG AGCTTAGTAG 1860 CTACTGTGAC TTCAGAGGAG GGTCAGTTAT TATACTGATG CATGCTTCTT CAAGTTCAAG 1920 ATTTTCGTCT TTTTGTTTTA TATTAGTGAA AAAAACTTAA AGATGAGATT TTTATATGAT 1980 TTTTGAAGTT TCATTTGGTG AAAATGAGAT CTGGGTACTT GTTATTTTCT ATTTTTGCTT 2040 TTTGTAATGG TTTTTTTTTA CTTGGTGGGT CTTCTATAGA ATCAAAAGAA GCTTTGAATA 2100 AATTAGGGTT TGAGTTTTAT TTTGTTTTCT TGGAAGTTGA ATTTTTAATC TTCTCAAGAA 2160 CTGACAAATA TTTTTTTTTG TTTTTGTGCG TGTGTGTTAA TAAAATATCC TTAAAACAAA 2220 ATTAAAGGAG CAACGTTGCT GGAGATTAAG AAGTCATTCA AAGAT GTGAA CAATGTTCTT 2280 TATGACTGGA CAACTTCACC TTCTTCGGAT TATTGTGTCT GGAGAGGTGT GTCTTGTGAA 2340 AATGTCACCT TCAATGTTGT TGCTCTGTAA GTTTCTTCAT TCCTTTAGAT TACTATTACA 2400 GTGGTTTTTG GTGTTCTTGT GGGAAAAAGT TGTAATTTGT TTTGTGTGTG TTTTCTATGT 2460 TTTGTAGTAA TTTGTCAGAT TTGAATCTTG ATGGAGAAAT CTCACCTGCT ATTGGAGATC 2520 TCAAGAGTCT CTTGTCAATG TAACTGTTTC AACATTCACT GTAGCATGAA ATAAAGTATC 2580 TTACTTTAAT TCTATTCCAC TCTCTGAGTT GTGACTTTTG TCTTCTGTTT TTTTCTAATG 2640 TAGTGATCTG CGAGGTAATC GCTTGTCTGG ACAAATCCCT GATGAGATTG GTGACTGTTC 2700 TTCTTTGCAA AACTTGTAAG AACAGTGATT GGTGTTATTC TACCATTAAA CTTTTGTTCA 2760 TAGAGGTTTT ATTTGATGAA GTGTGTTCAT GTTGTTTTTA ATTCAGAGAC TTATCCTTCA 2820 ATGAATTAAG TGGTGACATA CCGTTTTCGA TTTCGAAGTT GAAGCAACTT GAGCAGCTGT 2880 AAGTAGCTAG TTATTCTGCT ACTAGTCTTC ATATGTCATT GCTAAAAATA TACTCACCAT 2940 GTGGAATATG GATTTTTACT TTGTCCAGGA TTCTGAAGAA TAACCAATTG ATAGGACCGA 3000 TCCCTTCAAC ACTTTCACAG ATTCCAAACC TGAAAATTCT GTATGTTCCC CATGATTCTT 3060 ACATGTCTTA CTACTTTTAG CTATATAGGT GATCATACAT GTGTAATTTC AATTGCAGGG 3120 ACTTGGCACA GAATAAACTC AGTGGTGAGA TACCAAGACT TATTTACTGG AATGAAGTTC 3180 TTCAGTATCT GTAAGTGTCA ATGTTTTTTG AAGTCTGTCA ATGTCTCTTC ATTACCCGGT 3240 GATAATTGTT GTACTATGAT GAGCAGTGGG TTGCGAGGAA ACAACTTAGT CGGTAACATT 3300 TCTCCAGATT TGTGTCAACT GACTGGTCTT TGGTATTTGT GAGTCTTCTT GCACATCTGA 3360 ATAGTATGAT GAGTTCTTTT GTAAATATCA AATATCTGAC TTTGTTTTGA TATTGAATCA 3420 GTGACGTAAG AAACAACAGT TTGACTGGTA GTATACCTGA GACGATAGGA AATTGCACTG 3480 CCTTCCAGGT TTTGTATGTG CCTCTTTCTC TACTTCTAAA CATCATTACT GTAATTTGGG 3540 TTACTTAAGA AAATCTACTT AACTGGTTTG CTTATTACGA ACTCAGGGAC TTGTCCTACA 3600 ATCAGCTAAC TGGTGAGATC CCTTTTGACA TCGGCTTCCT GCAAGTTGCA ACATTGTTAG 3660 TTCTCACCTC TACTAATCTT TTGCTTTAAA TTTTGGCTAG CCTTTGTTTT CTTTTAAAGA 3720 AGATCATTTT CTTATCTTAG ATCATTGCAA GGCAATCAAC TCTCTGGGAA GATTCCATCA 3780 GTGATTGGTC TCATGCAAGC CCTTGCAGTC TTGTAAGTAC TTTTCTTCTA ATCAATGAAG 3840 CTACTTATAA CATTTTCATG AACTTAGGTT ATATGTTTTC TTTTACAGAG ATCTAAGTGG 3900 CAACTTGTTG AGTGGATCTA TTCCTCCGAT TCTCGGAAAT CTTACTTTCA CCGAGA AATT 3960 GTAATTCTTT ACCTGTTTGT TTTCAGTTTG GAGTCAAATG TCATACCATG TTAATGATAG 4020 TGATTTATCT TTTTGGCTTT ATCTCTAGGT ATTTGCACAG TAACAAGCTG ACTGGTTCAA 4080 TTCCACCTGA GCTTGGAAAC ATGTCAAAAC TCCATTACCT GTATGACCAA CCTTCTCTTC 4140 ACTTCTCTTT TTGCATACAG TCACTACTAA GTTGTGTTTC CTTATCAACT ATTTGTAAAA 4200 TATTCATAGG GAACTCAATG ATAATCATCT CACGGGTCAT ATACCACCAG AGCTTGGGAA 4260 GCTTACTGAC TTGTTTGATC TGTAAGTAGT TCTTCCTATG CTTGACATGT TTTGATGTTC 4320 TTATGCTTAT ATGAACTATG TACATATAGG AATGTGGCCA ACAATGATCT GGAAGGACCT 4380 ATACCTGATC ATCTGAGCTC TTGCACAAAT CTAAACAGCT TGTATGTATC TCTTTCTCTG 4440 AAAACTTCTC ACTTGAATGT TCAAGATTGG TGCTTTATAT GATTTTGTGT CTCATTAATG 4500 TAATGTAGAA ATGTTCATGG GAACAAGTTT AGTGGCACTA TACCCCGAGC ATTTCAAAAG 4560 CTAGAAAGTA TGACTTACCT GTAAGTATCG ACGCTGAGAA TTTCTCTAAT CTTATATAAT 4620 ATATAGTTCC ACAGCGTTTG TTTTTTCGAA TTTCAAGTCA TTAACTACTG AGTTTTTGGT 4680 TGCCTTTGAT TTATCGGTTC AACCAGTAAT CTGTCCAGCA ACAATATCAA AGGTCCAATC 4740 CCGGTTGAGC TATCTCGTAT CGGTAACTTA GATACATTGT AAGTGTTTCT TGTTTTCTGT 4 800 GAAGTATACA TCATTATATG TGCCTTGTCT CACATTTATT AAATTTAATG ACATTTGAAG 4860 GGATCTTTCC AACAACAAGA TAAATGGAAT CATTCCTTCT TCCCTTGGTG ATTTGGAGCA 4920 TCTTCTCAAG ATGTGAGCAT CCATAAGACC TCCAGTTTTA TTGTTTATTT CTAGCAAAAG 4980 ATGAAAATGG TTTGTGAACT CTTGCATTCT TGTTATAGGA ACTTGAGTAG AAATCATATA 5040 ACTGGTGTAG TTCCAGGCGA CTTTGGAAAT CTAAGAAGCA TCATGGAAAT GTAAGAAGTT 5100 AACTTCTATC TGCTTGGTTA GAGTTTTTTT CATTTATCTC AATTACTGTT CTGAATTTGT 5160 GTGTTTGTGG TTGCAGAGAT CTTTCAAATA ATGATATCTC TGGCCCAATT CCAGAAGAGC 5220 TTAACCAATT ACAGAACATA ATTTTGCTGT AAGCAATCTT CCTCTTATCC CTTCCAAGCT 5280 GTTAAGAAAT TGTTTTTGTA GAATGAAACT AAAACTCTGT ATACACAATA ATGAGGTCAC 5340 TATAGTGTGA TCCAGGAACA TGTATTGGGT TGGTGATCTA TCTAATGTTG TGTTTCTTAA 5400 AATTGCTTGC AGGAGACTGG AAAATAATAA CCTGACTGGT AATGTTGGTT CATTAGCCAA 5460 CTGTCTCAGT CTCACTGTAT TGTAAGTAGG CACCTTTGGT TCTGAAACAT TTTTTGTCCC 5520 TCTTTGTGCA TCTTTTGCTA AGAATATAAC CCTGCAATCT TCACTAACTC TTATAGGAAT 5580 GTATCTCATA ACAACCTCGT AGGTGATATC CCTAAGAACA ATAACTTCTC AAGATTTTCA 5640 CC AGACAGGT ATGGTAATTT AGCAGGTTTT GGTATTGTGC ATTTTGTTTT GTTTGCTAAT 5700 ATCTATGTTT ATGTTTTTGG ATAAAGCTTC ATTGGCAATC CTGGTCTTTG CGGTAGTTGG 5760 CTAAACTCAC CGTGTCATGA TTCTCGTCGA ACTGTACGAG GTGATTACAT TCTTCTAAAA 5820 GCTTCCATTC ACAAAACCTA AGATAATTAA AGCTCATGTT TCTATCCATG TTTTGTCTGC 5880 AGTGTCAATC TCTAGAGCAG CTATTCTTGG AATAGCTATT GGGGGACTTG TGATCCTTCT 5940 CATGGTCTTA ATAGCAGCTT GCCGACCGCA TAATCCTCCT CCTTTTCTTG ATGGATCACT 6000 TGACAAACCA GGTCTACTCT CCAAACCACT TTACGAATGT TCTTCACCTA CAATGTAATC 6060 CAATAGTTAA TCCTTAAATT TCCTGGTGAC ATCAGTAACT TATTCGACAC CGAAGCTCGT 6120 CATCCTTCAT ATGAACATGG CACTCCACGT TTACGAGGAT ATCATGAGAA TGACAGAGAA 6180 TCTAAGTGAG AAGTATATCA TTGGGCACGG AGCATCAAGC ACTGTATACA AATGTGTTTT 6240 GAAGAATTGT AAACCGGTTG CGATTAAGCG GCTTTACTCT CACAACCCAC AGTCAATGAA 6300 ACAGTTTGAA ACAGAACTCG AGATGCTAAG TAGCATCAAG CACAGAAATC TTGTGAGCCT 6360 ACAAGCTTAT TCCCTCTCTC ACTTGGGGAG TCTTCTGTTC TATGACTATT TGGAAAATGG 6420 TAGCCTCTGG GATCTTCTTC ATGGTAAGTC TCATCGCCAA ACATAGAAAA TTATTTGAAT 6480 CTTCTGTG AC ATAACAACTT GCTTGTGTGT TTTGTAAAGG CCCTACGAAG AAAAAGACTC 6540 TTGATTGGGA CACACGGCTT AAGATAGCAT ATGGTGCAGC ACAAGGTTTA GCTTATCTAC 6600 ACCATGACTG TAGTCCAAGG ATCATTCACA GAGACGTGAA GTCGTCCAAC ATTCTCTTGG 6660 ACAAAGACTT AGAGGCTCGT TTGACAGATT TTGGAATAGC GAAAAGCTTG TGTGTGTCAA 6720 AGTCACATAC TTCAACTTAC GTGATGGGCA CGATAGGTTA CATAGACCCC GAGTATGCTC 6780 GCACTTCACG GCTCACTGAG AAATCCGATG TCTACAGTTA TGGAATAGTC CTTCTTGAGT 6840 TGTTAACCCG AAGGAAAGCC GTTGATGACG AATCCAATCT CCACCATCTG GTTTGTTCTT 6900 TCTTGCCTAT CTCTCTCAGC TGCTCTGTTT AGGTCAAGTC CGTAATCTTG TTTTCATTGA 6960 TTCACTTACA TCAGATAATG TCAAAGACGG GGAACAATGA AGTGATGGAA ATGGCAGATC 7020 CAGACATCAC ATCGACGTGT AAAGATCTCG GTGTGGTGAA GAAAGTTTTC CAACTGGCAC 7080 TCCTATGCAC CAAAAGACAG CCGAATGATC GACCCACAAT GCACCAGGTG ACTCGTGTTC 7140 TCGGCAGTTT TATGCTATCG GAACAACCAC CTGCTGCGAC TGACACGTCA GCGACGCTGG 7200 CTGGTTCGTG CTACGTCGAT GAGTATGCAA ATCTCAAGAC TCCTCATTCT GTCAATTGCT 7260 CTTCCATGAG TGCTTCTGAT GCTCAACTGT TTCTTCGGTT TGGACAAGTT ATTTCTCAGA 7320 ACAGTGAGTA GTT TTTCGTT AGGAGGAGAA TCTTTAAAAC GGTATCTTTT CGTTGCGTTA 7380 AGCTGTTAGA AAAATTAATG TCTCATGTAA AGTATTATGC ACTGCCTTAT TATTATTAGA 7440 CAAGTGTGTG GTGTGAATAT GTCTTCAGAC TGGCACTTAG ACTTCCTATA AGTTCTTGCC 7500 TATCTAAGTT TTTCTAAATT GGGTTATTCT TGTAACATAT CTTAGATCTA GTACTCAACA 7560 CCACGTCACC ACCACAAAAG ATTTCTTATG CTCAAAAACA TATACATAGA AAGAACCTTC 7620 TAAACTACGA GAAACGTTTT GCTATGTAGT GTTATATGTC AACCACGTCT ATGAGAGTGC 7680 AAACGATAGG TTAATAAGTT TTCTCACTTG GCAATAAAAA TGATAAACAA ATATATTGTC 7740 TGATTAATTT ATTTTATATA GTTTTTTTAT AATTTCTTAT ATTAATTCGA ACTCATACAG 7800 CGCGTGAGAC TTTCTAGTTT AGTATAAAGT ACGTATTTTT GCAAAATCAA AATCGTAAAT 7860 ACATACATTT TAAAATGTTA AAAAAGATAA ATCCGTACAC CATTTAAAAA TGGCATTTTC 7920 CTAAGATTTT TTTCAAAAAA GGCATTTTAG ACAAGAACTA ATTACTACAA CTAAAATCTA 7980 CTAACTTTGG TTTTTATGTA TACATTTACG AGAGTCTACA CAAAAAAAAT ACATAAAAGA 8040 AGAAGTAGTA AATAATTAAA ACGTAAAAAA AAAGACTTTT CAAGAAGGCA GAAGAGTAGC 8100 ACTGTTGTGC GATTGTAAAA TCGTCTTGAT TGTTGTTTAT CCCACTGATA AGCCTACCCT 8160 TTTCAAAACT TGTTCTAAG T TTAAATTCTA TTTTTGAACA TGACATACAG TATAAGGCTT 8220 TTTAAAGATA TCATCTTGAT TTTGTTTCTT CCACAGGGAA GCCCTATCCT TTCTTACATA 8280 ATCTTTGTTA GATAATTTTT TATTATTTTC AAAAAAAATA AAATTGAACA TAAGTTTTCT 8340 CAAAGTAATA TGTTCTAACA ATAATAAACA TAATATCATT TTTTTGTTTT AAACTATAAA 8400 GGACTAACAT GGTAAAAAGT TGCAATATAT AAATGATAAT TTAAACTAAA AATTAGAATA 8460 TGGTAACTTT TTCTTCAACA ACATGCCACA TTCGGCTACA TGTCCACTAG GAAGTGTTAT 8520 TATAGAATCG TTAATGTTGG GTACGCTTAT GAAATTATCA ATGTTTGCTT AAATCTATGC 8580 TTAGAAAATT ACCAATATTA CCTTAAAACT ATATTTACGA ATGACCAATA TTGCTTAGAA 8640 CTATGCTTAT GAAATTACCA ATATTTTCTT AAAACTTAAA CACAAAACTC TTTAACAAAA 8700 AAAACTTTAT TTTTATTTTT ATTTTTTTGG CAAAAAAAAA AACTTTATTT ATAAAGTGAA 8760 AGTCTCCAGA TAATTTTGAA TTTCATTTTT CCAGTTTTTA TTTAGAATAA TTTTTCTTCA 8820 TTTACAAAAT AAAAGAAAAC CCTAGGGTTT AGGGTTTAGG GTTTAGGAAA AAGCGATGAT 8880 ATATTAATTG TTATGAAATG TTTTTTTAAA AATAGTTAAC CAAACATTTT TTTAAAGAGA 8940 GTTTAGTTTC ACAAGGCATT TGTAAATTAG AGTAATTATC AATAAAAATG GAAGACAATC 9000 TAATTATTAT TTAGCAAAAA CTAT ATTTAG GAAAATTAGT TAAAGTTTAG AAATATATCA 9060 TCATAGTGTC AAACTAATTA AAATTATTTA ATTTTGTGAT ATACGTGATC ATATAATTTT 9120 ATGAATATTT AATATTATGA TACATGTAAC TCAGTAAACC TAAATTTAGA AGAAAAGTCA 9180 AAATAATCAT AACCAATTTA GATTCAACTT CTACTTTTGT TCCAAGAAAA AAACACATGG 9240 TTTGTTTTGT GGGATACTAA TGACATCTAT CAAAATCTAT GAAACCAAAT CTAGA 9295 SEQ ID NO: 3 sequence length of 18 sequence types: nucleic acid strands Number: Single strand Topology: Linear Sequence type: Other nucleic acids Synthetic DNA Antisense: NO Sequence: TATCTAAAAA CGCAGTCG 18 SEQ ID NO: 4 Sequence length: 18 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: Other nucleic acids Synthetic DNA Antisense: YES Sequence: AAGATTCTCC TCCTAACG 18

[Brief description of drawings]

FIG. 1 is a restriction enzyme map of the morphological control gene clone obtained in the example.

FIG. 2 is a view showing a T-DNA insertion site on a chromosomal DNA fragment deduced from a restriction enzyme map and Southern analysis.

FIG. 3 is a diagram showing the positions of subclones in the sequence near the T-DNA insertion site.

Continued Front Page (72) Inventor Robert F. Whittier 2-1-6 Sengen, Tsukuba-shi, Ibaraki Tsukuba Research Support Center D-6 Mitsui Industry Co., Ltd.

Claims (6)

[Claims] 1. An activity of controlling plant morphogenesis,
It encodes a protein containing the amino acid sequence shown in SEQ ID NO: 1 or an amino acid sequence having substitution, deletion or insertion of one or more amino acid residues which does not affect the activity of controlling plant morphogenesis in this amino acid sequence. D
NA. 2. A DNA encoding an antisense RNA that suppresses the expression of the DNA according to claim 1. 3. The DNA according to claim 2, which has a base sequence substantially complementary to at least a part of the base sequence shown in SEQ ID NO: 1. 4. A plant transformed with the DNA of claim 1. 5. A plant transformed with the DNA according to claim 2 or 3. 6. A DNA that controls the expression of the DNA according to claim 1, which has at least a part of the sequence represented by base numbers 1 to 1752 of SEQ ID NO: 2.