MXPA97006128A - Expression of la sacarosa fosforil - Google Patents

Abstract

The introduction of the activity of sucrose phosphorylase to plants by transformation with a gene for the enzyme increases the rate of hydrolysis of sucrose, leading to increased levels of starch, oil and proteins, the preferred gene is Streptococcus mutans. Surprisingly, in potatoes transformed to express this gene in the tubers, the reduced susceptibility to decoration by slab and the increased uniformity of the starch deposition throughout the tuber were achieved.

Description

EXPRESSION OF Lfl SñCñROSñ FOSFORILflSñ DESCRIPTIVE MEMORY Recent advances in genetic enginery have provided the tools required to transform plants to contain foreign genes. Now it is possible to produce plants that have unique characteristics of agronomic importance and crop processing. 1 (1) Certainly, one of such advantageous features is the improved content and quality of starch and / or solids in various crop plants.Other is the improved content of acetyl and protein of the seeds of various crop plants. Sucrose is the carbon supply unit Ib that is transported from the source tissues of most of the plants to the collecting tissues. In the collecting tissues, it hydrolyzes and composes * es is used to form other un? Da < more complex supply, mainly starch, protein and oil. The hydrolysis is carried out mainly for sucrose without * sa that produces UDPglucose and fructose. The UDPglucose is converted to glucose 1-phosphate or to the U-glucosamine pyrophosphoplasin. The starch content of the harvest tissues of various crop plants has increased through the use of a gene "that encodes a bacterial Phosphosaphine phosphate. See PCT Application UO 91/19806 (equivalent to Mo. Series 08/120, r03, from the United States, Kishore, incorporated herein by reference). This enzyme cataLiza The production of IgGuellae from glucose-I-phosphate. It has also been noted that its expression during several stages of seed development can reduce the oil content which is believed to be due to the deviation of matter. the starch path with a concomitant reduction in its availability for oil production. The harvest < lura do papas is a phenomenon that occurs during production, handling and storage on a large scale. The marsh is seen as a dark spot mainly in the area of the tuber rind. The maize must lead to the loss of quality in the tuber, higher acceptance of the consumer part of potatoes and potato products, and the loss of processing of the tubers that have excessive levels of suction. It has been noted that potato varieties with a higher starch content have a higher susceptibility to maize. It would be desirable to reduce the level or incidence of slab and particularly desirable to do so while increasing the starch content of the tuber. A more uniform distribution of starch and potato tuber solids is also desirable. The heart or core of the potato has a general lower-solids content than the outer region or the crust. When long strips of the potato tuber are cut to make potato chips, the intermediate portions of these strips therefore have lower levels of solids than the ends and this is especially veniaclear of the strips cut from the center of the tuber. . Strips with lower solids content or regions with lower solids content require longer cooking times to achieve the same degree of consumer acceptability. These longer cooking times may result in overcooking of strips with higher solids content. Longer frying times may also result in higher fat absorption and therefore strips with less solids and those regions with lower solids content will have a higher fat content. Fry foods with a higher fat content are less nutty food. In the production of potato strips, the slices are cut through the potato tuber and the uneven distribution of the solids can result in a fried product with overcooked edges, poorly cooked centers and a higher content. of fat (especially in the center). The uneven distribution of solids in the tuber of the potato also results in disproportionate losses of the solids of the potato C of the bark during the process of peeling. The highest content of solids in the tomato is also desirable. The higher content of solids in the form of soluble solids (usually sugars and acids) and more soluble contribute to the efficiency of the process and the production of such items as tomato sauce., pasta, purees and hot sauce. These solids also contribute to the taste- and texture of the processed products. The higher solids content also contributes to the improved flavor of the fresh ji omat. The saccharose phospho- phase is a microbial enzyme that catalyzes the production of glucose-1-phosphate directly from the sucrose. It has observed its activity on a wide range of bacterial and fungal species, and the vain enzyme has been isolated from pots (Punentel et al., 1992).; Vandarnme et al., 1987). Genes of this enzyme have been isolated from Hgrobacterium spp. íFou nier et al., 1994 and references cited therein), Strept ococcus utans, called gt ffl, (Russell and others, Pe ry and others) and Leuconostoc m senteroí de, called spl (Kitao et al., 1992). The heterologous expression of the S. mutans gene in E. coli is described in the United States 4,888,170 (Ourtiss, 1989), incorporated herein by reference. The useful portion of the trans ormed microorganism is used as a vaccine with S. mutans. It is an object of this invention to provide improved means for increasing the starch content of various plants. It is also another objective to provide means to reduce the sucrose content of the seeds in the oilseed crops which result in a reduction in the level of undesirable carbohydrates -such as the + acjuiosa and the raphmosa, while the increases the carbon available for the production of oil and protein. It is also another objective to provide novel constructions of the PDN that are useful in the provision of said means. It is also another objective to provide potato tubers that exhibit an increased content of more uniform starch through the tubercle. In addition, another objective of this invention is to provide potato tubers with reduced susceptibility to maize. It is also an object of this invention to provide improved cereal crops, such as corn, rice, wheat and barley.

BRIEF DESCRIPTION OF THE INVENTION The present invention provides constructs of flüN encoding the enzyme sucrose phosphonlase CSF) and which are useful in the production of the improved starch content in plants. In or aspect of the present invention, seeds are provided < They have a reduced mielo of saca-osa and other carbohydrates, and that will result in an increased content of oil and protein, as a result of SF expression. In carrying out the foregoing, there is provided, in accordance with one aspect of the present invention, a method for modifying the carbohydrate content of the target tissues of the transgenic plants, < } These include the steps of: (a) Inserting a recornbi nan + e RDN molecule from the genome of a plant cell from < double chain, which contains in the sequence, (i) a promoter that functions in the cells of the tissue of a crop plant, (n) a structural sequence of RDN that originates the production of a sequence of RRN which encodes an enzyme sucrose phytoplan, (m) a sequence "RDNR untranslated at the 3 'end that functions in the cells of the plants that cause the termination of the transcript and the addition of poly-denatured nucleotides at the 3' end of the sequence of RRN; (b) obtaining transformed cells of the plant; and (c) regenerate from the transformed cells of the plant trans genetically engineered plants. In another aspect of the present invention there is provided a double stranded RDN recombinant molecule, which contains in the sequence: (i) a promoter - which functions in the cells of the tissue of a target plant, tu) a structural sequence of RDN that originates the production of a RRN sequence that encodes an enzyme pulls out fos fos fon 1 asa, (m) an untranslated RDN sequence at the end 3 'which functions in the cells of the odo plant which requires the termination of the transcript and the addition of polyhadelated mclotides at the 3' end of the RRN sequence. Also provided, in accordance with another aspect of the present invention, are transformed cells of the RDN-containing plant comprised in the aforementioned elements (i), (11) and (111). According to yet another aspect of the present invention, differentiated plants of potato, tomato and cereal are provided which have increased the starch content in the tubers, the fruit and the seeds, respectively, and provided for the cultivation of crops. of oil seed have reduced sucrose and oligosaccharides containing sucrose, such as stachyose and raryose, in the seeds. Methods have also been provided to increase * the starch content in plant organs for starch production, such as potato tuber and cereal seed, and reduce sucrose levels in crop plants with oilseed, such as soybean and cane, leading to increased content of oil and protein. In the realization of the method in the potato, it has unexpectedly been noted that there is a comparatively more uniform distribution of starch, between the heart and the tuber rind. In another aspect of the invention, a method is provided for providing potatoes that have reduced susceptibility to maize. An additional advantage: The activity of sucrose phospho- plas in the collector tissue, such as the tuber of the potato, is related to the provision of an increased, novel activity, which has a much lower Km. for the sucrose (1-25 mil) that for the enzymes of hi-lolization of saca-osa in the plants sucrose synthases and invertases, which have a Km in the range of 50-300 mi. This advantage is important in the establishment and resistance of such collecting tissues, resulting in the virtual improvement of production. DETAILED DESCRIPTION OF THE INVENTION Expression of the gene of a plant that exists in a double-stranded RDN form involves the transcription of the messenger RRN (RRNin) of an RDN chain of an RRN polymerase enzyme, and in the subsequent processing the master transcript of the RRNm within the core. This processing implies a smaller region on the former oar 3 '«| ue aqt' < < -? & nucleus gone from polyademlate to the 3 'end of the RRN. The expression of the RDN to the RRNrn is regulated in a region of the RDN referred to as the promoter.The promoter region contains a sequence of bases that sends signals to the RRN polymerase to associate with the RDN and initiate transcription of the RRN use One of the "Jenas" and the RDN is a model to be a corresponding complementary chain of the NRN A large number of promoters that are active in the plant cells have been described in the specialized forms. Promoters of Nopal i na synthase (NOS) and 00 + opine synthase (OCS) (Which are contained in the tumor-inducing plasmids of Rgrobactenum t urnefac1ens), caul irnoví rus promoters, such as promoters (CalV) 19S and 35S of cauliflower mosaic virus and 35S of the leafwort mosaic virus, the light-promoting promoter of the small unit of La p bulosa-L, 5-bis-phosphate-carboxylase (ssRURTSCO, a very abundant polypeptide in the s plants) and the promoter «Jel protein gene for the binding of chlorophyll a / b, etc. All of these promoters have been used to create various types of RDN constructions < Thu have been expressed in the plants; see, for example, PCT Publication UOB4 / 02913 (Rogers et al., Monsanto). Promoters < can be used in the present invention; What is known and what is noted is that they require the transcription of the RDN in the cells of the plants. Such promoters can be obtained from a variety of sources such as plants and plant viruses including, but not limited to, the promoter-enhanced CaMV35S and promoters isolated from plant genes such as the ssRU ISCO genes. As described below, it is preferred that the particular promoter selected be capable of causing sufficient expression that it results in the production of an effective amount of the enzyme sucrose phosphorylase (SF) that causes the desired increase in starch content. In addition, it is preferred to produce SF gene expression in specific tissues such as the root, tuber, seed, fruit, etc., and the promoter-chosen must have the tissue ^ 1 specific character of desired development. . Those skilled in the art will recognize that the amount of sucrose osfoplase needed to induce the desired increase in starch content may vary with the amount of plant weight already that the excessive activity of sucrose phosphoplase may be detrimental to the plant. Therefore, the function of the promoter must be optimized by selecting a promoter with the desired capacities of tissue expression and the approximate resistance of the promoter, selecting a transformant that will produce the desired activity of sucrose phosphoplasse in target tissues. This form of selection of the combination of transformants is routinely employed in the expression of heterologous structural genes in plants, since there is variation in the tranformants < They contain the same heterologous gene due to the site of insertion of the gene with the plant genorna (commonly referred to as "position effect"). It is preferred that the promoters used in the double-stranded RDN molecules of the present invention have relatively high expression in the tissues in which the increased content of starch and / or dry matter, such as the tuber, is desired. of the potato plant, the fruit of the tomato or the seed of corn, wheat, rice and barley. The expression of the double-stranded RDN molecules of the present invention will rarely be preferred by a cons ituti promoter, which expresses the RDN molecule in all or almost all the tissues of the plant and can be, in some cases, harmful to the growth of the plant. The patatma promoter of class T has shown to be both highly active and tuber specific (Bevan et al., 1986).; Jefferson et al., 1990). A sequence of a -1.0 b portion of the p atm promoter of the tuber-specific T-type for the expression of tuber-in the present invention is preferred. Other vain genes with specific or tuber-specific expression are known, including the potato tuber RDPGPP genes, both large and small subunits (Muí ler et al., 1990), saca-osa smtasa (Salanoubat and Belliard, 1987). , 1989), the main tuber proteins including protein proteases of 22 d and ios protemase inhibitors (Hannapel, L990), the granulated starch synthase gene (SRDG) (Ronde et al., 1990), and other patatins of classes T and T? (Rocha Sosa et al 1989; Mignery et al., 1988). Other promoters that are contemplated to be useful in this invention are those that exhibit improved and specific expression in potato tubers, which are promoters normally associated with expression of bio-genetic starch genes or modification enzymes, or which exhibit different models. of expression inside the tuber of the potato. Some examples of these promoters include those for the genes for granule and other starch enzymes, ramification enzymes (Kossmann et al., 1991; Blennow, A. and Johansson, G., 1991; UO 92/14827; UO 92/11375), enzyme deprotection (Takaha et al., 1993), enzymes, my Lasas, debranching phosphoresis starch (Nakano et al., 1989; Don et al., 1991), pectin esterases (Fbbelaar, and others, 1993), 40kD protein inhibitor, ubstatin, asphalt proteinase (Stukerlj et al., 19990), the carboxy peptide inhibitor adasa, tuberculin polyphenol oxidases (Sha ha ry otr-os, 1992, GenBankR Rccession Nurnbers M95196 and M951 7), putative trypsin inhibitor and other tubercle RDNcs (Stiekerna et al., 1988), and for ß-arnilasa and eeporarnins (from Ioenoea batata, Yoshida et al., 1992; Ohta et al., 1991). In addition, the promoters can be identified as tuber specimens by selecting a gene library for potato genes with respect to genes that are selected selectively or pri- vately in tubers and then determining the promoter regions to obtain the tuber selective promoters. improved tubing. Other promoters can also be used to express a sucrose phosphoplasse gene in specific tissues, such as seeds or fruits. Β-conglicin (also known as 7S protein) is one of the main supply proteins in soybean (Glycine rnax) (Tierney, 1987). Promoters of (3-congl? Ci and other specific promoters and seed, such as the Je napma and fasoolin promoters, can be used to oversease an SF gene specifically in the seeds, or would lead to a reduction in the sucrose content of the seeds, which would result in a reduction the undesirable oligosaccharides and potentially an increase in the content of oil and / or protein, which would be desirable in the seeds used for the production of oil and protein, such as soybeans, sugarcane, sepu rape oilseed, sunflower, safflower, etc. The SF gene will provide more raw material more quickly, from the regulatory mechanisms proper to the dynamic plants, unless they are influenced by other enzymes produced by heterologous genes, their use in the collected tissues. The zeins are a group of supply proteins that are found in the endosperm of corn. Genoin clones have been isolated for Ze genes (Pedersen, 1982), and the promoters of these genes, including the 15 D, 16 kD, 19 kD, 22 kD, 27 kD and gamma genes, could also be used. to express a gene «He SF the seeds of corn and other plants. Other promoters known to work in corn include the promoters of the following genes: Waxy, Frag11, Contracted 2, Branching T and TI enzymes, starch synthases, debranching enzymes, oleosms, glutelmas and sucrose synthases. A particularly preferred promoter for the expression of the maize endosperm of an SF gene is the promoter for a rice glutelm gene, most notably the Osgt-1 promoter (Zheng et al., 1993), If one wanted to increase the oil in the corn seed, instead of the starch, one would choose a promoter that would cause the expression of the SF gene during the deposition of oil. Activate-The such promoter "During the formation of the embryo of the [> lanfa Some examples of promoters active during the ernbpogenesis are the promoters to pair of the genes for globulin and the active proteins (lea) of the final embodiogenesis .. To some examples of suitable promoters for the expression of a gene of SF in Wheat includes those for the genes for the subdomains of RDPglucose pyrophosphoplasse (RDPGPP), for synthases for granules and other alidons, for branching and de-enrichment enzymes, pair-to proteins abundant in embryogenesis, pair -to the gliadins and to the glutelins. Examples of such promoters in rice include those for the genes for the sub-stocks of RDPGPP, for synphase purposes for granules and other starches, for the enzymes for fractionation, for sucrose and for glutamines.

A particularly preferred promoter is the promoter for rice glutelm, Osg -l. Some examples of such promoters for barley include those for genes for ADPGPP subunits, for smtasae for granules and other starches, for branching enzymes, for branching enzymes, for sucrose smtasas, for hordei as, for the embryo globulins and the specific proteins of aleurone. The solid content of the fruit of the tomato can be increased and expressed by a related SF gene (-on a specific fruit promoter.) The promoter of genomic clone 2A11 (Pear, 1989) will contract the expression of the ADPglucose pyrophosphoplasse in the fruit. The promoter E8 (Deikrnan, 1988) would also express the gene of the SF in the fruits of the joma.R "Jemas, the promoters that work during the green stage of the fruit of the tomatoes are described in the PCT Application. PCTUS94 / 07072, filed on June 27, 1994, which designates the United States, incorporated herein by reference, are designated TFM7 and TFM9, which is a fragment of the RDN, isolated from the tomato, of approximately 2.3 kb, which shows 1.4 kb of extrusion 3 'in SEQUENCE ID NO.3, which is a fragment of RDN of approximately 900 bp, of which 400 bp of the 3' end is shown in TD. SEQUENCE NO 4. It is now known also that the promoters It is from potato tubers that work in tomato plants to cause the specific expression of the fruit of an introduced gene. (see for example Serial No. 08 / 344,639, Barry et al., of the United States, RDPGPP of November 4, 1994, incorporated herein by reference). Such promoters include the promoters of the papain potato, the potato RDPGPP promoters and the promoters of the alpudon synthase "Jest ined to potato granules. A particularly preferred promoter for fruit expression is the promoter for the gene that includes a small subunit of RDPGPP in the potato. The solid content of the root tissue can be increased by expressing a SF gene behind a root-specific promoter. The promoter from the acid chitinase gene (Saac et al., 1990) would express the SF gene in the root tissue, the expression in the tissue of the root could also be carried out using the root specific subdorninios. of the promoter of CaMV35S that they have identified (Benfey yo ros, 1989)., The RRN? ro <; Juc? «Jo by an RDN construction of the present invention may also contain a 5 'non-translated main sequence. This sequence can be derived from the promoter selected to express the gene and can be modified specifically to increase or translate the mRNA. The 5'-untranslated regions can also be obtained by par * from the RNAs from suitable eukaryotic genes or from a synthetic sequence of genes. The present invention is not limited to constructs, such as are presented in the following examples, where the untranslated region is derived from the 5 'untranslated sequence that accompanies the promoter sequence. Rather, the untranslated master sequence can be derived from the unrelated promoter or the coding sequence as discussed above.

SIGNAL SEQUENCES FOR THE DETERMINATION OF OBJECTIVES An alternative method to increase the rate of The hydrolysis of the sucrose would direct the SF to the apoplast. Doing this requires that a signal peptide is required at the N 'end of the functional protein. A preferred example of a c-ue sequence encoding such a signal sequence is a signal sequence from the endoplasmic reticulum of a plant from proto-to PR-IB (Ohshirna et al., 1990). In this way, the SF would be active in the apoplast and would allow the sucrose to hydrolyze excellularly and take into account the rapid rnaß transfer of glucose to the cell. Another alternative is to direct the BF to the vacuolar space. Directing the SF to the vacuole of the cell of another plant requires information in addition to the signal peptide (Nakarnura and Matsuoka, 1993). A peptide can be fused at the amino terminus of the FT. directing the enzyme to the vacuole (Sonnewald et al., 1991). Alternatively, a termination sequence extension of carboxy could be combined with an ER signal sequence to direct the enzyme to the vacuole.

SACAROSA FOSFORILRSAS As used in The present, the term "saccharose phosphorylase" means an enzyme that catalyzes a reversible conversion of sucrose and inorganic phosphate to α-D-gl ucosa-1-phosphonate and D-fructose. It can be isolated - from many microbial sources, including S o p o co c c u s m u t n n, 01 ost ridium pasteurianum (Vandarnrne et al., 1 87) P se u d ome sacchar-pphila (Silverstem and otr-os), Pseudomonas put r? facie, Pul luían or pullulans, Acetobacter xylmurn (Vandarnrne and others, 1987), Rgrobacten urn sp. (Fournier et al., 1994) Leuconostoc mésent oí des. The gene per SF enzyme can be obtained by all known methods and has already been carried out from several organisms, such as Agrobactep urn sp. (Fournier and the others, 1 94) and Leuconostoc mesent er-oi «Jes (Kitao et al., 1992) .. The gene has been thickened from S. mutans in C. coli (Robeson et al., 1983), identifying the activity as glucosyl transferase) . The isolation of a gene from St reptococcus mutans is described later in the examples.

The sequence is given as TD. SEQUENCE No. 5. This gene can be used isolated by inserting it into a plant's expression venders suitable for the method of choice transformation as described below. The gene coding for SFORF 488) has been identified in the plasmids "le Ti de A obactepu vitus (formerly A. turne faciens Biotype 3). It has been reported about related sequences in the Ti-plasmids of other strains of R. turnefac ons, in particular pT? C58 (fournier et al., L994) It is likely that a gene encoding SF will be found in all strains. such pLasrni os. Purification of the SF enzyme from * from other bacterial and fungal sources (described above) has been demonstrated. The availability of such materials makes it easy to clone the sequence of the gene for this enzyme; The protein can be used as an immunoassay to raise antibodies that can be used to identify clones in gene libraries based on such expression as (Sarnbrook et al.); The sequences of the peptide at the N terminus. Such proteins can be obtained by the routine sequencing of proteins; and, following the well-established procedures of limited proteolysis, the sequences of the internal regions can also be determined. Such sequences can be used in the designs of nucleotide probes and primers that can be used to identify the genes of the cells. clones or to amplify the gene or portions of the gene from preparations of RRN, RDNc or RDN of the source organism. The detection of E. coli containing clones of saca-osa phosphoplasa is also possible by the minimum medium culture with sucrose as the sole carbon source (Ferreti and the others, 1988). You can find other microorganisms that use SF to hydrolyze * sucrose, examining organisms that can use sucrose or the only carbon source (Russell and others). The protein can be isolated following the activity in the fractions "using the known methods. FL gene that encodes the protein [Hiede isolate the effort co or just described. In this way, many different genes that encode a protein and which have a sucrose phospho- psase activity can be isolated and used in the present invention.

POLIADENYLATION SIGNAL The 3 'non-translated region of the plant chimeric gene contains a polyadenylation serial that functions in the plants to cause the addition of polyladenylate nucleotides to the 3' end of RRN. Some examples of suitable r-egions in 3 'are (1) the untranslated regions, transcribed in 3', 'which contain the pol ladeni lada signal of Arobactepurn of the tumor inducing casimir (Ti) genes, such as the nopalma synthase (NOS) gene, and (?) plant genes such as genes for the supply of soybean and the small safety of the pb-1, 5-b? sphosphonate-carboxy ? lasa (ss RUBISCO). An example of a preferred 3 'region is that of the ssRUBISCO gene of chicho also known as the 3' region of E9.

SYNTHETIC CONSTRUCTION OF GENES The SF gene from Strept ococ us utans is high in fl * T content, which may be adverse in high level expression in plant cells, although as shown below, the gene is expressed at sufficient levels to positively influence the starch content. If it is «Jesea, the gene sequence of the SF can be changed without changing the protein sequence in such a way that it will increase the expression and thus influence even more positively the starch content in the samples. transformed plants. The r-eglas to make changes in the gene sequence is established in UO 90/10076 (Fisehoff et al.). A gene synthesized following the rules set forth in the same can be introduced to the plants as described below and results in higher levels of expression of the SF enzyme. This can be *? A? particularly useful in the monocot monocle, such as corn, rice, wheat and barley.

COMBINATIONS OF OTHER TRANSGENES The effect of SF on transgenic plants can < It can be improved by combining it with other genes that positively influence the content of almulon and / or oil. For example, urges to increase the activity of HDPglucose pyrophosphoplasin (ADPGPP) in plants can be used in 00 combination with a SF gene to increase the starch. Such RDPGPP genes include the glgC gene of E. coli and its utencer glgCld. WO 91/19806 describes how to incorporate this gene into many plant species in order to increase the starch and / or the solids. Another gene that can be combined with SF to increase starch is a gene for sucrose phosphate synphase (SFS) which can be obtained from plants. UO 92/16631 describes one such gene and its use in the [t] ttrics. Another gene that can be combined with SF to increase the oil is a gene for acetyl-CoA-carboxyl, which can be obtained from plants. UO 93/1 1 243 describes one "Such is genes.

TRANSFORMATION / REGENERATION OF PLANTS Plants that can be made to have increased content of polysaccharides (eg, starch) by practicing the present invention include, but are not limited to, corn, wheat, rice, tomato, potato, sweet potato, peanut, barley, cotton, strawberry, raspberry and yucca. Plants that can be made to have modified carbohydrate content by practicing the present invention include, but are not limited to, corn, wheat, rice, tomato, potato, sweet potato, peanut, fat, ha, beet, sugar cane. , apple, pear, orange, grape, cotton, strawberry, raspberry and yucca. Plants that can be made to have reduced discoloration by slab by practicing the present invention include, but are not limited to, tri, potato, sweet potato, barley, beet, cane sugar, apple, pear, peach, orange, grape, banana, plantain and yucca. Plants that can be made to have uniform improved solids content by practicing the present invention include, but are not limited to, potato, sweet potato, banana, yanten and cassava. Plants that can be made to have < Increased production of harvested material by practicing the present invention include, but are not limited to, corn, wheat, rice, tomato, potato, sweet potato, peanut, barley, beet, sugar cane, apple, pear, orange, peach, banana, plantain, grape, cotton, strawberry, raspberry and yucca. The plants that can beMake them have sucrose r * e «Juc? Da that with« Juzcan to the increased content of oil or protein they include soy, corn, sugarcane and gi asol. A double-stranded RDN molecule of the present invention containing an SF gene can be inserted into the genome of a plant by any suitable method. The vectors suitable for transformation of plants include those derived from a plasmid of Ti from Agrobactepurn turnefaciens, as well as those described, as for example by Herrera-Estrella (1983), Bevan (1984), Klee (1985) and the EPO publication 120,516 (Schi lperoort and others). In addition to the vectors for the transfusion of derived plants from the Ti plasmids or root inducers (Ri) of Rgrohactep um, native alterative methods can be used to insert the RDN constructs of this invention into the cells of these plants. Such methods may include, for example, the use of liposomes, electroporation, chemical substances that increase free DNA uptake, the free delivery of RDN by bombardment of my croproyecti les and transformation using virus or pollen. An expression vector < It is suitable for the introduction of an SF gene into monocotyledoneas using microprojectile bombardment, it is composed of the following: a promoter * that is specific and improved for expression in starch supply tissues in monocotyledons, generally the endospepna, such as promoters of the genes of zeina found in the corn husks (Pedersen and others, 1982); an ion provides a splice site to facilitate expression of the gene such as the intron Hsp70 (PCT Publication U093 / 19189); and a 3 'polyadenylation sequence, such as the 3' sequence of nopalma smtase (NOS 3 '; Frley et al., 1983). This expression cassette can be assembled in high copying replicas suitable for the production of large quantities of RDN. A vector for the transformation of plants based on Agrobacter-iurn, particularly useful in the formation of dicotyledonous plants is the vector of the plasmid? M0N53ü (Rogers, S.G., 1987). The plasmid? MON530 is a derivative of ') K PMON505 prepared to transfer the StuT-H ndlll fragment from 2. 3 kb «Je pMON316 (Rogers, S.G., 1987) to pMON526. The plasmid PMON526 is a simple derivative of? MON505 where the Smal site is r * emov¡do by the digestion with Xmal, treatment with Klenow polymerase and ligation ,. The plasmid? MON530 r * et? All the properties of? MON505 and the expression cassette of CaMV35S-NOS now contains a single cutoff for Smal between the promoter and the polyadelation signal. The binary vector pMON5Q5 is a derivative of? MON? 00 (Rogers, SG, 1987) in which the region of plasmid homology of Ti, l IH, has been replaced with a segment and HindIII to Sinal of 3.8 kb of the plasmid rnim from RK2, (Schmidhauser &Helinski, 1985). This segment contains the origin of replica RK2, oriV, and the origin of the transfer, opT, for its conjugation to robac enurn using the procedure «Je copulation tpparental (Horsch R Klee, 1986). The plasmon pMONSGb contains all the important characteristics of? MON200 including the synthetic rnul t ißnlazador for the insertion of the DNA fragments, the NOS / NPTII '/ NOS chimeric gene for resistance to canarnicida in the cells of plants, resistance to the spectinomicida / streptomi ci to determínate for the selection in E. coli and A. turn faciens, intact gene of nopal i na smtasa for the phase and graduation of tranformants and inheritance in progeny and an origin «Je ? BR322 «He replication to more easily make large amounts of the vector in E. coli. The plasmid? MON505 contains a single boundary of T-DNA derived from the former right oar of the nopalma pT? T37 type T-DNA. Southern analyzes have shown that the plasmid? MON505 and any * DNA it contains are integrated into the genome of the plant, that is, the whole plasm is the RDM-1 that is inserted into the genome of the plant. One end of the integrated RDM is between the right boundary sequence and the nopa tassum gene and the other ex oar is between the boundary sequence and the PBR322 sequences. Another particularly useful plasmid or Ti cassette vector is? MON17227. This vector is described by Barry and others in UO 92/04449 (corresponding to U.S.S.N. 07 / 749,611, incorporated herein by reference) and contains a < It encodes an enzyme that confers resistance to glyphosate (called CP4) which is an excellent selection marker gene for many plants, including potato and tomato. The gene is fused to the peptide gone through the EPSPS chloroplast transit of arabuJopsis (CTP2) and is expressed from the FMV promoter as described herein. When the proper number of cells (or pro-oplasts) containing the SF or ADMc gene is obtained, the cells (or protoplasts) are regenerated to whole plants. The selection of the methodology for the regeneration step is not critical, with adequate protocols available for guests from legumes (alfalfa, soybeans, tr bol, etc.), umbelliferae (carrots, celery, parsnip), cruciferas (coL, radish, canola / rapeseed, etc.), cucurbit (melons and cucumber), grasses (wheat, barley, rice, corn, etc.), solanaceae (potato, tobacco, tomato, peppers), vain floral crops, such as sunflower, and trees that produce nuts, such as almonds, cashews, walnuts and pecans. See, for example, Rnmirate, 1984: Shunarnoto, 1989; Fro m, 1990; Vasil, 1990; Vasil, L992; Hayashi oto, 1989; Shimarnoto, 1989; and Datta, 1990. The following examples are provided to elucidate the practice of the present invention and should not be construed in any way to restrict the scope of the present invention. Those skilled in the art recognize that vain modifications, truncations, etc. can be made. to the methods and genes described herein while not deviating from the spirit and scope of the present invention. EXAMPLES EXAMPLE 1 All DNA manipulations such as PCR, agarose electrophoresis, restriction digestions, ligations and transformations of E. coli were performed by normal protocols as described in Sambrook et al. A sucrose-phospholase gene, gt A, was generated by PCR amplification from Stre tococcus rnutans cells. The gene was amplified using an oligonucleotide in 5 '5'OCCGGATOOATGGOAflTTRCAAATAAAAC (SEQ ID NO: 1) and the oligonucleotide in 3' b'GGGGRGCTCRCTCGRRGCTTRTJGTTTGRTCRTTTTCTG (SEQUENCE ID NO.2) The PCR cyclisation conditions were as follows: 94 ° C, 3 '; 55 ° C, 2 '; 7? ° C, 2 '(5 cycles); 94 ° C 1 '; 55ßC? '; 72 ° C 2 '(30 cycles). In the 1462 b PCR product, it was purified using the GeneClean purification system, digested with (BiolOl, Vista, California), and ligated to the BamHI and SacT sites of PUC119. The ligated RDN was synthesized to JM101 and a blue-white sieve was used to identify colonies for plasmid preparation and restriction digestion. Digestion with HindlII was used to screen the forms containing the gtfR. The clones were screened with the correct restriction patterns for phenotypic expression by the ability to use * sucrose as the sole carbon source as follows: the clones were ansformed to the strain of gal-F. coli, SK1592, and were cultured on the minimal medium containing raffinose (which is absorbed and hydrolyzed to galactose and sucrose) and a clonactivo was identified and named PMON17353. An expression cassette was constructed to take into account the constructive expression of gt A in plants. A fragment containing the improved 35S promoter (Kay, R. 1987), the 3 'region of nopaline synthase (Bevan, M. 1984), and the base structure of the pUC vector was prepared from ? MON999 (Rogers and others 1987a) by digestion of resorption with Bg T and Sacl. A fragment containing the coding region of gtfR was prepared from pMO 17353 by restriction digestion with BamHI and SacT. The correct fragments were separated by electrophoresis from the agarose gel and subjected to the GeneClean method. The fragments were ligated, transformed into E. coli JM101, and the plasmids r * ecornb? before putative were terminated by digestion resorption with NotT. A clone was identified and he was named LC) pMON17359. A second expression cassette was constructed to direct the expression "Je gtfA to the tuber of the map. A fragment was prepared containing the promoter of patat ma 1.0 (described above, the 3 'region of cactus ina smtasa and the base structure of the pUC reader from an intermediate vector by restriction digestion with BarnHT and Sacl. An expression cassette was also constructed to direct the expression of gtfA to the tomato fruit. A fragment containing the TFM7 promoter, the 3 'region of the nopaline smtasa and the base structure "Jel vector from pUC starting" Je pMON16987 (Application "Je PCT US94 / 07072, filed on June 27, 1994), which is derived from" Je pMON999 but contains the TFM7 promoter , by restriction digestion with Bg II and Cacl. The correct fragments were separated by electrophoresis of the agarose gel and purified with the GeneClean procedure. Each of these fragments is linked to the fragments "Je BamHI and Sacl from? R10N17353. The transformation and screening of the clones was as described above. The clones were designated as correct and they were not pMON17356 (Pa 1.0 / gt fR / NO) and pMON 17309 (TFM7 / gtfR / N0S). A third expression cassette was constructed to direct the expression of gtfA to the potato tuber using a 3.5 kb promoter from pata ina. The pap 3.5 promoter was obtained from the plasmid pBT240.7. (Bevan and others 1986) the major part of the 3.5 promoter was extruded from PBI240.7, from the HindlII site (a ~ .3500) to the Xbal site at -337 and combined with the rest of the promoter, from the Xbal site to the BglII site at + 22 (formerly a Dral site), in a triple ligation to a rector who provided a site of BgLII to form pl1ON17280. An intermediate vector was prepared by digestion of [> MON17353 with Ba HI / SacI and intersection of the fragment a [> BS. This vector was digested or later with EcoRI and Sacl. pMONl7280 was digested with EcoRI and Sacl resulted in a fragment containing the 3.5 promoter of patatin, the 3 'region of nogali to smtase and the base structure of the pUC vector. The fragments of the correct size were obtained by electrophoresis of the agarose gel and the GeneClean procedure. The fragments were ligated, transformed into E.coli and JM101, and screened by restriction digestion with HindIII. A clone was designated as correct and it was named PMON17495.

Fn pMON1 356, pr10N17359, pMON17389 and PMON17495, the promoter-, the gtfA gel and the 3 'region We can be isolated in a Noti restriction fragment. These fragments can then be inserted into a single site of any vector - MON17227 (described above) or PMON17320 for the transformation of glyphosate-selectable plants. [> MON17320 is a derivative of PMON17227 that contains a cassette "Je Ppatatm 1.0 / CTTP-glgCJ 6. The CTPl-glglCld fusion encodes a modified pyropholase ADPglucose as described by Kishor-e in UO 91/19806. A vector for the inspection of GtfA of the tomato was also constructed by combining the gtfA gene and the 3 'region of pMON17356 with the small subonity gene promoter of potato RDPglucose pyrophoplasse of -2.0 kp (see serial number). US Pat. No. 08 / 344,639, Barry et al., filed on November 4, 1994, incorporated in the present preference) into a plant transfusion vector to form pMON 17486. The RDN is prepared "read vector by digestion with No i followed for treatment with calf intestinal alkaline photase (FAIB). The fragments containing gt are prepared by * digestion with Noti, elect roforesis of the agarose gel and purification with GeneClean. The DNA of the insert vector is ligated, transformed into E.coli strain LE392 and the transformants were screened by digestion. The restriction was used to identify the clones that contained the expression cassettes of gtfA. The clones in which the transcript is from gtfA are in the same direction as the transcription of the selectable marker were assigned as correct and were named PMONL7357 (FMV / CP4 / E9, Patl .0 / gt A / NO), pMON17358 (Patl.0 / CTPl-glgC16 / E-: 9, Patl.Oigtf A / NOS, FMV (0P? / E9), pl * 1ON17360 (FMV / CP4 / E9, E35S / gt fA / NOS), pMONl 390 ( FMV / CP4 / L9, TFM7 / gtfA / N0S), pMON17392 (Patl./CTPl -glgC! 6 / r9, TFM7 / gt R / NOS, FMV / CP4 / E9), and pMON17496 (FMV / CP? / F9, Pat 3.5 / gt fR / NOS). A transformation vector was constructed to direct the expression of gtfA in the seed of naiz. The fragment containing the Osp-L promoter of glutelin, intron of Hsp70 (described above), the 3 'region of nopaline smtase, the resistance "Je cana icma and the base structure of pUC was prepared by restriction digestion, elect roforosis «Jel agarose gel and GeneClean ,. A fragment containing the coding region of gtfA was prepared with? MON1 359 by digestion "restriction with Neo I" and Notr.The fragments were ligated, transformed and screened by restriction digestion. MON 4502 (Osgtl / Hsp70 / gtfA / NOS) A transformation vector was constructed for the expression of gtfA in seeds of oleogenous seed cultures A fragment of Ba HI-EcoRi from PMON17353 was removed from BglII-EcoRI sites "An intermediate vector to give pMON26104 that placed the gtfA gel behind a TS promoter (discussed above) and used for the 3 'sequence of F.9. A fragment of NotT «| ue containing the promoter *« Je FMV, the fusion of CTP2 and the resistance gene of gil phosphate and a sequence «Je 3 'We linked to the site of NotT« Je? RON26J06 trust «Jar? R1ON26106 , vector transformation «Je limit plants EXAMPLE 2 The vector? MONJ7357 was transformed into the Russet Burbank potato callus by following the method described by Earry et al. In UO 94/28149 for the selection of glyphosate from the transformed lines. A line was obtained and the lines were evaluated in the field tests. The results of this test are shown in table 1. As you can see in the same one, there are several lines "} They contained higher levels of starch (measured as total solids) and some of those had re < luc? «Jo The majadura.

TABLE 1 I in P amage P ome io s of the Average of the SoluJos Line (%) Ma r a ry Indicator Control 21.9 3.399 1 72.9 3.798 * 3 22.2 3.479 4 23..3 7.899 6 71.8 2..79 F! 8 22.7 2.979 11 21.6 7, .968 12 22.0 3.383 14 22.3 3.218 1 155 2 .7 2, 979 17 77.3 3.394 18 21.7 3.394 19 22.4 3.713 22 22.7 3.503 Twelve-line tubers were examined for any change in the distribution of starch between the heart and the bark. This was done by molding the tubers, cutting them into strips similar to French fries and measuring the SÓIKJOS using «Jo a test« The comparison of flotation on saline. The average level of solids "The strips of the heart were subtracted" from the average level of solids from the strips of the cortex. In this way a difference in the level of solid solids is lower than that of the control (4.61% in this test) is an indication of uniform distribution of starch in the tuber, which is highly desirable. Results are shown in table 2. As you can see, the difference in the level of solidity between the heart and the heart was reduced to 10 of the 12 1 lines.

TABLE 2 Line Difference of Solids (%) 4.61 3 4.53 4 4"16 6 3.57 8 3.20 11 3. 4 12 4.39 14 4.67 15 3.56 1 7 2.61 18 3.79 19 5.19 22 3.92 Six of these lines were examined the following year in the field, four "He them in locations 6 multiple. (Line number eight was examined only one location). The absolute increase in the level of solids in these five lines, indicating an increase in starch content, was demonstrated again in each line. The TABLE 3 L i nea Inc re sol ting L 0.7 h 4 0.0 6 8 2.61 15 0.211 22 0.1 7 EXAMPLE 3 The expression of gtfA in maize introduces a novel catalytic activity which can facilitate the insertion of sucrose to the endosperrna creating a more inclined concentration ingredient, and conserve energy, since the equivalent of mol of RTP is normally required to convert sucrose to hexose as hexose phosphate. The vector MON24502 has been introduced to the cells of the corn by bornbar-deo with microprojections using different types of embryogeocal callus tissue for its transformation. It was jointly transformed with either (1)? MON19476 containing a selection cassette of the promoter- "improved Je 35S, of the HspTO mtron, the coding sequence of NPT11" for the resistance of cañamiel na and the sequence in 3 'of or (2) PM0N1 336 that contains the selection cassettes for the glyphosate resistance, each using the promoter of the rice aetma and the Hsp70 in, but one uses a tonifying glyphosate oxnJase. CP4 gLyphosphate resistance gene. (1) Immature embryos were isolated from the maize (genotypes "H99 H99) as described in FP 586 355 A. The bipogenic callus was obtained by culturing the immature embryos. 2 weeks on the medium described by Duncan (1985), Called medium D. After 2 weeks, callus (type 1) was obtained and kept subcultivating every 2-3 weeks on fresh medium, approximately four hours before of the bombardment, the callus in culture (middle cycle of subcult ivo) is placed on the medium with added rnanitol and sorbitol for its osmotic retreatment. Approximately 16-24 hours after bombardment with particles coated with? MON245Q2 and pMON19476, the tissue is placed on medium D without mannitol or sorbitol. Approximately two "lees later, the tissue is transferred to medium D containing paramornici na. The resistant tissue is transferred to the fresh medium with paranamomies at intervals of approximately three weeks. The regeneration of the plant is carried out on medium D with 6-benz? Lam? nopup na (without dica ba) during a "pulse" «Je 3-6 days, followed by its placement on the MS medium without hormones. (2) El callo de io El, derived and immature embryos of the genotype" Hi-TI " , it is used following the method and Dennohy, 1994. The structure of Ti or IT was previously treated on N6 1-100-75 medium containing 0.4M of rnanitol • sorbitol (0.2M of each) for four hours before of the bombardment with PMON24502 and pMON19336 and was on this or medium for 16 to 24 hours after the bombardment, the tissue was then transferred to N6 1-100-25 medium without anitol or sorbitol added. 1 - 3 rnM of glyphosate in the medium of N6 1-0-25 (which did not contain casammo acids) Corn fertile plants have been obtained for each method that their seeds have been examined. g fA by the analysis of Ue in blot using goat antibody cul- tivated against gt A expresses «Jo in E. coli. For their expression, 9 have been shown to express gtfA at approximately 0.05 to 0.5% of the total cellular protein. The biosynthetic regime of starch in corn endosperm tissue expressing gtfA (sucrose phosphoplasse) in vitro was omitted using a sugar feeding sample described above (Felker, 1990). Field-grown plants were screened by PCR to identify positive and negative segregants. Positive and control ears were harvested from the two tran lines formed by gtfA (Knowl and De) at 20 or 22 days after pollination, at the time of 1 linear grain filling. The endosperse sections were scanned and fed with L4C sucrose at concentrations of 50 to 200mM. The concentration "Je 200mM is the most physiologically applicable but" Jetudo at the lowest Krn of GtfA for sucrose than the endogenous enzymes, the lowest concentration (50rnM) was used to improve * the probability in issuing an effect of G fA. The points were converted in time to one and two hours after feeding the 140 and the radioactivity incorporated in the starch reaction was determined. The results with the two lines are summarized in the following table (data presented as average calculations incorporated to the starch reaction): TABLE 4A Feeding with Sucrose at 50mM Time of the Sample Torque Cont rol Knowi 1 hr * 9033 20895 2 hr * 15947 76695 with roll 1 hr 10909 10860 2 hr- 19193 2420 TABLE B Feeding with Sucrose at 200 M Sampling time Cont ol Knowl 1 hr 9880 12980 7 r 11175 71407 control 1 hr * 7703 74 1 7 hr * 11038 13007 The results show that endosperm maize tissues expressing GtfA can produce starch at a faster rate (twice) than controls. The differences in the amount of starch are more evident at lower concentrations of susate, essentially due to differences in the kinetics of susatos between GtfA and endogenous sucrose synthase. Differences were also noted when comparing the effects in the De and Knowl lines, exhibiting a more positive effect. The expression of GtfA is very high in Knowl, in the scale of 0.5% of the total protein, while the expression of GtfA in De is in the scale of 0.05%. The differences in the biosynthetic proportions of lmjon are probably a function of GtfA expression levels.

EXAMPLE 4 The vector? MON26106 has been introduced to soybeans and caye through the agrifood of Agrobactepum (Hmchee). After the selection of the transformed cells using glyphosate and the regeneration of whole plants, the seeds placed by the plants were channeled.

EXAMPLE 5 The vector-? MON? 4502 has been introduced into wheat cells by bombardment with icroproyectiies. Immature wheat embryos were isolated as described by Vasil (1993). Ernbpogenic callus was obtained by culturing the immature embryos for 4 to 7 days on a modified MS medium containing approximately 40 g / L of maltose and approximately 7 rngSL 2,4-D of 2,4-D. The callus was subjected to "Jeo with icroproyectiies coated with? MON24502 and a plasmid containing a gene for resistance to bialophos a day after the bombardment, the immature embryos were transferred to a culture dish containing the selective agent. biálofos. After seven days on the culture and the selective medium the medium derived from embryo? Nma «Juro was changed to a shoot-producing medium (modified MS medium without 2,4-D) containing bialophos and grown for 28-40 days . A PCR sample will be made to confirm that the gtfA gene is present in the outbreaks. The buds that contain the. gtFa gene will be rooted and taken to land. When the plants are transformed and cultivated until the last straw, their seeds will exhibit increased levels of starch.

EXAMPLE 6 The pMON? 4502 vector can be introduced into rice cells by bombardment with microprojects. After regeneration and selection, the transected plants will be examined for the expression of the gtfA gene and those that show high expr ession will be cultured until maturity. The seeds of mature plants will exhibit increased levels of starch. All publications and patents mentioned in this specification are hereby incorporated by reference as if they were specifically and individually expressed as a single publication or patent incorporated by reference. By the foregoing, it will be seen that the invention is well adapted to achieve the purposes and objectives set forth hereinbefore together with the advantages which are obvious and which are inherent in the invention. It is understood that certain features and sub-assemblies are useful and can be used without reference to other features or sub-assemblies. This is contemplated by and is within the scope of the claims. Since many modalities can be done pos ib Les de la invention s deviate from the scope of it, it is to be understood that any material exhibited therein or exhibited in the accompanying drawings is to be construed as illusive and not in a limiting sense.

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LIST OF SEQUENCES GENERAL FORMATION : (l) APPLICANT: (A) NAME: Monsanto Coinpany (B) STREET: 800 North L mdbergh Boulevard (C) CITY: St. Louis (ü) STATE: Missouri (E) PRIS: E.U.R. (F) CODTGO POSTAL ': 63167 (G) TELEPHONE: (314)694-3131 (H) TELEFAX: (314) 694-5 5 (n) TITLE OF THE NONVENTION: Expression of the Sucrose Fooplasa in the Plants (lll) NUMBER OF SEQUENCES '5 (iv) COMPUTER READABLE FORM: (A): TYPE OF MEDIUM: Flexible disk (B) COMPUTER: IBM compatible PC (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: Patent In Relay # 1.0, Version # 1.30 (EPO) (vi) PREVIOUS INFORMATION OF THE APPLICATION: (A) APPLICATION NUMBER: US 08 / 386,860 (B) DATE OF SUBMISSION: 10-FEE-1995 (2) INFORMATION FOR LR ID. TO SEQUENCE NO. 1: (l) CHARACTERISTICS OF THE SECUENCTR: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN:? n «Jl? ? al (D) TOPOLOGY: linear (ii) MOLECULE TTPO: different from ac? < Nucleic jo (R) DESCRIPTION: desc = "Synthetic RDN" (xi) DESCRIPTION OF LR SEQUENCE: IDENTIFICATION DF SEQUENCE NO. 1: CCCGGRTCCA TGGCAATTAC AAATAAAAC 29 (?) INFORMATION FOR THE ID. TO SEQUENCE NO. 2: (L) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 39 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: i dividual (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: different from neical acid DESCRIPTION: / desc = "synthetic DNA" (i) SEQUENCE DESCRIPTION: ID. OF SEQUENCE NO: 7: GGGGAGCTCA CTCGAAGCTT ATTGTTTGAT 39 (?) INFORMATION FOR LR ID. OF SEQUENCE NO. 3: (l) CHARACTERISTICS OF SFOUI-N01A: (R) LENGTH: 1478 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: double (D) TOPOLOGY: linear (n) TYPE OF MOLECULE: RDN (geno ico) (i) DESCRIPTION DF SEQUENCE: TD. DE SECUENCTR NO:: (? i) SEQUENCE DESCRIPTION: ID. TO SEQUENCE NO. 3: (xi) SEQUSNCE DESCRIPTIONt SEO. ID NO: 3: AGCCTTGTGT TAGCCGGTAT TCAAACCTTC TTTGACTGAA AATTTTATTA TTTATACATG 60 TTTAAAAtTA CTTTTTAATC TATATAtAAT AGATATCAAT CCTTCATTTA ATTGTATTTT 120 TGTATTAATT CTATAAATAT TAAATTACTT TATTAAAAAT TCTAATTCTG TCACTCGTCA 180 TTTCATAATA TTCTTGACGC TGATGGTAGT GATAATTACG TTGATTGGAC CCACATGGGC 240 CGCTA TTTT TAAAAGCATC AACCTTGGAA TGTAGTGAAT GTTGAGTCTC ATAGCTCACT 300 CACGtr'ACTCA ACAGCAAAAT CTCTCCTCTT TTTCCCTTCT CCAATTCACA TACTGTCACT 360 TGGACAAATA ATATTTGAAA ATTTTGGCCT AAAGTTAGCT TTGGAGCCGT ATGGTAATTT 420 GATACACAAA TTATTATATA ATTGATATAT C? GGTAT? TA TATCAAGTTG TCGCTTCTTC 480 GTTCATTGTT TCTCTCACTA AAATTTTCAA TTCACTTTTT AAAAAATCGA TAAATTTTTA 540 ATATAACTTT ACATAACATA TTCAAAATTA CAAAAATAAA GGATATTTTT ATATGTTTAT 600 TTTTAATGTA AGATTAAATA TTTAGAATTC TTTTTAAGAA CGGTACAAGC AAATTAAAAG 660 AGAGAAGGTA TATTAGTGGG CCTATGTATC TTTGATATCA TATGCCTCTC AAAGAGCATC 720 CT TGAGTC TATATATCTT TGTTGATAGT GATTTAACCA TTTATGTATG TACGTAGTAC 780 TAAGACATGT TAAATAAGAT CCTAGAGAAA GATTTTTGGA AAAGTGAAAA CAGCAATAAA 840 GAAAAGTCAT TTAAACACTT TCCAACAAAC ATTTGGTAAT CGATTTTAAT TACCCACTTA 900 AACAAAACTA TTTGTACGTA AAATGTTTAA GTAGAAAAGA GATTTTTTTA AAAAAAAAAA 960 GAAGGCAAGA GGTCATATAT CTGACCCTTC CTTAAATCCC CGCGTATAAC ACTTTCTTTT 1020 TTTTGTGTGT GTATGTTCAG GAACATTTGT ATTTTCTATT TGAAATTTCT CATTAAGTCA 1080 AATTCGAAAT CTTTTAAATA ATGTAGAGAA ATCTCATTAT? TTTAACAAT CCCACTTGAT 1140 • 5AATTCCTAA ACATTTTCTA TAAAATAACA CTAAATCTTT AATTATACAT ATTACATACC 1200 TAACTCAAGC AATCTTGTCG GAAAAATCAT TAGAAAAGAA TTGGAAATAG GGAAATAAAT 1260 AGACATATTT TGGTTAGTAT CTTTGTCTAT AAGAATGGGT GTGTTAAAGA GCTAGTGCCA 1320 TAGTGTACCA TTCTATTGGT AGCATTTGGC AAGAGTTATT CCCTCTCTCC ATACCAATGG 1380 AGAAGTTTAA TCTTGCTAGA GTCTTATTGT TGCTTCTTCA ACTTGGAACT TTGTTCATTG 1440 CCCATGCATG TCCTTATTGT CCATATCCTC CTTCCACC 1478 KO (?) INFORMATION FOR THE ID. TO SEQUENCE NO. 4: (l) CHARACTERISTICS OF LR SFCUT.NC1A: (A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (OR CHAIN TYPE: «Job le (O) TOPOLOGY: 11 ea L (n) TYPE OF MOLECULE: DNA (genoinic) (xi) SEQUENCE DESCRIPTION: ID. SEQUENCE NO: 4: AAATAAATAT TTCAAAGTAA ATTGTTACTC CCTCTATCCC ATACTCTTTT CTTTTTTTTAA 60 TCC? TTTCTT ACTCTAATTG AACTATTGCA GACAACTTAA ATGTAAATTT TTTTTTTCTT 120 TATCAAAATG ATTGGCTGCT ATATAAATAT CTAATGGTTA TTATACATAA ATTTTAATAT 180 TTTTTTATAAA AAAATATCGA GCTAAATCAT ATCGTTTAAA TATAGAGATG TGTTATTTAT 240 TTAAA? ATTA ATTTTAAAAA AGTCAATATT OTA? ATTAGG ATGAAAGAGT ATTATATTGG 300 TTCTCGCAGT ATAAATACCC TGCATGCCAT TACATTTGTT CAATCATCTT TGCAACGATT 360 TGTCTGCTTT AGCTTCCTTA CATAACATGG CTTCTATAAC TAAAGCCTCA TTACTTATCC 420 TTTTCCTCTC CTTGAATCTC CTTTTCTTCG 450 (2) PRIOR INFORMATION REPORT. OF SEQUENCE NO. 5: (l) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1446 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: «Job le (D) TOPOLOGY: l ineal di) TYPE OF MOLECULE: DNA (geno ico) (? .i.5 DESCRIPTION OF SEQUENCE: SEQUENCE ID NO: 5: ATGGCAATTA CAAATAAAAC AATGTTGATT ACTTACGCAG ACAGTTTGGG TAAAAATTTG 60 AAACAATTGA ATGAAAATAT TGAGAATTAT TTTCCAGATG CTGTTGGCßG TGTCCATTTG 120 CTGCCATTCT TTCCTTCCAC AGGTGATCGT GGCTTTGCAC CGATTGATTA CCATGAAGTT 180 GACTCTGCTT TTGGCCATTG GGATGATGTC AAACGTTTGG GTGAAAAATA TTACCTCATG 240 TTTCATTTCA TGATTAATCA TATTTCGCGT CAGTCTAAAT ATTATAAAGA TTACCAAGAA 300 AAGCATGAAG CAAGTCCTTA TAAAGATCTA TTTTTAAATT GGGATAAATT TTGGCCTAAA 360 AATCGCCCGA CACAAGAAGA TGTGGACCTG ATTTATAAGC GTAAGGATCG AGCACCTAAG 420 CAGGAAATCC AATTTGCAGA TGGCACTGTT GAACATCTCT GGAACACTTT TGGGGAGGAA 480 CAGATTGATC TTGACGTGAC TAAAGAAGTG ACTATGGATT TTATTCGCTC TACCATTGAA 540 AATTTAGCAG CCAACGGCTG TGATCTCATT CGTTTGCATG CCTTTTGCTTA TGCTGTTAAA 600 AAGCTAGATA CGAATGATTT CTTTGTTGAA CCTGAAATCT GGACTCTGCT AGATAAAGTT 660 CGTGATATAG CTGCTGTATC GOGTGCGGAA ATCTTGCCGG AAATTCATGA ACACTATACT 720 ATTCAATTTA AAATTGCAGA CCATGATTAC TATGTTTATG ATTTTGCCCT GCCTATGGTG 780 ACGCTCTACA GCCTATATTC GGGCAAGßTT GACCGTCTTG CCAAATßGGT GAAAATGAGT 840 CCGATGAAAC AGTTCACCAC CCTTGATACA CATGACGGTA TTCCTGTGGT TGATGTTAAG 900 GATATCCTGA CTGACGAAGA AATTACCTAT ACTTCTAATG AOCTTT? TAA GGTCGGTGCC 960 AATGTCAATC GTAAGTATTC AACTGCCGAA TATAATAACT TQGATATCTA TCAAATTAAT 1020 TCAACTTACT ATTCAGCACT TGGTGATGAT GATCAAAAAT ACTTTTTGGC CCGGTTGATA 1080 CAAGCTTTTG CTCCAGOTAT TCCACAGCTT TATTACGTTG ßCTTTTTAGC TGGCAAGAAT 1140 GATCTTGAAT TACTGGAAAG CACTA ?? GAA GGCCGCATTA TCAACCGTCA TTATTATACT 1200 AGTGAAGAAA TTGCTAAGCA ACTGAAGCGG CC? ßTTGTCA AGCCACTTTT AAATCTCTTT 1260 ACTTACCGCA TTCAGTCAGC AGCTTTTGAT TTGGATGGCC GTATTGAAGT GGAAACGCCA 1320 AATGAAGAGA ACATTGTCAT AßAACGTCAA AATAAAQATG ßCAGTCATAT CGCAACAGCA 1380 GAGATTAATC TCCAAGATAT GACATACAGA GTAACAGAAA ATGATCAAAC AATAAGCTTC 1440 GAGTGA 14 * 6

Claims (26)

')' - *. NOVELTY OF THE INVENTION CLAIMS

1. A method to produce a transgenic plant, "Comprising the steps of: (a) inserting into the genome of a plant cell a recornhmanto double-stranded DNA molecule comprising: (i) a promoter that functions in the cells of the target plant tissue, (11) ) a structural sequence of the DNA that originates the production of an RNA sequence encoding a sucrose phosphoplasse enzyme, (111) a 3 'non-derived DNA sequence that functions in the cells of the plants that cause the termination "Jel transcribed and the addition of polyadenylated nucleotides to the 3"end of the RNA sequence, (b) obtaining the transformed cells of the plants, and (c) regenerating from said transformed cells of the plants a genetically transformed plant, of which The genome contains said recombinant molecule "double-stranded DNA of step (a)

2. The method according to claim 1, further characterized in that said RDN sequence encoding the enzyme sucrose phospho- a is obtained to par-ti of Streptococcus mutans.

3. The method according to claim 2, characterized by "Jemas" because said sequence of the RDN coding for sucrose phosphonilase has a sequence shown in the ID. No. 5.

The method according to claim 1, further characterized wherein said genetically formed plant exhibits a property selected from the group "consisting of containing a modified carbohydrate content; unscrewed content of poly saccharides (e.g. starch); improved production; improved uniformity distribution of solids; and reduced susceptibility to discoloration by * majadura.

5. The method of conformity with claim 4, further characterized "or" that said property comprises a content rnod? f? ca «Jo of carbohydrates.

6. Method according to claim 5, further characterized by said modified carbohydrate content is an increase in the solids content. r

7. The method according to claim 6, characterized in that said genetically engineered plant is selected from the group "consisting of potato and tomato.

8. The method according to claim 4, further characterized in that said property is the uni form "J improved distribution of solids.

9. The method according to claim 8, further characterized in that said genetically trans formed plant is selected from the group "consisting of potato and sweet potato.

10. The method according to claim 4, characterized in that said? Rop? E «Ja« i is the suscep ili ad produced by maja «Jura.

11. The method according to claim LO, further characterized in that said genetically transformed plant is selected from the group consisting of potato, banana, apple, triangle, grape and peach.

12. The method according to claim 4, further characterized because "Jicha property is an increased content of pol i take two (for example starch).

13. The "Jo" in accordance with the claim 12, further characterized in that said genetically transformed plant is selected from the group consisting of corn, wheat, rice, tomato, potato, potato, peanut, barley, cotton, strawberry, raspberry and cassava.

14. The method according to claim 4, further characterized in that said property is the improved production.

15. The conforming method gives "J with claim 14, characterized in that said genetically transformed plant is selected from the group consisting of rnaLZ, wheat, rice, tomato, potato, sweet potato, peanut, barley, beet, sugar cane. , apple, pear, orange, peach, grape, cotton, strawberry, raspberry and yucca.

16. The method "Jo" according to claim 1, further characterized in that said plant cell is selected from the group consisting of maize plant cell, plant cell "Je rice, plant cell <.The wheat, cell of the tomato plant, cell of the plant "Je barley, plant of the plant" Je beet., Cell of the sweet potato plant, cell of the peanut plant, cell of the plant of sugarcane, cell «He the grape plant, cell« Je The plant of beast, cell of the plant «Je apple, cell of the orange plant, relula of the yucca plant, cell« Je the plant of banana, plantation plant cell, cell "Je la planta" Je cotton, cell of the strawberry plant, cell of the raspberry plant, and cell of the peach plant.

17. The method according to claim 16, further characterized in that said plant cell is cell of the potato plant.

18. The method according to claim 16, further characterized in that said cell "Je lan a is one of the corn plant.

19. The method according to claim 16, further characterized by said plant cell being selected from the group consisting of a cell of the wheat plant, a cell of the barley plant, a cell of the plant of ar-roz and a cell of the tomato plant.

20. A recombinant molecule of double-stranded DNA comprising in the sequence: (a) a promoter "functioning in the cells of the tissue" of the target plant; (b) a structural DNA sequence that results in the production of an RNA sequence which encodes a sucrose phosphoprotein enzyme; and (c) a 3 'untranslated region that functions in plant cells to cause transcript termination and the addition of lagged pol nucleotides to the 3"end of the RNA sequence, further characterized because the plant genetically transforms to which has in its genus «Jicha DNA molecule exhibits a property selected from the group consisting of containing modified carbohydrate content, increased content of poly saccharides, improved performance, improved uniformity of the distillation of solids, and Reduced susceptibility to bleaching by bleaching

21. The conformation DNA molecule "Ja" l with claim 20, further characterized because "Jicha DNA sequence coding for the enzyme sucrose fos on lasa is obtained from S1-reptococcus

22. The DNA molecule according to claim 21, characterizes further the said DNA sequence which contains the sucrose phosphoplasse. the sequence of the «Ja in ID. SEQUENCE No. 5.

23. The DNA molecule according to claim 20, further characterized in that said promoter is selected from the group consisting of a zein promoter, a palatin promoter, a glutamine promoter, rice, a promoter of soy 7s, a promoter for a subunit of ADP glucose pyrophosphonlase, the promoter of TFI17 and the promoter of TFM9.

24. A cell t rans form «The plant« has a molecule r'ecomt > ? nant of double-stranded RDN containing ^ n the sequence: (a) a promoter which functions in said plant cell; (b) a structural sequence of RDN that causes the production of a RRN sequence which encodes a sucrose phosphonase enzyme; and (c) a 3'-untranslated region which functions in the cells of the plants to originate * the termination of the transcript and the addition of the polyadenylated nucleotides to the 3 'end of the RNA sequence.

25. The plant cell according to claim 74, further characterized in that said RDN sequence encoding the enzyme sucrose fos fon lasa is from St reptococcus mutans.

26. The plant cell in accordance with the r? E? V? N «J? Cac? On 25, character- ized in addition because« Jicha frequency of RDN that encodes the sac-osa phosphon lasa have the sequence exhibited «Ja en the ID. TO SEQUENCE No. 5. 2 ?. The plant cell according to claim 24, further characterized in that said promoter is selected from the group consisting of a Je zein promoter, a patatm promoter, a rice glutelm promoter, a 7 soybean promoter , a promoter * of a second ADP glucose pyrophosphoplasse, the promoter * of TFM7 and the e TFM9 promoter. 28. The plant cell according to claim 24, further characterized in that said tire cell is selected from the group < It consists of potato plant cell, corn plant cell, La cell [rice rind, wheat plant cell, plant cell «Jel tomato, plant cell of the barley, beet plant cell, sweetpotato plant cell, plant cell «Jel peanut, cell of the sugarcane plant *, cell« He the plant «He the grape, plant cell of the pear, cell of the apple plant, cell of the [orange rim, cell of the yucca plant, cell of the banana plant, cell of the plant of the plant and cell of the plant of the peach.