DE19741375C2 - Transgenic plants, the above-ground parts of which ripen earlier and die off completely - Google Patents

Abstract

Transgenic plants are described, in particular potato plants, the above-ground parts of which ripen earlier and faster than in wild-type plants and die completely, without this affecting the yield of underground plant parts. These plants are characterized by a decrease in the activity of GDP-mannose pyrophosphorylase in the cells. DOLLAR A also describes processes for the production of such plants, and also nucleic acid molecules which encode plant GDP-mannose pyrophosphorylase.

Description

The present invention relates to transgenic plants whose Above-ground parts ripen earlier and faster than in game type plants and die completely without this Yield of parts of plants affected. The plants are characterized by a decrease in activity the GDP-mannose pyrophosphorylase in the cells. Furthermore be the invention relates to the use of nucleic acid molecules len encoding a GDP-mannose pyrophosphorylase, or of fragments of such molecules for the production of plants with a reduced activity of this enzyme. Likewise be the present invention relates to nucleic acid molecules that  vegetable proteins with the enzymatic activity of a Encode GDP-mannose pyrophosphorylase.

In production technology when growing a lot of agriculture Cultures are cultivated through the use of herbicides aims to kill vegetative plant parts above ground even ripening, d. H. the ripening and dying, to reach above-ground parts of plants. this makes possible easier harvesting of valuable organs or plants share and ensure better plant hygiene. So especially when growing cotton and the Kar toffel the plants with herbicides before ripening delt. In the case of cotton, this causes the leaves and the machine harvesting of the fibers is lightened. In the case of the potato, killing the kraut tes done for a variety of reasons:

Killing the herb causes the herb to diminish mass and causes the dams to dry better. This causes an increase in the shell strength and min damage caused during harvesting Potato tubers and thus the appearance of stock rot. Furthermore, the contaminations of the harvested tubers are thanks to better drying. It also allows Killing the herb a higher driving speed with the ro and thus a higher area coverage. The separation of Tuber and cabbage are more effective, which means that any nourishment loss of tea can be reduced. The phytosanitary aspects of Herb killing is a reduction in tuberous infections tion with Phytophthora and a variety of potato viruses. Especially when growing in late ripening economic locations harvesting is made possible by killing herbs. Killing and removing the potato herb with herbicides is usually supported by the use of an herb racket, or performed exclusively with it. This Processes are labor and cost intensive and would be in the event that the No need for herbs.  

In view of the increasing importance that of the people tion of the use of herbicides in agriculture in is attributed recently, it is one of the tasks of biotechnological research to adapt the To strive plants to the requirements of the cultivation technology, so that if necessary ver on the use of herbicides can be waived.

It therefore appears desirable to have plants available places that are on schedule and with full yield fully ripen so as to use chemical or me chanic process for killing the potato herb mini be able to lubricate or if necessary completely ver to be able to do.

The present application is therefore based on the task To provide plants, their aerial parts earlier and faster than with corresponding wild-type plants ripen and die completely without a reduction in yield other, especially underground, plant parts point.

This task is by be in the claims Drawn embodiments solved.

The present application thus relates to transgenic plants containing plant cells that are genetically modified, the genetic modification to reduce the acti vity of an endogenous GDP occurring in the plant cell Mannose pyrophosphorylase (EC 2.7.7.22) leads in comparison to corresponding non-genetically modified cells sol plants.

In a preferred embodiment, the GDP man-made pyrophosphorylase, the activity of which is reduced, is a vegetable protein which is encoded by a nucleic acid molecule selected from the group consisting of:

• a) Nucleic acid molecules that encode a protein that the under Seq ID No. 2 includes amino acid sequence indicated;
• b) Nucleic acid molecules which are listed under Seq ID No. 1 attached include given coding region;
• c) nucleic acid molecules that encode a protein that the Amino acid sequence encompassed by the insertion in the Plasmid pGMPase (DSM 11746) is encoded;
• d) nucleic acid molecules encompassing the coding region sen in the insertion of the plasmid pGMPase (DSM 11746) is included;
• e) nucleic acid molecules, which with a strand of a Nucleic acid molecule according to (a), (b), (c) and / or (d) hy bridify; and
• f) nucleic acid molecules whose sequence is due to the Dege generation of the genetic code from the sequence of a Nucleic acid molecule deviates according to (e).

For the definition of the term "hybridization" referenced below.

It has now surprisingly been found that Kartof felp plants in which the expression and thus the activi the GDP-mannose pyrophosphorylase due to the express sion of a corresponding antisense RNA is reduced, the Above-ground parts (the herb) rip off much faster than with corresponding unmodified wild-type Plants. Furthermore, they ripen completely in a relatively short time dig from. However, this will increase the tuber yield of the plants not affected.

The term "corresponding wild-type plants" means that it is plants that are used as starting material served for the production of the plants according to the invention, d. H. whose genetic information apart from the one performed genetic modification of an inventive Plant corresponds.  

The term "ripening" means ripening and destroying ben of the aerial parts of the plants. Preferably be ripening and dying indicate the occurrence darker Stains on stems first, the appearance of lesions Scroll in the form of small black dots, as well as the ver dry the leaves. The sheet preferably dries up progressing from the tip to the stem. About that in addition, the leaves of the plant preferably dry up first, the bottom of the stem, and the drying of the Leaves progressively settle on the plant from below continue up until finally all aerial plants parts have matured.

That the genetically modified plants mature faster means that on the one hand the death of the above ground parts, in particular of the leaves, stems and given if necessary, fruit starts earlier than with corresponding ones Wild-type plants and / or that the whole process of wiping fens runs much faster than the corresponding ones Wild-type plants and completed earlier.

That the process starts "earlier" means preferentially evidence that the ripening and death of the aerial parts at least one week, preferably at least 2 to 3 weeks and particularly preferably starts about 4 weeks earlier than at corresponding wild-type plants.

That the ripening process means "faster" preferably that the process of ripening and abster bens of the aerial parts of the plants as a whole It runs before than with corresponding wild-type plants moves at least a week faster and especially before moves at least two weeks faster.

That the process of ripening and dying ended earlier sen means that the aerial parts of the er plants according to the invention preferably at least 1 to 4 weeks chen, preferably at least 2 to 4 weeks and especially be preferably died at least 3 to 4 weeks earlier than the above-ground parts of corresponding wild-type plants.  

That the process of ripening is completed earlier is important tet preferably also that the aerial parts of the he plants according to the invention have already died completely are more appropriate before the above-ground parts Wild-type plants have started the process of ripening.

In particular, the aerial parts die according to the invention moderate plants about 4 weeks earlier than that of the wild-type plants if the plants are among the following Conditions are kept: in 16-piece pots with potting soil (Prickly soil) / sand mixture 3: 1; Humidity 60%; Tem temperature 22 ° C during the day / 15 ° C at night; 16 h light / 8 h dark rhythm mus ,; Light intensity approx. 150 µE.

That the plants according to the invention mature completely indicates that all aerial parts of the plant are dying, especially that these parts die completely.

Such plants according to the invention have, for example Advantage that ver completely on the use of herbicides can be waived in order to ripen evenly the upper to reach earthly plant parts. Related to for example, with potato plants that mature in themselves there is now the possibility to change them in such a way that her herb ripens and dies earlier and completely.

Furthermore, there is generally an advantage that the Herb of the plants according to the invention dies completely. Even with the early hitherto used in agriculture The problem with ripening potato plants is that Herb never dies completely and therefore in any case the one set of herbicides and herb beaters is required. By growing plants where the herb is complete dig dies, could use these measures fully come to be waived.

The term "transgenic" means that cells of the invention plants according to their genome a foreign DNA sequence ent keep that in appropriate non-genetically modified Cells of corresponding wild-type plants do not occur. "Foreign DNA sequence" means a DNA sequence that  either naturally not at all in the corresponding plant cells occurs, d. H. the heterologous too the cells is like that in the concrete spatial arrangement does not occur naturally in the plant cells or located in the genome of the plant cell on which it does not naturally occur. Is preferred the foreign DNA sequence is a recombinant molecule that consists of different elements, its combination or specific spatial arrangement naturally in plants cells does not occur.

The existence, the location and the spatial location order of the foreign DNA sequence in the genome of the cells invented Plants according to the invention can be, for example, by Detect Southern blot analysis.

The term "genetically modified" means that the plant cell by introducing a foreign nucleic acid molecule is changed in their genetic information and that the Presence or expression of the foreign nucleic acid leads to a phenotypic change. Phenoty This change preferably means a measurable one Change one or more functions of the cells. The genetically modified plant cells of the invention Plants show due to the presence or with express sion of the introduced foreign nucleic acid molecule a ver decrease in the activity of an endogenous plant mend GDP-mannose phosphorylase. The enzyme GDP-Mannose Py rophosphorylase (EC 2.7.7.22) catalyzes the conversion of GTP and Mannose-1-Phosphate to GDP-Mannose and Pyrophosphate. The term "reduction in activity" means in Within the scope of the present invention, a reduction in Ex pression of endogenous genes that constitute a GDP-mannose pyrophospho encode rylase, a reduction in the amount of GDP mannose Pyrophosphorylase in the cells and / or a decrease the enzymatic activity of GDP-Mannose Pyrophosphory read in the cells.  

The reduction in expression can be determined, for example are determined by measuring the amount of GDP-Mannose Pyrophospho transcripts encoding rylase, e.g. B. by Northern blot Analysis. A "reduction" preferably means a reduction in the amount of transcripts compared to corresponding non-genetically modified cells by min at least 10%, preferably by at least 30%, in particular by at least 50%, particularly preferably at least 70% and in particular by at least 90%.

Reducing the amount of GDP-mannose pyrophosphorylase can be determined, for example, by Western blot ana lysis. A "reduction" preferably means one Reduction in the amount of GDP-Mannose Pyrophosphorylase-Pro tein compared to corresponding non-genetically modifi decorated cells by at least 10%, preferably by at least 30%, in particular by at least 50%, particularly preferred by at least 70% and in particular by at least 90%.

The reduction in the enzymatic activity can, for example can be determined wisely by converting GTP and Man nose-1-phosphate to GDP-mannose and pyrophosphate.

The activity of GDP-mannose pyrophosphorylase can be for example, as described in Asua et al. (Biochim. Biophys. Acta 128 (1966), 582-585) or in Szumilo et al. (1993, J. Biol. Chem. 268: 17943-17950). A "Ver Reduction "of the enzymatic activity means before preferably a reduction compared to the corresponding non-genetically modified cells by at least 10%, be preferably by at least 30%, in particular by at least 50%, particularly preferably by at least 70% and in particular by at least 90%.

The genetic modification using a foreign nuclein acid molecule that enters the cells of the invention Plants was introduced and its presence or ex pression to reduce GDP-mannose pyrophosphorylase Activity leads can be of different types.  

On the one hand, the genetic modification can consist in that an endogenous gene present in the plant cell, the encoded a GDP-mannose pyrophosphorylase by insertion of a foreign nucleic acid molecule is changed in such a way that its expression is reduced or completely suppressed is. A reduction preferably means a Ver reduction in transcription by at least 10%, preferably by at least 30%, especially at least 50%, especially preferably by at least 70%, in particular at least 90% compared to non-genetically modified cells. This can e.g. B. be determined by Northern blot analysis. This Reducing transcription preferably causes one corresponding reduction in the activity of GDP-Mannose Py rophosphorylase. Such a genetic modification can for example in an insertion or deletion in the code the area or promoter region of a GDP mannose Pyrophosphorylase gene exist.

Furthermore, the genetic modification can be the synthesis of a GDP-Mannose Pyrophosphorylase cause that no enzyma table activity has more. This can e.g. B. on mutation are based in the coding region of the gene leading to a Loss of enzymatic activity will result in or faulty splicing of the mRNA.

Process to make genetic modifications in genes or their introduce regulatory areas to reduce or lead to loss of expression of the gene are the Known specialist. These include e.g. B. Transposon mutagenesis or gene tagging.

The introduction of a foreign nucleic acid is also conceivable remolecule, which is an inhibitor of GDP-mannose pyrophospho rylase encoded, e.g. B. an antibody whose binding to the Enzyme leads to inhibition of the enzymatic activity.

The genetic modification preferably consists of the one introduction of a foreign nucleic acid molecule into the genome of the Plant cell, the expression of which inhibits the expression sion of endogenous genes that leads to GDP-mannose pyrophospho  encode rylase, especially on the transcription or at the translation level.

Preferred are nucleic acid molecules that have a ge suitable antisense RNA, a suitable ribozyme or a ge encode suitable cosuppression RNA.

With an antisense effect, a corresponding DNA Se expressed in antisense orientation, so that in the Cells an antisense RNA is synthesized, which leads to Trans scripting of GDP-mannose pyrophosphrylase genes or to Tei len such transcripts is homologous. This preferably has a length of at least 30 nucleotides, preferably of min at least 50 nucleotides and particularly preferably of minde at least 100 nucleotides. The expressed antisense RNA should a high homology to the endogenously expressed in the plant transcripts that code GDP-mannose pyrophosphorylase ren, have. The homology is preferably at least 90%, preferably at least 95% and particularly preferably minde at least 99%. In the case of a ribozyme effect, an RNA is expressed miert, the specific transcripts of a GDP-Mannose Pyro phosphorylase gene can cleave. The expression of Ribozy to reduce the activity of certain enzymes in Cells are also known to the person skilled in the art and is exemplary Wisely described in EP 0 321 201 B1. The expression of Ri bozymen in plant cells was e.g. B. described in Feyter et al. (Mol. Gen. Genet. 250 (1996), 329-338). Of the Cosuppression effect is based on the expression of a sense RNA, the expression of endogenous GDP-Mannose Pyrophos suppressed phorylase mRNA. The execution of these techniques are known to the person skilled in the art. The process of cosuppression is described for example in Jorgensen (Trends Biotechnol. 8: 340-344 (1990), Niebel et al. (Curr.Top. Microbiol. Immunol. 197: 91-103 (1995), Flavell et al. (Curr. Top Microbiol. Immunol. 197 (1995), 43-46), Palaqui and Vaucheret (Plant. Mol. Biol. 29 (1995), 149-159). Vaucheret et al. (Mol. Gen. Genet. 248 (1995), 311-317), de Borne et al. (Mol. Gen. Genet. 243 (1994), 613-621) and an their sources.  

In a particularly preferred embodiment of the present invention, the reduction in the activity of a GDP-mannose phosphorylase in the cells is achieved in that the foreign nucleic acid molecule comprises the following elements:

• a) a promoter that translates into vegetable Cells enabled; and
• b) ver in an antisense orientation with this promoter linked DNA sequence, whose transcripts in whole or in part are complementary to transcripts of an endogenous in Plant cells present gene that is a GDP mannose Encoded pyrophosphorylase.

Promoters that ge expression in plant cells Guarantees are described on a large scale. For a con stitutive expression is suitable for. B. the 35S promoter of CaMV (Franck et al., Cell 21 (1980), 285-292) or the Pro motors of the polyubiquitin genes (Christensen et al., Plant Mol. Biol. 18: 675-689 (1992). For an expression in pho To synthetically active tissue is suitable for. B. the ST-LS1- Promoter (Stockhaus et al., EMBO J. 8 (1989), 2445-2451; Stockhaus et al., Proc. Natl. Acad. Sci. USA 84 (1987), 7943-7947).

Nucleic acid molecules, the vegetable GDP-Mannose Pyrophos encode phorylase are by the present invention made available. The in Seq ID No. 1 shown Nucleic acid sequence encodes a GDP-mannose pyrophospho rylase from potato. With the help of this sequence it is Expert possible, using common molecular biological measurement corresponding sequences from other plants or Isolate plant species (see below).

In a preferred embodiment, for the express sion of an antisense RNA he described below nucleic acid molecules according to the invention from potato or Parts of it used.  

As a rule, all cells or essentially all of them Cells of the plants according to the invention have the genetic modification cation on. On the other hand, it is not absolutely necessary that all cells decrease the activity of GDP mannose Have pyrophosphorylase. In a first embodiment it is intended that all or essentially all cells the plant has a reduced GDP-mannose pyrophosphorylase Have activity.

In a preferred embodiment, only cells have upper terrestrial parts of plants, preferably only cells in the Leaves or just cells in the leaves and stems one decreased GDP-mannose pyrophosphorylase activity on up due to the expression of the foreign nucleic acid molecule.

In principle, it can be the case with the plants according to the invention is a plant of any plant species, d. H. both monocot and dicot plants. It is preferably an agricultural one Useful plant, particularly preferred around cotton or potatoes and especially around late ripening potato plants.

The present invention also relates to transgenic plants cells by the introduction of a foreign nucleic acid molecules are genetically modified, the presence of which or the expression of this nucleic acid molecule the verrin an endogenous activity in the plant cell upcoming GDP-mannose pyrophosphorylase.

Regarding the genetic modification of the cells that already above in connection with the invention Plant what has been said.

Such plant cells according to the invention can rain ration of whole plants can be used.

The present invention also relates to a process for the production of transgenic plants, the above-mentioned parts of which ripen earlier and faster than in corresponding wild-type plants and die off completely

• a) a plant cell is genetically modified by the introduction of a foreign nucleic acid molecule and the genetic modification to reduce activity activity of an endogenously occurring in plant cells GDP-mannose pyrophosphorylase leads; and
• b) a plant is regenerated from the cell; and give otherwise
• c) who produces plants from the plant according to (b) the.

For the one introduced into the plant cell according to step (a) Modification is the same as what is already related above was explained with the plants according to the invention.

The regeneration of the plants according to step (b) can be started methods known to the person skilled in the art.

The production of further plants according to step (c) of Ver driving z. B. done by vegetative propagation (at for example via cuttings, tubers or via callus culture and regeneration of whole plants) or through sexual ver increase. Sexual reproduction takes place preferentially controlled instead of d. H. selected plants are included certain characteristics crossed and increased.

In a preferred embodiment, the method for Production of transgenic potato plants used.

In a further preferred embodiment, this enables introduced foreign nucleic acid molecule the synthesis of a antisense RNA leading to transcripts from GDP-Mannose Pyrophos phorylase genes is complementary.

The present invention also relates to the invention plants available according to the method according to the invention.  

The present invention also relates to propagation material plants according to the invention and that according to the invention Transgenic plants produced according to the method, which he contains genetically modified cells according to the invention. Of the The term propagation material includes those components the plant that is suitable for the production of offspring vegetatively or sexually. For vegetative ver For example, cuttings, calluskul are suitable structures, rhizomes or tubers. Other propagation material around holds fruit or seeds, for example. Preferably acts the propagation material is seeds and especially preferably around tubers.

The present invention also relates to the use of nucleic acid molecules that form a GDP-mannose pyrophospho encode rylase, of their complements or parts of sol Cher molecules to reduce GDP-mannose pyrophospho rylase activity in plant cells, for the production of Plant cells with reduced GDP-Man activity nose pyrophosphorylase, for the production of plants, ent tend such plant cells and / or to produce trans plants whose above-ground parts mature more quickly and die off than in corresponding wild-type plants and who die completely. For the nucleic acid molecules it is preferably the described below benen nucleic acid molecules of the invention.

The present invention further relates to nucleic acid molecules which encode a vegetable protein, the mature form of which has the enzymatic activity of a GDP-mannose pyrophosphorylase, selected from the group consisting of:

• a) Nucleic acid molecules that encode a protein that the under Seq ID No. 2 includes amino acid sequence indicated;
• b) Nucleic acid molecules which are listed under Seq ID No. 1 attached include given coding region;  
• c) nucleic acid molecules that encode a protein that the Amino acid sequence encompassed by the insertion in the Plasmid pGMPase (DSM 11746) is encoded;
• d) nucleic acid molecules encompassing the coding region sen in the insertion of the plasmid pGMPase (DSM 11746) is included;
• e) nucleic acid molecules, which with a strand of a Nucleic acid molecule according to (a), (b), (c) and / or (d) hy bridify; and
• f) nucleic acid molecules whose sequence is due to the Dege generation of the genetic code from the sequence of a Nucleic acid molecule deviates according to (e).

The invention also relates to the respective complementary Strands of the molecules described above.

The in Seq ID No. 1 is the nucleic acid sequence shown a complete cDNA containing a GDP mannose pyrophospho Coded rylase from potato (Solanum tuberosum). A Plas mid containing this cDNA was deposited as DSM 11746. With the help of this sequence or this molecule it is Those skilled in the art can now make homologous sequences from other plants isolate zen and plant species. For example using conventional methods such as screening cDNA or genomic banks with suitable hybridization samples are taken. In this way it is possible, for example to identify and isolate nucleic acid molecules, those with the Seq ID No. 1 specified sequence hybridi and encode a GDP-mannose pyrophosphorylase.

The nucleic acid molecules according to the invention can be in principle a GDP-mannose pyrophosphorylase from any encode a plant organism, preferably one from a higher plant, especially a monocot or a dicotyledonous plant. This is particularly preferably encoded Nucleic acid molecule a protein from an agricultural Chen crop, especially from a cereal plant  (e.g. barley, rye, oats, wheat etc.), or from corn, Rice, pea, flax etc. In a particularly preferred way In the embodiment, the nucleic acid molecule codes out a protein Potato or cotton.

The term "hybridization" means in the context of this invention a hybridization under conventional Hybridization conditions, preferably under stringent ones Conditions such as those described in Sambrock et al. Molecular Cloning, A Laboratory Manual, 2nd ed. (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY) are written. Nucleic acid molecules with the inventions hybridize according to the invention, can principally come from any plant that ent speaking proteins expressed. Nucleic acid molecules that can hybridize with the molecules according to the invention e.g. B. isolated from genomic or from cDNA libraries become. The identification and isolation of such Nucleic acid molecules can use the invent nucleic acid molecules according to the invention or parts of these molecules or the reverse complement of these molecules take place, for. B. by means of hybridization according to standard procedures (see e.g. Sambrook et al., 1989, Molecular Cloning, A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY) or by amplification using PCR. As a hybridization sample, e.g. B. Nucleic acid molecules are used that exactly the or essentially the un ter Seq ID No. 1 specified nucleotide sequence or parts of the have this sequence. In the ver as hybridization sample fragments used can also be synthetic frag act with the help of common synthetic techniques were prepared and their sequence essentially with the of a nucleic acid molecule according to the invention. Has one identified and isolated genes that are related to the inventions? hybridize according to the invention, should a determination of the sequence and an analysis of the properties th of the proteins encoded by this sequence  determine if it is a GDP-Mannose Pyrophospho rylase acts. Homology is particularly suitable for this compare at the level of the nucleic acid or amino acid sequence and the determination of the enzymatic activity. The hybridi with the nucleic acid molecules of the invention Molecules in particular include fragments, derivatives and allelic variants of the nucleic acid described above remolecules that encode a vegetable protein with the Activity of a GDP-mannose pyrophosphorylase. The expression In this context, derivative means that the sequences of these molecules differ from the sequences described above Nucleic acid molecules at one or more positions differ and have a high degree of homology to these Have sequences. Homology means a sequence Identity over the entire length of at least 60%, especially in particular an identity of at least 70%, preferably over 80%, particularly preferably over 90% and in particular of at least 95%. The deviations from those described above benen nucleic acid molecules can z. B. by Dele tion, addition, substitution, insertion or recombination have arisen.

Homology also means that functional and / or struc tural equivalence between the relevant nucleic acid mo lekulen or the proteins encoded by them. At the nucleic acid molecules homologous to those described above are molecules and represent derivatives of these molecules len, these are usually variations of these mo readings that represent modifications that are the same biologi perform a specific function. It can be both natural Variations that usually occur are, for example, Sequences from other plants, especially other crops plants or varieties, or for mutations, these Mu tations may have occurred naturally or through targeted mutagenesis. Furthermore, it can the variations on synthetically produced sequences act zen. With the allelic variants it can be act both naturally occurring variants, as well  for synthetically produced or by recombinant DNA Techniques generated variants.

The of the different variants of the invention Proteins encoded nucleic acid molecules have certain ge common characteristics. For this, e.g. B. biological Activity, molecular weight, immunological reactivity, Conformation, etc., as well as physical properties such as B. the running behavior in gel electrophoresis, chromato graphic behavior, sedimentation coefficient, soluble speed, spectroscopic properties, stability; pH opti mum, optimum temperature etc.

Important characteristics of a GDP-mannose pyrophosphorylase are (i) their location in the cytosol of plant cells, (ii) their ability to synthesize GDP mannose under Ver use of GTP and mannose-1-phosphate as substrate. This Activity can be determined, for example, according to the in Asua et al. (op. cit.) or in Szumilo et al. (loc. cit.) be written methods.

The molecular weight derived from the amino acid sequence the GDP-mannose pyrophosphorylase from potatoes 39,578 daltons. The derived molecular weight of an invented protein according to the invention is therefore preferably in the range from 35 kDa to 45 kDa, preferably in the range from 38 kDa to 42 kDa and particularly preferably around 40 kDa. The calculated nete isoelectric point of the protein from potato lies at 7.21.

The nucleic acid molecules according to the invention can be any Be nucleic acid molecules, especially DNA or RNA moles cool, for example cDNA, genomic DNA, mRNA etc. you can be naturally occurring molecules, or by gen Mo or technical or chemical synthesis processes read. They can be single-stranded molecules that either of the coding or the non-coding strand ent hold, or double-stranded molecules.  

The present invention further relates to nucleic acid moles cool of at least 15, preferably more than 50 and especially ders preferably more than 200 nucleotides in length, the speci fish with at least one nucleic acid mo according to the invention hybridize lekül. Hybridizing specifically means here in that these molecules hybridi with nucleic acid molecules sieren, which encode a protein according to the invention, however not with nucleic acid molecules that encode other proteins Ren. Hybridization preferably means hybridization under stringent conditions (see above). In particular concerns the invention is such nucleic acid molecules that with Trans scripts of nucleic acid molecules according to the invention hybridi be able to prevent their translation. Sol che nucleic acid molecules that are specific to the Invention appropriate nucleic acid molecules can hybridize at for example components of antisense constructs or Ri be bozymen or can be used as primers for the amplification be used by PCR.

The invention further relates to vectors, in particular Plasmids, cosmids, viruses, bacteriophages and others in the Genetic engineering common vectors that he described above contain nucleic acid molecules according to the invention.

In a preferred embodiment, those in the vectors contained nucleic acid molecules in sense orientation linked to regulatory elements that drive expression in prokaryotic or eukaryotic cells most. The term "expression" can be used as a transcription also mean transcription and translation.

The expression of the nucleic acid molecules according to the invention in prokaryotic cells, for example in Escherichia coli, enables, for example, a more precise characterization of the enzymatic activities of the encoded proteins. In addition, it is possible to introduce various types of mutations into the nucleic acid molecules according to the invention by means of common molecular biological techniques (see, for example, Sambrook et al., Loc. Cit.), Which leads to the synthesis of proteins with possibly changed biological properties. It is possible to generate deletion mutants in which nucleic acid molecules are generated by progressive deletions from the 5 'or 3' end of the coding DNA sequence, which lead to the synthesis of correspondingly shortened proteins. It is also possible to introduce point mutations at positions that have an influence, for example, on the enzyme activity or the regulation of the enzyme. In this way, e.g. B. mutants who who have a changed K _m value or are no longer subject to the regulatory mechanisms normally present in the cell via allosteric regulation or covalent modification. Furthermore, mutants can be produced which have an altered substrate or product specificity. Furthermore, mutants can be produced which have a changed activity-temperature profile. Genetic engineering manipulation in procokaontic cells can be carried out according to methods known to the person skilled in the art (cf. Sambrook et al., Loc. Cit.).

For the expression of a nucleic acid mole according to the invention A lot of promotions are available in plant cells gates available. In principle, everyone can in the for Transformation selected plants functional promoter ver be applied. The promoter can be homologous or heterologous in relation to the plant species used. Suitable is for example the 355 promoter of the Cauliflower mosaic virus (Odell et al., Nature 313 (1985), 810-812) who published a consti Ensure tutive expression in all tissues of a plant stet and the promo described in WO / 9401571 gate construction. Another example are the promoters of Maize polyubiquitene genes (Christensen et al., Supra). It however, promoters can also be used that are only available a point in time determined by external influences fourth (see for example WO / 9307279). By beson  their interest can be promoted by heat-shock Be proteins that allow easy induction. Further the promoters can be used in a be tuned tissue of the plant for expression switch sequences (see e.g. Stockhaus et al., EMBO J. 8 (1989), 2445-2451), for example the ST-LS1-Pro motor that only works in photosynthetically active tissue tiv is (Stockhaus et al., Proc. Natl. Acad. Sci. USA 84 (1987), 7943-7947). Also worth mentioning are promoters that in the starch-storing organs to be transformed Plants are active. These are e.g. B. in maize the corn kernels, while the potatoes are tubers. To the Überex pression of the nucleic acid molecules according to the invention in the Potato, for example, the bulb-specific B33 Promoter (Rocha-Sosa et al., EMBO J. 8 (1989), 23-29) ver be applied. Seed-specific promoters are already for different plant species have been described. So z. B. the USP promoter from Vicia faba, which is a seed-specific Ex guaranteed in V. faba and other plants (Fiedler et al., Plant Mol. Biol. 22 (1993), 669-679; Bäumlein et al., Mol Gen. Genet. 225: 459-467 (1991)). In Maize, for example, ensures promoters of the zein genes a specific expression in the endosperm of the maize kernels (Pedersen et al., Cell 29 (1982), 1015-1026; Quattrocchio et al., Plant Mol. Biol., 15: 81-93 (1990).

Regulatory sequences for expression in prokaryotic Organisms, e.g. B. E. coli are sufficient in the literature described, as well as those for expression in yeast, ins special in Saccharomyces cerevisiae.

Termination signals for transcription in plant Cells are described and are arbitrarily different from each other exchangeable. For example, the terms can be used tion sequence of the octopine synthase gene from Agrobacterium tumefaciens.

In a further embodiment, the invention relates Host cells, especially prokaryotic or eukaryotic  cells with a nucleic acid described above molecule or a vector were transformed, as well as Zel len derived from such host cells and the be contain written nucleic acid molecules or vectors. The host cells can be bacterial (e.g. E. coli) or fungal cells (e.g. yeast, especially S. cerevisiae), and plant or animal cells. The term "trans formed "means that the cells of the invention genetically with a nucleic acid molecule according to the invention are modified in that they are in addition to their natural Lichen genome at least one nucleic acid according to the invention molecule included. This can be freely given in the cell if present as a self-replicating molecule, or it can be stably integrated into the genome of the host cell gene.

Furthermore, the present invention relates to methods for the manufacture position of a GDP-mannose pyrophosphorylase, in which he host cells according to the invention are cultivated under conditions those that allow expression of the protein and the pro tein from culture, d. H. from the cells and / or the Kul turmedium is obtained.

The invention also relates to proteins which are invented by the inventors nucleic acid molecules according to the invention are encoded, or the are obtainable by a method according to the invention.

The present invention further relates to antibodies which specifically recognize a protein according to the invention. This can e.g. B. be monoclonal or polyclonal antibodies. It can also be fragments of antibodies, the fiction recognize appropriate proteins. Methods for the preparation of such Antibodies or fragments are known to the person skilled in the art.

In a preferred embodiment, the fiction According to host cells, transgenic plant cells, which are due to the presence of an introduced nuclein according to the invention  acid molecule a GDP-mannose pyrophospho according to the invention express rylase.

By providing the nucleic acid of the invention Molecules now have the ability to plant cells to change by means of genetic engineering methods, that they are a new or an additional herbal GDP-Man express nose pyrophosphorylase.

Such transgenic plant cells differ from non-transformed cells in that the introduced Nucleic acid molecule either heterologous to the transform is th cell, i. H. geno from one cell with another mix background, or by the fact that the imported Nucleic acid molecule if it is homologous to the transformed Plant species is located in one place in the genome, on which it is naturally found in non-transformed cells does not occur. The introduced nucleic acid mole cool either under the control of his natural promo tors stand or with regulatory elements of foreign genes be linked.

Furthermore, the plant cells according to the invention differ len from corresponding non-transformed plant cells preferably by inventing an increased amount of have protein according to the invention. This can be done with the subject determine known detection methods such. B. Western Blot analysis. An increased amount preferably means an increase of at least 5%, preferably at least 10% and particularly preferably by at least 20%.

There is basically the possibility that the syntheti protein in any compartment of the plant Lichen cell can be localized. To localize in to reach a certain compartment of the cell optionally ver the coding region with DNA sequences be linked, the localization in the respective com ensure division. Such sequences are known (See, for example, Braun et al., EMBO J. 11 (1992), 3219- 3227; Wolter et al., Proc. Natl. Acad. Sci. USA 85 (1988), 846-850; Sonnewald et al., Plant J. 1 (1991), 95-106).  

The invention further relates to transgenic plants which contain transgenic plant cells described above. In the plant, the plant cells according to the invention ent holds, it can in principle be any plant deln. It is preferably a monocot or dicot Plant, especially an agricultural crop, for example a starch-storing plant, such as B. Ge cereal plants (rye, barley, oats, wheat etc.), rice, Corn, legumes (e.g. pea), cassava or potato, or an ornamental plant.

Cereal plants include monocotyledon plants zen understood that to order Poales, preferably those belonging to the Poaceae family. examples for this are the plants belonging to the genera Avena (oat), Triticum (Wheat), secale (rye), oryza (rice), panicum, Pennisetum, Setaria, sorghum (millet), Zea (corn) etc. belong ren. Hordeum (barley) is particularly preferred. Starch spit breeding legumes are e.g. B. some species of the genus Pisum (e.g. Pisum sativum), Vicia (e.g. Vicia faba), Cicer (e.g. Cicer arietinum), Lens (e.g. Lens culinaris), Phaseolus (e.g. Phaseolus vulgaris and Phaseolus coccineus), etc.

The invention also relates to propagation material from transgenic plants according to the invention, for example seeds, Fruits, cuttings, tubers, rhizomes, calli, cell culture structures etc., this propagation material described above bene contains transgenic plant cells.

The present invention further relates to transgenic plants cells in which the activity of the invention Protein is decreased due to the inhibition of trans description or translation of endogenous nucleic acid molecules len encoding such a protein. This can happen with can be achieved, for example, that a fiction moderate nucleic acid molecule or a part thereof in the ent speaking plant cells in antisense orientation expri  is lubricated and because of an antisense effect ver decrease in the amount of protein according to the invention comes. A another way to reduce the amount of inventions protein according to the invention in plant cells consists in the Expression of suitable ribozymes that specifically trans cleave scripts of the DNA molecules according to the invention. Possible is also the expression of molecules that have both a antisense as well as a ribozyme effect in combination to practice. Alternatively, reducing the amount of inventions protein according to the invention in the plant cells also by a Cosupression effect take place.

Such methods are known to the person skilled in the art (see above).

The reduction in the activity of the Pro according to the invention Teins can be determined as described above.

The invention further relates to transgenic plants which contain transgenic plant cells described above.

The invention also relates to propagation material of the first described transgenic plants, for example Sa men, fruits, cuttings, tubers, rhizomes, calli, Cell cultures, etc., this described above contains transgenic plant cells.

For the introduction of DNA into a plant host cell a variety of techniques are available. This Techniques include transforming plant cells with T-DNA using Agrobacterium tumefaciens or Agrobacterium rhizogenes as a transformation agent, the Fusion of protoplasts, injection, electroporation of DNA, the introduction of DNA by means of biolistic Method as well as other possibilities.

The use of Agrobacterium-mediated transformation of plant cells has been intensively examined and sufficient in EP 0 120 516 A1; Hoekema, In: The Binary Plant Vector System Offsetdrukkerij Kanters B.V., Alblasserdam (1985), Chapter  V; Fraley et al., Crit. Rev. Plant. Sci., 4, 1-46 and An et al. EMBO J. 4 (1985), 277-287. For the Transformation of potato, see e.g. B. Rocha Sosa et al. (1989, EMBO J. 8: 29-23).

The transformation of monocotyledonous plants by means of Agrobacterium-based vectors have been described (Chan et al., Plant Mol. Biol. 22: 491-506 (1993); Hiei et al., Plant J. 6 (1994), 271-282, Deng et al., Science in China 33 (1990), 28-34; Wilmink et al. Plant Cell Reports 11 (1992), 76-80; May et al., 1995, Bio / Technology 13: 486-492; Conner and domisse; Int. J. Plant Sci. 153: 550-555 (1992); Ritchie et al., Transgenic Res. 2: 252-265 (1993). alternative Systems for transforming monocotyledonous plants are the Transformation using the biolistic approach (Wan and Lemaux, Plant Physiol. 104: 37-48 (1994); Vasil et al., Bio / Technology 11: 667-674 (1992); Ritala et al., Plant Mol. Biol. 24: 317-325 (1994); Spencer et al., Theor. Appl. Genet. 79 (1990), 625-631), the protoplast transformation, the electroporation of partially permeabilized cells, the introduction of DNA using glass fibers. especially the Transforntation of maize is used several times in the literature (see e.g. WO95 / 06128, EP 0 513 849 A1; EP 0 465 875 A1, EP 0 292 435 A1; Fromm et al., 1990 Biotechnology 8: 833-844; Gordon-Kamm et al., Plant Cell 2 (1990), 603-618; Koziel et al., Biotechnology 11 (1993), 194-200; Moroc et al., Theoretical Appl. Genet. 80: 721-726 (1990)).

Also the successful transformation of other types of grain has already been described, e.g. B. for barley (Wan and Lemaux, s. O.; Ritala et al., P. O.; Krens et al., Nature 296 (1982), 72-74) and for wheat (Nehra et al., Plant J. 5 (1994), 285- 297).

Another object of the present invention is Use of the nucleic acid molecules according to the invention or Parts of these molecules or their reverse complement Molecules for the identification and isolation of homologous moles  cool, the proteins with the enzymatic activity of a Encode GDP-mannose pyrophosphorylase.

The Plas manufactured within the scope of the present invention mid pGMPase was registered as an international recognized German collection of microorganisms (DSM) in Braunschweig, Federal Republic of Germany in accordance with the requirements of the Budapest Treaty on September 15 tember 1997 under the deposit number DSM 11746 behind sets.

Fig. 1 shows schematically the plasmid pGMPase. The fine line corresponds to the sequence of pBluescript II SK (-). The strong line represents the cDNA encoding the GDP mannose pyrophosphorylase from Solanum tuberosum. Restriction sites of the insertion are given. The cDNA insert is ligated between the EcoRI and XhoI sites of the polylinker of the plasmid. The DNA sequence of the cDNA insertion is under Seq ID No. 1 specified.

Figure 2 shows schematically the plasmid p355-antiGMPase.

Construction of the plasmid:
A = fragment A: caMV35S promoter of cauliflower moss virus, 540 bp.
B = fragment B: cDNA from Solanum tuberosum encoding GDP-mannose pyrophosphorylase; Orientation to the promoter: antisense.
C = fragment C: ocs terminator of the octopine synthase gene from Agrobacterium tumefaciens.

Fig. 3 shows a comparison of the amino acid sequences of GDP-mannose pyrophosphorylases from Saccharomyces cerevisiae (lower row and Solanum tuberosum (upper line, respectively).

Fig. 4 shows a Northern blot analysis of the expression of the GMPase in tubers of GDP-mannose pyrophosphorylase "antisense" plant (lines 6, 28 and 37) in comparison to the same corresponding non-transformed wild-type plants (WT).

FIG. 5 shows a comparison of 3-month-old transgenic GMPase "antisense" plants (right and middle) and an identical wild-type plant (left).

Fig. 6 shows the appearance of dark spots on stems and leaves of 3 month old transgenic GMPase "antisense" plants and the progressive drying of the leaves.

The following examples illustrate the invention.

example 1 Identification, isolation and characterization of a cDNA, which is a GDP-mannose pyrophosphorylase from Solanum encoded tuberosum

To identify a cDNA encoding a potato GDP-mannose pyrophosphorylase, 6.5 × 10 ⁵ phage gene plaques from a cDNA library prepared from leaves of potato plants (Koßmann et al., Planta 188 (1992), 7-12 ), screened for fragments contained therein that hybridize specifically with the probe used. The probe was derived from an EST (expressed sequence tag) from Arabidopsis thaliana, the sequence of which has homology to a yeast mannose-1-phosphate guanyl transferase (EC 2.7.7.13) (Gen Bank accession number T 21688).

For this purpose, phage plaques were transferred to Hybond N nylon membranes (Amersham, UK) and the DNA was denatured by NaOH treatment, the filters were neutralized and the DNA was fixed on the filters by UV treatment. The filters were prehybridized in 0.25 M NaHPO ₄ , pH 7.2, 0.25 M NaCl, 7% SDS, 1 mM EDTA, 25% formamide, 10% PEG for 2 hours at 42 ° C. The filters were then placed in 0.25 M NaHPO ₄ , pH 7.2, 0.25 M NaCl, 7% SDS, 1 mM EDTA, 25% formamide, 10% PEG after addition of the corresponding radio-labeled probe for 10 h at 42 ° C hybridizes. The filters were then washed for 30 min in 1 × SSC, 0.5% SDS and for 15 min in 0.5 × SSC, 0.5% SDS each at 42 ° C. and exposed on X-ray films. Positive phage clones were further purified using standard procedures. Using the in vivo excision method, E. coli clones were obtained from positive phage clones which contain a double-stranded pBluescript plasmid with the respective cDNA insert. After checking the size and restriction pattern of the inserts, a suitable clone, cGMPase, was further analyzed.

The plasmid pGMPase ( FIG. 1) was isolated from the clone and its cDNA insertion was determined by standard methods using the dideoxy method (Sanger et al., Proc. Natl. Acad. Sci. USA 74 (1977), 5463-5467).

The insertion is 1524 kb long and represents a complete cDNA. A part of the nucleotide sequence (1478 bp) is under Seq ID No. 1 specified. The deduced amino acid sequence is in Seq ID No. 2 specified. Sequence comparisons showed that this DNA sequence and the deduced amino acid sequence showed, in different areas, strong homology to the cDNA from Saccharomyces cerevisiae, which encodes a GDP man-made pyrophosphorylyase, or to the corresponding amino acid sequence (see FIG. 3).

Example 2 Construction of the plasmid p35S-antiGMPase and introduction of the Plasmids in the genome of potato plants

The plasmid pGMPase was converted using the restrictions donucleases Asp718 and BamHI an approx. 1.5 kb DNA fragment ment isolates the coding region for the GDP mannose  Pyrophosphorylase includes, and in those with Asp718 and BamHI cut vector pBinAR (DSM 9505) ligated.

The insertion of the cDNA fragment creates an expression cassette which is constructed from fragments A, B and C as follows ( FIG. 2):

Fragment A (540 bp) contains the 35 S promoter from the Cauliflower mosaic virus.

In addition to flanking areas, fragment B contains the protein-coding region of a GDP-mannose pyrophosphorylase from Solanum tuberosum. This was made up as described above pGMPase isolated and in antisense orientation at the 35 S Promoter fused in pBinAR.

Fragment C (215 bp) contains the polyadenylation signal of Octopin synthase gene from Agrobacterium tumefaciens.

The size of the plasmid p35S-antiGMPase is approximately 13.5 kb. The plasmid was placed in potato using agrobacteria plants transferred.

In addition 100 small leaves were wounded with the scalpel a potato sterile culture (Solanum tuberosum, L.cv. Desiree) in 10 ml MS medium (Murashige & Skoog Physiol. Plant. 15 (1962), 473) with 2% Sachcarose, which 50 µl one under selection grown Agrobacterium tumefaciens-Über night culture included. After shaking for 3-5 minutes another incubation for 2 days in the dark. Thereupon the leaves were used for callus induction on MS medium 1.6% glucose, 5 mg / l naphthylacetic acid, 0.2 mg / l benzylami nopurin, 250 mg / l claforan, 50 mg / l kanamycin and 0.8% bacto Agar placed. After a week's incubation at 25 ° C and 3000 The leaves were used for induction of shoots on MS medium 1.6% glucose, 1.4 mg / l zeatin ribose, 20 µg / l naphthylacetic acid acid, 20 µg / l giberellic acid, 250 mg / l claforan, 50 mg / l Kanamycin and 0.8% Bacto Agar. From the transformier Whole plants were regenerated in the cells.

The transgenic potato plants obtained were kept in the greenhouse under the following conditions:
Light period: 16 h at 25000 lux and 22 ° C
Dark period: 8 h at 15 ° C
Air humidity: 60%.

As a result of the transformation, transgenic potato plants showed a decrease in the activity of a GDP-mannose pyrophosphorylase and a decrease in the GDP-mannose pyrophosphorylase mRNA (see FIG. 4). This was demonstrated by Northern blot analysis.

For this purpose, RNA was extracted from leaf tissue according to standard protocols Plants isolated. 50 µg RNA were on an agarose gel separated (1.5% agarose, 1x MEN buffer, 16.6% formaldehyde hyd). After electrophoresis, the RNA was exposed to 20 × SSC using capillary blot on a nylon membrane Hybond N (Amersham, UK). The RNA was on the membrane fixed by UV radiation.

The membrane was in phosphate hybridization buffer (Sambrook et al., Op. Cit.) Prehybridized for 2 h and on closing by adding the radiolabelled probe for Hybridized for 10 h.

Transgenic plants showed no differences in their appearance compared to untransformed plants both in sterile culture and up to two months after transfer to greenhouses. After two months, dark spots formed on the stem. Lesions in the form of small black dots were visible on the leaves ( FIG. 6). In addition, the leaves of the plants started to dry up. The tip of the terminal leaflet of a leaf first dried up until the entire leaf was brown and dried up to the stem. This ripening of the leaves continued, progressing from bottom to top, until finally all above-ground parts of the plants had ripened ( Fig. 5).

Transgenic GDP-mannose antisense plants differ in her tuber yield not from untransformed potato potted plants. Tubers from 15 weeks old untransformed Potato plants weighed an average of 169.7 g each Plant. The returns for three independently transformed Li on average, 168.6 g (line 37) and 162.5 g (Line 28) or 160.7 g (line 6).  

SEQUENCE LOG

Claims (29)

1. Transgenic plant containing plant cells, the gene are modified table, the genetic modifications tion to reduce the activity of an endogenous in the Plant cells occurring GDP-Mannose Pyrophospho rylase (EC 2.7.7.22) leads in comparison to match the non-genetically modified cells. 2. The transgenic plant of claim 1, wherein the GDP-mannose pyrophosphorylase, the activity of which is reduced, is a vegetable protein that is encoded by a nucleic acid molecule selected from the group consisting of

• a) Nucleic acid molecules which encode a protein which has the properties listed under Seq ID No. 2 includes amino acid sequence indicated;
• b) Nucleic acid molecules which are listed under Seq ID No. 1 indicated coding region;
• c) nucleic acid molecules encoding a protein comprising the amino acid sequence encoded by the insertion in plasmid pGMPase (DSM 11746);
• d) nucleic acid molecules comprising the coding region contained in the insertion of the plasmid pGMPase (DSM 11746);
• e) nucleic acid molecules which hybridize with a strand of a nucleic acid molecule according to (a), (b), (c) and / or (d); and
• f) nucleic acid molecules whose sequence deviates from the sequence of a nucleic acid molecule according to (e) due to the degeneration of the genetic code.

3. Transgenic plant according to claim 1 or 2, the upper parts compared to those of a corresponding one mature non-genetically modified plants faster and die.   4. Transgenic plant according to one of claims 1 to 3, where in the genetic modification is that a foreign nucleic acid molecule in the genome of the in the Plant contained cell is introduced, the express sion to inhibit the expression of endogenous genes which encode a GDP-mannose pyrophosphorylase. 5. The transgenic plant according to claim 4, wherein the foreign nucleic acid molecule encodes a molecule selected from the group consisting of

• a) antisense RNAs which is complementary to transcripts of GDP-mannose pyrophosphorylase genes or to parts of such transcripts;
• b) ribozymes which can specifically cleave transcripts of a GDP man-made pyrophosphorylase gene; and
• c) cosuppression RNAs to an mRNA encoding a GDP-nose pyrophosphorylase.

6. Transgenic plant according to one of claims 1 to 5, where at the activity of GDP-mannose pyrophosphorylase in Cells in all parts of the plant are reduced. 7. Transgenic plant according to one of claims 1 to 5, where at the activity of GDP-mannose pyrophosphorylase in Cells of the aerial plant parts is reduced. 8. Transgenic plant according to one of claims 1 to 7, where in the activity of GDP-mannose pyrophosphorylase is reduced by at least 10% compared to the share vity in corresponding wild-type plants. 9. Transgenic plant according to one of claims 1 to 8, the is a potato plant. 10. Transgenic plant cell by introducing a foreign nucleic acid molecule is genetically modified, the presence or expression of the nuclein  acid molecule reducing the activity of an endo gene in the plant cell, GDP-mannose pyro phosphorylase causes compared to corresponding non-genetically modified wild-type cells. 11. Propagation material of a transgenic plant after a of claims 1 to 9 containing plant cells according to An saying 10. 12. Propagation material according to claim 11, which is a tuber or is a seed. 13. Process for the production of transgenic plants, the above-ground parts of which ripen and die earlier and faster than in corresponding wild-type plants, where

• a) a plant cell is genetically modified by the introduction of a foreign nucleic acid molecule, and the genetic modification leads to a reduction in the activity of an endogenously occurring GDP-mannose pyrophosphorylase occurring in plant cells; and
• b) a plant is regenerated from the cell; and if necessary
• c) further plants are produced from the plant according to (b).

14. The method of claim 13, wherein the plant a Kar potato plant is. 15. The method according to claim 13 or 14, wherein the one foreign nucleic acid molecule led to the synthesis of a antisense RNA to transcripts from GDP-Mannose Pyrophos phorylase genes allowed. 16. Use of nucleic acid molecules that a GDP-Man encode pyrophosphorylase from fragments of such Molecules or of nucleic acid molecules that lead to such  Molecules are complementary to reduce GDP Mannose pyrophosphorylase activity in plant cells len. 17. Use of nucleic acid molecules that a GDP-Man encode pyrophosphorylase from fragments of such Molecules or of nucleic acid molecules that lead to such Molecules are complementary to the production of plants cells with a reduced activity of the GDP mannose Pyrophosphorylase. 18. Use of nucleic acid molecules that a GDP-Man encode pyrophosphorylase from fragments of such Molecules or of nucleic acid molecules that lead to such Molecules are complementary to the production of plants containing plant cells as defined in claim 17. 19. Use of nucleic acid molecules that a GDP-Man encode pyrophosphorylase from fragments of such Molecules or of nucleic acid molecules that lead to such Molecules are complementary to produce transge plants whose above-ground parts mature faster and die as the above-ground parts of correspond wild-type plants. 20. Nucleic acid molecule encoding a vegetable protein, the mature form of which has the enzymatic activity of a GDP-mannose pyrophosphorylase, selected from the group consisting of

• a) Nucleic acid molecules which encode a protein which has the properties listed under Seq ID No. 2 includes amino acid sequence indicated;
• b) Nucleic acid molecules which are listed under Seq ID No. 1 indicated coding region;
• c) nucleic acid molecules encoding a protein comprising the amino acid sequence encoded by the insertion in plasmid pGMPase (DSM 11746);
• d) nucleic acid molecules comprising the coding region contained in the insertion of the plasmid pGMPase (DSM 11746);
• e) nucleic acid molecules which hybridize with a strand of a nucleic acid molecule according to (a), (b), (c) and / or (d); and
• f) nucleic acid molecules whose sequence deviates from the sequence of a nucleic acid molecule according to (e) due to the degeneration of the genetic code.

21. Vector containing a nucleic acid molecule according to claim 20th 22. The vector of claim 21, wherein the nucleic acid molecule ver in sense orientation with regulatory sequences that ties the transcription into prokaryotic or eukaryotic cells. 23. Host cell that is genetically modified with a Nucleic acid molecule according to claim 20 or with a Vek Gate according to claim 21 or 22. 24. Process for the preparation of a GDP-Mannose Pyrophospho rylase, wherein a host cell according to claim 23 under Be conditions are cultivated, the expression of the Pro allow teins, and the protein from the cultured Cells and / or the culture medium is isolated. 25. Protein encoded by a nucleic acid molecule according to An claim 20 or obtainable by a method according to An Proverb 23 26. Antibody, which is specifically a protein according to claim 25 recognizes.   27. Transgenic plant cell containing a nucleic acid mole kül is genetically modified according to claim 20 and the compared to a non-genetically modified one Plant cell increased activity of GDP-Mannose Py has rophosphorylase. 28. Transgenic plant containing plant cells according to An Proverbs 27 29. Plant propagation material according to claim 28.