Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se
Reexamination Certificate
2000-11-06
2004-01-13
Bui, Phuong T. (Department: 1637)
Multicellular living organisms and unmodified parts thereof and
Plant, seedling, plant seed, or plant part, per se
C800S278000, C800S279000, C435S006120, C435S069100, C435S070100, C435S320100, C435S410000, C435S418000, C435S419000, C536S023100, C536S023600, C536S024100, C530S370000
Reexamination Certificate
active
06677505
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to a set of plant promoters which can be induced by biotic or abiotic stresses, in particular by pathogens, to their use, to expression vectors comprising said promoters and a gene of interest, and to cells and/or plants transformed with said vectors. The invention also relates to methods for obtaining said cells or plants, said transformed plants exhibiting improved resistances to said pathogens.
BACKGROUND OF THE INVENTION
Diseases, whether they are of fungal, bacterial or viral origin, are the major problem in viticulture, both in terms of the quality of the musts and of the wines produced (for example
Botrytis cinera,
which is a grey mould agent which attacks the berries at grape-harvesting and leads to bad tastes in the wines), drops in production (for example foliar diseases such as grape downy mildew or grape powdery mildew, attacks by Botrytis on flowers or Fan leaf disease of the grapevine linked to the presence of the virus G.F.L.V., Grape Fan Leaf Virus), or even to that of the survival of the vineyard (for example wood diseases such as eutypiosis or esca syndrome). The conventional control arsenal ranges from simple prophylaxis to plant-protection treatments, biological control to date being used very little.
Chemical control is of course the most widely used method, even though treatments are increasingly being tempered (models for forecasting risks of disease for grape downy mildew for example). With regard to fungicides for example, the French vineyard, which represents approximately 10% of the cultivated land in France, each year uses close to 40% of the fungicides consumed in this country. On a European level, on close to 4 million hectares of vineyard, the 9 to 10 treatments which are carried out each year to control these diseases lead to the use of 120,000 tonnes of fungicidal products.
To cite just the problem of grey mould, it is estimated that over the years 1992 to 1993, 25 to 30% of the 3.7 million hectares of European vineyard were concerned, for a cost of plant-protecting products of 97 and 69 million German Marks (DM), respectively.
The use of these products is not without consequence for the environment (it is the case, for example, for the soil fumigants used to destroy nematodes, which are vectors of the grape fan leaf virus). It also sometimes poses technological problems, with difficulties which can occur during fermentations (the use of sterol biosynthesis inhibitors can block yeast growth at the end of fermentation) and commercial difficulties (procymidone, an anti-Botrytis product, which is sometimes found in wines, hence the American dispute in the years 1990 to 1991).
Moreover, the use of these products has already led to the appearance of resistant strains. This phenomenon has been particularly marked in Champagne and, for some years, the “Comité Interprofessionnel des Vins de Champagne” (CIVC) [Interprofessional Committee of the Wines of Champagne] has recommended not treating against Botrytis.
To overcome these drawbacks, it is imperative to balance the use of plant-protecting products by developing novel methods of control in order to considerably decrease the amounts of products spread over vineyards.
Two approaches are currently envisaged:
Reinforce prophylaxis and decrease the amount of products used (cultivation methods and preventive control, models for forecasting risks of disease, novel spreading materials, novel, more degradable molecules, etc.).
Improve the resistance of the grape varieties to disease.
For this second approach, conventional genetic improvement via the sexual pathway (hybridization with tolerant varieties) is impossible according to French legislation on Appellations d'Origine Contrôlée (A.O.C.) (registered designations of origin) which imposes the grapevine varieties which are to be used for a given appellation (designation). In addition, technically, grapevine, which is a ligneous plant, would require several tens of years to integrate one or more novel resistance properties while at the same time conserving the biochemical and aromatic properties of the grapevine varieties, which are factors of the organoleptic quality of the wines produced.
The control of the regeneration and of the genetic transformation of the grapevine, which has been carried out by the research team of the laboratories of the applicant company since 1988-1990, has made it possible to envisage using the modern techniques of cellular and molecular biology to increase the tolerance of grapevine varieties to fungal diseases.
Henceforth, it is possible, on the one hand, to integrate, into the genome of the grapevine, one or more homologous or heterologous genes which enable the overexpression or expression of a molecule of interest, which is of protein or other nature, and/or the opening of a new biosynthetic pathway and, on the other hand, to regenerate a plant which is more tolerant to one or more diseases, i.e. which has defense mechanisms which are reinforced with respect to the pathogen(s) in question.
There are several different mechanisms of this type in plants. Some can be regarded as being passive and are linked to the physicochemical properties of the cells, epidermal tissues and/or organs of the plant (for example the cuticle or the morphological properties of the grape cluster). Others belong to the dynamics of gene/gene interactions (plant resistance genes and pathogen avirulence genes, mechanisms of host/parasite interactions). These interactions can lead to the development of hypersensitivity reaction (rapid death of the cells of the plant around the point of infection in order to block the colonization of the plant by the fungus, bacterium or virus), but also to the synthesis and to the accumulation of a whole series of compounds. Among these, some can be parietal constituents which are involved in the formation of a “physical” barrier around the point of infection (callose, lignin, hydroxyprolinerich protein, etc.), and others are molecules having antimicrobial functions which are more or less well defined (phytoalexins, pathogen-associated proteins: PR proteins (pathogenesis-related protein), etc.).
The overexpression of these molecules which have antimicrobial functions or which are involved in the formation of a physical barrier around the point of infection can provide plants with a “natural” resistance in response to stresses, in particular stresses of microbial type.
However, constitutive overexpression of this type of protein cannot be envisaged without drawbacks for the plant (energetic cost, slowing down of growth, etc.), which is why it is necessary to envisage the use of inducible promoters and in particular of promoters which are inducible by the stress itself. This is precisely the subject of the present invention.
The present invention relates to a nucleic acid sequence chosen from the group comprising:
a) the IND S1 sequence,
b) any sequence corresponding to a fragment of the IND S1 sequence and having a promoter sequence effect in plants.
The invention also relates to promoters in plants, chosen from the group comprising:
a) the promoter PMs PR10-1 corresponding to the IND S1 sequence,
b) a promoter of PMs PR10-1 type corresponding to a sequence according to the invention.
Said promoters of PMs PR10-1 type are preferred which exhibit at least 80% homology with the IND S1 sequence. Those which exhibit at least 90% or 95% homology with said sequence are particularly preferred. The promoter sequences in plants which are characterized in that they comprise at least one sequence which is identical to those of the abovementioned promoters are also included in the present invention.
The subject of the invention relates most particularly to the use of the promoters according to the invention for the tissue-specific or non-tissue-specific expression of a gene in a way which is inducible in plants by a biotic or abiotic stress.
Among said biotic stresses according to the invention, the biotic stresses engendered by the attack of a parasite such as a
Boulay Michel
Breda Colette
Buffard Dominique
Coutos-Thevenot Pierre
Esnault Robert
Bui Phuong T.
Champagne Moer and Chandon
Connolly Bove & Lodge & Hutz LLP
Spiegler Alexander H.
LandOfFree
Promoter inductible in plants, sequence incorporating same... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Promoter inductible in plants, sequence incorporating same..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Promoter inductible in plants, sequence incorporating same... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3235976