RNAi FOR THE CONTROL OF FUNGI AND OOMYCETES BY INHIBITING SACCHAROPINE DEHYDROGENASE GENE
The present invention relates to control of plant pathogens, particularly fungi or oomycetes, by inhibiting one or more biological functions, particularly by inhibiting saccharopine dehydrogenase gene(s) using RNA interference. The invention provides methods and compositions using RNA interference of plant pathogens target genes for such control. The invention is also directed to methods for making transgenic plants tolerant to said plant pathogens, and to transgenic plants and seeds generated thereof.
1 . A dsRNA molecule comprising 1) a first strand comprising a sequence substantially identical to at least 18 contiguous nucleotides of a fungus or oomycete saccharopine dehydrogenase gene and ii) a second strand comprising a sequence substantially complementary to the first strand.
2 . A dsRNA molecule according to claim 1 wherein the fungus or oomycete gene is selected from the group consisting of:
a) a polynucleotide comprising a sequence as set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43;
b) a polynucleotide encoding a polypeptide having a sequence as set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44;
c) a polynucleotide having at least 70% sequence identity to a polynucleotide having a sequence as set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43:
d) a polynucleotide encoding a polypeptide having at least 70% sequence identity to a polypeptide having a sequence as set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44;
e) a polynucleotide hybridizing under stringent conditions to a polynucleotide having a sequence as set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43;
and
f) a polynucleotide hybridizing under stringent conditions to a polynucleotide encoding a polypeptide having at least 70% sequence identity to a polypeptide having a sequence as set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44.
3 . A composition comprising at least a dsRNA molecule according to claim 1 .
4 . A composition according to claim 3 , characterised in that it further comprises an agriculturally acceptable support, carrier, filler and/or surfactant.
5 . A composition according to claim 3 comprising further a phytopharmaceutical or plant growth promoting compound.
6 . A micro-organism producing a dsRNA molecule according to claim 1 .
7 . A genetic construct which comprises at least one DNA sequence as well as heterologous regulatory element(s) in the 5′ and optionally in the 3′ positions, characterized in that the DNA sequence(s) is able to form a dsRNA molecule according to claim 1 .
8 . A cloning and/or expression vector, characterized in that it contains at least one genetic construct according to claim 7 .
9 . A transgenic plant cell capable of expressing at least a dsRNA molecule according to claim 1 .
10 . A transgenic plant, seed or part thereof, comprising a transgenic plant cell according to claim 9 .
11 . The transgenic plant cell according to claim 9 or transgenic plant, seed or part thereof comprising said transgenic plant cell, wherein said plant is a soybean, oilseed, rice or potato plant.
12 . A method of making a transgenic plant cell capable of expressing a dsRNA that inhibits a fungus or oomycete saccharopine dehydrogenase gene, said method comprises the steps of transforming a plant cell with a genetic construct according to claim 7 .
13 . A method of controlling a plant pathogen, particularly a fungus or oomycete, comprising providing to said pathogen a dsRNA molecule according to claim 1 , or a composition comprising at least a dsRNA molecule according to claim 1 .
14 . A method for controlling a plant pathogen, particularly a fungus or oomycete, characterized in that an effective quantity of a dsRNA molecule according to claim 1 or a composition comprising at least a dsRNA molecule according to claim 1 is applied to the soil where plants grow or are capable of growing, to the leaves and/or the fruit of plants or to the seeds of plants.
15 . A method of controlling a plant pathogen, particularly a fungus or an oomycete, comprising providing in the host plant of said plant pathogen a transformed plant cell according to claim 9 .
16 . A method for inhibiting the expression of a plant pathogen gene, comprising the following steps
i) transforming a plant cell with a genetic construct according to claim 7 ,
ii) placing the cells thus transformed under conditions that allow the transcription of said construct,
iii) bringing the cells into contact with the plant pathogen.
17 . The method according to claim 13 wherein said plant pathogen is Magnaporthe grisea, Phytophthora infestans. Sclerotinia sclerotinium or Phakopsora pachyrhizi.
18 . The method according to claim 12 wherein said plant is soybean, oilseed, rice or potato.