Rhg1 mediated resistance to soybean cyst nematode
Methods of increasing the resistance of plants, in particular soybeans, to nematodes, in particular soybean cyst nematodes, are provided herein. The methods include increasing the expression of Glyma18g02580, Glyma18g02590 and/or Glyma18g2610 in cells of a plant and in particular in root cells of a plant to increase the resistance of the plant and plant cells to nematodes. The methods include increasing the expression using constitutive promoters or by increasing the copy number of the polynucleotides. Constructs for expressing these polypeptides, transgenic cells, transgenic plants and methods of generating the same are also provided. Methods of screening plant cells for resistance or susceptibility to nematodes are also provided.
1. A method of increasing resistance of plant cells to nematodes comprising:
introducing three or more copies of polynucleotides that encode
a polypeptide according to SEQ ID NO: 1, or a polypeptide with at least 98% identity to SEQ ID NO: 1,
a polypeptide according to SEQ ID NO: 2, SEQ ID NO: 5, or SEQ ID NO: 6, or a polypeptide with at least 95% identity to SEQ ID NO: 2, SEQ ID NO: 5, or SEQ ID NO: 2, and
a polypeptide according to SEQ ID NO: 3,
wherein said plant cells have increased resistance to nematodes relative to plant cells without increased copy number of said polynucleotides,
and wherein said plant cells are soybean, Arabidopsis or potato.
2. The method of claim 1 , wherein four or more copies of polynucleotides encoding said polypeptides are introduced in the plant cells.
3. The method of claim 1 , wherein at least ten copies of polynucleotides encoding said polypeptides are introduced in the plant cells.
4. The method of claim 1 , wherein the plant cells comprise a plant, and wherein increased nematode resistance comprises the plant having one or more of:
(a) a lower percentage of invading nematodes that develop past the J2 stage,
(b) a lower rate of cyst formation on the roots,
(c) reduced nematode female index of a plant exposed to nematodes,
(d) reduced SCN egg production within cysts,
(e) reduced overall SCN egg production per plant, or
(f) greater yield of soybean seeds on a per-plant basis or a per-growing-area basis, as compared to a plant without increased copy number of said polynucleotides grown in a similar growth environment.
5. The method of claim 1 , further comprising increasing expression of said polynucleotides in the plant cells,
wherein said plant cells have increased resistance to nematodes relative to plant cells without increased expression of said polynucleotides.
6. The method of claim 5 , wherein expression of one or more polynucleotides is increased in plant cells in the root of the plant.
7. The method of claim 5 , wherein expression of one or more polypeptides is increased by increasing expression of one or more native polynucleotides.
8. The method of claim 5 , wherein expression of said polypeptides is increased by introducing a construct comprising a promoter operably linked to a polynucleotide encoding the polypeptide into the plant cells.
9. The method of claim 5 , wherein expression of said polypeptides are increased by incorporation of a construct comprising a promoter operably linked to a polynucleotide encoding one of said polypeptides in the plant cells.
10. The method of claim 5 wherein the plant cell exhibits increased expression of each of
a polypeptide according to SEQ ID NO: 1,
a polypeptide according to SEQ ID NO: 2, SEQ ID NO: 5, or SEQ ID NO: 6, and
a polypeptide according to SEQ ID NO: 3.
11. A transgenic plant cell comprising:
three or more copies of polynucleotides encoding polypeptides capable of increasing resistance to nematodes, the polypeptides comprising:
a polypeptide according to SEQ ID NO: 1, or a polypeptide with at least 98% identity to SEQ ID NO: 1,
a polypeptide according to SEQ ID NO: 2, SEQ ID NO: 5, or SEQ ID NO: 6, or a polypeptide with at least 95% identity to SEQ ID NO: 2, SEQ ID NO: 5, or SEQ ID NO: 2, and
a polypeptide according to SEQ ID NO: 3,
wherein said plant cell has increased resistance to nematodes relative to plant cells
without increased copy number of said polynucleotides, and wherein said plant cells are soybean, Arabidopsis or potato.
12. The transgenic cell of claim 11 , wherein the polynucleotides are present in at least four copies in the cell.
13. The transgenic cell of claim 11 , wherein the polynucleotides are present in at least ten copies in the cell.
14. A seed comprising the transgenic cell of claim 11 .
15. A plant grown from the seed of claim 14 .
16. A transgenic plant comprising the cell of claim 11 .
17. A part, progeny or asexual propagate of the transgenic plant of claim 16 .
18. The transgenic plant cell of claim 11 , further comprising increasing expression of the polynucleotides encoding said polypeptides capable of increasing resistance to nematodes,
wherein said plant cells have increased resistance to nematodes relative to plant cells without increased expression of said polynucleotides.
19. The transgenic plant cell of claim 18 , wherein the polynucleotides encode at least
a polypeptide according to SEQ ID NO: 1,
a polypeptide according to SEQ ID NO: 2, SEQ ID NO: 5, or SEQ ID NO: 6, and
a polypeptide according to SEQ ID NO: 3.
20. A method of generating a transgenic plant with increased resistance to nematodes comprising introducing into a plant cell or its progeny three or more copies of exogenous polynucleotides encoding one or more polypeptides capable of increasing resistance to nematodes, the polypeptides comprising:
a Glyma18g02580 polypeptide having at least 98% identity to SEQ ID NO: 1,
a Glyma18g02590 polypeptide having at least 95%; identity to SEQ ID NO: 2, 5 or 6, and
a Glyma18g02610 polypeptide according to SEQ ID NO: 3, wherein said plant has increased resistance to nematodes relative to a plant without increased copy number of said polynucleotides, and wherein said plant is soybean, Arabidopsis or potato.
21. The method of claim 20 , wherein the polynucleotides are present in at least four copies in the plant.
22. The method of claim 20 , further comprising increasing expression of the polynucleotides encoding polypeptides capable of increasing resistance to nematodes,
wherein said plant has increased resistance to nematodes relative to a plant without increased expression of said polynucleotides.
23. The method of claim 22 , wherein the polynucleotides encode at least a Glyma18g02610 polypeptide according to SEQ ID NO:3, a Glyma18g02590 polypeptide according to SEQ ID NO:2, 5 or 6 and a Glyma18g02580 polypeptide according to SEQ ID NO:1.