31-08-2010 дата публикации
Номер:
Контакты:
Номер заявки: 53-00-201024
Дата заявки: 16-12-2008
[1]Patents Granted (Contd.)
[2](51) International Classification: Cl2Ν15/82(2006.01)
[4]Genotypes, Alleles and Molecular Markers asscociated with Asian soybean rust, as well as methods, processes and uses thereof
[6]The present invention relates to screening methods for rust resistance or
[7]tolerance, in particular, Asian soybean rust (ASR - Phakopsora pachyrhizi). In
[8]addition, the present invention relates to the use of molecular markers for the
[9]Glycine genus, in particular, for the Glycine max species. The present
[10]invention further relates to a method for identifying loci with quantitative and/
[11]or qualitative traits associated with nist resistance or tolerance in plants by
[12]means of molecular markers. Said markers can be used for assisted screening
[13]in improvement programs directed to selecting disease-resistant or -tolerant
[14]plants. The present invention also relates to gene pyramiding related to rust
[15]resistance. The markers of the present invention are also usefiil for the
[16]positional cloning of rust-resistant or -tolerant genes. Also disclosed are a
[17]method for obtaining disease-resistant or - tolerant cultivars, a process for
[18]obtaining a plant population and a method for controlling diseases in a plant
[19]population. Another object of the present invention is the use of species from
[20]the Glycine genus as a source of resistance for obtaining ASR-resistant or
[22](56) Documents Cited : MEESANG Ν et al MONTEROS M.J. et al N.D. YOUNG: "QTL MAPPING AND QUANTITATIVE DISEASE RESISTANCE IN PLANTS" ANNU. REV. PHYTOPATHOL, vol. 34, 1996, pages 829-836 D.L. ΗΥΤΕΝ et al
[23]FORM 25 (12) PATENT (19) ΑΡ
[24](11) Patent No : ΑΡ 3062 (73) Applicant(s)
[25](21) Application No : AP/P/2010/005324 TMG - TROPICAL MELHORAMENTO Ε GENETICA LTDA Rodovia Celso Garcia CID, KM 87, Caixa Postal 387, (Parke
[26](22) Filing Date : 16.12.2008 Industrial), Cambe-PR, 86183-600, Brazil
[27](24) (45) Date of Grant 8c Publication : 23.12.2014
[29](33) Country (31) Number (32) Date (72) Inventors
[30]BR PI0704999-4 21.12.2007 HARADA Arlindo, Brazil
[31]CALVO SANCHES Cberson, Brazil
[32]SOUZA KIIHL Romeu, Afonso de, Brazil
[34](84) Designated State (74) Representative
[35]GM GH ΜΖ SL SD UG ΖΜ ZW FISHER CORMACK & BOTHA
[36]The present invention relates to screening methods for rust resistance or tolerance, in particular, Asian soybean rust (ASR—Phakopsora pachyrhizi). In addition, the present invention relates to the use of molecular markers for the Glycine genus, in particular, for the Glycine max species. The present invention further relates to a method for identifying loci with quantitative and/or qualitative traits associated with rust resistance or tolerance in plants by means of molecular markers. Said markers can be used for assisted screening in improvement programs directed to selecting disease-resistant or -tolerant plants. The present invention also relates to gene pyramiding related to rust resistance. The markers of the present invention are also useful for the positional cloning of rust-resistant or -tolerant genes. Also disclosed are a method for obtaining disease-resistant or -tolerant cultivars, process for obtaining a plant population and a method for controlling diseases in a plant population. Another object of the present invention is the use of species from the Glycine genus as a source of resistance for obtaining ASR-resistant or tolerant cultivars.
1. A method for identifying and selecting individuals carrying a phenotypic trait of interest from a biological population, characterized by comprising:
(a) associating a molecular marker to the phenotypic trait of interest;
(b) segregating individuals from the biological population based on the presence or absence of at least one molecular marker associated to said phenotypic trait, wherein said phenotypic trait is attributable to one or more genetic loci;
(c) indicating the markers which show the profile of the segregating individuals of (b); and
(d) determining the patterns of the markers of (c) the genetic loci associated with the phenotypic trait of interest.
2. A method according to claim 1 , characterized in that said biological population is a plant population.
3. A method according to claim 1 or 2 , characterized in that said biological population comprises at least 11 individuals.
4. A method according to any one of claims 1 to 3 , characterized in that said biological population is composed of plants of the Glycine genus.
5. A method according to claim 4 , characterized in that said biological population is composed of plants of the Glycine max species.
6. A method according to any one of claims 1 to 5 , characterized in that said phenotypic trait is selected from the group consisting of yield, resistance to diseases and tolerance to diseases.
7. A method according to claim 6 , characterized in that said disease is Asian soybean rust.
8. A method according to any one of claims 1 to 7 , characterized in that said phenotypic trait is conferred by a locus or QTL.
9. A method according to any one of claims 1 to 8 , characterized in that said marker exhibits at least slight variations in the sequences of nucleotides between individuals of the biological population.
10. A method according to any one of claims 1 to 9 , characterized in that said marker is a DNA, cDNA or RNA fragment.
11. A method according to any one of claims 1 to 10 , characterized in that said marker is selected from RFLPs, RAPDs, AFLPs, SSRs, SNPs, ESTs and RGA probes.
12. A method according to any one of claims 1 to 11 , characterized in that said marker is positioned in a region of a linkage group.
13. A method according to claim 12 , characterized in that the linkage group represents all the loci which may be directly or indirectly connected by linkage relations.
14. A method according to claim 12 or 13 , characterized in that the linkage group is part of a chromosome or a complete chromosome.
15. A method according to any one of claims 12 to 14 , characterized in that the marker is any marker mapped at 10 cM or less from the locus or QTL of the phenotypic trait of interest.
16. A method according to any one of claims 12 to 15 , characterized in that the linkage group is selected from the group consisting of linkage group G, linkage group J and linkage group N of the soybean consensus linkage map.
17. A method according to claim 16 , characterized in that the linkage group is linkage group G.
18. A method according to claim 17 , characterized in that said marker is one or more among Satt12, A816—1, A890—1, Sat—164, Satt503, Satt517, Sat—143, Mng273—2, Satt288, A121—2, A885—1, Satt612, K493—1, T005—2, bac1F11Rhnd and OP_M02a.
19. A method according to claim 16 , characterized in that the linkage group is linkage group J.
20. A method according to claim 19 , characterized in that said marker is one or more among Satt215, Sat—361, Sct—001, Sat—093, A109—9, Sat—366, Satt621, Satt620, Sat—350, RGA—3, RGA—2a, Satt244 and Satt431.
21. A method according to claim 16 , characterized in that the linkage group is linkage group N.
22. A method according to claim 21 , characterized in that said marker is one or more among A426—2, Sle—003, i4—2, Sat—084, Satt393, Satt584, Satt485, Sat—166, Sat—208, BLT049—1, Bng095—2, OP_F13, Satt125, Sat—275, Sle—3, RGA—6b, OP_U09b, mO128—1, Sat—280, Satt080 and Sat—266.
23. A method according to any one of claims 1 to 22 , characterized in that the markers are selected from genotypes having different phenotypes.
24. A method according to claim 23 , characterized in that said genotypes are selected from soybean genotypes PI 459025, PI 230970, PI 200456, PI 224270, PI 200526, PI 200487 and PI 471904 or from genotypes having Asian soybean rust resistance genes or even alleles of these PIs.
25. A method according to any one of claims 1 to 24 , characterized in that the identification of the markers is made by BSA linkage analysis followed by genetic mapping.
26. A method of pyramiding genes associated with phenotypic traits of interest in a plant population, characterized by comprising:
(a) associating one or more molecular markers to the phenotypic trait of interest in one or more plants or plant varieties;
(b) identifying the marker(s) for each gene to be incorporated in step (c);
(c) incorporating the genes from the plant population by crossbreeding between said plants or plant varieties; and
(d) monitoring the incorporation of genes into the plant population through the marker(s).
27. A method according to claim 26 , characterized in that step (c) consists of incorporating the largest possible number of favorable genes for a given trait of interest into a single genotype.
28. A method according to claim 26 or 27 , characterized in that said plant population is composed of plants of the Glycine genus.
29. A method according to claim 28 , characterized in that said population is composed of plants of the Glycine max species.
30. A method according to any one of claims 26 to 29 , characterized in that said phenotypic trait is selected from the group consisting of yield, resistance to diseases and tolerance to diseases.
31. A method according to claim 30 , characterized in that said disease is Asian soybean rust.
32. A method according to any one of claims 26 to 31 , characterized in that the marker is any marker mapped at 10 cM or less from the locus or QTL of the phenotypic trait of interest.
33. A method according to any one of claims 26 to 32 , characterized in that the marker is positioned in a region of a linkage group and is selected from the group consisting of linkage group G, linkage group J and linkage group N of the soybean consensus linkage map.
34. A method according to claim 33 , characterized in that the linkage group is linkage group G.
35. A method according to claim 34 , characterized in that said marker is one or more among Satt12, A816—1, A890—1, Sat—164, Satt503, Satt517, Sat—143, Mng273—2, Satt288, Al21—2, A885—1, Satt612, K493—1, T005—2, bac1F11Rhnd and OP_M02a.
36. A method according to claim 33 , characterized in that the linkage group is linkage group J.
37. A method according to claim 36 , characterized in that said marker is one or more among Satt215, Sat—361, Sct—001, Sat—093, A109—9, Sat—366, Satt621, Satt620, Sat—350, RGA—3, RGA—2a, Satt244 and Satt431.
38. A method according to claim 33 , characterized in that the linkage group is linkage group N.
39. A method according to claim 38 , characterized in that said marker is one or more among A426—2, Sle—003, 14—2, Sat—084, Satt393, Satt584, Satt485, Sat—166, Sat—208, BLT049—1, Bng095—2, OP_F13, Satt125, Sat—275, Sle—3, RGA—6b, OP_U09b, mO128—1, Sat—280, Satt080 and Sat—266.
40. A method according to any one of claims 26 to 39 , characterized in that said plants or plant varieties are selected from soybean genotypes PI 459025, PI 230970, PI 200456, PI 224270, PI 200526, PI 200487 and PI 471904, or from genotypes having Asian soybean rust resistance genes or even alleles of these PIs.
41. A method according to any one of claims 26 to 39 , characterized in that the soybean genotypes are PI 200456, PI 224270, PI 200526, PI 471904 or alleles of these PIs.
42. A method according to claim 41 , characterized in that the alleles are selected from locus rpp2, located in linkage group J, and from locus Rpp5/rpp5, located in linkage group N.
43. A method according to claim 42 , characterized in that the markers of claim 39 are used for gene Rpp5/rpp5 in genotypes PI 200456, PI 200487 and PI 200526, the markers of claim 37 are used for gene rpp2 in genotype PI 224270, and the markers of claim 35 are used for gene Rpp4 in genotype PI 459025.
44. A method for isolating and identifying genes associated with phenotypic traits of interest in a plant population by positional cloning, cha-racterized by comprising:
(a) preparing a genomic library;
(b) selecting cloned fragments from said library by using molecular markers;
(c) using the selected cloned fragments to create a contig of cloned DNA inserts; and
(d) identifying the gene(s) in one or more clones residing in the contig.
45. A method according to claim 44 , characterized in that said plant population is composed of plants of the Glycine genus.
46. A method according to claim 45 , characterized in that said population is composed of plants of the Glycine max species.
47. A method according to any one of claims 44 to 46 , characterized in that said phenotypic trait is selected from the group consisting of yield, resistance to diseases and tolerance to diseases.
48. A method according to claim 47 , characterized in that said disease is Asian soybean rust.
49. A method according to any one of claims 44 to 48 , characterized in that the marker is any marker mapped at 10 cM or less from the locus or QTL of the phenotypic trait of interest.
50. A method according to any one of claims 44 to 49 , characterized in that the marker is positioned in a region of a linkage group selected from the group consisting of linkage group G, linkage group J and linkage group N of the soybean consensus linkage map.
51. A method according to claim 50 , characterized in that the linkage group is linkage group G.
52. A method according to claim 51 , characterized in that said marker is one or more among Satt12, A816—1, A890—1, Sat—164, Satt503, Satt517, Sat—143, Mng273—2, Satt288, Al21—2, A885—1, Satt612, K493—1, T005—2, bac1F11Rhnd and OP_M02a.
53. A method according to claim 50 , characterized in that the linkage group is linkage group J.
54. A method according to claim 53 , characterized in that said marker is one or more among Satt215, Sat—361, Sct—001, Sat—093, A109—9, Sat—366, Satt621, Satt620, Sat—350, RGA—3, RGA—2a, Satt244 and Satt431.
55. A method according to claim 50 , characterized in that the linkage group is linkage group N.
56. A method according to claim 55 , characterized in that said marker is one or more among A426—2, Sle—003, i4—2, Sat—084, Satt393, Satt584, Satt485, Sat—166, Sat—208, BLT049—1, Bng095—2, OP_F13, Satt125, Sat—275, Sle—3, RGA—6b, OP_U09b, mO128—1, Sat—280, Satt080 and Sat—266.
57. A method according to claim 56 , characterized in that said marker is used for the gene Rpp5/rpp5 located in linkage group N.
58. A method according to claim 57 , characterized in that contig WmContig404, WmContig562, WmContig5258 or WmContig6713, or any contig having a marker closer than 5 cM from the gene is used.
59. A method for obtaining disease-resistant or -tolerant cultivars, characterized by comprising:
(a) hybridizing a source carrying one or more genes and/or alleles which are resistant or tolerant to a disease with a line or cultivar which is susceptible to said disease but agronomically adapted to the region for which the cultivar is to be obtained;
(b) generation advancement using at least one genetic improvement method;
(c) using at least one among the method of identification and selection as defined in any one of claims 1 to 25 , the method of gene pyramiding as defined in any one of claims 26 to 43 , and the method of isolation and identification of genes as defined in any one of claims 44 to 58 , to obtain the disease-resistant or -tolerant cultivar.
60. A method according to claim 59 , characterized in that said cultivar belongs to the Glycine genus.
61. A method according to claim 60 , characterized in that said cultivar is of the Glycine max species.
62. A method according to any one of claims 59 to 61 , characterized in that said disease is Asian soybean rust.
63. A method according to any one of claims 59 to 62 , characterized in that said at least one genetic improvement method is selected from Bulk, SSD, MSSD and Backcrossing.
64. A method according to any one of claims 59 to 63 , characterized in that, in step (a), the source was PI 635026 and the disease-susceptible line was TMGLM-3219.
65. A method according to any one of claims 59 to 64 , characterized in that the cultivar obtained is selected from among CB06-953, CB06-954, CB06-955, CB06-956, CB06-957, CB06-958, CB06-959, CB06-960, CB06-961, CB06-962, CB06-963 and CB06-964.
66. A process for obtaining a plant population having a phenotypic trait of interest, characterized by comprising at least one among the method of identification and selection as defined in any one of claims 1 to 25 , the method of gene pyramiding as defined in any one of claims 26 to 43 , the method of isolating and identifying genes as defined in any one of claims 44 to 58, and the method for obtaining disease-resistant or -tolerant cultivars as defined in any one of claims 59 to 65 .
67. A plant population, characterized by being obtained through a process comprising at least one among the method of identification and selection as defined in any one of claims 1 to 25 , the method of gene pyramiding as defined in any one of claims 26 to 43 , the method for isolating and identifying genes as defined in any one of claims 44 to 58 , and the method for obtaining disease-resistant or -tolerant cultivars as defined in any one of claims 59 to 65 .
68. A plant population according to claim 67 , characterized by being selected from among seeds, grains, shoots, cultivars and adult plants.
69. Use of soybean genotypes PI 459025, PI 230970, PI 200456, PI 224270, PI 200526, PI 200487 and PI 471904, characterized in that they are used as the source of a phenotypic trait of interest in a process as defined in claim 66 .
70. The use according to claim 69 , characterized in that said phenotypic trait is selected from the group consisting of yield, resistance to diseases and tolerance to diseases.
71. The use according to claim 70 , characterized in that said disease is Asian soybean rust.
72. The use according to any one of claims 69 to 71 , characterized in that the soybean genotypes are PI 200456, PI 224270, PI 200456, PI 200526 and PI 471904 or PIs having the resistance or tolerance gene at the same loci as them.
73. A method for controlling diseases in a plant population, characterized by comprising the genetic control of said disease by using at least one among the method of identification and selection as defined in any one of claims 1 to 25 , the method of gene pyramiding as defined in any one of claims 26 to 43 , the method for isolating and identifying genes as defined in any one of claims 44 to 58 , and the method for obtaining disease-resistant or -tolerant cultivars as defined in any one of claims 59 to 65 .
74. A method according to claim 73 , characterized in that said plant population is composed of plants of the Glycine genus.
75. A method according to claim 74 , characterized in that said population is composed of plants of the Glycine max species.
76. A method according to any one of claims 73 to 75 , characterized in that said disease is Asian soybean rust.
77. A method according to any one of claims 73 to 76 , characterized by comprising the chemical control of said disease by applying at least one fungicide to said plant population.
78. A method according to claim 77 , characterized in that said at least one fungicide is selected from triazoles, strobilurins, benzimidazoles, prothioconazole, protilcnazole and mixtures thereof.