PATENTSCOPE will be unavailable a few hours for maintenance reason on Tuesday 19.11.2019 at 4:00 PM CET
Search International and National Patent Collections
Some content of this application is unavailable at the moment.
If this situation persists, please contact us atFeedback&Contact
1. (WO2007002267) NUCLEOTIDE SEQUENCES MEDIATING PLANT MALE FERTILITY AND METHOD OF USING SAME
Note: Text based on automatic Optical Character Recognition processes. Please use the PDF version for legal matters

What is claimed is:
1. A method of maintaining a homozygous recessive condition of a male sterile plant, the method comprising:
(a) providing a first plant comprising homozygous recessive alleles of the Ms26 gene and is male sterile;
(b) introducing a construct into a second plant, the second plant comprising homozygous recessive alleles of the Ms26 gene, the construct in the hemizygous condition, the construct comprising:
(i) a first nucleotide sequence comprising the Ms26 nucleotide sequence, that when expressed in the first plant would restore male fertility;
(ii) a second nucleotide sequence that when expressed inhibits the function or formation of viable male gametes in the second plant, such that viable male gametes are produced in the second plant containing the recessive alleles of Ms26 and that do not contain the construct; and
(c) fertilizing the first plant with the male gametes of the second plant to produce progeny which maintain the homozygous recessive condition of the first plant.
2. The method of claim 1 , wherein the first nucleotide sequence is operably linked to a third nucleotide sequence directing expression preferentially to male plant cells.
3. The method of claim 2, wherein the third nucleotide sequence functions only in the presence of an inducing substance or condition.
4. The method of claim 2, wherein the third nucleotide sequence is selected from the group consisting of the male tissue regulatory sequence of 5126, Ms26 and

Ms45.
5. The method of claim 1 , wherein the second nucleotide sequence is operably linked to a fourth nucleotide sequence, the fourth nucleotide sequence directing expression preferentially to male gametes.
6. The method of claim 1 , wherein the second nucleotide sequence is the selected from the group consisting of the nucleotide sequence of the DAM methylase gene, Zea mays alpha amylase gene, and a cytotoxin encoding gene.

7. The method of claim 6, wherein the second sequence is operably linked to a fourth nucleotide sequence directing expression to male gametes.
8. The method of claim 5, wherein the fourth nucleotide sequence is selected from the group consisting of the regulatory region of the polygalacturonase 47 gene, Zm13 gene, pectin methylesterase gene, calmodulin binding protein gene, actin depolymerizing factor gene, prolfilin gene, and sulphated pentapeptide phytosulphokine gene.
9. The method of claim 1 , further comprising a fifth nucleotide sequence encoding a product, the expression of which is capable of being used for selection of plant cells having the construct.
10. The method of claim 9 wherein the fifth nucleotide sequence is selected from the group consisting of red fluorescent gene, cyan fluorescent protein gene, yellow fluorescent protein gene, luciferase gene, green fluorescent protein gene, anthocyanin p1 gene and phosphinothricin acetyltransferase encoding gene.
11. The method of claim 9 wherein the fifth nucleotide sequence is operably linked to a regulatory region selected from the group consisting of the polygalacturonase 47 gene, Zm13 gene, pectin methylesterase gene, calmodulin binding protein gene, actin depolymerizing factor gene, prolfilin gene, and sulphated pentapeptide phytosulphokine gene.
12. The method of claim 1 , further comprising selecting for said second plant by identifying plants having said construct.
13. A method of maintaining a homozygous recessive condition of a first plant when crossing the first plant to a second plant, the method comprising:
(a) providing a first plant comprising homozygous recessive alleles, of the Ms26 gene, the expression of which results in male sterility;
(b) introducing a construct into a second plant, the second plant comprising homozygous recessive alleles of the Ms26 gene, the construct in a hemizygous condition comprising:
(i) a first nucleotide sequence comprising the Ms26 nucleotide sequence, that when expressed in the first plant would restore male fertility;
(ii) a second nucleotide sequence selected from the group consisting of the sequence of the DAM methylase gene, Zea mays alpha amylase gene, and cytotoxin encoding gene;

(iii) a third nucleotide sequence operably linked to the first nucleotide sequence, selected from the group consisting of the male tissue regulatory region of 5126, Ms26 or Ms45, directing expression preferentiallty to male plant tissue;
(iv) a fourth nucleotide sequence operably linked to the second nucleotide sequence directing expression to plant male gametes, such that viable male gametes are produced in the second plant containing the recessive alleles, and that do not contain the construct; and
(c) fertilizing the first plant with the male gametes of the second plant to produce progeny which is male sterile and maintain the homozygous recessive condition of the first plant.
14. The method of claim 13, further comprising a fifth nucleotide sequence encoding a product capable of being used for selection of plant cells containing the construct.
15. The method of claim 14 wherein the fifth nucleotide sequence is selected from the group consisting of red fluorescent gene, cyan fluorescent protein gene, yellow fluorescent protein gene, luciferase gene, green fluorescent protein gene, anthocyanin p1 gene and phosphinothricin acetyltransferase encoding gene.
16. The method of claim14 wherein the fifth nucleotide sequence is operably linked to a regulatory region selected from the group consisting of the polygalacturonase 47 gene, Zm13 gene, pectin methylesterase gene, calmodulin binding protein gene, actin depolymerizing factor gene, prolfilin gene, and sulphated pentapeptide phytosulphokine gene.
17. The method of claim 1 wherein the first nucleotide sequence is selected from the group consisting of:
a) a sequence encoding any of the amino acid sequences of
SEQ ID NO: 2, 4 or 18;
b) the sequence of any of SEQ ID NO: 1 , 3, 7 or 17;
c) a sequence having at least 90% identity to any of the
foregoing sequences;
d) a sequence which hybridizes to any of the foregoing
sequences under highly stringent conditions of a wash of
0.1 SSC, 0.1% (w/v) SDS at 65°C; and e) a fragment of any of the foregoing sequences which
fragment is a functional sequence critical to male fertility in a
plant.
18. The method of claim 17, wherein the first nucleotide sequence is operably linked to a third nucleotide sequence directing expression preferentially to male plant cells.
19. The method of claim 18, wherein the third nucleotide sequence functions only in the presence of an inducing substance or condition.
20. The method of claim 18, wherein the third nucleotide sequence is selected from the group consisting of the male tissue regulatory sequence of 5126, Ms26 and Ms45.
21. The method of claim 17, wherein the second nucleotide sequence is linked to a fourth nucleotide sequence, the fourth nucleotide sequence directing expression preferentially to male gametes.
22. The method of claim 17, wherein the second nucleotide sequence is selected from the group consisting of the nucleotide sequence of the DAM methylase gene, Zea mays alpha amylase gene, and cytotoxin encoding gene.
23. The method of claim 22, wherein the second sequence is operably linked to a third nucleotide sequence directing expression preferentially to plant male gametes.
24. The method of claim 21 , wherein the fourth nucleotide sequence is selected from the group consisting of the regulatory region of the polygalacturonase 47 gene, Zm13 gene, pectin methylesterase gene, calmodulin binding protein gene, actin depolymerizing factor gene, prolfilin gene, and sulphated pentapeptide phytosulphokine gene.
25. The method of claim 17, further comprising a fifth nucleotide sequence encoding a product capable of being used for selection of plant cells having the construct.
26. The method of claim 25 wherein the fifth nucleotide sequence is selected from the group consisting of red fluorescent gene, cyan fluorescent protein gene, yellow fluorescent protein gene, luciferase gene, green fluorescent protein gene, anthocyanin p1 gene and phosphinothricin acetyltransferase encoding gene.
27. The method of claim 25 wherein the fifth nucleotide sequence is operably linked to a regulatory region selected from the group consisting of the polygalacturonase 47 gene, Zm13 gene, pectin methylesterase gene, calmodulin binding protein gene, actin depolymerizing factor gene, prolfilin gene, and sulphated pentapeptide phytosulphokine gene.
28. The method of claim 17 further comprising selecting for said second plant by identifying plants having said construct.
29. A method of producing seed from a plant having female and male gametes, the method comprising:
(a) introducing into a male sterile plant comprising homozygous recessive alleles of the Ms26, a construct in the hemizygous condition comprising:
(i) a first nucleotide sequence comprising the Ms26 nucleotide sequence;

(ii) a second nucleotide sequence, that when expressed inhibits the function or formation of male gametes in the plant, such that viable male gametes are produced in the plant that do not contain the construct;
(b) self fertilizing the plant; and
(c) producing seed which contain the construct.
30. The method of claim 29, wherein the first nucleotide sequence is operably linked to a third nucleotide sequence directing expression preferentially to male plant cells
31. The method of claim 29, wherein the second nucleotide sequence is operably linked to a fourth nucleotide sequence, the fourth nucleotide sequence directing expression preferentially to male gametes.
32. The method of claim 29, further comprising a fifth nucleotide sequence encoding a product capable of being used for selection of plant cells containing the construct.
33. The method of claim 29, further comprising identifying plants having said construct.
34. The method of claim 29 wherein the first nucleotide sequence is selected from the group consisting:
a) a sequence encoding any of the amino acid sequences of SEQ ID NO: 2, 4 or 18;
b) the sequence of any of SEQ ID NO: 1 , 3, 7 or 17;
c) a sequence having at least 90% identity to any of the foregoing sequences;

d) a sequence which hybridizes to any of the foregoing sequences under highly stringent conditions of a wash of 0.1 SSC, 0.1% (w/v) SDS at 65°C; and
e) a fragment of any of the foregoing sequences, which fragment is a sequence critical to male fertility in a plant.
35. A method of maintaining a homozygous recessive condition of a plant, the method comprising:
(a) providing a first plant comprising homozygous recessive alleles of a gene required for the phenotypic expression of a plant trait;
(b) introducing a construct into a second plant, the second plant comprising homozygous recessive alleles of a gene required for the phenotypic expression of a plant trait, the construct in the hemizygous condition comprising:
(i) a first nucleotide sequence that when expressed in the first plant functionally compliments the homozygous recessive plant trait;
(ii) a second nucleotide sequence, the expression of which inhibits the function or formation of male gametes in the second plant; such that male gametes are produced in the second plant containing the recessive alleles and that do not contain the construct; and
(c) fertilizing the first plant with the male gametes of the second plant to produce progeny which maintain the homozygous recessive condition of the first plant.
36. The method of claim 335, wherein the first nucleotide sequence is operably linked to a third nucleotide sequence directing expression preferentially to male plant cells.
37. The method of claim 35, wherein the second nucleotide sequence is operably linked to a fourth nucleotide sequence, the fourth nucleotide sequence directing expression preferentially to male gametes.
40. The method of claim 35, further comprising a fifth nucleotide sequence encoding a product capable of being used for selection of plant cells having the construct.
41. The method of claim 35, further comprising selecting for said second plant by identifying plants having said construct.
42. The method of claim 35, wherein the trait is male fertility.

43. A method of producing seed from a plant having female and male gametes, the method comprising:
(a) introducing a construct into a first plant, the plant comprising homozygous recessive alleles of a gene required for phenotypic expression of a plant trait, the construct in a hemizygous condition comprising:
(i) a first nucleotide sequence, the sequence that when expressed in a second plant having homozygous recessive alleles of a gene required for male fertility and is male sterile, would restore male fertility;
(ii) a second nucleotide sequence operably linked to the first nucleotide sequence which inhibits the function or formation of male gametes in the plant, such that viable male gametes not having the first nucleotide sequence are produced in the plant;
(b) self fertilizing the plant; and
(c) producing seed which contain the construct.
44. The method of claim 43, wherein the first nucleotide sequence is operably linked to a third nucleotide sequence directing expression preferentially to male plant cells.
45. The method of claim 43, wherein the second nucleotide sequence is operably linked to a fourth nucleotide sequence, the fourth nucleotide sequence directing expression to male gametes.
46. The method of claim 43, further comprising a fifth nucleotide sequence encoding a product capable of being used for selection of plant cells having the construct.
47. The method of claim 43, further comprising identifying plants having said construct.
48. The method of claim 43, wherein the trait is male fertility.
(ii) 49. The method of claim 2 wherein the third nucleotide
sequence is selected from the group consisting of:
(a) SEQ ID NO: 5;
(b) SEQ ID NO: 6;
(c) bases 1-1088 of SEQ ID NO: 7;
(d) bases 830 to 962 of SEQ ID NO: 7;
(e) bases 830 to 914 of SEQ ID NO: 7;
(f) bases 917 to 962 of SEQ ID NO: 7;

(g) bases 875 to 954 of SEQ ID NO: 7;
(h) bases 935 to 954 of SEQ ID NO: 7;
(i) bases 875 to 924 of SEQ ID NO: 7;
0) SEQ ID NO: 19;
(k) SEQ ID NO: 20; and
(I) a fragment of any of the foregoing sequences,
which functional fragment is essential for male
tissue expression of a linked sequence.

50. The method of claim 49, wherein the second nucleotide sequence is operably linked to a fourth nucleotide sequence, the fourth nucleotide sequence directing expression preferentially to male gametes.
51. The method of claim 49, wherein the second nucleotide sequence is selected from the group consisting of the nucleotide sequence of the DAM methylase gene, Zea mays alpha amylase gene, and a cytotoxin encoding gene.
52. The method of claim 51 wherein the second sequence is operably linked to a fourth nucleotide sequence directing expression to plant male gametes.
53. The method of claim 50, wherein the fourth nucleotide sequence is selected from the group consisting of the regulatory region of the polygalacturonase 47 gene, Zm13 gene, pectin methylesterase gene, calmodulin binding protein gene, actin depolymerizing factor gene, prolfilin gene, and sulphated pentapeptide phytosulphokine gene.
54. The method of claim 49, further comprising a fifth nucleotide sequence encoding a product capable of being used for selection of plant cells having the construct.
55. The method of claim 49, further comprising selecting for said second plant by identifying plants having said construct.
56. The method of claim 30 wherein the third nucleotide sequence isselected from the group consisting of:
(a) SEQ ID NO: 5;
(b) SEQ ID NO: 6;
(c) bases 1-1088 of SEQ ID NO: 7;
(d) bases 830 to 962 of SEQ ID NO: 7;
(e) bases 830 to 914 of SEQ ID NO: 7;

(f) bases 917 to 962 of SEQ ID NO: 7;
(g)bases 875 to 954 of SEQ ID NO: 7;
(h) bases 935 to 954 of SEQ ID NO: 7;
(i) bases 875 to 924 of SEQ ID NO: 7;
G) SEQ ID NO: 19;
(k) SEQ ID NO: 20; and
(I) a fragment of any of the foregoing sequences
which functional fragment is essential for male
tissue expression of a linked sequence,
such that viable gametes are produced in the first
plant that do not contain the construct.
57. The method of claim 56, wherein the second nucleotide sequence is operably linked to a fourth nucleotide sequence, the fourth nucleotide sequence directing expression preferentially to male gametes.
58. The method of claim 56, further comprising a fifth nucleotide sequence encoding a product capable of being used for selection of plant cells having the construct.
59. The method of claim 56, further comprising identifying plants having said construct.
60. A method of restoring male fertility in a male sterile plant, the male sterile plant comprising homozygous recessive alleles of Ms26, the method comprising introducing into said plant a Ms26 nucleotide sequence that is the functional compliment of the homozygous recessive condition.
61. A method of restoring male fertility in a male sterile plant, the male sterile plant comprising homozygous recessive alleles of a gene critical to male fertility, the method comprising introducing into said plant a nucleotide sequence that is the functional compliment of the homozygous recessive condition, wherein the nucleotide sequence is selected from the group consisting of:
a) a sequence encoding any of the amino acid sequences of SEQ
ID NO: 2, 4 or 18;
b) the sequence of any of SEQ ID NO: 1 , 3, 7 or 17;
c) a sequence having at least 90% identity to any of the foregoing sequences;

d) a sequence which hybridizes to any of the foregoing sequences under highly stringent conditions of a wash of 0.1 SSC, 0.1%
(w/v) SDS at 65°C; and
e) a fragment of any of the foregoing sequences, which functional fragment is a sequence critical to male fertility in a plant;
62. An isolated nucleotide sequence comprising SEQ ID NO: 26.
63. An expression cassette comprising SEQ ID NO: 26.
64. A plant cell comprising a heterologous nucleotide sequence comprising SEQ ID NO: 26.
65. A plant comprising a heterologous nucleotide sequence comprising SEQ ID NO: 26.