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Last Updated: October 18, 2019

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Claims for Patent: 5,880,331

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Summary for Patent: 5,880,331
Title: Use of anthocyanin genes to maintain male sterile plants
Abstract:This invention is directed to a method to maintain male sterility in corn by the genetic linkage via genetic engineering of a homozygous male-fertility gene with a color linked restorer and plant comprising said genes. Color genes disclosed as useful in this invention include those involved in anthocyanin biosynthesis, which in corn are under the control of as many as 20 or more genes. In particular, the use of the anthocyanin color genes R, B and C1, and specific regulatory elements sequences are taught.
Inventor(s): Krebbers; Enno (Houston, TX), Williams; Mark (Gent, BE), Leemans; Jan (Deurle, BE)
Assignee: Plant Genetic Systems, N.V. (Gent, BE)
Application Number:08/485,139
Patent Claims:1. A process for maintaining a male-sterile corn line, said process comprising:

(1) crossing:

(a) a male-sterile corn line to be maintained comprising male-sterile parent plants which comprise a homozygous male-sterility genotype at a first genetic locus and which do not comprise a functional regulatory gene required for anthocyanin production in corn seeds; with

(b) a maintainer corn line comprising male-fertile parent plants which comprise said homozygous male-sterility genotype at said first genetic locus, and which further comprise, at a second genetic locus which segregates independently from said first genetic locus, foreign DNA comprising:

I) a restorer gene the expression of which prevents the phenotypic expression of said homozygous male-sterility genotype; and

II) at least one gene selected from the group of a first gene that is expressed in corn seeds to produce a first active regulatory protein having the same activity as the protein encoded by a functional R or B gene, and a second gene that is expressed in corn seeds to produce a second active regulatory protein having the same activity as the protein encoded by a functional C1 gene, wherein said foreign DNA is heterozygous at said second genetic locus; wherein said male-sterile plants produce seeds producing anthocyanin, as well as seeds not producing anthocyanin, said anthocyanin being produced only in seeds comprising said at least one gene in said foreign DNA;

2) harvesting seeds from said male-sterile parent plants; and

3) selecting from the harvested seeds, those seeds which do not produce anthocyanin, said selected seeds being capable of growing into a new generation of male-sterile plants.

2. The process of claim 1, in which said at least one gene in said foreign DNA is expressed at least in the aleurone of corn seeds.

3. The process of claim 2, in which said male-sterile parent plants do not contain a gene that is functionally expressed in the aleurone of corn seeds and encodes an active regulatory protein having the same activity as the protein encoded by a functional R or B gene, and wherein said foreign DNA in said male-fertile parent plants comprises said first gene.

4. The process of claim 3, in which said gene comprises a DNA encoding the B-peru protein which is under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between position 447 and 1061 or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

5. The process of claim 4, in which said first gene does not comprise an intron.

6. The process of claim 2, in which said male-sterile parent plants do not contain a gene that is functionally expressed in the aleurone of corn seeds which gene encodes an active regulatory protein having the same activity as a functional C1 gene, and wherein said DNA in said male-fertile parent plants comprises said second gene.

7. The process of claim 6, in which said second gene comprises a DNA encoding the C1 protein which is under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

8. The process of claim 2, in which said male-sterile parent plants do not contain a gene that is functionally expressed in the aleurone of corn seeds and encodes an active regulatory protein having the same activity as the protein encoded by a functional R or B gene, and do not contain a gene that is functionally expressed in the aleuron of corn seeds and encodes an active regulatory protein having the same activity as the protein encoded by a functional C1 gene, and wherein said foreign DNA is said male-fertile parent plants comprises said first gene and said second gene.

9. The process of claim 8, in which the said first gene comprises a DNA encoding a B-peru protein under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG; and in which said second gene comprises a DNA encoding the C1 protein under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

10. The process of claim 1, in which said first genetic locus in said male-sterile parent plants and said male-fertile parent plants is a foreign genetic locus and comprises a male-sterility gene comprising a DNA encoding barnase under control of a first promoter which directs expression selectively in specific stamen cells of corn plants, and wherein said male-sterility gene is homozygous at said first genetic locus; and in which said restorer gene at said second genetic locus in said male-fertile parent plant comprises a DNA encoding barstar under control of a second promoter which directs expression at least in said specific stamen cells.

11. The process of claim 10, in which said promoter is a TA29 promoter or a CA55 promoter.

12. The process of claim 10, in which said second promoter is identical to said first promoter.

13. The process of claim 10, in which said second promoter is a TA29 promoter or a CA55 promoter.

14. The process of claim 11, in which said second promoter is identical to said first promoter.

15. The process of claim 1, in which said first genetic locus is an endogenous male-sterility locus comprising a recessive allele in homozygous condition, and in which said restorer gene is the dominant allele at said endogenous male-sterility locus.

16. A male-fertile parent plant for maintaining a male sterile corn line comprising male-sterile parent plants which comprise a homozygous male-sterility genotype at a first genetic locus, and which male-sterile parent plants do not comprise a functional regulatory gene required for anthocyanin production in corn seeds, wherein said male-fertile parent plants comprise said homozygous male-sterility genotype at said first genetic locus and further comprise, at a second genetic locus which segregates independently from said first genetic locus, foreign DNA comprising:

I) a restorer gene, the expression of which prevents the phenotypic expression of said homozygous male-sterility genotype to render said plant male-fertile; and

II) at least one gene selected from the group of a first gene that is expressed in corn seeds to produce a first active regulatory protein having the same activity as the protein encoded by a functional R or B gene of corn, and a second gene that is expressed in corn seeds to produce a second active regulatory protein having the same activity as the protein encoded by a functional C1 gene of corn,

wherein said foreign DNA is heterozygous at said second genetic locus,

wherein said male-fertile parent plant can be crossed to said male-sterile parent plants to produce, on said male-sterile parent plants, seeds which produce anthocyanin, said anthocyanin being produced only in seeds containing said at least one gene in said foreign DNA.

17. The plant of claim 16, in which said at least one gene in said foreign DNA is expressed at least in the aleurone of corn seeds.

18. The plant of claim 17, which comprises said first gene in said foreign DNA, and which does not otherwise contain a gene that is functionally expressed in the aleurone of corn seeds which gene encodes an active regulatory protein having the same activity as the protein encoded by a functional R or B gene.

19. The plant of claim 18, in which said first gene comprises a DNA encoding a B-peru protein which is under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 and a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

20. The plant of claim 19, in which said first gene does not comprise an intron.

21. The plant of claim 17, which comprises said second gene in said foreign DNA, and which plant does not otherwise contain a gene that is functionally expressed in the aleurone of corn seeds which gene encodes an active regulatory protein having the same activity as the protein encoded by a functional C1 gene.

22. The plant of claim 21, in which said second gene comprises a DNA encoding the C1 protein which is under control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

23. The plant of claim 17, which comprises said first gene and said second gene in said foreign DNA, and which plant does not otherwise contain 1) a gene that is functionally expressed in the aleurone of corn seeds which gene encodes an active regulatory protein having the same activity as the protein encoded by a functional R or B gene, and 2) a gene that is functionally expressed in the aleurone of corn seeds which gene encodes an active regulatory protein having the same activity as the protein encoded by a functional C1 gene.

24. The plant of claim 23, in which said first gene comprises a DNA encoding the B-peru protein under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG; and in which said second gene comprises a DNA encoding the C1 protein which is under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 and a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

25. The plant of claim 16, in which said first genetic locus is a foreign genetic locus and comprises a male-sterility gene comprising a DNA encoding barnase under control of a first promoter which directs expression selectively in specific stamen cells of corn plants, and wherein said male-sterility gene is homozygous at said first genetic locus; and in which said restorer gene at said second genetic locus comprises a DNA encoding barstar under control of a second promoter which directs expression at least in said specific stamen cells.

26. The plant of claim 25, in which said first promoter is a TA29 promoter or a CA55 promoter.

27. The plant of claim 25, in which said second promoter is identical to said first promoter.

28. The plant of claim 25, in which said second promoter is a TA29 promoter or a CA55 promoter.

29. The plant of claim 26, in which said second promoter is identical of said first promoter.

30. The plant of claim 16, in which said first genetic locus is an endogenous male-sterility locus comprising a recessive allele in homozygous condition, and in which said restorer gene is the dominant allele at said endogenous male-sterility locus.

31. A kit for maintaining a male-sterile corn line, said kit comprising:

(a) a male-sterile corn line comprising male-sterile parent plants which comprise a homozygous male-sterility genotype at a first genetic locus and which male-sterile parent plants do not comprise a regulatory gene required for anthocyanin production in corn seeds; and

(b) a maintainer corn line comprising male-fertile parent plants which comprise said homozygous male-sterility genotype at said first genetic locus, which further comprise, at a second genetic locus which segregates independently from said first genetic locus, foreign DNA comprising:

I) a restorer gene, whose expression prevents phenotypic expression of said homozygous male-sterility genotype; and

II) at least one gene selected from the group of a first gene that is expressed in corn seeds to produce a first active regulatory protein having the same activity as the protein encoded by a functional R or B gene, and a second gene that is expressed in corn seeds to produce a second active regulatory protein having the same activity as the protein encoded by a functional C1 gene,

wherein said foreign DNA is heterozygous at said second genetic locus,

wherein said male-sterile and male-fertile parents can be crossed to produce, on said male-sterile parent plants, seeds which produce anthocyanin, said anthocyanin being produced only in seeds comprising said at least one gene in said foreign DNA.

32. The kit of claim 31, in which said at least one gene in said foreign DNA is expressed at least in the aleurone of corn seeds.

33. The kit of claim 32, in which said male-sterile parent plants do not contain a gene that is functionally expressed in the aleurone of corn seeds which gene encodes an active regulatory protein having the same activity as the protein encoded by a functional R or B gene, wherein said foreign DNA in said male-fertile parent plants comprises said first gene.

34. The kit of claim 33, in which said first gene comprises a DNA encoding the B-peru protein which is under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

35. The kit of claim 34, in which said first gene does not comprise an intron.

36. The kit of claim 32, in which said male-sterile parent plants do not contain a gene that is functionally expressed in the aleurone of corn seeds which gene encodes an active regulatory protein having the same activity as a functional C1 gene, and wherein said foreign DNA in said male-fertile parent plants comprises said second gene.

37. The kit of claim 36, which said second gene comprises a DNA encoding the C1 protein which is under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

38. The kit of claim 32, in which said male-sterile parent plants do not contain a gene that is functionally expressed in the aleurone of corn seeds which gene encodes an active regulatory protein having the same activity as the protein encoded by a functional R or B gene, and do not contain a gene that is functionally expressed in the aleuron of corn seeds which gene encodes an active regulatory protein having the same activity as the protein encoded by a functional C1 gene, and wherein said foreign DNA in said male-fertile parent plants contains said first gene and said second gene.

39. The kit of claim 38, in which said first gene comprises a DNA encoding the B-peru protein under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 or a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG; and in which said second gene comprises a DNA encoding the C1 protein under the control of a promoter of the B-peru gene, a promoter comprising the nucleotide sequence of SEQ ID No. 6 between positions 1 and 188, a 35S promoter, a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 and a promoter comprising the sequence of SEQ ID No. 1 between positions 447 and 1061 in which TGCAG between positions 935 and 939 is changed to TTAGG.

40. The kit of claim 31, in which said first genetic locus in said male-sterile plants and said male-fertile parent plants is a foreign genetic locus which comprises a male-sterility gene comprising a DNA encoding barnase under control of a first promoter which directs expression selectively in specific stamen cells of corn plants, and wherein said male-sterility gene is homozygous at said first genetic locus; and in which said restorer gene at said second genetic locus in said male-fertile parent plants comprises a DNA encoding barstar under control of a second promoter which directs expression at least in said specific stamen cells.

41. The kit of claims 40, in which said first promoter is a TA29 promoter or a CA55 promoter.

42. The kit of claim 40, in which said second promoter is identical to said first promoter.

43. The kit of the claim 40, in which said second promoter is a TA29 promoter or a CA55 promoter.

44. The kit of claim 41, in which said second promoter is identical to said first promoter.

45. The kit of claims 31, in which said first genetic locus is an endogenous male-sterility locus comprising a recessive allele in homozygous condition, and in which said restorer gene is the dominant allele at said endogenous male-sterility locus.

Details for Patent 5,880,331

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Schering INTRON A interferon alfa-2b VIAL 103132 001 1986-06-04   Start Trial Plant Genetic Systems, N.V. (Gent, BE) 2016-03-09 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 002 1986-06-04   Start Trial Plant Genetic Systems, N.V. (Gent, BE) 2016-03-09 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 003 1986-06-04   Start Trial Plant Genetic Systems, N.V. (Gent, BE) 2016-03-09 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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