CA2392722A1

CA2392722A1 - Male tissue-preferred regulatory region and method of using same

Info

Publication number: CA2392722A1
Application number: CA002392722A
Authority: CA
Inventors: Marc C. Albertsen; Timothy W. Fox; Carl W. Garnaat; Gary Huffman; Timmy L. Kendall
Original assignee: Individual
Current assignee: Pioneer Hi Bred International Inc
Priority date: 2000-02-15
Filing date: 2001-02-13
Publication date: 2001-08-23
Anticipated expiration: 2021-02-13
Also published as: AU3817701A; HUP0301053A3; HUP0301053A2; NZ520704A; WO2001060997A3; EP1255825A2; CA2392722C; WO2001060997A2

Abstract

The present invention relates to an isolated nucleic acid sequence encoding the Ms45 male tissue-preferred regulatory region. In one aspect this invention relates to the use of this male tissue-preferred regulatory region in mediating fertility. An example of such use is the production of hybrid seed such as in a male sterility system. The Ms45 male tissue-preferred regulatory region can be operably linked with exogenous genes, such as those encoding cytotoxins, complementary nucleotidic units and inhibitory molecules. This invention also relates to plant cells, plant tissue and differentiated plants which contain the regulatory region in this invention.

Claims

1. A male tissue-preferred regulatory region comprising nucleotide sequences essential for initiating transcription of the MS45 gene.

2. The regulatory region of claim 1 comprising nucleotide sequences of about contiguous base pairs from about -38 and higher upstream of the TATA box of the MS45 gene.

3. The regulatory region of claim 1 comprising sequences of about 100 base pairs which hybridize to a promoter region of the MS45 gene which is -38 or higher upstream of the TATA box.

4. An isolated nucleic acid that is a male tissue specific regulatory element comprising a fragment of the nucleotide sequence of SEQ ID NO. 1 or SEQ ID NO. 2 or those nucleotide sequences which hybridize to either of SEQ ID NO. 1 or NO.2 under conditions of high stringency wherein the regulatory element is essential for initiating transcription of SEQ ID NO. 1 or 2.

5. The isolated DNA molecule of claim 4 wherein the fragment is SEQ ID No. 3, 4, 5 or 6 or those nucleotide sequences which hybridize to any SEQ ID Nos. 3, 4, 5 or 6 under conditions of high stringency.

6. A regulatory region comprising the sequences of -38 to -195 upstream of the TATA
box of SEQ ID NO. 1 or 2 or those nucleotide sequences which hybridize to said sequences under conditions of high stringency

7. The regulatory region of claim 6 comprising sequences -152 to -181 upstream of the TATA box of SEQ ID NO. 1 or 2 or those nucleotide sequences which hybridize to said sequences under conditions of high stringency.

8. The regulatory region of claim 6 comprising sequences -72 to -111 upstream of the TATA box of SEQ ID No. 1 or 2 or those nucleotide sequences which hybridize to said sequences under conditions of high stringency

9. An expression vector comprising a promoter that is operably linked with the male tissue specific regulatory element of claim 1.

10. The expression vector of claim 9 further comprising a exogenous gene, wherein the exogenous gene is operably linked to the promoter.

11. The expression vector of claim 9 wherein the promoter is selected from any one of CaMV35S, SGB6, MS45 or 5126.

12. The expression vector of claim 10 wherein the product of the exogenous gene disrupts the function of male tissue.

13. A chimeric male tissue specific regulatory cassette comprising the nucleotide sequence of claim 1.

14. The isolated DNA of claim 1 wherein the DNA fragment comprises the nucleotide sequences selected from the group consisting essentially of AGGATACCTA
CTCCCAAACA ATCCATCTTA CTCATGCAAC.

15. Plant cells comprising the vector of claim 9.

16. A method of mediating male fertility in a plant comprising introducing into a plant the expression vector of claim 10 wherein the exogenous gene impacts male fertility of the plant and the regulatory element in conjunction with the promoter control expression of the exogenous gene.

17. The method of claim 16 wherein the exogenous gene disrupts function of male tissue of the plant causing the plant to be male sterile.

18. The method of claim 16 wherein the regulatory element in conjunction with the promoter is inducible.

19. The method of claim 18 wherein the plant is constitutively sterile when the promoter and regulatory element are not induced and is fertile when the promoter and regulatory element are induced.

20. The method of claim 17 further comprising cross-fertilizing the male sterile plant with a second plant, the second plant comprising a second exogenous gene, the product of the second gene preventing disruption of the male tissue by the first heterologous gene, producing a male fertile hybrid plant.

21. A method of producing hybrid seeds comprising: (a) producing a first parent plant comprising nucleotide sequences essential for initiating transcription of the MS45 gene operably linked with an exogenous gene impacting male fertility of the plant such that the plant is male sterile; (b) producing a second parent plant which is male fertile;(c) cross-fertilizing the first parent plant and the second parent plant to produce hybrid seeds.

22. The method of claim 21 wherein the gene impacting male fertility is dominant and further comprising growing the hybrid seed to produce a third male sterile parent plant;
producing a fourth parent plant comprising one or more genes controlling a desired gene trait and cross-fertilizing the third and fourth parent plants to produce second hybrid seed.