1991 Fiscal Year Final Research Report Summary
Cloning DNA markers, mapping and transformation in the tribe Triticeae crops
| Project/Area Number |
01480038
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Breeding science
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| Research Institution | Tottori University |
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Principal Investigator |
YASUMURA Yoshimasa Tottori U., Fac. of Agric., Prof., 農学部, 教授 (50026374)
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| Co-Investigator(Kenkyū-buntansha) |
TOMITA Motonori Tottori U., Fac. of Agric., Res. Assoc., 農学部, 助手 (70207611)
NAKATA Noboru Tottori U., Fac. of Agric., Assoc. Prof., 農学部, 助教授 (40032312)
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| Project Period (FY) |
1989 – 1991
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| Keywords | Triticale / Rye genome / Genome-specific repeated sequences / RFLP / DNA marker |
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| Research Abstract |
Rye genome- and D genome-specific repeated sequences were cloned for the use of chromosome identification in newly reconstructed triticale genomes.
The strategy for the cloning method of rye genome- or rye chromosome specific repeated DNA sequences was based on the selective reannealing of rye specific sequences by using rye chromosome addition wheat lines as follows : Rye chromosome addition wheat genomic DNA was digested with Ybo I and mixed with five times of sheared wheat genomic DNA, and then, the DNA mixture was denatured and annealed in phenol emulsion. So that only reannealed cohesive fragments enriched in rye specific sequences were legated into pUCi9 Bav HI sites. Recombinants of 145 and 77 clones were obtained from 5R and 6R chromosome addition wheat lines, respectively. Reassociation rates of Afbo I fragments were estimated to be 0.1-1.2%. Fourteen highly repetitive rye-specific clones were selected by differential dot blot hybridization of total rye and wheat DNA to Ybo I library. These clones were classified into six different rye genome-specific repeated sequences by southern hybridization. Clone pTS5R-l from 5R could probe a tandemly repeated 380bp-sequence family which was defined by Eco 0109I and consisted of various numbers of polymers caused by the mutation at the ends of Eco 0109I palindrome sites. RFLPs among inbred rye lines (S. cerea-l&) were detected with three rye specific clones.
A tandemly repeated 2010bp-sequence family was cloned by Kpn I from Ae. Sq, va. rrosa. AB genomic DNA fragments was subtracted from Ybo I-digested common wheat genomic DNA by hybridizing sheared Tdzirus genomic DNA, and the rests were cloned to pUC19 Rain HI sites. Among 253 recombinants seven D genome-specific repetitive sequences were obtained.
Above genome-specific repeated clones are useful DNA markers which identify chromosome fragments of R and D genomes.
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