2000 Fiscal Year Final Research Report Summary
Development of DNA markers for selection in soybean breeding
| Project/Area Number |
09356001
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| Research Category |
Grant-in-Aid for Scientific Research (A).
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Breeding science
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| Research Institution | Chiba University |
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Principal Investigator |
HARADA Kyuya Faculty of Horticulture, Chiba University, Professor, 園芸学部, 教授 (70011913)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAMATSU Mitsuo Nagano Chushin Agricultural Experiment Station, Researcher, 畑作育種部, 研究員
ISHIKAWA Keiko Graduate School of Science and Technology, Chiba University, Assistant professor, 大学院・自然科学研究科, 助手 (20212839)
KOBA Takato Faculty of Horticulture, Chiba University, Associate professor, 園芸学部, 助教授 (40170302)
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| Project Period (FY) |
1997 – 2000
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| Keywords | soybean / marker-assisted selection / DNA marker / RFLP / microsatellite / cDNA / QTL / recombinant inbred lines |
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| Research Abstract |
1. About 500 random cDNA clones from a library derived from fully-expanded green leaves of Norin No.2, were used as probes and RFLPs of six pairs of 11 soybean varieties and one wild soybean line, were analyzed using eight 6-base recognition enzymes. The polymorphism frequencies extended from 34.4% to 69.0% and were found to be much higher than those reported before.
2. Using a variety Norin No.2, sheared genomic fragments with 300-600bp were constructed by pipetting and DNase I treatment followed by size separation by agarose gel electrophoresis. Blunting and reparing of fragment ends were performed by T4 DNA polymerase and T4 polynucleotide kinase. Following ligation of EcoRI adapters to the resulting fragments, they were inserted into a Lamda ZAP II vector to construct genomic libraries. From the libraries, positive clones were screened using(CT)10, (GAA)6, (GTG)6 as probes. Nucleotide sequences of positive clones for CT and GAA core repeats were determined and primer pairs were designed. As a result about 70 SSR markers were developed.
3. A soybean genetic linkage map was constructed using 190 F2 plants derived from a cross between the soybean varieties, Misuzudaizu and Moshidou Gong 503, based on known markers and RFLP markers developed from cDNA clones and new SSR markers. This linkage map has 503 markers and 21 linkage groups covering 2,908.7cM of the soybean genome.
4. Using this linkage map, four QTLs for flowering time were identified and almost all phenotypic variations of flowering time were explained by detected QTLs, indicating that this map coves a large region of the soybean genome.
5. From the same parents, 156 recombinant inbred lines(F8)were developed. A linkage map constructed from these lines has 232 markers and 31 linkage groups and consists with a map made from the F2 population.
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