| Project/Area Number |
24651234
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied genomics
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| Research Institution | Okayama University |
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Principal Investigator |
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
|
| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
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| Keywords | イチゴ / 黒斑病 / DNAマーカー / 次世代シーケンス / 病害罹病性 / 病害抵抗性 / 国際情報交換 |
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| Research Abstract |
To develop DNA maker for isolation of disease specificity gene, we have combined AFLP (amplified fragment length polymorphism) bulk tagging method, NGS (next generation sequencing) and sequence clustering by CD-HIT-EST. Both bulks of genomic DNA derived from disease susceptible and resistant genotype were prepared according to AFLP method. Determined sequences were clustered into 89,531 by CD-HIT-EST. Total of 3,679 sequence clusters was filtered by read count bias (a ratio of read count derived from susceptible bulk / resistant bulk). To check availability of these sequence clusters as linked makers, PCR primers were designed through mapping of sequences on Fragaria vesca genome. Currently, of tested 231 primer combinations, 126 primer sets showed polymorphic amplification. These results reveal that along with our methods was proven to be useful for development of DNA makers; we have succeeded to develop at least 126 linked makers to disease specificity locus (loci).
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