| Project/Area Number |
17580134
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
林学・森林工学
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| Research Institution | Forest Tree Breeding Center |
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Principal Investigator |
WATANABE Atsushi Forest Tree Breeding Center, Breeding department, Researcher (10360471)
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| Co-Investigator(Kenkyū-buntansha) |
ISODA Keiya Forest Tree Breeding Center, Breeding department, Researcher (60391702)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | SNP marker / Isolation of gene / gene associated with defence / pine wood nematode / pathogen related protein / EST / SNP / アカマツ / クロマツ / 連鎖地図 / サブトラクションライブラリー |
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| Research Abstract |
We have established how to develop SNP marker based on ESTs derived from Pinus taeda in previous year. A major goal of our project is to detect any DNA makers or genes linked to resistance against pine wood nematode using the consensus map. However any genes associated with defense response of Japanese pine trees have been not clear. In present work, we attempted to isolate genes derived from Japanese black pine infected with pine wood nematode. To compare differentially expression between resistant and susceptible trees in early defense response, two subtractive libraries were constructed using suppression subtractive hybridization method (SSH). Some genes isolated were converted to SNP marker to locate on the consensus map.
Approximately 500 ESTs were obtained from a SSH library, and the EST with moderate to high similarity scores based on Blast X were 272. Among genes identified, 70 were Thaumatin-like protein and this gene which is related to the defense response of plants to pathogen attack was the most abundant representing 15.8% of ESTs. A cDNA library from RNA expressed only in the susceptive clone infected was constructed to estimate accuracy of the subtractive library and only four of 81 EST were associated with Thuamtin-like protein. Two genes, Antimicrobial peptide and Metallothionein-like protein, which were related to plant defense and stress, were frequent. In next step, expression of these genes must be analyzed further using Northern blot or real time PCR. When a reverse subtraction, R-RI was constructed, metallothionein-like protein was frequent. We have developed 20 SNPs markers.
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