| Project/Area Number |
17580053
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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 |
Plant nutrition/Soil science
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| Research Institution | Kagoshima University |
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Principal Investigator |
SAKAI Masao Kagoshima University, Faculty of Agriculture, Associate Prof., 農学部, 助教授 (20225775)
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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 |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2006: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2005: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Soil mice oorganisms / PNA / rhizosphere / 植物根菌 |
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| Research Abstract |
Contamination with plastid small-subunit (SSU) rDNA is a major drawback when analyzing the microflora of plant rhizosphere using culture-independent methods. In this study, PNA oligomers, Mit1492 and Pla1492, were designed and tested to specifically amplify the SSU rDNA of various bacterial species without amplifying the SSU rDNA of plant mitochondria and plastids. To confirm how useful the analysis of rhizosphere microflora is using Mit1492 and Pla1492, the terminal restriction fragment length polymorphism (T-RFLP) method was performed with cucumber and spinach root samples. Using the standard T-RFLP method, a large T-RF peak of plant mitochondria and plastids SSU rDNA interfered with the rhizosphere microflora analysis. In contrast the T-RFLP method using the PNA-mediated PCR clamping was able to detect the bacterial DNA while directly eliminating the influence of the plant-derived DNA extracted from the plant roots. PNA-mediated PCR clamping might be a useful technique for the culture-independent analysis of rhizosphere microflora in plant roots using SSU rDNA.
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