| Project/Area Number |
17380048
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | Tokyo University of Agriculture and Technology |
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Principal Investigator |
TOYODA Koki Tokyo University of Agriculture and Technology, Institute of Symbiotic and Technology, Assistant Professor (30262893)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥14,130,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥930,000)
Fiscal Year 2007: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2006: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥6,600,000 (Direct Cost: ¥6,600,000)
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| Keywords | Pratylenchus penetrans / Meloidogyne incognita / Globodera rostochiensis / Heterodera glycines / denaturing gradient gel electrophoresis / Baermann / soil compaction / nematode diagnosis / キタネグサレセンチュウ / 自活性線虫 / PCR-DGGE / リアルタイムPCR / 定量PCR / 群集構造 / 微生物 / ダイコン |
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| Research Abstract |
Specific primers in real-time PCR were developed for the quantitative detection of the root-lesion nematode Pratylenchus penetrans the root-knot nematode Meloidogyne incognita, the potato-cystnematode Globodera rostochiensis and the soybean-cyst nematode Heterodera glycines. Then, this detection method using real-time PCR was combined with adirect DNA extraction from soil. We found that their detection was significantly enhanced by extracting DNA after heavily compaction (1.4g cm^<-3> in case of volcanic ash soils). In general method, it takes more than 3days to count the number of the plant-parasitic nematodes. However, our method using a combination of soil compaction and real-time PCR enabled their detection within 4hours.
We established a PCR-DGGE method to evaluate community structure of nematodes extracted from soil using the conventional method and found that nematode community structure, predominantly consisted of free-living nematodes, might be involved in estimating damages by the root-lesion nematode.
In 4years field survey, relationship between damage by the root-lesion nematode and initial density of the nematode was studied. No damage was observed in the radishes in fields containing P. penetrans at a density of less than 2.5 individuals 20g^1 soil. In contrast, severe damage was always observed in fields at a density of more than 30 individuals 20g^1 soil. In fields containing P. penetrans at a density of 3.4 to 12 individuals 20g^1 soil, damage by the nematode was quite variable, but could be estimated by the difference in the nematode community structure.
This study demonstrated that the soil compaction and real-time PCR assay for the four kinds of plant-parasitic nematodes provides a sensitive and reliable means for the rapid quantification of these the major pests in Japan.
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