Development of a novel diagnosis method for nematode diseases based on soil metagonome
| 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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Report
(4 results)
Research Products
(34 results)
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[Journal Article] Suppressive effect of anaerobically digested slurry on the root lesi on nematode Pratylenchus penetrans and its potential mechanisms2007
Author(s)
Min, Y. Y., Sato, E., Shirakashi, T., Wada, S., Toyota, K., Watanabe, A.
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Journal Title
Japanese Journal of Nematology 37(2)
Pages: 93-100
Description
「研究成果報告書概要(和文)」より
Related Report
Peer Reviewed
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[Journal Article] Detection of the root-lesion nematode, Pratylenchus penetrans, from a nematode community using real-time PCR2007
Author(s)
Sato, E., Min, Y. Y., Shirakas hi, T., Wada, S., Toyota, K.
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Journal Title
Japanese Journal of Nematology 37(2)
Pages: 87-92
Description
「研究成果報告書概要(和文)」より
Related Report
Peer Reviewed
-
[Journal Article] Suppressive effect of anaerobioally digested slurry on the root lesion nematode Pratylenchus penetrans and its potential mechanisms2007
Author(s)
Min, Y. Y, Sato, E., Shirakashi, T., Wada, S., Toyota, K., Watanabe, A.
-
Journal Title
Japanese Journal of Nematology 37(2)
Pages: 93-100
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Journal Article] Detection of the root-lesion nematode, Pratylenchus penetrans, from a nematode community using real-time PCR2007
Author(s)
Sato, E., Min, Y. Y, Shirakashi, T., Wada, S., Toyota, K.
-
Journal Title
Japanese Journal of Nematology 37(2)
Pages: 87-92
Description
「研究成果報告書概要(欧文)」より
Related Report
-
[Journal Article] Detection of the root-lesion nematode, Pratylenchus penetrans, from a nematode community using real-time PCR2007
Author(s)
Sato, E., Min, Y. Y., Shirakashi, T., Wada, S., Toyota, K.
-
Journal Title
Japanese Journal of Nematology 37(2)
Pages: 87-92
Related Report
Peer Reviewed
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