|Table of Contents|

Development and Application of a Real-Time Quantitative PCR Assay for Detection of Fusarium spp. in Soil(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2010年04期
Page:
655-658,662
Research Field:
Publishing date:

Info

Title:
Development and Application of a Real-Time Quantitative PCR Assay for Detection of Fusarium spp. in Soil
Author(s):
WEI Wei1; 2; 3;  XU Yan-li1;  LIU Jin-bo1; 3;  LI Chun-jie1;  HAN Xiao-zeng1;  LI Wen-bin2; LI Shu-xian4
1.Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences (CAS), Harbin 150081, Heilongjiang;
2.Key Laboratory of Soybean Biology of Ministry of Education, Harbin 150030, Heilongjiang;
3.Graduate School, CAS, Beijing 100049，China;
4.Crop Genetics Research Unit (CGRU), United States Department of Agriculture-Agricultural Research Service (USDA-ARS), Stoneville 38776, USA
Keywords:
Fusarium spp.; Real-Time QPCR; Absolute quantitation
PACS:
S154.3
DOI:
10.11861/j.issn.1000-9841.2010.04.0655
Abstract:
Base on the internal transcribed spacers (ITS) of Fusarium, a pair of specificity primers, TS and TR, were designed and synthesized for Real-time quantitative PCR (Real-Time QPCR) system of Fusarium spp. and then its optimal reaction system was established. Using this reaction system, the masses of genomic DNA of Fusarium spp. in soybean filed，which were extracted from three fertilization management, including no fertilizer (NF), chemical N and P (NP) and chemical N, P and manure (NPM), were quantified absolutely. TS and TR have possessed a good specificity to Fusarium spp.. In the amplification curve of this Real-Time QPCR reaction, the spacings of Cycle Threshold (Ct) of standard substance under every concentration gradient were uniform. The melting curve showed a simple peak. The correlation coefficient (R2) and slope of the equation from the standard curve were respective 0.993 and -0.2927. At soybean seedling stage, the mass of genomic DNA of Fusarium spp. per gram of soil under NF, NP and NPM fertilization management were 18.33, 44.61 and 140.83 pg， respectively. The mass in NPM measure was higher than NF and NP measure significantly(P＜0.01).

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