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Studies on Relationships between the Diversity of Bacterial Community, Ketone Synthase Genes (KS) in Different Cucumber Rhizosphere Soil and Fusarium Wilt

Author: HuYingChang
Tutor: LiuZhiHeng；LiuQiu
School: Shenyang Agricultural University
Course: Plant Pathology
Keywords: Fusarium wilt of cucumber Rhizosphere soil Actinomycetes species Bacterial populations Keto synthase Diversity
CLC: S436.421
Type: Master's thesis
Year: 2010
Downloads: 64
Quote: 0
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Abstract


In this paper, cucumber rhizosphere soil bacterial populations and keto synthase (KS) gene diversity and wilt incidence relations were studied. Keto test isolated and cultured using traditional methods, PCR-DGGE technology and molecular cloning techniques, for the cucumber rhizosphere microbial population of the soil samples and encoding polyketide synthase (KS) 'genetic diversity of both Fusarium disease. Analysis of Test of microbial populations in the incidence varying degrees of cucumber rhizosphere soil samples and genetic diversity of KS. PCR-DGGE technology to analyze the diversity of microbial populations in the extent of the disease of cucumber rhizosphere soil samples, results showed that the diseased cucumber rhizosphere soil bacterial population was significantly higher than the incidence cucumber rhizosphere soil, which in the past The reported incidence cucumber rhizosphere soil microbial populations richer conclusion is not consistent. Cloning and sequence of the target gene fragment was used to analyze the KS gene the incidence varying degrees of cucumber rhizosphere soil diversity, the results show that the diseased cucumber rhizosphere soil richer KS gene. This result is consistent with the results of the analysis of the soil bacterial diversity of the population. Test design their own different soil samples of biological genomic DNA extraction method - alkaline lysis method, and were compared with the other three DNA extraction methods. The results showed that the alkaline lysis method using potassium acetate and magnesium chloride to remove impurities such as humic acid, can obtain high-quality, large fragment, the high integrity of the DNA, for the latter experiments thus provide a good 'DNA template for PCR amplification and subsequent the molecular operation is more suitable. Test proved that the alkaline lysis method the independent innovation DNA extraction method, which has been applied for a patent (Application No.: 2010101032923). Test using the conventional separation methods, actinomycetes were isolated and cultured from the incidence varying degrees cucumber rhizosphere soil samples. The difference in the number of different types of soil samples isolated antagonistic actinomycetes bacteria were compared, results showed that the type of antagonistic actinomycetes in the rhizosphere soil infected with Fusarium wilt soil more than uninfected. Test using PCR-DGGE method, special primers with \Before planting, inoculation, morbidity, incidence of changes in bacterial populations in cucumber rhizosphere soil samples, the results show that the diseased cucumber rhizosphere soil bacterial populations richer. Type I polyketide synthase gene degenerate primers, extracted diseased and unaffected rhizosphere soil total DNA for PCR amplification and were cloned and sequenced, a total of 205 KS gene, all submitted to GenBank and granted access No. for similar studies. Statistical analysis showed that the number of gene sequencing KS, from diseased soil samples obtained more positive clones, showing a more extensive KS genetic diversity, the results of PCR-DGGE diverse bacterial populations in the soil analytical results of the same trend.

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CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Horticultural Crops Pest and Disease Control > Vegetable pests > Melons, pests and diseases > Cucumber pests and diseases
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