中国生物防治学报2011，Vol.27Issue(2)：233-240,8.
用于土壤中生防芽孢杆菌B006和BH1检测的SCAR标记及其特异性
Identification of the SCAR Markers for Detection of Biocontrol Bacillus Strains B006 and BH1 in Soil
摘要
Abstract
Two Bacillus strains, B006 and BH1, are highly suppressive against Fusarium spp., the causal agents of plant root diseases. Discrimination of these strains from native soil bacteria was critical to study their population dynamics and behaviors in soil. Polymorphism in genomic DNA of the strains B006 and BH1 as well as 25 references including Bacillus spp., Sphingosine spp. And Pseudomonas spp. Were analyzed with 16 random primer pairs by RAPD. Sequence-specific fragments with 730 bp and 658 bp could be obtained from the strains B006 and BH1 by PCR with single specific primers B19 and C01, respectively, which were absent in other strains, hi order to increase the amplification specificity and stability, 6 and 12 SCAR marker primer pairs were designed on the base of the nucleotide sequence information of RAPDs closely linked to the strains B006 and BH1, respectively, and their specificity were confirmed by amplification of specific DNA bands of the strains B006 and BH1. Two primer sets SCAR-B006 and SCAR-BH1 were selected and the amplification products with 523 bp and 658 bp were transformed into more-specific SCAR markers, respectively. Specificity of these markers was further tested in natural soil. Only the soil amended with B006 or BH1 gave specific SCAR bands. This PCR-based method is useful to track the strains B006 and BH1 in soil samples rapidly and efficiently.关键词
SCAR分子标记/RAPD分子标记/芽孢杆菌/生物防治Key words
SCAR/ RAPD/ Bacillus spp./ biological control分类
农业科技引用本文复制引用
耿文义,郭荣君,李世东,许修宏..用于土壤中生防芽孢杆菌B006和BH1检测的SCAR标记及其特异性[J].中国生物防治学报,2011,27(2):233-240,8.基金项目
国家大宗蔬菜产业技术体系建设项目(nycytx-35) （nycytx-35）
中央级公益性科研院所基本科研业务费专项资金资助项目(2010YW04) （2010YW04）
