马建伟,王曦茁,汪来发,王源,苟大平,王艳菊.淡紫拟青霉T-DNA插入突变体的表型分析[J].林业科学研究,2014,27(2):219-224
淡紫拟青霉T-DNA插入突变体的表型分析
Phenotypes Analysis for T-DNA Insertional Mutants of Paecilomyces lilacinus
投稿时间:2013-12-27  
DOI:
中文关键词:  淡紫拟青霉  插入突变  生物学性状  致病力
英文关键词:Paecilomyces lilacinus  insertional mutagenesis  biological characteristics  pathogenicity
基金项目:国家自然科学基金项目(No. 31100477)
作者单位E-mail
马建伟 中国林业科学研究院森林生态环境与保护研究所, 国家林业局森林保护学重点实验室 北京 100091  
王曦茁 中国林业科学研究院森林生态环境与保护研究所, 国家林业局森林保护学重点实验室 北京 100091 wangxz@caf.ac.cn 
汪来发 中国林业科学研究院森林生态环境与保护研究所, 国家林业局森林保护学重点实验室 北京 100091  
王源 中国林业科学研究院森林生态环境与保护研究所, 国家林业局森林保护学重点实验室 北京 100091  
苟大平 中国林业科学研究院森林生态环境与保护研究所, 国家林业局森林保护学重点实验室 北京 100091  
王艳菊 中国林业科学研究院森林生态环境与保护研究所, 国家林业局森林保护学重点实验室 北京 100091  
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中文摘要:
      研究淡紫拟青霉T-DNA插入突变体,鉴定突变体的表型特征,为淡紫拟青霉致病机制分析及致病相关的基因克隆奠定基础。PCR检测筛选的14株T-DNA插入突变体,观察和测定菌落形态,菌落生长速率,产孢量,孢子萌发率和菌丝干质量等生物学性状,并比较突变体致病力。结果表明突变体均含有beta-tubulin基因序列,T-DNA已整合进入野生型菌株20-7的基因组中。通过与野生型菌株对比,发现突变体菌落形态发生了不同程度的改变。菌落生长速率明显增大的菌株占全部菌株的85.71%,只有BN-11菌株的菌落生长速率降低。产孢量显著变大的菌株有BN-11和BN-12,为菌株总数的14.29%。孢子萌发率发生改变的菌株占78.57%。菌丝干质量发生明显增大和减少的菌株都占全部突变体的21.43%。筛选获得了1株致病力增强的突变体BN-11和11株致病力显著降低的突变体。
英文摘要:
      The T-DNA insertional mutants of Paecilomyces lilacinus were studied and the phenotypes were identified. It may contribute to understanding the pathogenic mechanism of P. lilacinus and the clone genes related to pathogenicity. All of 14 mutants generated by T-DNA insertion were tested with PCR for their insertional sequence. The morphology, colony growing rate, spore production, conidia germination rate, dry weight of mycelium and pathogenicity against Meloidogyne incongnita of phenotypes of mutants were analyzed by comparing with wild-type strain 20-7. The results demonstrated that the genomes of these mutants contained the sequence of beta-tubulin gene and T-DNA had been integrated into wild-type genome of P. lilacinus. Compared with wild-type strain 20-7, the morphologies of these colonies changed at different degrees. The colony growing rate of 85.71% strains speeded up obviously, only the strain BN-11 slowed down its colony growing rate distinctly. The strain BN-11 and BN-12, accounting for 14.29% of the total, increased the spore production. 78.57% mutants changed their conidia germination rate significantly. Both the rates of mutants that dry weight of mycelium reduced and increased were 21.43%. One mutant with increased pathogenicity was obtained, and the pathogenicity of 11 mutants decreased enormously.
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