摘要
在利用球孢白僵菌Beauveria bassiana有性态改良球孢白僵菌菌株的研究中,菌株的选择非常重要,其中利用交配型基因选择菌株是必要手段。而且虫生真菌的有性态能够使菌株间产生基因交流,促进其进化的过程。为更好地研究球孢白僵菌的遗传规律,以获得避免菌种衰退的方法,本研究建立了分别以柞蚕蛹为活体寄主和糙米培养基为基质的2种人工培养有性世代繁殖体系。结果表明,交配型为MAT1-1-1型和MAT1-2-1型混合型菌株BbDPSD5,经过自交在2种基质上均能够产生有性态。而具有2种单一交配型的菌株BbOFDH1-5和BbDPSD2进行配伍试验,BbOFDH1-5×BbDPSD2、BbOFDH1-5×BbDPSD5和BbDPSD2×BbDPSD5在2种基质上均没有有性态产生。对获得的有性态子囊孢子经无性繁殖产生的菌株进行r DNA-ITS片段以及交配型基因检测,并对测序结果进行比对分析。结果显示,扩增得到的r DNA-ITS序列和MAT1-2-1交配型基因与亲本序列同源性为100%,而没有检测到MAT1-1-1型。2种培养方式获得的结果完全一致。对有性态子代子囊孢子产生的株系进行毒力测试结果表明,所有子代菌株与原始菌株BbDPSD5的毒力差异不显著,子代交配型基因虽然缺失了MAT1-1-1,但是毒力没有衰退,未出现菌种退化现象。
Strains selected by improved and typical mating practice are very important to enhance sexual reproduction of Beauveria bassiana.Gene flow and evolution can occur during the stage of teleomorph.To study the inheritance of B.bassiana and solve the recession-related problem,we developed two novel methods for cultivating sexual generation,Cordyceps bassiana,using two kinds of media,pupae of Antheraea pernyi and brown rice.The results showed that BbDPSD5 inbred strain of B.bassiana which possessed two mating type genes,MAT1-1-1 and MAT1-2-1,could develop into C.bassiana through these two media.No fruit bodies,however,were developed in other combinations,i.e.BbOFDH1-5×BbDPSD2,BbOFDH1-5×BbDPSD5,and BbDPSD2×BbDPSD5,on the two media.Mating type genes and partial segments of rDNA-ITS gene were amplified and sequenced from the cloning strains of ascospores.The results indicated that the r DNA-ITS gene and MAT1-2-1 gene from strains of ascospores had 100% homologous with their parental type,and no MAT1-1-1 gene was detected.The results were consistent between the two media treatments.There was no significant difference on mortality between the ascospore strains and Bb DPSD5 strain in toxicity tests.The results also indicated that there was no decrease in toxicity and no recession on parental strain.
引文
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