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不同基质和培养时间对球孢白僵菌孢子耐逆性研究

Study on stress tolerance of Beauveria bassiana conidia in different substrates and culture times

  • 摘要: 球孢白僵菌孢子是应用最广泛的真菌杀虫剂关键因子,为了解不同培养基和培养时间对球孢白僵菌孢子抗逆能力的影响,从采集自全国不同温区30株球孢白僵菌菌株中筛选出5株生长快、产孢量大的菌株——RCEF5609、RCEF5746、RCEF5688、RCEF6200和RCEF6857。采用营养丰富度不同的常用培养基(PDA、SDAY和MEA)和不同培养时间两个方面研究菌株耐热和抗紫外性,并分析相关基因转录水平上的变化。结果表明,菌株RCEF6200和RCEF5688分生孢子发芽率最高,抗逆性最强,RCEF5609孢子萌发率最低,抗逆性最弱;营养贫瘠的PDA较营养丰富的SDAY和MEA产生的孢子更有利于耐热和抗紫外,培养7 d的分生孢子抗逆性更强。进一步分析抗逆性最强的菌株RCEF6200和抗逆性最弱的菌株RCEF5609抗逆基因转录水平,结果显示,菌株RCEF6200和RCEF5609逆境胁迫后与对照组相比,在PDA培养基上和7 d的培养条件下,较其他处理中耐热性和抗紫外线相关基因表达水平呈不同程度增加,可见抗逆相关基因转录分析结果进一步验证上述结果。研究为生产抗逆性强的球孢白僵菌菌剂提供一定理论基础。

     

    Abstract: The conidia of Beauveria bassiana is the key factor of the fungal insecticides used most widely. To know the effects of different media and culture times on the stress resistance of B. bassiana conidia. Five strains with fast growth and high spore production from 30 strains collected from different temperature zones in China(RCEF5609, RCEF5746, RCEF5688, RCEF6200 and RCEF6857) were selected. The heat resistance and UV resistance of strains were studied by using commonly used culture media with different nutritional richness degrees in the laboratory(PDA, SDAY and MEA) and different cultivation times, and the changes in transcription levels of related genes were analyzed. The results showed that the germination rate of conidia of strains RCEF6200 and RCEF5688 was the highest and with the strongest stress resistance, while RCEF5609 had the lowest germination rate and the weakest stress resistance. It was shown that the conidia produced by poor nutrient culture medium PDA were stronger tolerance for heat resistance and UV resistance than those produced by rich nutrient SDAY and MEA, and the conidia cultured for 7 d had stronger stress resistance. Further analysis of the transcription levels of stress resistant genes in the strongest stress resistant strain RCEF6200 and the weakest stress resistant strain RCEF5609 showed that compared with the control group, the expression levels of heat resistance and UV resistance related genes in the strains RCEF6200 and RCEF5609 increased to varying degrees on PDA medium and under 7 d of cultivation conditions compared to other treatments.The transcriptional analysis results of stress resistance related genes further confirmed the above results. A theoretical basis for the production of strong stress resistant fungicide of B. bassiana was provided.

     

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