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谷祖敏,戈婉儿,纪明山.2株蜡蚧轮枝菌耐热性差异分析[J].华中农业大学学报,2018,37(5):40-44
2株蜡蚧轮枝菌耐热性差异分析
Difference analysis of thermotolerance between two Lecanicillium lecanii strains
投稿时间:2018-04-20  
DOI:
中文关键词:  蜡蚧轮枝菌  高温胁迫  热激蛋白
英文关键词:Lecanicillium lecanii  high temperature stress  heat shock protein
基金项目:辽宁省自然科学基金项目(2014027008)
作者单位E-mail
谷祖敏 沈阳农业大学植物保护学院沈阳 110866 guzumin1212@163.com 
戈婉儿 沈阳农业大学植物保护学院沈阳 110866  
纪明山 沈阳农业大学植物保护学院沈阳 110866  
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中文摘要:
      采用菌丝生长速率法和孢子萌发法比较蜡蚧轮枝菌不同菌株VL17和VL18菌株菌丝和分生孢子耐热性的差异,喷雾接菌法测定两菌株高温胁迫下的杀蚜毒力,qPCR法研究高温胁迫后热激蛋白sHsp和Hsp70基因的表达量,初步探索热激蛋白抗高温胁迫的作用机制。结果显示,VL18菌株菌丝和分生孢子对高温的耐受性显著好于VL17菌株,34℃时VL18菌株的菌落直径比VL17大63.16%,37℃时VL18分生孢子萌发率比VL17高103%;温度高于31℃时两者的杀蚜活性差异显著,37℃时VL18对蚜虫的校正死亡率比VL17高166.67%;42℃时胁迫相同时间,VL18的sHsp和Hsp70基因表达量均高于VL17,胁迫120 min时VL18的Hsp70和sHsp基因表达量分别是VL17的16.97倍和63.74倍。两菌株sHsp基因的表达量均明显高于Hsp70基因。通过比较分析认为小分子热激蛋白sHsp对蜡蚧轮枝菌抗高温胁迫起重要作用。
英文摘要:
      By comparing the difference of thermotolerance and the relationship between thermotolerance and heat shock protein (Hsp) mRNA level of two Lecanicillium lecanii strains,the mechanism of Hsp on heat stress tolerance was studied. The thermotolerance of mycelia and conidia between L. lecanii strain VL17 and VL18 were compared using mycelium growth rate method and spore germination method. Spray inoculation method was adopted to estimate the virulence of strain VL17 and VL18 against Lipaphis erysimi under high temperature stress. Real time quantitative PCR (qRT PCR) was used to compare expression of the sHsp and Hsp70 genes. The results showed that the thermotolerance of both mycelia and conidia from strain VL18 were significantly higher than those from strain VL17. The colony diameter of strain VL18 was 63.16% bigger than that of VL17 at 34℃ and the conidia germination rate of VL18 was 103% higher than that of VL17 at 37℃. The insecticidal activities of the two strains against L. erysimi were significantly different when the temperature was above 31℃. The corrected mortality of L. erysimi inoculated with VL18 was 166.67% higher than that with VL17 at 37℃. The qRT PCR analysis showed that the sHsp and Hsp70 mRNA of VL18 were higher than that of VL17 at 42℃ when the stress time was same. After a 120 min stress,expression of the Hsp70 and sHsp genes of strain VL18 were 16.97 and 63.74 folds as those of strain VL17,respectively. Furthermore,the expression of sHsp in both strains were significantly higher than those of Hsp70.In conclusion,sHsp played an important role in the response of L. lecanii to heat stress.
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