交配和黑光灯处理对棉铃虫Helicoverpa armigera生物钟基因cryptochromes mRNA表达的影响

Effect of Mating and Black Light on mRNA Expression of Clock Gene Cryptochromes of Helicoverpa armigera

[目的]明确交配和黑光灯处理对棉铃虫生物钟基因cryptochromes mRNA表达的影响。[方法]应用实时定量PCR(SYBR Green)技术检测不同条件下棉铃虫cryptochromes(cry1和cry2)基因的表达。提取棉铃虫头部总RNA,经DNase I消化后进行反转录合成cDNA,并采用特异性引物分别对cry1、cry2和EF-1α基因进行实时定量PCR扩增。[结果]黑光灯处理棉铃虫2h,其cry1mRNA的表达量明显降低;cry2mRNA的表达量小于对照,但两者之间差异性并不显著。交配对棉铃虫cry1和cry2mRNA表达均存在显著影响,并且雌、雄两性cry1和cry2mRNA表达量在交配后随时间延长呈下降趋势。[结论]该结果对进一步研究cry基因的功能以及棉铃虫防治具有重要意义。

[Objective]The paper was to confirm the effect of mating and black light on mRNA expression of clock gene cryptochromes of Helicoverpa armigera.[Method]Quantitative real-time PCR（SYBR Green） technique was applied to detect the expression of cryptochromes gene（cry1 and cry2） of H.armigera under different conditions.Total RNA was extracted from the head of H.armigera,and carried out reverse transcription to synthesize cDNA after digested by DNase I.Specific primers were used to carry out quantitative and real-time PCR on cry1,cry2 and EF-1α gene,respectively.[Result] The expression of cry1 mRNA of H.armigera significantly decreased after exposure to black lamps for 2 h,the mRNA expression of cry2 was smaller than control but without significant difference.Mating had significant effect on mRNA expression of cry1 and cry2 of H.armigera,and the mRNA expression of cry1 and cry2 of male and female showed decrease trend with the prolongation of time after mating.[Conclusion]The result had important significance for further study on function of cry gene and the control of cotton bollworm.