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为探究松材线虫抗逆态幼虫形成的分子机理,本文对抗逆态基因daf-6进行了鉴定和基因沉默研究。应用BLAST比对技术,在松材线虫基因组数据中鉴定得到秀丽线虫daf-6的同源基因,命名为Bx-DAF6。应用PCR技术进行Bx-DAF6完整蛋白编码区(CDS)扩增。采用RNAi技术进行Bx-DAF6沉默,分析该基因的沉默对松材线虫抗逆态幼虫形成的影响。PCR扩增得到CDS区全长2 700 bp,Bx-DAF6编码的蛋白质具有固醇传感结构域。Bx-DAF6沉默抑制了2龄幼虫转变为抗逆态幼虫,表明Bx-DAF6在2龄幼虫转变为抗逆态幼虫生理过程中起正向调控作用。
In order to explore the molecular mechanism of pine wood nematode anti-converse larva formation, the anti-stress gene daf-6 was identified and the gene silencing was studied. The homologous gene daf-6 of C. elegans was identified and identified as Bx-DAF6 using BLAST technology. The Bx-DAF6 complete protein coding region (CDS) was amplified by PCR. The Bx-DAF6 silencing was performed using RNAi technique and the effect of silencing of this gene on the formation of pine bugs in response to laxative larvae was analyzed. The total length of CDS region was 2 700 bp by PCR, and the protein encoded by Bx-DAF6 had a sterol sensing domain. Bx-DAF6 silencing inhibited the transformation of 2nd instar larvae into resistant larvae, indicating that Bx-DAF6 plays a positive regulatory role in the physiological process of 2nd instar larvae transformed into resistant larvae.