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本研究对棉铃虫丝氨酸蛋白酶抑制剂(Helicoverpa armigera serine protease inhibitor,Haspi)基因的基本特征、分布情况及功能进行了初步探究。通过与已报道的家蚕丝氨酸蛋白酶抑制剂基因同源比对,利用RT-PCR技术成功克隆到3条与家蚕丝氨酸蛋白酶抑制剂基因相似性高的Haspi基因,分别命名为:Haspi-5、Haspi-6和Haspi-10。实时定量PCR结果表明3个基因在棉铃虫各组织部位均有转录,但在不同组织部位处的表达水平差异显著。Haspi-5基因在表皮中的表达水平最高;Haspi-6基因在头部的转录水平远远高于其他组织部位;Haspi-10基因在前肠中表达水平最高,中肠的转录水平也相比其他组织较高。构建表达Haspi-5、Haspi-6基因的ds RNA的载体,用表达ds RNA的HT115菌液喂食,其中Haspi-5基因在棉铃虫的转录水平明显下降,约降低了6.5倍,而Haspi-6基因转录水平略有下降但并不明显,约降低了1.2倍,但是没有观察到明显的表型变化。上述研究为Haspi基因的功能及棉铃虫防治研究提供科学依据。
In this study, the basic characteristics, distribution and function of Helicoverpa armigera serine protease inhibitor (Haspi) gene were studied. Three homologous genes of silkworm serine protease inhibitor Haspi were successfully cloned by RT-PCR, which were named Haspi-5, Haspi-5, 6 and Haspi-10. The results of real-time quantitative PCR showed that all the three genes were transcribed in the tissues of H. armigera, but the expression levels in different tissues were significantly different. Haspi-5 gene expression in the epidermis of the highest level; Haspi-6 gene transcription in the head is much higher than other tissues; Haspi-10 gene expression in the foregut the highest level of midgut transcription level Other organizations are higher. The vector expressing dsRNA of Haspi-5 and Haspi-6 genes was constructed and fed with HT115 strain expressing dsRNA. The expression level of Haspi-5 gene in Helicoverpa armigera was reduced by about 6.5-fold, while the expression of Haspi-6 The gene transcription level slightly decreased but not obvious, about 1.2 times lower, but no obvious phenotypic change was observed. These studies provide a scientific basis for the function of Haspi gene and the control of cotton bollworm.