杀虫活性化合物杠柳新苷P对东方粘虫中肠细胞上结合蛋白的影响

Effect of Active Insecticidal Periplocoside P on Mythimna separata Midgut Cell Binding Proteins

杠柳新苷P是从杠柳根皮提取物中分离得到的一种甾体糖苷类化合物,对东方粘虫表现出较好的杀虫活性。为了进一步研究其杀虫机理,探究杠柳新苷P作用于东方粘虫中肠疑似作用位点,通过对杠柳新苷P进行结构修饰,与东方粘虫幼虫中肠刷状缘膜囊泡(BBMV)组织体外孵育,利用其结构修饰后增强的紫外吸收特性,经蛋白分离、纯化,进而得到杠柳新苷P作用于粘虫中肠BBMV上的结合蛋白。结果表明:在粘虫中肠BBMV上存在杠柳新苷P的疑似结合蛋白,且经分离鉴定得到5种疑似结合蛋白,分别为类异翅盘蛋白、氨肽酶、二硫键异构酶、苯基贮存蛋白以及精氨酸激酶;通过进一步的分析推测,氨肽酶可能是杠柳新苷P作用于东方粘虫幼虫中肠BBMV上的结合蛋白。初步明确杠柳新苷P作用于东方粘虫中肠的疑似作用位点,但仍需进一步的验证工作进行确认,为今后研究杠柳新苷类化合物的作用机理及靶标提供依据。

The pe Periploca sepium riplocoside P is a kind of steroidal glycoside compound isolated from the root bark of Bunge, which showed good insecticidal activity to Mythimna separata larvae. In order to study the insecticidal mechanism of periplocoside P, and research the suspected action site for the periplocoside P in the midgut of Mythimna separate, this paper modified the structure of periplocoside P and then incubated it with the BBMV in vitro. Though the enhanced UV absorption value, the binding protein is separated and purified from the BBMV in the midgut of Mythimna separata larvae. The results showed that suspected binding protein of periplocoside P exists in the BBMV in the midgut of Mythimna separata larvae. After the isolation, purification and identification, five kinds of suspected binding proteins was got, which is abnormal wing disc-like protein, aminopeptidase N, protein disulfide isomerase, arylphorin and arginine kinase-like protein, respectively. Through further analysis, we speculated that aminopeptidase N may be the most likely binding protein of the periplocoside P. In this paper, we preliminary make clear the suspected action site for the periplocoside P in the midgut of Mythimna separate,but further validation research still needs. This study provides a foundation and basis for further study on insecticidal mechanism and target location of the periplocosides compounds.