CAJ | 학술논문

茶尺蠖是茶园中一种重要的鳞翅目害虫，主要以幼虫取食危害茶树叶片，给茶叶生产带来了严重威胁。茶尺蠖在寻找配偶、产卵地以及觅食活动中其嗅觉系统发挥了非常重要的作用。本研究以茶尺蠖Ectropis obliqua Prout嗅觉系统中的一个普通气味结合蛋白--EoblGOBP2为研究对象，在克隆得到的EoblGOBP2基因基础上，通过对 EoblGOBP2进行原核表达、分离纯化后，利用1-NPN 的荧光竞争结合法研究了体外重组EoblGOBP2与茶树挥发物的结合生理功能。Scatchard方程显示 EoblGOBP2与1-NPN的解离常数 KD为2.310μmol·L-1。在荧光竞争结合实验中，7种待测配基均能使1-NPN 的相对荧光强度降低到50%以下，表明与EoblGOBP2均有较强的结合能力，而其中邻苯二甲酸二丁酯结合能力最强， KD 值达到4.353μmol·L-1。本研究7种配基均为茶树叶片挥发物，表明EoblGOBP2在茶尺蠖嗅觉系统对茶树挥发物的信息识别结合功能中发挥了重要作用。
다척확시다완중일충중요적린시목해충，주요이유충취식위해다수협편，급다협생산대래료엄중위협。다척확재심조배우、산란지이급멱식활동중기후각계통발휘료비상중요적작용。본연구이다척확Ectropis obliqua Prout후각계통중적일개보통기미결합단백--EoblGOBP2위연구대상，재극륭득도적EoblGOBP2기인기출상，통과대 EoblGOBP2진행원핵표체、분리순화후，이용1-NPN 적형광경쟁결합법연구료체외중조EoblGOBP2여다수휘발물적결합생리공능。Scatchard방정현시 EoblGOBP2여1-NPN적해리상수 KD위2.310μmol·L-1。재형광경쟁결합실험중，7충대측배기균능사1-NPN 적상대형광강도강저도50%이하，표명여EoblGOBP2균유교강적결합능력，이기중린분이갑산이정지결합능력최강， KD 치체도4.353μmol·L-1。본연구7충배기균위다수협편휘발물，표명EoblGOBP2재다척확후각계통대다수휘발물적신식식별결합공능중발휘료중요작용。
As one main Lepidopteran pest in tea garden, the larvae of tea geometrid damages mainly the tea leaves, which severely reduce the yield of tea products. The olfactory system of tea geometrid plays a very important role in terms of seeking host and food. Therefore, a general odorant binding protein-EoblGOBP2, existing in the olfactory system of tea geometrid, the function binding with tea plant volatiles was studied in vitro here. Based on the cloned EoblGOBP2 gene by RT-PCR, the recombinant protein EoblGOBP2 was successfully expressed in an optimized prokaryotic prokaryotic expression system, then the binding characteristics of the purified EoblGOBP2 with 1-NPN was measured by fluorescence spectrometry. Scatchard plot analysis indicated that the dissociation constant between EoblGOBP2 and 1-NPN was 2.310μmol·L-1. In the competitive binding assays, to the seven candidate ligands, all of the relative fluorescence intensity of 1-NPN decreased more than 50%, which suggested that all of the tested ligands have strong binding capability with EoblGOBP2. The compound of dibutyl phthalate showed the maximum binding capability with EoblGOBP2, and their dissociation constant was 4.353μmol·L-1. As all of the candidate chemical ligands belong to the tea volatiles secondary metabolites, indicating that EoblGOBP2 plays an important role in the odor recognition process of olfactory identification in the olfactory system of tea geometrid.