近零磁场下干扰磁响应关键基因对褐飞虱寿命的影响  

Effects of the Interference of Key Magnetic Response Genes on the Longevity of Brown Planthopper (Nilaparvata lugens) Under Near-Zero Magnetic Field

作　　者：贺静澜 张明 刘瑞莹 万贵钧 潘卫东[2] 陈法军[1] HE JingLan;ZHANG Ming;LIU RuiYing;WAN GuiJun;PAN WeiDong;CHEN FaJun(College of Plant Protection,Nanjing Agricultural University,Nanjing 210095;Beijing Key Laboratory of Bioelectromagetics, Institute of Electrical Engineering,Chinese Academy of Seiences,Beijing 100190)

机构地区：[1]南京农业大学植物保护学院,南京210095 [2]中国科学院电工研究所生物电磁学北京市重点实验室,北京100190

出　　处：《中国农业科学》2019年第1期45-55,共11页Scientia Agricultura Sinica

基　　金：国家自然科学基金(31670855);国家自然科学基金青年科学基金(31701787);江苏省自然科学基金青年基金(BK20160717);中央高校基本科研业务费青年科技创新基金(KJQN201820)

摘　　要：【目的】隐花色素(cryptochrome, Cry)和铁硫簇蛋白IscA(iron-sulfur cluster assembly,即MagR)是生物体内潜在的磁受体蛋白,本研究通过RNA干扰(RNAi)技术,分别敲减褐飞虱(Nilaparvata lugens)体内的磁响应关键基因NlCry1、NlCry2和NlMagR,旨在探明近零磁场(near-zero magnetic field,NZMF)环境下,以上3种基因在褐飞虱寿命调节过程中的作用,从而间接探讨这3种基因对磁场的响应情况。【方法】采用RNAi技术,以实验室正常磁场环境下稳定饲养的短翅初羽化褐飞虱雌雄成虫为材料,通过向其体内注射双链RNA(dsRNA)分别抑制磁响应关键基因NlCry1、NlCry2和NlMagR,随后立即分别放入正常磁场(geomagnetic field,GMF)和近零磁场中,于每日相同时间观察记录试虫寿命。同时于注射后的1、2和3 d通过RNAiso Plus法提取GMF中褐飞虱雌成虫总RNA,反转录合成第一链DNA,后采用实时荧光定量PCR(RT-qPCR)技术检测该基因的表达情况,以确定基因干扰效率。【结果】注射ds NlCry1后,褐飞虱雌雄成虫寿命在近零磁场和正常磁场间均无显著差异。注射ds NlCry2后,近零磁场中褐飞虱雌雄成虫寿命比正常磁场分别显著延长27.78%和50.04%;此外,与注射ds GFP处理相比,正常磁场下注射ds NlCry2的雌成虫寿命缩短,而近零磁场下注射ds NlCry2的雌成虫寿命延长,但二者差异均不显著;近零磁场和正常磁场下注射ds NlCry2的雄成虫寿命均缩短(25.41%和10.73%),且正常磁场下差异显著。近零磁场中,注射ds NlMagR的雌成虫寿命较注射ds GFP的寿命显著缩短了16.48%,而雄成虫寿命在磁场间、干扰处理间的差异均不显著。【结论】磁场变化下褐飞虱雌雄成虫体内3种磁响应关键基因对其寿命的调节功能存在差异。其中,NlCry2对磁场变化存在敏感响应,表现为敲减该基因与磁场变化的互作显著地影响雌雄成虫寿命,且表现出"性二型性";NlMagR也可对磁场变化产生明显响应,但该响【Objective】Cryptochrome(Cry) and iron-sulfur cluster protein IscA(iron-sulfur cluster assembly, MagR) are potential magnetic receptor proteins in organisms. In this study, key magnetic response genes of the brown planthopper(Nilaparvata lugens) were knocked-down by RNA interference(RNAi), including NlCry1, Nl Cry2 and Nl Mag R. The objective of this study is to investigate the role of these three magnetic response genes in the longevity mediation of N. lugens in near-zero magnetic field(NZMF). Thus, the response of these three genes to magnetic field could be studied indirectly.【Method】Newly emerged brachypterous female and male adults of N. lugens fed in the lab magnetic field were chosen as the experimental material, and RNAi technology was used to inhibit the key magnetic response genes’(NlCry1, Nl Cry2 and NlMagR) expression by injection of double stranded RNA, respectively. Then the RNAi treated adults were immediately transformed into the geomagnetic field(GMF) and NZMF respectively to observe their longevity. The total RNA of the RNAi treated adults under GMF was extracted by using the RNAiso Plus method on the 1 st, 2 nd and 3 rd day after the microinjection, respectively. And then the gene expressions of NlCry1, NlCry2 and Nl Mag R were measured by using the RT-qPCR(real-time quantitative polymerase chain reaction) after the reverse transcription synthesis of first strand DNA in order to test the efficiency of RNAi. 【Result】There was no significant difference in the longevity of female and male adults after the injection of ds NlCry1 between the treatments of NZMF and GMF, while after the injection of ds Nl Cry2, the longevity of female(27.78%) and male(50.04%) adults under NZMF was significantly longer than that of the individuals under GMF, respectively. Moreover, the longevity of female adults injected with ds NlCry2 was shorter under GMF while longer under NZMF than that of individuals injected with ds GFP, even if the difference was not significant. The longevity of male adults injecte

关 键 词：磁场强度变化 褐飞虱 磁响应基因 RNA干扰 成虫寿命 磁生物学效应 

分 类 号：S435.112.3[农业科学—农业昆虫与害虫防治]