隐花色素蛋白和铁硫簇蛋白相互作用及其在褐飞虱发育与生殖中的作用
Interaction between cryptochrome and iron-sulfur cluster assembly 1 protein and their roles in the development and reproduction of the brown planthopper, Nilaparvata lugens
张英超；曾路影；万贵钧；江幸福；陈法军；潘卫东
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DOI：10.7679/j.issn.2095-1353.2021.007
作者单位：1. 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；2. 中国科学院大学，北京 100049；3. 南京农业大学植物保护 学院昆虫系昆虫信息生态研究组，南京 210095；4. 中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室，北京 100193
中文关键词：隐花色素蛋白；铁硫簇蛋白；生物磁感受；褐飞虱
英文关键词：cryptochrome; iron-sulfur cluster assembly 1; magnetoreception; Nilaparvata lugens
中文摘要：
【目的】 在真核生物中验证隐花色素蛋白（Cryptochrome）/铁硫簇蛋白（Iron-sulfur cluster assembly 1）的相互作用，研究2种蛋白在褐飞虱Nilaparvata lugens发育和生殖过程中的作用，为二者在磁感受机制中的协同作用及磁场对昆虫生理状态的调控机制提供依据和新的研究方向。【方法】 通过酵母双杂交系统对褐飞虱Ⅰ型隐花色素蛋白/Ⅱ隐花色素蛋白与铁硫簇蛋白的相互作用进行验证。在褐飞虱3龄若虫和羽化第1天成虫中建立Ⅰ型隐花色素蛋白和铁硫簇蛋白基因RNAi（RNA interference）沉默模型，并采用RT-qPCR检测2个基因沉默效率，统计沉默虫体的若虫历期和产卵量。【结果】 褐飞虱Ⅰ型隐花色素蛋白与铁硫簇蛋白存在相互作用，而Ⅱ隐花色素蛋白与铁硫簇蛋白不存在相互作用。褐飞虱3龄若虫和羽化第1天成虫中成功建立Ⅰ型隐花色素蛋白和铁硫簇蛋白基因RNAi沉默模型，在注射dsRNA后第2天、第4天和第6天时，2个基因的沉默效率均能达到70%以上。Ⅰ型隐花色素蛋白和铁硫簇蛋白基因沉默虫体的若虫历期无明显变化，但是产卵量受到显著性抑制。【结论】 褐飞虱Ⅰ型隐花色素蛋白和铁硫簇蛋白存在相互作用，二者在褐飞虱生殖过程中发挥重要作用。
英文摘要：
[Objectives] To verify the interaction between cryptochrome (Cry) and iron-sulfur cluster assembly 1 (IscA1) protein in eukaryotes, and to study the role of these two proteins in the development and reproduction of brown planthopper, Nilaparvata lugens, so as to provide a basis for investigating the synergy between these proteins in magnetoreception and the physiological regulation of the magnetic field in insects. [Methods] The interaction between N. lugens Cry1/Cry 2 and IscA1 was verified using yeast, two hybrid technology. The RNAi (RNA interference) silencing model of Cry1/IscA1 gene was established in 3rd instar nymphs and 1-day-old adults and the silencing efficiency of these two genes detected by RT-qPCR. The duration of the nymph period and the oviposition rate of silenced individuals were measured. [Results] There is an interaction between Cry1 and IscA1 in N. lugens but not between Cry2 and IscA1. The RNAi silencing model of the Cry1/ IscA1 gene was successfully established in 3rd instar nymphs and 1-day-old adults; the silencing efficiency of these two genes was > 70% on the 2nd, 4th and 6th day after dsRNA injection. There was no change in the duration of the nymph period after Cry1/IscA1 gene silencing but oviposition was significantly inhibited. [Conclusion] Cry1 interacts with IscA1 in N. lugens and plays an important role in the reproduction of this species.