昆虫学报 ›› 2019, Vol. 62 ›› Issue (12): 1351-1358.doi: 10.16380/j.kcxb.2019.12.001

• 研究论文 •    下一篇

RNAi和取食不同品种水稻后白背飞虱多铜氧化酶4基因的表达差异及生物学效应

贾浩康, 刘玉娣*, 侯茂林   

  1. (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)
  • 出版日期:2019-12-20 发布日期:2019-12-12

Differential expression of multicopper oxidase 4 gene and biological effects after RNAi and feeding on different rice varieties in Sogatella furcifera (Hemiptera: Delphacidae)

JIA Hao-Kang, LIU Yu-Di*, HOU Mao-Lin   

  1.  (State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China)
  • Online:2019-12-20 Published:2019-12-12

摘要: 【目的】分析RNAi及取食不同品种水稻后白背飞虱Sogatella furcifera多铜氧化酶4(multicopper oxidase 4, MCO4)基因的表达差异和生物学效应,以期为该基因的功能研究提供理论依据。【方法】通过生物信息学技术方法分析白背飞虱MCO4基因的核苷酸和氨基酸序列;利用RNAi技术沉默白背飞虱3龄若虫的MCO4基因,检测显微注射法和饲喂法的沉默效率,研究该基因被干扰后对白背飞虱3龄若虫存活率的影响;白背飞虱成虫取食不同抗性水稻品种24 h后,采用RT-qPCR检测其体内MCO4基因表达量的变化,并统计其体重。【结果】白背飞虱MCO4基因的核苷酸序列全长为2 166 bp,编码721个氨基酸。显微注射法进行MCO4基因的RNAi可成功沉默白背飞虱的MCO4基因,且沉默效率远高于饲喂法。MCO4基因被RNAi沉默后白背飞虱3龄若虫存活率显著低于正常若虫的。与取食水稻感虫品种TN1相比,取食水稻抗虫品种IR26的成虫MCO4基因表达量显著增加,且成虫体重显著降低。【结论】显微注射法可以更好地干扰白背飞虱MC04基因的表达。沉默MCO4基因对白背飞虱3龄若虫的存活率有显著影响,我们推测MCO4基因在白背飞虱取食水稻中具有重要的作用。

关键词: 白背飞虱, 唾液, 多铜氧化酶4, RNAi, 显微注射法, 饲喂法

Abstract: 【Aim】 The study aims to analyze the expression difference of multicopper oxidase 4 gene and the biological effects after RNAi and feeding on different rice varieties in Sogatella furcifera, so as to provide a theoretical basis for fundamental function research of this gene. 【Methods】 The nucleotide and amino acid sequences of MCO4 gene were analyzed by bioinformatic methods. The MCO4 gene in the 3rd instar nymphs of S. furcifera was silenced by RNAi, the silencing efficiencies of microinjection method and feeding method were compared, and the effect of RNAi of MCO4 gene on the survival rate of the 3rd instar nymphs was studied. After feeding on rice varieties with different insect resistance for 24 h, the changes in the expression level of MCO4 in S. furcifera adults were detected by RT-qPCR, and their body weight was detected. 【Results】 The full-length cDNA of MCO4 gene is 2 166 bp, encoding 721 amino acids. RNAi by microinjection method successfully silenced the MCO4 gene of S. furcifera, with much higher silencing efficiency than that by feeding method. The survival rate of the 3rd instar nymphs with silenced MCO4 was significantly lower than that of the normal nymphs. Compared with the adults fed on the susceptible cultivar TN1, the adults fed on the resistant variety IR26 had significantly higher expression level of MCO4 and significantly reduced body weight. 【Conclusion】 Microinjection method can more efficiently silence the MCO4 gene of S. furcifera. Silence of MCO4 gene has significant effects on the survival rate of S. furcifera, and we speculate that MCO4 plays an important role in feeding of S. furcifera on rice.

Key words: Sogatella furcifera, saliva, multicopper oxidase 4, RNAi, microinjection method, feeding method