Function Analysis of Ribophorin Ⅱ Gene in the Brown Planthopper (Nilaparvata lugens)
WANG Sai-Nan1, ZHANG Chuan-Xi1,2, LU Jia-Bao1,*
1 Institute of Plant Virology State/Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products/Key Laboratory of Biotechnology in Plant Protection of Ministry of Agriculture and Rural Affairs and Zhejiang Province, Ningbo University, Ningbo 315211, China; 2 Institute of Insect Science, Zhejiang University, Hangzhou 310058, China
Abstract Ribophorin Ⅱ (RⅡ) is a crucial subunit of the oligosaccharide transferase, and studies have shown that RⅡ is associated with cancer occurrence in humans (Homo sapiens), but there is currently limited research on the function of RⅡ in insects. In this study, a gene encoding RⅡ was identified in the brown planthopper (Nilaparvata lugens, BPH), a rice (Oryza sativa) pest, and is named N. lugens ribophorin Ⅱ (NlRⅡ)(GenBank No. XP_022202517.1). To investigate the function of NlRⅡ in BPH, the temporal expression pattern of NlRⅡ using transcriptomic data was first obtained, and the result revealed the expression pattern of NlRⅡ changed periodically with molting and reached a peak in each mid-instar stage. Then, qRT-PCR was used to detect the tissue-specific expression of NlRⅡ, the result showed that the expression level of NlRⅡ was highest in the ovary. Finally, RNA interference (RNAi) technique was used to silence NlRⅡ in the 2nd instar nymphs, and no significant effect was found on the survival of BPH. However, silencing NlRⅡ in the 5th instar resulted in female adult abdominal swelling, malformed ovary development, and almost no egg production. Additionally, significant decreases in egg number and hatchability were observed in wild-type female adults mated with male adults with extremely significantly constricted vas deferens (P<0.01). This study indicated that NlRⅡ could affect the reproduction of BPH, and provide a potential new target gene for RNAi-based BPH control.
WANG Sai-Nan,ZHANG Chuan-Xi,LU Jia-Bao. Function Analysis of Ribophorin Ⅱ Gene in the Brown Planthopper (Nilaparvata lugens)[J]. 农业生物技术学报, 2023, 31(12): 2556-2567.
[1] 毕崇尧. 2020. 通过下调RPN2 (N-寡糖转移酶复合物)诱导细胞凋亡抑制结肠癌的迁移和侵袭的研究[J]. 中国医药导报, 40(13): 11-19. (Bi C Y.2020. A study of inhibition of colon cancer migration and invasion by downregulating RPN2 (N-oligosaccharide transferase complex) induced apoptosis[J]. China Medical Herald, 40(13): 11-19.) [2] 兰斓. 2009. 稻瘟病菌假定Rpn2蛋白的生物信息学分析及基因敲除研究[D]. 硕士学位论文, 福建农林大学, 导师: 周洁, pp. 7-37. (Lan L.2009. Bioinformation and gene knock-out analysis of putative protein Rpn2 in Magnaporthe grisea[D]. Thesis for M.S., Fujian Agriculture and Forestry University, Supervisor: Zhou J, pp. 7-37.) [3] 黎丹, 官展文, 黄诗韵, 等. 2018. 褐飞虱卵巢特异性基因Nl10847的功能初探[J]. 环境昆虫学报, 40(03): 497-504. (Li D, Guo Z W, Huang S Y, et al.2018. Functional analysis of ovarian specific gene Nl10847 in the Brown planthopper Nilaparvata lugens[J]. Journal of Environmental Entomology, 40(03): 497-504.) [4] 韦亚东, 牛淼淼, 刘子瑜, 等. 2011.昆虫与哺乳动物蛋白质N-糖基化修饰异同的比较及其潜在的应用意义[J]. 中国蚕业, 32(02): 8-13. (Wei Y D, Niu M M, Liu Z Y, et al.2011. Comparison of similarities and differences between insect and mammalian protein N-glycosylation modifications and their potential applications[J]. China Sericulture, 32(02): 8-13.) [5] 赵征, 张克难, 孙思, 等. 2018. 核糖体结合糖蛋白2在脑胶质瘤中的表达及其在预后评估中的作用[J]. 中华神经外科杂志, 34(11): 1155-1160. (Zhao Z, Zhang K N, Sun S, et al.2018. RPN2 expression in glioma and its role in prognostic assessment[J]. Chinese Journal of Neurosurgery, 34(11): 1155-1160.) [6] 张士俊, 王延民, 吴焕成, 等. 2022. miR-142-5p靶向调控RPN2对胶质瘤细胞增殖、侵袭的影响及其机制[J]. 山东医药, 62(13): 45-49, 55. (Zhang S J, Wang Y M, Wu H C, et al.2022. Effects of miR-142-5p regulating RPN2 on proliferation and migration of glioma cells[J]. Shandong Medical Journal, 62(13): 45-49, 55.) [7] Aebi M.2013. N-linked protein glycosylation in the ER[J]. Biochimica et Biophysica Acta, 1833(11): 2430-2437. [8] Burda P, Aebi M.1999. The dolichol pathway of N-linked glycosylation[J]. Biochimica et Biophysica Acta, 1426(2): 239-257. [9] Esmail S, Manolson M F.2021. Advances in understanding N-glycosylation structure, function, and regulation in health and disease[J]. European Journal of Cell Biology, 100(7-8): 151186. [10] Fernndez-Ponce C, Geribaldi-Dold N N.2021. The role of glycosyltransferases in colorectal cancer[J]. International Journal of Molecular Sciences, 22(11): 5822. [11] Fu J, Pirozzi G, Sanjay A, et al.2000. Localization of ribophorin Ⅱ to the endoplasmic reticulum involves both its transmembrane and cytoplasmic domains[J]. European Journal of Cell Biology, 79(4): 219-228. [12] Jeanmougin F, Thompson J D, Gouy M, et al.1998. Multiple sequence alignment with Clustal X[J]. Trends in Biochemical Sciences, 23(10): 403-405. [13] Kelleher D J, Gilmore R.2006. An evolving view of the eukaryotic oligosaccharyltransferase[J]. Glycobiology, 16(4): 47-62. [14] Kelleher D J, Kreibich G, Gilmore R.1992. Oligosaccharyl transferase activity is associated with a protein complex composed of ribophorins Ⅰ and Ⅱ and a 48 kd protein[J]. Cell, 69(1): 55-65. [15] Knauer R, Lehle L.1994. The N-oligosaccharyltransferase complex from yeast[J]. FEBS Letters, 344(1): 83-86. [16] Kreibich G, Czakó-Graham M, Grebenau R, et al.1978a. Characterization of the ribosomal binding site in rat liver rough microsomes: Ribophorins Ⅰ and Ⅱ, two integral membrane proteins related to ribosome binding[J]. Supramolecular Struct Cell Biochemistry, 8(3): 279-302. [17] Kreibich G, Ulrich B L, Sabatini D D.1978b. Proteins of rough microsomal membranes related to ribosome binding. Ⅰ. Identification of ribophorins Ⅰ and Ⅱ, membrane proteins characteristics of rough microsomes[J]. The Journal of Cell Biology, 77(2): 464-487. [18] Kreil G.1984. Occurrence, detection, and biosynthesis of carboxy-terminal amides[J]. Methods in Enzymology, 106(1): 218-223. [19] Kumar S, Stecher G, Tamura K.2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets[J]. Molecular Biology and Evolution, 33(7): 1870-1874. [20] Letunic I, Khedkar S, Bork P.2021. SMART: Recent updates, new developments and status in 2020[J]. Nucleic Acids Research, 49(D1): D458-D460. [21] Livak K J, Schmittgen T D.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C(T)) Method[J]. Methods (San Diego, Calif), 25(4): 402-408. [22] Mar T, Liu W, Wang X.2014. Proteomic analysis of interaction between P7-1 of Southern rice black-streaked dwarf virus and the insect vector reveals diverse insect proteins involved in successful transmission[J]. Proteomics, 102(6): 83-97. [23] Mohanty S, Chaudhary B P, Zoetewey D.2020. Structural insight into the mechanism of N-linked glycosylation by oligosaccharyltransferase[J]. Biomolecules, 10(4): 624. [24] Sanihaliru B, Rafii M Y.2020. Recent strategies for detection and improvement of brown planthopper resistance genes in rice: A review[J]. Plants (Basel), 9(9): 1202. [25] Shen Y, Chen Y Z, Zhang C X.2021. RNAi-mediated silencing of ferritin genes in the brown planthopper Nilaparvata lugens affects survival, growth and female fecundity[J]. Pest Management Science, 77(1): 365-377. [26] Sugiyama Y, Nishimura A, Ohno S.2008. Symmetrically dividing cell specific division axes alteration observed in proteasome depleted C. elegans embryo[J]. Mechanisms of Development, 125(8): 743-755. [27] Wilson C M, Roebuck Q, High S.2008. Ribophorin I regulates substrate delivery to the oligosaccharyltransferase core[J]. Proceedings of the National Academy of Sciences, 105(28): 9534-9539. [28] Xu H J, Xue J, Lu B, et al.2015. Two insulin receptors determine alternative wing morphs in planthoppers[J]. Nature, 519(7544): 464. [29] Ye Y X, Zhang H H, Li DT, et al.2021. Chromosome-level assembly of the brown planthopper genome with a characterized Y chromosome[J]. Molecular Ecology Resources, 21(4): 1287-1298. [30] Zhang W, Xiao Z Y, Chun S H.2011. Ribophorin Ⅱ is a substrate of spermatid-specific E3 ligase RNF148[C] //, Zhuang X H, Sun B L(eds). Proceedings of the Chinese Society of Cell Biology General Assembly and the 12th Academic Conference. Yang J, Shanghai, pp. 69-70.
