东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂过程中nce-miR-10660及其靶基因表达谱

doi:10.16380/j.kcxb.2023.02.007

昆虫学报 ›› 2023, Vol. 66 ›› Issue (2): 181-189.doi: 10.16380/j.kcxb.2023.02.007

东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂过程中nce-miR-10660及其靶基因表达谱

张凯遥^1,#, 张文德^1,#, 王紫馨¹, 胡颖¹, 钱加珺¹, 王思懿¹, 赵浩东¹, 顾小雨¹, 牛庆生³, 付中民^1,2, 陈大福^1,2,*, 郭睿^1,2,*

(1. 福建农林大学动物科学学院(蜂学学院), 福州 350002; 2. 福建省蜂疗研究所, 福州 350002; 3. 吉林省养蜂科学研究所, 吉林 132000)

出版日期:2023-02-20 发布日期:2023-04-07

Expression profiles of nce-miR-10660 and its target genes in Apis mellifera ligustica workers during the infection of Nosema ceranae

ZHANG Kai-Yao^1,#, ZHANG Wen-De^1,#, WANG Zi-Xin¹, HU Ying¹, QIAN Jia-Jun¹, WANG Si-Yi¹, ZHAO Hao-Dong¹, GU Xiao-Yu¹, NIU Qing-Sheng³, FU Zhong-Min^1,2, CHEN Da-Fu^1,2,*, GUO Rui^1,2,*

(1. College of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2. Apitherapy Research Institute of Fujian Province, Fuzhou 350002, China; 3. Apiculture Science Institute of Jilin Province, Jilin 132000, China)

Online:2023-02-20 Published:2023-04-07

摘要/Abstract

摘要： 【目的】本研究旨在为探究nce-miR-10660调控东方蜜蜂微孢子虫Nosema ceranae侵染的作用机制提供理论和实验依据。【方法】采用Stem-loop RT-PCR对前期鉴定到的东方蜜蜂微孢子虫nce-miR-10660进行表达验证，再通过Sanger测序验证nce-miR-10660的序列。利用相关生物信息学软件预测和分析nce-miR-10660的靶基因。通过RT-qPCR检测nce-miR-10660及其靶基因在东方蜜蜂微孢子虫侵染意大利蜜蜂Apis mellifera ligustica工蜂过程中中肠中的表达谱。【结果】Stem-loop RT-PCR和Sanger测序结果分别证实了nce-miR-10660在东方蜜蜂微孢子虫孢子中的表达和真实存在。靶向预测结果显示nce-miR-10660共靶向RRDRP和RCDP 42等9个基因；分别有2和6个靶基因可被分别注释到KEGG数据库中的4条通路和GO数据库中的23个条目。RT-qPCR结果显示，相较于东方蜜蜂微孢子虫侵染后1 d时意大利蜜蜂工蜂中肠中nce-miR-10660的表达量，东方蜜蜂微孢子虫侵染后2 d时意大利蜜蜂工蜂中肠中nce-miR-10660的表达量显著上调，在东方蜜蜂微孢子虫侵染后4, 6和8 d时意大利蜜蜂工蜂中肠中nce-miR-10660的表达量均显著下调；东方蜜蜂微孢子虫侵染后4, 6和8 d时意大利蜜蜂工蜂中肠中靶基因RRDRP的表达量均显著下调表达；东方蜜蜂微孢子虫侵染后2, 4, 6和8 d时意大利蜜蜂工蜂中肠中靶基因RCDP 42的表达量显著下调。【结论】nce-miR-10660在东方蜜蜂微孢子虫孢子中真实存在和表达；nce-miR-10660及其靶向的RRDRP和RCDP 42在东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂过程中差异表达；nce-miR-10660通过正调控RRDRP和RCDP 42表达潜在参与调节东方蜜蜂微孢子虫侵染。

关键词: 东方蜜蜂微孢子虫, 意大利蜜蜂, nce-miR-10660, 靶基因, 表达谱

Abstract: 【Aim】This study aims to provide theoretical and experimental bases for exploring the mechanism underlying Nosema ceranae infection regulated by nce-miR-10660.【Methods】Stem-loop RT-PCR was used to confirm the expression of previously identified nce-miR-10660 in N. ceranae and Sanger sequencing was performed to validate the nce-miR-10660 sequence. Related bioinformatic software was employed to predict and analyze the target genes of nce-miR-10660. RT-qPCR was used to detect the expression profiles of nce-miR-10660 and its target genes in the midguts of Apis mellifera ligustica workers during the infection process of N. ceranae. 【Results】Stem-loop RT-PCR and Sanger sequencing results confirmed the expression and true existence, respectively, of nce-miR-10660 in N. ceranae spores. Targeting prediction showed that nce-miR-10660 can target nine genes including RRDRP and RCDP 42, among which two and six target genes can be annotated to four pathways of KEGG database and 23 terms of GO database, respectively. RT-qPCR results indicated that the expression level of nce-miR-10660 in the midgut of A. m. ligustica workers at 2 d post infection by N. ceranae was significantly up-regulated, while those at 4, 6 and 8 d post infection by N. ceranae were significantly down-regulated, the expression levels of the target genes RRDRP in the midguts of A. m. ligustica workers at 4, 6 and 8 d post infection by N. ceranae and RCDP 42 in the midguts of A. m. ligustica workers at 2, 4, 6 and 8 d post infection by N. ceranae were significantly down-regulated, as compared to those in the midguts of A. m. ligustica workers at 1 d post infection by N. ceranae.【Conclusion】nce-miR-10660 truly exists and is expressed in N. ceranae spores. nce-miR-10660 and its target genes RRDRP and RCDP 42 are differentially expressed in A. m. ligustica workers during the infection of N. ceranae. nce-miR-10660 potentially participates in modulation of N. ceranae infection via positive regulation of the expression of RRDRP and RCDP 42.

Key words: Nosema ceranae; Apis mellifera ligustica, nce-miR-10660, target gene, expression profile

张凯遥, 张文德, 王紫馨, 胡颖, 钱加珺, 王思懿, 赵浩东, 顾小雨, 牛庆生, 付中民, 陈大福, 郭睿. 东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂过程中nce-miR-10660及其靶基因表达谱[J]. 昆虫学报, 2023, 66(2): 181-189.

ZHANG Kai-Yao, ZHANG Wen-De, WANG Zi-Xin, HU Ying, QIAN Jia-Jun, WANG Si-Yi, ZHAO Hao-Dong, GU Xiao-Yu, NIU Qing-Sheng, FU Zhong-Min, CHEN Da-Fu, GUO Rui. Expression profiles of nce-miR-10660 and its target genes in Apis mellifera ligustica workers during the infection of Nosema ceranae[J]. Acta Entomologica Sinica, 2023, 66(2): 181-189.

[1]	王紫馨, 许雅静, 张文德, 张凯遥, 吴鹰, 刘佳美, 朱乐冉, 牛庆生, 赵红霞, 陈大福, 郭睿. ame-miR-14在意大利蜜蜂工蜂幼虫肠道发育过程的调控作用[J]. 昆虫学报, 2023, 66(1): 1-10.
[2]	张泽麟, 嵇杰, 徐文雨, 刘卓, 范晓春, 邓昌鑫, 何倩毓. miR-252-5p通过靶向调控保幼激素酸甲基转移酶基因JHAMT的表达而影响果蝇变态发育[J]. 昆虫学报, 2022, 65(7): 799-806.
[3]	范小雪, 隆琦, 孙明会, 郭意龙, 赵浩东, 宋岳梅, 康育欣, 顾小雨, 陈大福, 郭睿. 意大利蜜蜂工蜂中肠piRNA的鉴定与分析[J]. 昆虫学报, 2022, 65(6): 684-694.
[4]	祝智威, 王杰, 隆琦, 许雅静, 冯睿蓉, 刘佳美, 赵浩东, 朱乐冉, 侯海青, 陈大福, 郭睿. 过表达和敲减ame-miR-13b对意大利蜜蜂幼虫肠道内基因表达的影响[J]. 昆虫学报, 2022, 65(4): 460-468.
[5]	刘倩, 罗迪, Roman JASHENKO, 季荣. 意大利蝗海藻糖酶基因克隆及卵低温驯化下的表达模式[J]. 昆虫学报, 2022, 65(2): 157-166.
[6]	胡颖, 张文德, 王思懿, 张凯遥, 任中民, 吉挺, 蔺哲广, 赵红霞, 陈大福, 郭睿. 意大利蜜蜂工蜂幼虫肠道中ame-miR-bantam对靶基因USP和P300的表达调控[J]. 昆虫学报, 2022, 65(11): 1401-1410.
[7]	蔡宗兵, 王思懿, 王紫馨, 赵萧, 张凯遥, 郭意龙, 姚雨彤, 钱加珺, 胡颖, 付中民, 陈大福, 郭睿. 过表达和敲减ace-miR-3720对中华蜜蜂工蜂幼虫肠道的靶基因表达和重量的影响[J]. 昆虫学报, 2022, 65(10): 1247-1255.
[8]	张凯遥, 刘佳美, 张文德, 胡颖, 康育欣, 王紫馨, 王思懿, 钱加珺, 赵萧, 张佳欣, 陈大福, 郭睿, 付中民. ame-miR-79负调控意大利蜜蜂工蜂幼虫肠道中靶基因CYP450和FG的表达[J]. 昆虫学报, 2022, 65(10): 1256-1265.
[9]	祝智威, 付中民, 隆琦, 杜宇, 张文德, 胡颖, 赵萧, 史小玉, 徐细建, 陈大福, 郭睿. 三种微小RNA在意大利蜜蜂工蜂蛹期发育过程中的表达谱及潜在功能[J]. 昆虫学报, 2022, 65(1): 53-62.
[10]	候梦赏, 邱园妹, 赵必安, 蔚添添, 梁立强, 苏松坤, 李志国. 亚致死剂量吡虫啉胁迫对意大利蜜蜂工蜂嗅觉学习行为的影响及其脑部转录组分析[J]. 昆虫学报, 2021, 64(7): 817-827.
[11]	候梦赏, 赵必安, 邱园妹, 万昆林, 梁立强, 李志国, 苏松坤. 球囊菌侵染对意大利蜜蜂幼虫肠道免疫与解毒相关基因表达及蛋白质、脂质和糖类含量的影响[J]. 昆虫学报, 2021, 64(5): 574-584.
[12]	马田田, 杨宗霖, 常美玲, 霍春月, 阚云超, 李丹丹. 豌豆蚜翅分化相关miRNA及其预测靶基因对蜕皮激素的应答及miR-92a-1-p5靶基因的验证[J]. 昆虫学报, 2021, 64(4): 419-427.
[13]	蔚添添, 邱园妹, 候梦赏, 王天宝, 苏松坤, 李志国. RNAi沉默王浆主蛋白1(Mrjp1)基因对意大利蜜蜂工蜂学习和记忆的影响[J]. 昆虫学报, 2021, 64(10): 1145-1152.
[14]	李琼艳, 陈安利, 荀利杰, 刘增虎, 廖鹏飞, 杨伟克, 董占鹏. 琥珀蚕丝腺转录因子基因AaSGF-1的克隆、原核表达及组织表达分析[J]. 昆虫学报, 2021, 64(1): 41-50.
[15]	宋月芹, 宋智煜, 董钧锋, 陈庆霄, 孙会忠. 双委夜蛾气味结合蛋白AdisOBP6的原核表达抗体制备及表达谱分析[J]. 昆虫学报, 2021, 64( 2): 149-157.

东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂过程中nce-miR-10660及其靶基因表达谱

Expression profiles of nce-miR-10660 and its target genes in Apis mellifera ligustica workers during the infection of Nosema ceranae

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0