학술논문
东方蜜蜂微孢子虫侵染过程中nce-miR-15338及候选靶基因的表达谱 / Expression patterns of nce-miR-15338 and candidate target genes during the infection process of Nosema ceranae
Document Type
Academic Journal
Author
王思懿; 高旭泽; 赵浩东; 荆欣; 刘小玉; 冯佩林; 宋宇轩; 刘彩珍; 陈大福; 郭睿; WANG Siyi; GAO Xuze; ZHAO Haodong; JING Xin; LIU Xiaoyu; FENG Peilin; SONG Yuxuan; LIU Caizhen; CHEN Dafu; GUO Rui
Source
福建农林大学学报（自然科学版） / Journal of Fujian Agriculture and Forestry University(Natural Science Edition). 53(1):89-95
Subject
Language
Chinese
ISSN
1671-5470
Abstract
[目的]本研究旨在为探究nce-miR-15338 调控东方蜜蜂微孢子虫(Nosema ceranae)侵染意大利蜜蜂(Apis mellifera li-gustica)工蜂的功能及作用机制提供参考和依据.[方法]采用Stem-loop RT-PCR和Sanger测序验证nce-miR-15338 的存在和表达.利用相关软件预测nce-miR-15338 的靶基因并进行数据库注释.通过RT-qPCR检测nce-miR-15338 及其候选靶基因在东方蜜蜂微孢子虫侵染过程中的表达谱.[结果]nce-miR-15338 在东方蜜蜂微孢子虫孢子中真实存在和表达;nce-miR-15338 共靶向LCFAL 2 和20sPS等 16 个基因;分别有 16、7、8、3 个靶基因可注释到Nr、Swiss-Prot、GO和KEGG数据库;相较于接种后 1 d,nce-miR-15338 的表达量在 2d极显著上调,在 4、6、8 d均极显著下调;LCFAL 2 的表达量在接种 2、4、6、8d后皆为极显著下调;20sPS在接种 2、4、6、8 d的表达水平均为极显著下调.[结论]研究结果证实了nce-miR-15338 的真实存在和表达,并明确了东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂过程中nce-miR-15338 及其候选靶基因LCFAL 2 和 20sPS的表达规律.
[Objective]This study aims to provide reference and basis for exploring the function and action mechanism underlying the nce-miR-15338 regulated Nosema ceranae infection of Apis mellifera ligustica workers.[Method]Stem-loop RT-PCR and Sanger se-quencing were firstly applied to verify the existence and expression of nce-miR-15338.After the prediction of its target genes and da-tabase annotation by related software,the expression patterns of nce-miR-15338 and its candidate target genes during N.ceranae′s in-fection of A.mellifera ligustica workers were determined by RT-qPCR.[Result]The results proved the existence and expression of nce-miR-15338 in N.ceranae spores.Nce-miR-15338 potentially targeted 16 genes,including LCFAL 2,20s PS,etc.,and up to 16,7,8 and 3 target genes were annotated to Nr,Swiss-Prot,GO and KEGG databases,respectively.Compared with its expression level 1 day post inoculation(1 d),the expression level of nce-miR-15338 was significantly up-regulated at 2 d,while significantly down-regulated at 4,6 and 8 d;the expression levels of LCFAL 2 and 20sPS were both significantly down-regulated at 2,4,6 and 8 d.[Conclusion]The study verified the authentic existence of nce-miR-15338 in N.ceranae,and clarified the expression patterns of nce-miR-15338 as well as its candidate target genes of LCFAL 2 and 20sPS during N.ceranae′s infection in A.m.ligustica workers.
[Objective]This study aims to provide reference and basis for exploring the function and action mechanism underlying the nce-miR-15338 regulated Nosema ceranae infection of Apis mellifera ligustica workers.[Method]Stem-loop RT-PCR and Sanger se-quencing were firstly applied to verify the existence and expression of nce-miR-15338.After the prediction of its target genes and da-tabase annotation by related software,the expression patterns of nce-miR-15338 and its candidate target genes during N.ceranae′s in-fection of A.mellifera ligustica workers were determined by RT-qPCR.[Result]The results proved the existence and expression of nce-miR-15338 in N.ceranae spores.Nce-miR-15338 potentially targeted 16 genes,including LCFAL 2,20s PS,etc.,and up to 16,7,8 and 3 target genes were annotated to Nr,Swiss-Prot,GO and KEGG databases,respectively.Compared with its expression level 1 day post inoculation(1 d),the expression level of nce-miR-15338 was significantly up-regulated at 2 d,while significantly down-regulated at 4,6 and 8 d;the expression levels of LCFAL 2 and 20sPS were both significantly down-regulated at 2,4,6 and 8 d.[Conclusion]The study verified the authentic existence of nce-miR-15338 in N.ceranae,and clarified the expression patterns of nce-miR-15338 as well as its candidate target genes of LCFAL 2 and 20sPS during N.ceranae′s infection in A.m.ligustica workers.