雌性黑腹果蝇不育基因cheerio的鉴定与研究  

Identification and study of female sterile gene cheerio in Drosophila melanogaster

作　　者：田应雪 杨敏 罗俊鹏 严江松 安云飞 任思佳 魏微 张庆海 TIAN Yingxue;YANG Min;LUO Junpeng;YAN Jiangsong;AN Yunfei;REN Sijia;WEI Wei;ZHANG Qinghai(Department of Biology,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;School of Biology and Engineering,Guizhou Medical University,Guiyang 550025,Guizhou,China;School of Clinical Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;Multimedia Laboratory of Morphology,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;Research Center for Basic Sciences,Guizhou Medical University,Guiyang 550025,Guizhou,China;Key Laboratory of Medical Insect,Guizhou Medical University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院生物学系,贵州贵阳550025 [2]贵州医科大学生物与工程学院,贵州贵阳550025 [3]贵州医科大学临床医学院,贵州贵阳550025 [4]贵州医科大学基础医学院多媒体形态学实验室,贵州贵阳550025 [5]贵州医科大学基础医学科学研究中心,贵州贵阳550025 [6]贵州医科大学医学昆虫重点实验室,贵州贵阳550025

出　　处：《贵州医科大学学报》2025年第3期361-369,共9页Journal of Guizhou Medical University

基　　金：国家自然科学基金(31601898,31960128);贵州省科学技术基金(黔科合基础[2017]1158,黔科合基础[2020]1Y082)。

摘　　要：目的鉴定雌性黑腹果蝇不育品系mf949和mf2150的突变基因,并研究其基因功能。方法以野生型(wide-type,WT)品系为对照,通过遗传杂交检测mf949和mf2150的育性,解剖卵巢并观察形态学变化;Hts-RC免疫荧光染色检测成虫卵巢的变化;利用反向PCR(iPCR)鉴定mf949和mf2150的突变基因;通过杂交实验从突变品系的基因组中跳除转座子,并再次检测育性及卵巢形态,验证筛选结果,排除遗传背景的影响。结果mf949和mf2150通过育性检测观察到产生前后轴和背侧肢变短的卵,确定为雌性不育品系;mf949与mf2150的幼虫性腺和蛹期卵巢都无明显变化;Hts-RC免疫荧光染色结果表明,mf949和mf2150的环状管只在卵室第4阶段之前存在,后面阶段未被标记;mf949与mf2150互补后代符合基因型的雌蝇,经Hts-RC染色其卵巢也是第4阶段后不能显示环状管;测序结果显示,mf949和mf2150是物质运输中果蝇丝素蛋白基因cheerio不同位点的新突变体;mf949和mf2150转座子跳除后果蝇育性和表型均恢复正常,证实mf949和mf2150的表型和不育均由cheerio突变所致。结论本研究鉴定出雌性黑腹果蝇2个cheerio不同位点的新突变体,为研究cheerio基因功能提供了新的遗传资源。Objective To identify the mutant genes of female sterile lines mf 949 and mf 2150 of Drosophila melanogaster,and research their gene functions.Methods The wild-type(WT)strain was used as the control,and the fertility of mf 949 and mf 2150 was detected by genetic hybridization.The ovary was dissected and morphological changes were observed.Hts-RC was used as primary antibody to determine the changes of adult ovaries by immunofluorescence staining.Reverse PCR(iPCR)was used to identify the mutant genes of mf949 and mf2150.Finally,the transposon was removed from the genome of the mutant strain,in order to test the fertility and ovarian morphology of the revertant fly and exclude the influence of genetic background.Results mf949 and mf2150 were determined to be female sterile lines after fertility test.The ovary was dissected and both anterior-posterior axes and dorsal limbs became shorter.Further dissection of larvae and pupae revealed that there was no significant change in the larval gonad and pupal ovaries of mf949 and mf2150.Hts-RC immunofluorescence staining showed that the ring canals of mf949 and mf2150 only existed before stage 4 of the egg chamber,and were not labeled in the subsequent stages.Sequencing results demonstrated that mf949 and mf2150 were new mutants of cheerio involved material transport.After transposon skipping,the fertility and phenotype of mf949 and mf2150 returned to normal,confirming that the phenotype and sterility of mf949 and mf2150 were caused by cheerio mutation.Conclusion In this study,two new mutants at different sites of cheerio were identified,which provides the study of the function of cheerio genes with a new genetic resource.

关 键 词：黑腹果蝇 卵巢 环状管 cheerio 卵室形成 生殖发育 

分 类 号：R394[医药卫生—医学遗传学] Q963[医药卫生—基础医学]