纯雌系苦瓜花经AgNO_3诱雄后的雄蕊发育和基因表达  

Stamen Development and Gene Expression of Flower of Male-induced Bitter Melon Gynoecious Line

作　　者：王日升[1,2,3] 李杨瑞[2] 周生茂 张曼 方锋学 杨丽涛[1] 

机构地区：[1]广西大学广西亚热带生物资源保护利用重点实验室,南宁530005 [2]广西作物遗传改良生物技术重点开放实验室,南宁530007 [3]广西农业科学研究院蔬菜研究中心,南宁530007

出　　处：《细胞生物学杂志》2009年第4期547-552,共6页Chinese Journal of Cell Biology

基　　金：国家科技支撑计划项目(No.2007BAD68B03);公益性行业(农业)科研专项(No.nyhyzx07-007-1);广西青年基金项目(No.桂科青0991077);广西自然科学基金项目(No.桂科自0728065);广西作物遗传生物技术重点开放实验室开放课题(No.桂科能0815011-6-1-14)资助~~

摘　　要：为了研究AgNO3诱导下纯雌系苦瓜花蕾发育过程中的差异基因表达,本文运用形态学观察、cDNA-AFLP、RT-PCR和反向Northern杂交等技术在形态学和转录组学水平上对花蕾转雄进行了研究。结果显示:(1)花蕾在AgNO3处理后第6天出现雄蕊,第14天形成两性花,而且两性花中的花粉具有完全生活力,第22天后恢复全雌花;(2)从128对引物组合中筛选出可获得丰富清晰条带的引物组合36对,它们从AgNO3处理的花蕾中分离到11个高度表达的差异片段,经克隆、测序和在线比对,有2个未知基因,9个已知基因,其中1个与植物性别分化有关的CYP450家族基因有高度同源性,命名为McCYP72A1;(3)McCYP72A1在AgNO3处理后2～12h内的花蕾中表达,而且在第8h时表达丰度最高,但在处理的根、茎和叶以及对照的各个器官中不表达。结果表明,AgNO3处理可诱导纯雌系苦瓜花蕾中与雄性分化相关基因表达,进而可能导致处理后14天内雌花发育为两性花。To elucidate the differential gene expression during the development of stamens in female flower buds of bitter melon gynoecious line induced with AgNO3, in the present study, it was studied at the levels of morphology and transcription genomics using morphologic observations, cDNA-AFLP, RT-PCR and reverse-Northern hybridization. The results showed as follows： （1） After treated with AgNO3, stamens were generated at the 6th day, and bisexual flowers with viable pollen grains were observed at the 14th day, and gynoecious flowers were recov- ered since the 22th day. （2） Thirty-six primer combinations could amplify many distinct bands, among them, 11 bands which were strongly expressed only in the AgNO3-treated buds were cloned, sequenced and aligned on line, resulting in discovery of 2 unknown genes and 9 known genes. The sequence showing high similarity with CYP450 gene super family which related to plant sex differentiation was named as McCYP72A1. （3） McCYP72A1 was expressed in the flower buds during the period of 2-12 h after treated with AgNO3, and expressed most strongly at 8 h, but not expressed in the roots, stems, leaves of treatment and the mixed samples of buds, roots, stems, leaves of the control. It suggested that genes related to male differentiation in the flower buds of bitter melon gynoecious line were expressed after treated with AgNO3, which might make female flowers develop into bisexual flowers within 14 days.

关 键 词：苦瓜 纯雌系 诱雄处理 cDNA—AFLP 差异基因表达 

分 类 号：S642.2[农业科学—蔬菜学]