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Cloning of genes preferentially expressed in early stage of parthenocarpic and pollinated fruit and stabilization of fruit set

单性结实和授粉果实早期优先表达基因的克隆及坐果稳定

基本信息

批准号：
13460013
负责人：
YAMAKI Shohei
金额：
$ 9.79万
依托单位：
NAGOYA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460013/
关键词：
cDNA library cDNA cloning pollination fruit set eggplant sweet pepper subtraction parthenocarpy オーキシン

项目摘要

The purpose of this project is to make clear the genes preferentially relating to parthenocarpy and fruit setting for its stabilization.It was obserbed using fluorescence microscopy that pollen tube elongated after pollination of eggplant flower and the tip reached the ovules in ovary within 1.5 to 2 days after pollination. Therefore, three experimental conditions of pollination, unpollination, and unpollination + auxin treatments were set after emasculation on the flower opening. And RNAs from each sample were extracted after 2 or 3 days growth. Regarding the expression on 2 days after pollination, subtracted cDNA library was prepared by the subtraction between pollinated and unpollinated fruit. We picked up randumly 384 cDNA clones and spotted them on the membrane for macroarray assay. Thirty two candidates of cDNA clones for genes expressed higher in pollinated fruit than unpollinated one were selected and used as probes for Nothern blot analysis. Eleven of these 32 cDNA clones show … More ed that hybridization signals with mRNA from pollinated fruit were stronger ttian that from unpollinated fruit. mRKAs of three genes having similarity to P450, Unknown protein, and Hypothetical protein were specifically expressed in both pollinated and auxin treated fruit from 2 to 4 days after anthesis and then reduced rapidly. On the other hand, regarding the expression on 3 days after pollination, there were 10 cDNA clones expressed preferentially, in which histone, carleticulin, polyphenol oxidase, cell wall structural protein, cell division relating protein, molecular chaperon were contained. However, almost all mRNAs continued to be expressed clearly ever after 3 days. Thus, it was suggested that the 3 genes expressed on 2 days related to the initiation of fruit set.In sweet pepper fruit, also, 7 EST genes preferentially expressed in pollinated fruit on 2 days after pollination were isolated using the same way to eggplant fruit. Embryomic flower 2, which functions the regulation of flowering, was prominently expressed by pollination. Less

本研究以茄子为材料，通过荧光显微镜观察，发现授粉后1.5 ~ 2d内，花粉管伸长，顶端到达子房胚珠。因此，在开花期去雄后，设置了授粉、不授粉和不授粉+生长素处理三种试验条件。并在生长2或3天后从每个样品中提取RNA。关于授粉后2天的表达，通过在授粉和未授粉果实之间进行减法来制备消减cDNA文库。随机挑取384个cDNA克隆，点样于膜上进行宏阵列分析。选择了32个在授粉果实中表达高于未授粉果实的基因的cDNA克隆作为探针，进行了Nothingblot分析。这32个cDNA克隆中有11个显示 ...更多信息艾德认为，授粉果实的mRNA杂交信号比未授粉果实的强。与P450、Unknown protein和Hypothetical protein具有相似性的3个基因的mRKA在授粉和生长素处理的果实中均在花后2 ~ 4天特异表达，随后迅速下降。另一方面，在授粉后第3天的表达中，有10个cDNA克隆得到了优先表达，这些克隆中含有组蛋白、carleticulin、多酚氧化酶、细胞壁结构蛋白、细胞分裂相关蛋白、分子伴侣。然而，几乎所有的mRNA在3天后继续清楚地表达。在甜椒果实中，采用与茄子果实相同的方法，分离出7个在授粉后2d的授粉果实中优先表达的EST基因。胚体花2在授粉过程中显著表达，具有开花调控功能。少