Cellular and molecular genetic approaches and control of differentiation on reproductive system, apomixes in crops

1.部分作物无融合性形成的细胞和分子遗传学分析研究项目在三家机构进行。采用Nomarski差示干涉对比显微镜（DLCM）观察Apospory Initial Cell（AIC）的表达过程。根据发育阶段对候选基因进行了定位克隆。利用图谱克隆和微阵列技术进行表达分析。对于这组标记，至少确定了9个BAC克隆池。并在制备转化体系后，尝试用三种二元载体对ASG-1候选基因进行诱导。现在正在进行这些转基因植物的再生。采用以下方法对韭菜（Allium tuberosum）的双孢子进行了研究；分离mRNA，克隆候选基因的差异表达，然后对200种不同表达进行微阵列分析。不幸的是，到目前为止，还没有关于它们的同步性和非同步性表达的研究。生殖系统的分化、控制与杂交致死。了解杂交种的种间致死性不仅对克服育种障碍，而且对控制生殖系统的细胞分化具有重要意义。通过控制胚胎生长过程中的温度，研究了致死率的表达。在种间杂交死亡过程中，不同程度地发生了细胞核退化、核小体单元核DNA断裂和细胞核断裂。细胞表达程序性死亡、凋亡。它不与细胞质因子有关，而是位于染色体上的某个基因位点上，与乙烯的形成有关。

1.Cellular and molecular genetic analysis on apomixes formation in some cropsResearch project was performed at three institutions in guineagross apospory. The process of the expression of Apospory Initial Cell(AIC) was observed by Nomarski differential interference-contrast microscope(DLCM). According to developmental stages we investigated the positional cloning for the candidate genes. Map-based cloning and micro array were also tried for expression analysis. For those group of markers, at least nine BAC clone pools were specified. And after the preparation of transformation system ASG-1 candidate gene was tried to induction by three kinds of binary vectors. Now the regeneration of those transgenic plants is undertaken.Diplospory of Chinese chive(Allium tuberosum) was also investigated by the following methods ; the isolation of mRNA, gene cloning of differential expression of candidate genes, and then to analyze of micro arrays for 200 different expressions. Unfortunately there are nothing concerned with them on the synchronism to apomictic expression to now.2.Differentiation Control and hybrid lethality for reproductive system.The understanding of inter-specific hybrid lethality is useful tool not only for overcoming breeding barriers, but also to control of cellular differentiation on reproductive system. The expression of lethality was investigated by controlling temperature during growth of embryo. During the process of inter-specific hybrid death, degeneration of nucleus, breakage of nuclear DNA at nucleosome unite and fragmentation of nucleus have occurred in different degrees. Cells expressed programmed cell death, apoptosis. It doesn't concerned on cyrtoplasmic factors, but located on a certain gene locus in chromosome and to be concerned to ethylene formation.