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

1.部分作物无融合生殖形成的细胞和分子遗传学分析用Nomarski微分干涉相差显微镜观察脱孢子起始细胞（AIC）的表达过程。根据发育阶段的不同，对候选基因进行了定位克隆。图位克隆和微阵列也被尝试用于表达分析。对于那些标记物组，指定了至少9个BAC克隆库。并在转化体系制备完成后，尝试用三种双元载体对ASG-1候选基因进行诱导。本研究还对韭菜（Allium tuberosum）二倍体孢子生殖进行了研究，通过mRNA的分离、差异表达候选基因的基因克隆和200个差异表达基因的微阵列分析，获得了200个差异表达基因。2.生殖系统的分化控制和杂种致死性对种间杂种致死性的认识不仅有助于克服育种障碍，而且有助于控制生殖系统的细胞分化。通过控制胚生长过程中的温度来研究致死率的表达。在种间杂种死亡过程中，不同程度地发生了细胞核的变性、核小体处DNA的断裂和细胞核的碎裂。细胞表达程序性细胞死亡、凋亡。它与胞质因子无关，但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.