应用微室饲养培养系统诱导水稻和小麦合子及早期原胚的生长

Growth of Zygotes and Early Proembryos in Rice and Wheat Using Microculture System with Feeder Cells

从两个水稻品种、 4种外植体来源的愈伤组织中 ,筛选出幼胚愈伤组织 ,通过反复继代和悬浮培养 ,建立了具旺盛分裂能力的胚性悬浮细胞系。胚性悬浮细胞为圆球形、单个或聚集成多细胞团 ;细胞以多方向分裂 ;并具有长期的分裂能力。以胚性悬浮细胞系作为饲养细胞 ,采用微室饲养培养系统诱导了水稻和小麦合子及授粉后 1～ 2 d早期原胚的生长。结果表明 :水稻合子分裂成多细胞团 ;1d原胚发育成球形胚 ;2 d原胚通过器官发生途径再生植株。小麦合子和 2～ 8胞原胚能分裂多次 ,形成细胞团。无饲养细胞的微室培养不能诱导合子和早期原胚分裂 ;

In order to induce the growth of zygotes and the early proembryos of rice ( Oryza sativa L.) and wheat ( Triticum aestivum L.), an efficient rice suspended cell line as a feeder system was established. For this purpose, calli were induced from two cultivars and four kinds of explants, i.e. anther, young panicle, young embryo and mature embryo. The calli originated from young embryos were selected as the best to build up an embryonic suspended cell line with vigorous division through continuous subculture and cell suspension culture. The embryonic cells were spherical shaped, single or gathered into multicellular clusters. Zygotes and early proembryos of rice and wheat 1～2 days after pollination were isolated and inoculated in Millicell (microchamber) and co cultivated with the feeder cells in Petri dishes. Zygotes of both rice and wheat could divide into multicellular structures. The 1 day old early proembryos of rice with 4 32 cells could develop into globular embryos, while the 2 day old proembryos could develop and regenerate plantlets via the way of organogenesis. Using suspended feeder cell system of rice, we were also able to induce 2～8 celled proembryos of wheat to divide into multicellular structures. Without feeder cells, the zygotes and early proembryos cannot divide in Millicell culture. The division frequency of 2 day old proembryos in rice was higher in co cultivation with suspended embryonic feeder cells than in that with small granular calli.