盐肤木体细胞胚胎发生及植株再生

以盐肤木(Rhus chinensis Mill.)幼胚为外植体,研究不同植物生长调节剂组合对其愈伤组织诱导及体细胞胚胎发生的影响,以建立盐肤木体细胞胚胎发生及植株再生体系。结果表明,最适愈伤组织诱导培养基为MS+6-BA 0.2 mg/L+2,4-D 1.0 mg/L,诱导率为84.57%,诱导出的初代愈伤组织白色或淡黄色,质地疏松,表面光滑,为非胚性愈伤。初代愈伤组织转移到1/2 MS+6-BA 2 mg/L+NAA 0.5 mg/L培养基上培养1个月后,长出淡黄色质地紧密的胚性愈伤组织,诱导率高达100%,在此培养基上胚性愈伤组织增殖倍数为854.73%。所获得的胚性愈伤组织转接到1/2 MS+6-BA 2 mg/L+NAA 0.5 mg/L+蔗糖4%的培养基上培养1个月后可诱导体细胞胚胎发生,诱导率可达32.67%。诱导得到的体细胞胚胎经历球形胚、心形胚、鱼雷胚、子叶胚进一步分化发育成苗。无菌苗炼苗后栽种到泥炭土∶蛭石∶珍珠岩为2∶1∶1的生长基质上,能100%稳定成活。经过细胞学观察分析,体细胞胚的发育与合子胚相似。

Somatic Embryogenesis and Plant Regeneration of Rhus chinensis

Somatic embryogenesis and plant regeneration system can provide effective technical means for seedling breeding and lay the foundation for genetic transformation system. In order to establish the embryogenesis and plant regeneration system of Rhus chinensis, the immature zygotic embryos of Rhus chinensis were selected as explants in this study, and the optimal conditions for the induction of primary callus, the induction and proliferation of embryonic callus as well as the induction of somatic embryo were expiored from the combination of plant growth regulators, the type of basic medium and the concentration of sucrose. Our results showed that the optimal medium for callus induction was MS+6-BA 0.2 mg/L+2,4-D 1.0 mg/L, with an induction rate of 84.57%. The newly induced callus was white or light yellow, with loose texture and smooth surface. When the callus was transferred to 1/2MS+6-BA 2 mg/L+NAA 0.5 mg/L medium for one month, the embryogenic callus with light yellow texture generated, with the induction rate up to 100%, and the multiplication of embryogenic callus on this medium was 854.73%. The embryogenic callus were transferred to MS+6-BA 0.2 mg/L+NAA 0.5 mg/L+sucrose 4% medium for one month to induce somatic embryogenesis with an induction rate of 32.67%. The somatic embryos developed into plantlets through globular embryo, heart-shaped embryo, torpedo embryo and cotyledon embryo. After trainin, sterile seedlings can be planted to the growth matrix (peat: vermiculite:perlite= 2:1:1), with survival rate of 100%. Taken together, a somatic embryogenesis and plant regeneration system of Rhus chinensis was established in this study, and the survival rate of the regenerated seedlings obtained was high.