盐肤木体细胞胚胎发生及植株再生
投稿时间:2023-03-08  修订日期:2023-07-12  点此下载全文
引用本文:杨庆敏,尹飞飞,龚清,等.盐肤木体细胞胚胎发生及植株再生[J].西北植物学报,2023,43(8):1369~1377
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杨庆敏三峡区域植物遗传与种质创新重点实验室三峡大学/三峡大学生物技术研究中心 460408915@qq.com 
尹飞飞三峡区域植物遗传与种质创新重点实验室三峡大学/三峡大学生物技术研究中心 956096936@qq.com 
龚清三峡区域植物遗传与种质创新重点实验室三峡大学/三峡大学生物技术研究中心 1289274241@qq.com 
杨洋三峡区域植物遗传与种质创新重点实验室三峡大学/三峡大学生物技术研究中心 2293633648@qq.com 
桑子阳五峰博翎种业有限公司 554635334@qq.com 
刘文* 三峡区域植物遗传与种质创新重点实验室三峡大学/三峡大学生物技术研究中心 liuwen@ctgu.edu.cn 
基金项目:宜昌市科技研究与开发项目(A21-4-013)、五峰土家族自治县林业科学研究所委托项目(SDHZ2021081)
中文摘要:体细胞胚胎发生及植株再生体系可为种苗繁育提供有效的技术手段,并为遗传转化体系奠定基础。为建立盐肤木(Rhus chinensis Mill.)体细胞胚胎发生及植株再生体系,本研究以盐肤木幼胚为外植体,从植物生长调节剂组合、基本培养基类型和蔗糖浓度等方面探索初代愈伤组织诱导、胚性愈伤组织诱导和增殖、以及体细胞胚胎诱导的最佳条件。实验结果显示,最佳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1/2MS+6-BA 2 mg/L+NAA 0.5 mg/L培养基上培养一个月后,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MS+6-BA 0.2 mg/L+NAA 0.5 mg/L+蔗糖4%的培养基上培养一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。本研究建立了盐肤木体细胞胚胎发生及植株再生体系,所获得的再生苗成活率高。
中文关键词:盐肤木  愈伤组织  体细胞胚胎  细胞学观察
 
Somatic Embryogenesis and Plant Regeneration of Rhus chinensis
Abstract: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.
keywords:Rhus chinensis  callus  somatic embryos  cytological observation
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