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草业学报 ›› 2024, Vol. 33 ›› Issue (7): 172-181.DOI: 10.11686/cyxb2023294

• 研究论文 • 上一篇    

兰州百合多倍体离体诱导及其分子细胞鉴定

符勇耀1(), 蔡莉1, 李丰耀1, 杨文俊1, 徐文姬1, 姜思佳2(), 杨利平1()   

  1. 1.长江师范学院现代农业与生物工程学院,重庆 408100
    2.黑龙江省森林植物园,黑龙江 哈尔滨 150040
  • 收稿日期:2023-08-29 修回日期:2023-10-26 出版日期:2024-07-20 发布日期:2024-04-08
  • 通讯作者: 姜思佳,杨利平
  • 作者简介:yangliping1962@126.com
    E-mail: zwyjsj@yeah.net
    符勇耀(1982-),男,四川眉山人,副教授,博士。E-mail: 34331107@qq.com
  • 基金资助:
    重庆市涪陵区科技局技术创新与应用发展项目(2023AAN1001);重庆市教委科学技术研究项目(KJQN202001418);重庆市科技局自然科学基金项目(cstc2020jcyj-msxmX0492);国家林业和草原局科技发展中心项目(KJZXSA202211)

Induction and molecular and cellular identification of polyploid Lilium davidii var. unicolor

Yong-yao FU1(), Li CAI1, Feng-yao LI1, Wen-jun YANG1, Wen-ji XU1, Si-jia JIANG2(), Li-ping YANG1()   

  1. 1.School of Advanced Agriculture and Bioengineering,Yangtze Normal University,Chongqing 408100,China
    2.Forest Botanical Garden of Heilongjiang Province,Harbin 150040,China
  • Received:2023-08-29 Revised:2023-10-26 Online:2024-07-20 Published:2024-04-08
  • Contact: Si-jia JIANG,Li-ping YANG

摘要:

兰州百合是我国最重要的可食用甜百合。为培育优质的百合新种质,用不同浓度的秋水仙素离体诱变兰州百合小鳞茎,通过不定芽诱导获得变异幼苗。利用染色体倍性分析、幼苗早期形态观察及ISSR分子标记对其进行鉴定。结果表明,用0.15%秋水仙素处理小鳞茎48 h诱导效果最好,变异率为58.06%。选择2个形态变异且生长较快的幼苗进行流式细胞仪分析和根尖压片计数,发现其中1个变异株系含36条染色体的细胞数比例达到了80%,接近纯合三倍体。形态学观察显示,2个变异幼苗的叶片、鳞茎及根系与对照相比存在显著差异,且气孔保卫细胞大于对照,气孔密度降低。ISSR标记检测表明,2个诱变株系与对照相比均发生遗传变异,变异率分别为19.64%和20.91%。研究结果可为获得兰州百合三倍体新种质及秋水仙素诱变育种提供重要参考依据。

关键词: 兰州百合, 多倍体, 秋水仙素, ISSR标记

Abstract:

Lilium davidii var. unicolor is the most important edible sweet lily in China. To cultivate new, high-quality germplasm of L. davidii var. unicolor, bulblets growing in vitro were treated with different concentrations of colchicine, and mutagenic seedlings were obtained via adventitious bud induction. The variant lines were further identified by chromosome ploidy level, morphological, and inter simple sequence repeat (ISSR) molecular marker analyses. The results indicated that treatment of the bulblets with 0.15% colchicine for 48 h was the optimal induction method, leading to the maximum mutation rate of 58.06%. Two fast-growing seedlings with stable morphological variations were selected for chromosome ploidy analysis by flow cytometry and tableting in the root tip. The proportion of cells containing 36 chromosomes in the root tip of one induced line was 80%, indicative of a homozygous triploid. Compared with the control, the seedlings of the mutated lines showed significant differences in the leaf, bulblet, and roots. The guard cells were larger and the stomatal density was significantly reduced in the mutated lines compared with the control. Analyses of ISSR markers revealed genetic variations in the two mutagenic lines compared with the control, with mutation rates of 19.64% and 20.91%, respectively. Together, these results provide an important basis for obtaining triploid germplasm of L. davidii var. unicolor, and for breeding of new lily materials using colchicine treatment.

Key words: Lilium davidii var. unicolor, polyploid, colchicine, ISSR marker