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中国农学通报

中国农学通报 ›› 2020, Vol. 36 ›› Issue (10): 69-78.doi: 10.11924/j.issn.1000-6850.casb18120105

• 林学 园艺 园林 • 上一篇    下一篇

国内外8个不同种源地莲(Nelumbo)的成熟胚离体培养比较

秦密1,2,3(), 刘凤栾2,3, 刘青青2,3, 曹建国1, 陈煜初4, 田代科2,3()   

  1. (1)上海师范大学生命科学学院,上海 200234
    (2)上海辰山植物园/中国科学院上海辰山植物科学研究中心,上海 201602
    (3)上海市资源植物功能基因组学重点实验室,上海 201602
    (4)浙江人文园林股份有限公司/杭州天景水生植物园,杭州 310000
  • 收稿日期:2018-12-25 修回日期:2019-02-19 出版日期:2020-04-05 发布日期:2020-03-19
  • 基金资助:
    上海市绿化和市容管理局攻关项目“荷花不同育种方法比较及杂交性状遗传规律探讨”(G162402);上海市绿化和市容管理局攻关项目“中国野生莲资源的多样性及保护”(G182412)

Comparis on In Vitro Culture of Mature Embryos of Nelumbo from Eight Provenances Worldwide

Mi Qin1,2,3(), Fengluan Liu2,3, Qingqing Liu2,3, Jianguo Cao1, Yuchu Chen4, Daike Tian2,3()   

  1. (1)College of Life Sciences, Shanghai Normal University, Shanghai 200234
    (2)Shanghai Chenshan Botanical Garden/ Shanghai Chenshan Plant Science Research Center of Chinese Academy of Sciences, Shanghai, 201602
    (3)Shanghai Key Laboratory of Plant Functional Genomics and Resources, Shanghai 201602
    (4)Zhejiang Humanities Landscape Co., Ltd./ Hangzhou Tianjing Aquatic Botanical Garden, Hangzhou 310000
  • Received:2018-12-25 Revised:2019-02-19 Online:2020-04-05 Published:2020-03-19

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摘要：

建立高效获取无菌莲胚外植体的消毒体系,了解不同种源地莲的离体培养生长差异,为今后构建莲的优良离体培养及遗传转化体系提供可靠的材料和评估方法。使用不同浓度植物保护混合剂(plant preservative mixture,PPM)预培养法确定适宜的莲子消毒体系和无菌莲胚获取方式;在此基础上获得国内外9个种源地莲的成熟胚离体培养幼苗,通过观测其在不同培养时期的形态指标,利用隶属函数法综合评价其离体条件下的生长差异。研究发现,莲子比较理想的消毒体系为75%酒精消毒30 s + 2% PPM消毒2 h + 0.05% PPM浸泡催芽3~5天。其中,8个参试种源地莲的离体生长差异主要体现在培养后期,表现在展开叶数、茎节长度和粗度、根数及侧芽数等指标上。在相同培养条件下,越南莲和泰国莲的生长势最好、繁殖率较高。使用PPM浸泡成熟莲子进行预培养以获取无菌莲胚的方式,克服了以往莲子消毒不理想的技术难题。热带、亚热带分布的越南莲和泰国莲的离体无性系生长势强,是构建莲再生体系和遗传转化体系的良好原材料。

关键词: 莲属, 野生型莲, 莲胚, 消毒, 植物保护混合剂, 离体培养, 隶属函数

Abstract:

Establishing a feasible disinfection system for obtaining the germfree mature lotus (Nelumbo) embryos as explants and understanding the differences of in vitro growth of lotus from different provenances is the basis for material selection and evaluation method to construct an efficient in vitro culture and genetic transformation system. Plant preservative mixture (PPM) at different concentrations was used in the pre-culture stage to develop a suitable disinfection system of lotus seeds and a method of acquiring the germ-free lotus embryos. Through this sterilization approach, the in vitro cultured seedlings of lotus from 9 worldwide provenances were successfully obtained. Based on the morphological indexes at different culture periods, the membership function method was used to evaluate the growth differences under the in vitro culture conditions. The results showed that the ideal disinfection system for lotus seeds was 75% alcohol 30 s + 2% PPM 2 h + 0.05% PPM 3-5 d soaking during germination. The in vitro growth differences of the seedlings from the tested lotuses of 8 provenances were mainly observed in the later culture stage, such as the number of leaves, the length and thickness of rhizomes, the number of roots and lateral shoots. The seedlings of lotus from Vietnam and Thailand had the highest growth potential and higher reproduction rate under the same culture conditions. The pre-culture method by soaking lotus seeds with PPM to successfully obtain germ-free lotus embryos has solved the technique problem in disinfecting lotus seeds in the past. The in vitro clones of lotus embryos from the tropical or subtropical regions in both Vietnam and Thailand have stronger growth capacity and therefore are the materials more ideal for establishing regeneration and genetic transformation system of Nelumbo.

Key words: Nelumbo, wild type lotus, lotus embryo, sterilization, plant preservative mixture, in vitro culture, membership function

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