三叶半夏种质资源扩繁及保存研究  

Propagation and Preservation of Germplasm Resources in Pinellia ternata(Thunb.)Breit.

作　　者：贾明良[1] 方荷芳 张本厚 胡燕花 周安佩 李同建[1] 金洪光[1] 韩兴杰[1] 文锋[1] JIA Ming-liang;FANG He-fang;ZHANG Ben-hou;HU Yan-hua;ZHOU An-pei;LI Tong-jian;JIN Hong-guang;HAN Xing-jie;WEN Feng(Jiu Jiang University,Jiujiang 332000,China;Dafeng Marine Industrial Institute,Nanjing Tech University,Yancheng 224100,China)

机构地区：[1]九江学院,九江332000 [2]南京工业大学大丰海洋产业研究院,盐城224100

出　　处：《中国生物工程杂志》2022年第11期18-26,共9页China Biotechnology

基　　金：江西省自然科学基金面上项目(20212BAB205024);江西省教育厅科技项目(GJJ201824);江西省中医药管理局科技计划(2022B1019)资助项目。

摘　　要：目的:通过对三叶半夏扩繁和保存方法的研究,为三叶半夏种质资源的保育和开发利用提供技术支撑。方法:首先,通过灭菌条件筛选、愈伤诱导及丛生芽诱导等进行扩繁条件探索;其次,通过设置不同基本培养基、蔗糖浓度、不同温度和光照强度等条件探索合适的种质保存条件;最后,探索了休眠块茎唤醒方式和丛生芽再诱导方法,为种质资源再扩繁提供基础。结果:以叶柄为外植体,升汞灭菌12 min后,接种至添加2,4-D的培养基中可诱导疏松愈伤组织。外植体接种至MS+2.0 mg/L 6-BA+0.1 mg/L NAA+蔗糖30 g/L+琼脂6.0 g/L,pH 5.8中可进行丛生芽诱导。合适的种质保存基本培养基为MS和N6培养基;合适的蔗糖浓度为30 g/L,过低或过高均不利于种质保存;在丛生块茎诱导时需要提供足够的温度和光照,低温及避光条件不利于种质保存。待丛生芽自然倒苗休眠后可进行长达1~2年的离体保存。对休眠块茎进行唤醒诱导时可采用切割块茎后再接种的方式。萌发出来的叶柄直接进行丛生芽诱导增殖,而块茎则接种至MS+6-BA 1.0 mg/L+NAA 0.2 mg/L+蔗糖30 g/L+琼脂6.5 g/L,pH 5.8中进行再分化诱导丛生芽。诱导出来的丛生芽进行规模化扩繁或继续进行离体保存。结论:通过对三叶半夏的种质扩繁,筛选合适的条件得到休眠丛生块茎,可对三叶半夏进行长时间的离体保存,而后唤醒和再次诱导的相关研究,为其种质资源保存、利用提供技术基础。Objective:To provide technical support for the conservation,development and utilization of Pinellia ternata(Thunb.)Breit.(P.ternata)germplasm resources by studying the propagation and preservation methods of P.ternata.Methods:Firstly,the propagation conditions were explored by screening sterilization conditions,callus induction and cluster buds induction.Secondly,the suitable conditions for germplasm preservation were explored by setting different basic media,sucrose concentration,temperature and light intensities.Finally,the methods of awakening dormant tubers and reinducing clumping buds were explored to provide a basis for the re-propagation of germplasm resources.Results:The petiole explants were sterilized with mercuric chloride for 12 minutes and inoculated into the medium supplemented with 2,4-D to induce loose callus.The explants were inoculated to MS+2.0 mg/L 6-BA+0.1 mg/L NAA+30 g/L sucrose+6.0 g/L agar,pH 5.8 for cluster buds induction.MS and N6 medium was the appropriate medium for germplasm preservation.The appropriate sucrose concentration is 30 g/L,and either too low or too high sucrose concentration was not conducive to germplasm preservation.It is necessary to provide sufficient temperature and light during tuber induction.Low temperature and light avoidance conditions are not conducive to germplasm preservation.It can be stored in vitro for up to 1-2 years after the sprout tumble of cluster buds.The wake-up induction of dormant tubers can be induced by cutting tubers and then inoculating them.The germination of the petiole can be directly induced by cluster buds proliferation,while the tubers can be inoculated into MS+6-BA 1.0 mg/L+NAA 0.2 mg/L+sucrose 30 g/L+agar 6.5 g/L,pH 5.8 to induce cluster buds re-differentiation.The cluster buds can be propagated or preserved in vitro.Conclusion:Through the propagation of germplasm of P.ternata,the dormant cluster tubers can be obtained by screening appropriate conditions,which can be preserved in vitro for a long time,and then awakened and induced again

关 键 词：三叶半夏 种质资源 组培快繁 种质保存 

分 类 号：Q819[生物学—生物工程]