Huang He-Ping, Gao Shan-Lin, Chen Lan-Lan, Wei Kun-Hua
Department of Genetics and Breeding, China Pharmaceutical University, Nanjing, 211198, R.P. China.
Pharmacogn Mag. 2010 Jan;6(21):51-6. doi: 10.4103/0973-1296.59966. Epub 2010 Feb 13.
This article describes an efficient colchicine-mediated technique for the in vitro induction of tetraploids in Dioscorea zingiberensis and its confirmation by flow cytometry. Buds immersed in 0.2% colchicine solution for 36 hours prior to culture induced as high as 35.6% tetraploid plants. Colchicine-induced tetraploids remained stable after six months in soil. Leaf characteristics of diploids and tetraploids in D. zingiberensis were compared. It was determined that the leaf sizes of glasshouse-grown plants and stomatal sizes of both in vitro and glasshouse-grown plants were suitable parameters for identifying putative tetraploids in D. zingiberensis. Besides generating tetraploids, this technique generated mixoploids in D. zingiberensis. Calli derived from mixoploid leaves were induced to form buds and shoots. Individual shoots were classed as diploid, mixoploid, and tetraploid by flow cytometry. This callus-based technique could be employed when a genome-doubling agent generated mixoploids, but no tetraploids.
本文描述了一种用秋水仙碱介导的高效技术，用于在体外诱导盾叶薯蓣四倍体，并通过流式细胞术对其进行确认。在培养前将芽浸入0.2%秋水仙碱溶液中36小时，诱导出高达35.6%的四倍体植株。秋水仙碱诱导的四倍体在土壤中生长六个月后仍保持稳定。比较了盾叶薯蓣二倍体和四倍体的叶片特征。确定温室种植植株的叶片大小以及体外和温室种植植株的气孔大小是鉴定盾叶薯蓣假定四倍体的合适参数。除了产生四倍体，该技术还在盾叶薯蓣中产生了混倍体。源自混倍体叶片的愈伤组织被诱导形成芽和枝条。通过流式细胞术将单个枝条分类为二倍体、混倍体和四倍体。当基因组加倍剂产生混倍体但不产生四倍体时，可以采用这种基于愈伤组织的技术。
