高山杜鹃离体叶片高效再生及其再生途径研究

为建立高效的高山杜鹃植株再生体系,以6种(‘Percy Wiseman’、‘Cunnigham’s White’、‘Halfdem Lem’、‘Madame Masson’、‘Scintillation’、‘XXL’)高山杜鹃组培苗叶片为材料,研究高山杜鹃离体叶片分化的基因型效应和外源激素效应,并通过形态学和组织细胞学研究其再生途径。结果表明,TDZ诱导6种高山杜鹃离体叶片再生的效果好于ZT,0.04~0.08 mg/L TDZ诱导离体叶片的分化率可达100%,叶片平均分化不定芽数为5.67~13个/片。6种高山杜鹃均可作为遗传转化的候选材料,其中‘Scintillation’、‘Halfdem Lem’和‘Cunnigham’s White’的再生能力更强。6种高山杜鹃离体叶片的再生途径主要为器官直接再生,其中‘Percy wiseman’和‘Madame Masson’除器官直接发生途径外,在TDZ诱导下少量叶片存在愈伤组织间接再生途径分化不定芽。试验结果可为研究高山杜鹃离体叶片再生芽机理,提高其植株再生频率和遗传转化效率提供技术支持。

Efficient Regeneration Method and Pathway of in Vitro Leaves of Rhododendron

In vitro seedling leaves of six Rhododendron cultivars (‘Percy Wiseman’, ‘Cunningham's White’, ‘Halfdem Lem’, ‘Madame Masson’, ‘Scintillitation’ and ‘XXL’) were took as materials to establish efficient plant regeneration system. The effects of genotypes and exogenous hormones on the differentiation of in vitro leaves of six Rhododendron cultivars were studied, and the regeneration pathway was studied by morphology and histocytology. The results showed that TDZ was better than ZT in inducing the regeneration of in vitro leaves for the six Rhododendron cultivars. The differentiation rate of in vitro leaves induced by TDZ 0.04-0.08 mg/L was 100% and the average number of adventitious buds was 5.67-13 per leaf. The six Rhododendron cultivars could be used as candidate materials for genetic transformation, among which ‘Scintillition’, ‘Halfdem Lem’ and ‘Cunnigham’s White’ had stronger regeneration ability. Direct organ regeneration was the main regeneration pathway for the six Rhododendron cultivars, besides, there was also a few of leaves with callus indirect regeneration pathway to differentiate adventitious buds in ‘Percy Wiseman’ and ‘Madame Masson’ under TDZ induction. The results of the study could provide technical support and an experimental basis for the research on the mechanism of in vitro leaves differentiation of Rhododendron, and improve its plant regeneration frequency and genetic transformation efficiency.