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茶树组织培养是开展茶树次生代谢及调控研究重要的研究平台。在建立茶树愈伤组织培养体系的基础上,开展了茶树含不同儿茶素含量的细胞系筛选方法的研究。结果表明,在H、S和B 3种诱导培养基中,只有B培养基适合于诱导生长迅速、组织疏松、质地均一且具有一定儿茶素合成能力的愈伤组织。从外植体方面考虑,种子诱导出的愈伤组织具有较强的植株再生能力;幼茎和叶片诱导出的愈伤组织,其生长速度快、组织疏松,但再生能力弱。在此基础上,采用目视法和二分法,结合HPLC分析,对B培养基诱导的细胞系“yunjing63Y”进行了筛选,得到含不同儿茶素含量的细胞系“yunjing63Y”和“yunjing63X”。分析结果显示,细胞系“yunjing63Y”和“yunjing63X”的儿茶素组分与鲜叶相似,且二者儿茶素含量(干重)差异较大,前者为7.77 mg·g-1,后者为0.13 mg·g-1。
Tea tree tissue culture is an important research platform for secondary metabolism and regulation of tea. Based on the establishment of tea tree callus culture system, the screening of cell lines with different catechins content in tea tree was carried out. The results showed that only B medium was suitable for inducing callus with rapid growth, loose tissue, uniform texture and certain catechin synthesis ability among three kinds of H, S and B induction medium. In terms of explants, the callus induced by seeds had strong plant regeneration ability. The callus induced by young stems and leaves had fast growth and loose tissue but weak regenerative ability. On this basis, using the visual method and the dichotomy, combined with HPLC analysis, the cell line “yunjing63Y” induced by B medium was screened to obtain cell lines containing different catechin contents “yunjing63Y” and “yunjing63X ”. The results showed that the catechin components of the cell lines “yunjing63Y” and “yunjing63X” were similar to fresh leaves, and the content of catechins (dry weight) was quite different. The former was 7.77 mg · g- 1, the latter is 0.13 mg · g-1.