小叶杨原生质体培养植株再生及其同工酶的变化

PLANTS REGENERATED FROM PROTOPLASTS OF POPULUS SIMONII AND VARIATIONS IN ITS ISOENZYME

从小叶杨（Ｐｏｐｕｌｕｓｓｉｍｏｎｉｉ）胚性悬浮细胞分离得到大量有活力的原生质体，产量高达３．８×１０７ｇ－１ＦＷ。高密度（３×１０６／ｍＬ）液体浅层培养可促进小叶杨原生质体的分裂和生长；以葡萄糖为唯一碳源的Ｋｍ８ｐ培养基，有利于原生质体的持续分裂；在培养基中添加有机氮，有助于提高原生质体的植板率。原生质体形成细胞团后，逐步降渗和分皿培养，能有效促进细胞团的持续分裂。愈伤组织在不同分化培养基上的分化频率存在着明显的差异，同工酶分析说明，分化能力强的愈伤组织，其ＴＣＡ循环和磷酸戊糖途径较为活跃，ＧＯＴ、ＥＳＴ和ＰＥＲＯＸ的活性比较高。当芽伸长到２—３ｃｍ时，从基部切下转移到无激素的１／２ＭＳ培养基上诱导生根并形成完整植株。移入盒内，在温室生长良好。

A large number of Populus simonii vigorous protoplasts were isolated from embryonic cell suspension culture. The yield of these protoplasts amounted to 3.8 × 107g-1FW. The protoplasts were cultured in a Km8p liquid medium containing 2, 4-D 3.0mg/L, NAA 0.2mg/L, and KT 0.2mg/L, and in higher density (3 × 106/mL) and thinner layer, which have been proved to be favourable to the division and growth of the protoplasts. When glucose was used in the medium as the only carbon source, it was of benefit to the sustained division of protoplasts. The protoplast plating effeciency increased when the medium contained organic nitrogen. It could be favourable for further division to decrease osmotic pressure and seperate mass into several dishes when cell colonies formed from protoplast. There is a evident difference in differentiation frequency of the calli on various differentiation media. The isoenzyme analysis results showed that the phosphopentose pathway and TCA cycle were more active in the calli with high ability of shoot formation, and the activities of GOT, EST, and PEROX were higher. The shoots of 2 - 3cm in height were cut off from the calli and rooted on 1 / 2 MS medium.After transplantation into pots, the regenerated plants grew vigorously in greenhouse.