摘要
华北落叶松(Larix principis-Rupprechtii)是我国北方中高山地区重要的针叶速生用材树种,进行其体细胞胚胎发生和植株再生的研究,在针叶树无性快速繁殖及基因工程育种上有其特殊的用途,既可为针叶树无性系林业提供产业化途径,也可作为目的基因遗传转化实验系统。针叶树的基因转化相对较难,再生更属不易,Lelu等报道过杂种落叶松与欧洲落叶松体细胞胚胎发生方面的研究;而我国尚未见有落叶松体细胞胚胎发生的研究报道。
Larix principis-Rupprechtii is one of the superior afforestation forest trees growing in north China. Embryogenic cultures were initiated from immature zygotic embryos of Larix principis-Rupprechtii on S culture medium containing 2, 4-D 0-2. 2mg/L, KT and BA each at 0 - 0. 8mg/L. Embryogenic calli were subcultured and multiplicated on S+B culture medium containing dropping off each hormone concentration. We set up 33 steady-going embryogenic cell lines; We studied on the growth stage and genotype differences of every embryogenic cell lines ; and Finded more than 10 high-frequency somatic embryogen-esis cell lines such as 2K,2T,2I,2J,3C etc. . The number of 2T somatic embryos reaches 314/ per gram of embryogenic tissue and the number of 3C
somatic embryos is 185/per gram of embryogenic tissue. The re-induction method of Larix principis-Rupprechtii from somatic embryos was used to produce renewable embryogenic cultures and steady-going embryogenic cell lines effectively. Mature somatic embryos can germinate and develop further into plantlets when they are isolated and cultured on a hormone-free WPM culture medium. The regeneration plantlets were obtained. Furthermore, the transformation with a truncated gene of Bacillus thuringensis (B. t) were carried out, the PCR showed positive results, because of this, embryogenic cell line of Larix principis-Rupprechtii can be used for transformation experiments to support further breeding in forestry.
出处
《实验生物学报》
CSCD
2000年第4期354-365,共12页
Acta Biologiae Experimentalis Sinica
基金
国家自然科学基金(批准号:No 39870632)
国家林业局"948"项目(批准号:No.98-4-04-02)
关键词
华北落叶松
体细胞
胚胎发生
遗传转化
Larix principis-Rupprechtii. Somatic embryogenesis. Embryogenic cell Lines. Re-in- duction
