落叶松体细胞胚胎发生诱导多倍体的研究

Study of Polyploid Induction by Somatic Embryogenesis of Larix ssp.

通过检测细胞系增值率及细胞分裂指数确定细胞系分裂最旺盛时期,利用秋水仙素和氨磺灵诱导落叶松多倍体体细胞胚胎的发生,揭示了两种抗微管物质对落叶松体细胞胚胎发生的影响。秋水仙素在浓度为500 mg·L^-1,浸泡处理36 h时能使多倍体体细胞胚胎发生的比例达到85.2%,体细胞胚发生率达到351.1个/g;氨磺灵在1-5 mg·L^-1的浓度下处理后的细胞系几乎失去体细胞胚分化能力,无法大量获得多倍体体细胞胚。通过细胞压片及去壁-低渗涂片对比观察发现氨磺灵诱导后的细胞系除了染色体数目加倍外还有部分细胞发生程序性死亡。该研究初步证明了秋水仙素较适合落叶松多倍体体细胞胚的诱导,氨磺灵对落叶松毒害明显,不适合其多倍体的诱导。

Vigorous growth period of Larix cell lines were investigated by cell mitotic index,and two antimicrotubule agents colchicine and oryzalin were evaluated for their ability on inducing polyploidy of Larix by somatic embryogenesis. Colchicine was found to be the most effective on inducing polyploidy embryos at the concentration of 500 mg·L^-1 for 36 h, the inductive frequency was 85.2% and somatic embryogenic ratio was 351.1 /g. However, the oryzaline could not induce polyploidy of Larix in normal somatic embryos under the concentration from 1 mg·L^-1 to 5 mg·L^-1. Using the squash method,wall degradation and hypotonic treatment, both chromosomes and cytological characters change of the cell lines were induced. The results showed that the chromosome number were double in some cells after induced by colchicines solution, and apoptotic bodies were found in most of the cells induced by oryzalin.