陆地棉体细胞胚胎发生过程的cDNA-AFLP分析

cDNA-AFLP Analysis of Somatic Embryogenesis at Early Stage in TM-1(Gossypium hirsutum L.)

以陆地棉标准系TM-1未经诱导的下胚轴、下胚轴经诱导40 d的愈伤组织和胚性愈伤组织为材料,利用cDNA-AFLP差异显示技术对陆地棉体细胞胚胎发生过程中的cDNA差异表达进行了初步分析.经反转录获得3个不同时期的cDNA,利用180对引物组合进行AFLP分析,结果表明,在总共显示的约3 000条谱带中,其中38条为胚性愈伤组织中所特有的差异谱带.对这些差异片段进行克隆、序列测定和同源性分析,其中12个差异片段同已知陆地棉、雷蒙德氏棉和亚洲棉不同发育时期器官的EST序列高度同源,其相似性达到90%以上.结果说明,棉属在形态建成早期(胚胎发育阶段),其特定器官的相关基因已经表达.

We used cDNA-AFLP technique to analyze gene expression profiles during somatic embryogenesis of hypocotyls section （explant） ,callus after forty days induction on medium of MS and somatic embryo in cotton （Gossypium hirsutum L. ） TM-1. cDNA-AFLP was analyzed in double strand cDNA after reverse transcription from the three different development stages tissues of TM-1. One hundred and eighty combinations of AFLP primers produced about 3 000 fragments. 38 specific bands were identified in the somatic embryo. These cDNA fragments were further cloned and sequenced, and sequence analysis demonstrated that there were twelve sequence-similar to cDNA sequences with the different tissues of Gossypium raimondii L.. Gossypium hirsutum L. and Gossypium arboretum L. in GenBank among 38 sequenced fragments. Their identities were above ninety percent. The results indicated that the gene expression of whole plant had already occured at the early stage of embryogenesis.