摘要
The study has established the F2 offspring obtained by crossing salt-tolerant with salt-sensitive alfalfa, and appraised the salt-tolerant F2 offspring seedling was evaluated in pot culture. With the F2 segregated population, the research has obtained a molecular marker linked with salt-tolerant genes of alfalfa using the improved BSA combined with RAPD. The RAPD PCR products were excised from the agarose gel and purified using a kit, then were mixed with pMD-18T vector and sequenced. Sequencing result indicated the RAPD marker was 1 438 bp in length. Similarity researches using blast in Genbank indicated that the nucleotide sequence of the RAPD marker showed 93% and 91% similarity with mth2-6el8 gene fragment （347 bp） and ruth2-33122 gene fragment （334 bp） of Medicago truncatula respectively. Medicago truncatula is a close relative of alfalfa and Mth2-6el8 is a molecular marker of the gene coding for a cysteine protease which was saltinducible in some plants. These results indicated the RAPD marker was possibly related to cysteine protease genes in alfalfa.
The study has established the F2 offspring obtained by crossing salt-tolerant with salt-sensitive alfalfa, and appraised the salt-tolerant F2 offspring seedling was evaluated in pot culture. With the F2 segregated population, the research has obtained a molecular marker linked with salt-tolerant genes of alfalfa using the improved BSA combined with RAPD. The RAPD PCR products were excised from the agarose gel and purified using a kit, then were mixed with pMD-18T vector and sequenced. Sequencing result indicated the RAPD marker was 1 438 bp in length. Similarity researches using blast in Genbank indicated that the nucleotide sequence of the RAPD marker showed 93% and 91% similarity with mth2-6el8 gene fragment （347 bp） and ruth2-33122 gene fragment （334 bp） of Medicago truncatula respectively. Medicago truncatula is a close relative of alfalfa and Mth2-6el8 is a molecular marker of the gene coding for a cysteine protease which was saltinducible in some plants. These results indicated the RAPD marker was possibly related to cysteine protease genes in alfalfa.
基金
This research was funded by the National High Tech R&D Program of China(863 Program,2002AA241101).
