AFLP Analysis And SCAR Marker Of Short Catkins Sport Of Chinese Chestnut (Castanea Mollissima Bl.)

Chestnut (Castanea mollissima Bl.) is one important kind of popular fruit tree in China. The sport of short catkins chestnut is a new germplasm resource, and is remarkable significance for chestnut breeding study; so far it has been hardly reported in recent years. In this study, original cultivar and sport line was both used, and AFLP and SCAR marker putatively related with this character of short catkins were detected. The results as followed:(1)Optimization of AFLP and related techniqueIn this study, AFLP and related technique were optimized. High quality DNA was one important factor in AFLP, the ratio of OD260/OD280 of the genomic DNA extracted by the modified CTAB method were 1.80, it was compatible to AFLP analysis. The silver staining was optimized, which produced the best quality of the image.(2)AFLP analysis on the bud mutation of short catkins of chestnutAFLP was used to compare the sport of short catkins and its mother plant in chestnut. A total of 72 selective primer pairs amplified 5130 clear bands. The Nei's association coefficient and genetic distance between them were 0.9904 and 0.0096 respectively. The molecular weight range of the polymorphic fragments was mainly 300~900 bp. The results indicated that there were some molecular-level differentiation between chestnut and its sport of short catkins. Some of the specific segments had been considered as markers of short catkins in chestnut.(3)Cloning, Sequencing and SCARSpecific fragments using E-ACT/M-CTT primer combinations were cloned and sequenced, the results of sequencing showed that there was 44.1% alignments to rice gene, the similar sequence has a character of retrotransposon protein, it could promote the study of functional genomes, while the SCAR marker transformed from AFLP markers could be used to forecast the pure seedlings of short catkins sport.