高粱抗丝黑穗病SCAR标记的建立

摘要

The DNA of the sorghum Head Smut disease-resistant restorer lines and susceptible-resistant restorer lines dwarf four,Head Smut resistance and susceptibility was extracted by CTAB method and marked with molecular marker to sorghm gene by RAPD in this paper.In the optimal RAPD reaction system,100 random primers were used to screen the markers linked to the resistance gene,88 pairs amplified products.The results show that S405 of 88 pairs of primers between disease-resistant and susceptible-resistant cultivars are amplified differences bands,further segregation analysis the resistance gene to Head Smut was 11.7 %.S405 amplified polymorphic bands were cloned and sequenced with the length of 320 bp.According to the nucleotide sequences of differences bands to design specific primer was designed for perform PCR,the RAPD markers were succeeded conversing to SCAR marker,the marker named S405-320 ,to lay a foundation for the selection and cultivation of improved varieties of sorghum.%选用高粱丝黑穗病感病恢复系矮四、抗病恢复系2381R 以及二者 F2代抗感个体为试验试材,采用CTAB法提取高粱基因组 DNA,并应用 RAPD 筛选技术对高粱基因进行分子标记,选取了100对 RAPD随机引物对其进行扩增,有88对引物扩增出条带。分析结果表明：88对引物中 S405在抗感品种间扩增出差异谱带。进一步对抗感群体进行分离分析,得出抗丝黑穗病基因重组率为11.7%。将 S405扩增出的差异片段进行克隆测序,差异谱带的片段长度为320 bp。根据差异谱带的碱基排列顺序,设计特异性引物,然后进行序列扩增,将 RAPD 分子标记转化为SCAR分子标记,并将该标记命名为 S405-320,为高粱优良品种的筛选和培育奠定一定的基础。