酿酒酵母糠醛耐受性状相关微卫星标记的筛选

Screening of Microsatellite Markers Associated with Furfural Tolerance of Saccharomyces cerevisiae

筛选两株表型差异单倍体菌株作为亲本,二者杂交获得F2群体共200株,利用具有多态性的微卫星位点对表型分布在两极端的菌株(39株)进行PCR扩增,运用t检验分析不同位点的基因型在两基因池中的差异显著性,以此获得与糠醛耐受性相关的微卫星位点。结果发现两个与糠醛耐受性有关的微卫星位点,其中2P3位点在两基因池中达到显著差异(P=0.039<0.05),而位点2P5达到极显著差异(P=0.000<0.01);对于微卫星位点2P5,耐受亲本P1在该位点的基因型(333 bp)在子代耐受群体中出现率为72.2%,而敏感亲本P2的基因型(318 bp)在子代敏感群体中出现率达100%(333 bp出现率为零)。上述结果表明微卫星位点2P5的基因型在群体分离中达到极显著差异可能与控制该性状的基因相连锁,在分子设计育种方面可用来选择有利等位基因,提高育种的准确性和预见性。

Two phenotypically different haploid strains were screened as parents and were crossed to obtain two hundred segregants. The polymorphic microsatellite markers were used to amplify offspring yeasts （ 39 strains ）, and t-test was used to analyze the significance of difference among the genotypes of different sites in two gene pools, which is aimed to have microsatellite markers associated with furfuraL tolerance. The results revealed that two mierosatellite markers associated with furfural tolerance were identified. Among them the marker 2P3 had significant differences （ P = 0.039 〈 0.05 ） and marker 2P5 had extremely significant differences （ P = 0.000 〈 0.01 ） in the 2 gene pools. Furthermore, 72.2% of high tolerant individuals contained the marker 2P5 （ 333 bp ） derived from P1, the high tolerant parent ; 100% of low tolerant individuals contained the marker 2P5 （ 318 bp ） derived from P2, the low tolerant parent. These results suggested that the significant difference of 2P5 genotype in population segregation may be linked with the gene controlling this trait and can be used to select the favorable allele in molecular design and breeding so as to improve the accuracy and predictability of breeding.