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利用同交育种中产生的回交群体,结合前人的研究结果构建了Pil基因区域的局部分子标记连锁图,通过BC1F2家系的接种结果判断其基因型。将Pil定位在RFLP标记RZ536与SSR标记RM144之间,图距分别为9.7cM、6.8 cM,从而建立了一套完整的以PCR为基础的分子标记辅助选择体系。通过分子标记和抗性验证两种选择方式相结合,经过三代回交将Pil区段快速导入受体亲本珍汕97B中。在BC3F1中利用15条ISSR引物扩增的167条随机分布在基因组中的多态性带筛选背景,得到4个背景较好的单株。经过纯合筛选及抗性验证后共得到17个带有抗性基因Pil的改良珍汕97株系。试验表明微卫星标记在正向选择、负向选择及背景选择中都起到极大的作用。
Using the backcross population generated from the crossbred breeding, the molecular linkage map of the Pil gene region was constructed based on the previous studies and the genotypes were determined by the inoculation results of the BC1F2 pedigree. Pil was located between the RFLP marker RZ536 and the SSR marker RM144 at 9.7 cM and 6.8 cM intervals respectively, thus establishing a complete PCR-based molecular marker-assisted selection system. Through the combination of molecular marker and resistance verification, the Pil segment was quickly introduced into the recipient parent Zhenshan 97B after three generations of backcrossing. In BC3F1, 167 random amplified polymorphic bands (ISSRs) amplified by 15 ISSR primers were screened for the presence of four clones with good background. After homozygous selection and resistance verification, a total of 17 Zhenshan 97 strains with resistance gene Pil were obtained. Experiments show that microsatellite markers play a significant role in forward selection, negative selection and background selection.