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通过PCR引物检测人参DNA特异性片段,快速、准确的评价低龄人参种质质量,加快人参育种进程。该研究基于个体之间的基因差异,设计7对引物,通过比对不同人参DNA的扩增图谱与皂苷含量相关性,成功得到一对特异性引物GC1,并采用L16(45)正交试验优化其25μL PCR体系,各组分最佳配比为:DNA 2.60 mg·L-1,Mg2+1.44 mmol·L-1、d NTP 0.19 mmol·L-1、引物0.32μmol·L-1、Taq 0.076 U·μL-1。对GC1引物进行验证,在24株供试人参中有2株扩增出1 200 bp特异性条带阳性样品,2株阳性样品中9种单体皂苷及其加和值含量均较高。该特异性条带序列与甘油-3-磷酸脱氢酶同源性达99%。GC1可以作为一种高皂苷人参株系早期筛选鉴定方法的特异性引物,有利于加快人参育种进程。
The PCR primers were used to detect DNA-specific fragments of ginseng to evaluate the quality of young ginseng germplasm quickly and accurately and accelerate the process of ginseng breeding. Based on the genetic differences among individuals, seven pairs of primers were designed and a pair of specific primers GC1 was successfully obtained by comparing the amplification profiles of different ginseng DNAs with the content of saponin. The optimized L16 (45) orthogonal test The 25 μL PCR system showed that the best proportion of each component was DNA 2.60 mg · L-1, Mg2 + 1.44 mmol·L-1, dNTP 0.19 mmol·L-1, primer 0.32 μmol·L-1, Taq 0.076 U · μL-1. GC1 primers were validated. Two of the 24 tested ginseng samples were positive for 1 200 bp band, and 9 monomeric saponins were positive for 2 positive samples. This specific band sequence shares 99% homology with glycerol-3-phosphate dehydrogenase. GC1 can be used as a specific primer for early screening and identification of high saponins ginseng strain, which is good for accelerating ginseng breeding.