麦芽品种多重荧光SSR标记指纹数据库的构建

Construction of multiplex fluorescence SSR marker database for malt varieties

为提高麦芽品种及纯度的鉴定效率,降低检测成本,利用SSR(simple sequence repeats,简单重复序列)荧光标记对24个麦芽品种进行核心引物筛选、多重PCR和指纹数据分析,构建适合麦芽品种鉴定的多重荧光检测体系。首先从前期筛选的多态性引物中选择12对引物进行组合设计,通过优化引物浓度、退火温度进行多重PCR体系构建,3个引物组合扩增成功。按照最少引物鉴定最多品种的指纹图谱构建原则,将引物HVM68、EBmag0007和Bmac755确定为品种区分的核心引物,每对引物平均产生8.3个等位基因。利用3对核心引物构建麦芽品种指纹数据库,通过指纹编码可实现全部24个麦芽品种的快速区分。该研究建立了一种准确、快速、高通量的麦芽品种鉴定检测方法,为啤酒企业在麦芽采购和酿造配方研究上提供技术支持和质量保障。

In order to improve the identification efficiency of malt varieties and purity,thus reduce the cost of detection,SSR(simple sequence repeats)fluorescent markers were used to perform core primer screening,multiplex PCR and fingerprint data analysis on 24 malt varieties.A suitable multiple fluorescence detection system for malt identification was constructed.12 pairs of primers were selected from the polymorphic primers screened in the previous stage for combination design.The multiplex PCR system was constructed by optimizing the primer concentration and annealing temperature,and 3 primer combinations were successfully amplified.According to the principle of fingerprint identification,HVM68,EBmag0007 and Bmac755 were identified as the core primers for cultivar differentiation,with each pair producing an average of 8.3 alleles.Using 3 pairs of core primers to construct a malt species fingerprint database,fingerprint coding could achieve rapid differentiation of all 24 malt varieties.This study established an accurate,rapid and high-throughput method for the identification and detection of malt varieties,providing technical support and quality assurance for beer companies in malt procurement and brewing formulations.