Cereal variety identification using MALDI-TOF mass spectrometry SNP genotyping

Correct identification of cereal varieties is important to food quality, safety and authenticity and recently, molecular markers have been applied to cereal varietal identification. Two major factors restricting the wider application of molecular markers in crop plant genetics have been throughput and cost per data point. However, in recent years, novel molecular tools, the re-invention/re-application of mature technologies, and miniaturisation of technology has both increased throughput and significantly reduced these costs. Here we have applied Sequenom^® MassARRAY^® MALDI-TOF mass spectrometry using a collection of recently developed SNPs to facilitate cereal varietal identification. We used a multiplexed Sequenom^® MassARRAY^® mass spectrometry SNP assay targeting 45 loci to genotype a collection of Australian barley varieties. Of the 45 loci screened, 33 were informative and were used to generate a unique barcode of SNPs for each variety tested. Only one variety could not be distinguished from two others due to a high level of varietal heterogeneity. This assay format provided a flexible, cost-effective, robust and moderate throughput SNP genotyping method well suited to varietal identification and purity analysis in cereals.     

蔗糖合成酶(SUS)基因的SNP标记开发及其与淀粉性状的关联分析

通过直接测序法获得玉米亲本(H21和紫糯96-619)sus1基因全长序列并进行亲本间序列比对,得到120个SNP位点,通过高分辨率熔解曲线(HRM)对300个近等基因系H21 BC5F2:3进行SNP位点分型,并将后代的基因型与群体的直链淀粉含量和淀粉糊化特性进行关联分析。结果表明,marker 31与直链淀粉含量和淀粉崩解值、峰值时间、糊化特性均连锁紧密;marker 17与直链淀粉含量和淀粉的峰值时间、淀粉最终黏度均连锁紧密;marker12与直链淀粉含量、淀粉的峰值时间连锁紧密。