基于SSR分子标记的‘汇丰2号’辣椒杂交种子纯度和真实性鉴定

Identification of Purity and Authenticity of Hybrid Seeds of‘Huifeng 2’Pepper Based on SSR Molecular Markers

【目的】‘汇丰2号’辣椒为杂交一代新品种,具有中早熟、抗病性强、耐储运和品质佳等特性,深受农户和消费者喜爱。为保障种子质量,拟建立一套快速、简便的种子纯度和真实性鉴定方法。【方法】基于辣椒基因组已公开的SSR分子标记资源,以‘汇丰2号’父、母本为材料,筛选多态明显、条带清晰且扩增稳定的SSR标记,对‘汇丰2号’杂交种子进行纯度鉴定,并结合田间形态鉴定验证分子标记鉴定的准确性。此外,利用筛选出的SSR标记对‘粤红1号’、‘汇丰1号’、‘汇丰2号’和‘汇丰5号’4个品种进行真实性鉴定。【结果】筛选出2个SSR标记L0143和L0622,其多态明显,且条带清晰、扩增稳定。其中,标记L0143在母本W2280中PCR扩增条带为149 bp,在父本W2102中的扩增条带为141 bp;标记L0622在母本和父本中的扩增条带分别为208 bp和196 bp。2个标记单独使用时均可以准确鉴定‘汇丰2号’种子纯度,且2个标记同时使用可以将‘粤红1号’、‘汇丰1号’、‘汇丰2号’和‘汇丰5号’4个品种区分开,即可鉴定它们的真实性。【结论】筛选出的SSR分子标记L0143和L0622能够快速、准确、低成本、有效的对‘汇丰2号’杂交种子进行纯度和真实性鉴定,该鉴定方法操作简单,能够替代传统杂交种子纯度和真实性鉴定的方法,具有较高的商业应用前景。

【Objective】‘Huifeng 2’is an F1 hybrid variety which is highly favoured by farmers and consumers due to its characteristics of mid-early maturity,strong disease resistance,storage and transportation resistance and good quality.To ensure the quality of seeds,it intends to establish a set of quick and easy methods for seed purity and authenticity identification.【Method】Based on the publicly available SSR molecular markers of pepper genome,SSR markers with obvious polymorphisms,clear bands and stable amplification were screened by using the male and female parents of‘Huifeng 2’as materials.Then,the purity of the hybrid seeds of‘Huifeng 2’was identified,and the accuracy of the molecular marker was verified by using morphology identification in the field.In addition,the authenticity of four varieties,including‘Yuehong 1’,‘Huifeng 1’,‘Huifeng 2’and‘Huifeng 5’,was identified with the screened markers.【Result】Two SSR markers,L0143 and L0622,were selected for their obvious polymorphism and clear bands with stable amplification.The PCR amplification bands of marker L0143 were 149 bp in the female parent W2280 and 141 bp in the male parent W2102,and the amplification bands of marker L0622 were 208 bp and 196 bp in the female parent and male parent,respectively.When used alone,the two markers could accurately identify the purity of‘Huifeng 2’seeds,and the simultaneous use of the two markers could distinguish the four varieties of‘Yuehong 1’,‘Huifeng 1’,‘Huifeng 2’and‘Huifeng 5’,which could be used to identify their authenticity.【Conclusion】The SSR molecular markers L0143 and L0622 selected in this study can identify the purity and authenticity of‘Huifeng 2’hybrid seeds rapidly,accurately,and effectively with low-cost,which are simple to operate and can replace the traditional method of identifying the purity and authenticity of hybrid seeds.It has a high commercial application prospect.