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荞麦13S球蛋白是荞麦种子中的一类主要贮藏蛋白。本研究选用荞麦属植物甜荞栽培种及其野生类型、苦荞栽培种及其野生类型、毛野荞、左贡野荞、细柄野荞和硬枝万年荞6个种共44份材料,进行PCR特异性扩增、测序得到荞麦13S球蛋白基因的保守片段序列。对序列进行差异分析,结果发现44份供试材料13S球蛋白基因片段的285个排列位点中不变位点为24个,多态性位点S为261个(含简约信息位点数198个和单型可变位点63个),序列总突变位点Eta为503个。野生甜荞种内13S球蛋白基因序列差异明显高于栽培甜荞,但野生苦荞种内13S球蛋白基因序列差异仅稍高于栽培苦荞。推测其一方面可能与繁殖方式有关,另一方面可能与荞麦驯化过程中通常只有少数野生型群体被驯化有关。系统聚类分析发现栽培甜荞与野甜荞亲缘关系近,与左贡野荞亲缘关系次之;栽培苦荞与野苦荞亲缘关系近,与毛野荞亲缘关系次之;细柄野荞和硬枝万年荞的13S蛋白基因片段序列差异较小,说明其亲缘关系较近。上述结果为荞麦属种间遗传多样性与进化关系研究提供了理论依据。
Buckwheat 13S globulin is a type of major storage protein in buckwheat seeds. In this study, 44 varieties of sweet buckwheat cultivars, wild type, tartary buckwheat cultivars and their wild type, Mao wild buckwheat, Zuo Gong wild buckwheat, PCR specific amplification, sequencing to obtain a conservative fragment of buckwheat 13S globin gene sequence. The results of differential analysis showed that the number of invariant loci in 285 loci of 44 13S globin gene fragments was 24, and 261 polymorphic loci (including 198 simple information loci) And 63 single haplotypes), and the total number of Eta was 503. The sequence differences of 13S globulin gene in wild buckwheat was significantly higher than that in cultivated buckwheat, but the sequence difference of 13S globulin in wild buckwheat was only slightly higher than that of cultivated buckwheat. It is speculated that on the one hand, it may be related to the mode of reproduction, on the other hand, it may be related to the domestication of only a few wild-type populations in the process of domestication of buckwheat. The results of phylogenetic tree analysis showed that the genetic relationship between cultivated sweet buckwheat and wild sweet buckwheat was close, followed by that of Zuo Gong wild buckwheat, that of cultivated tartary buckwheat was close to that of wild buckwheat, followed by that of wild buckwheat, The 13S protein gene fragments of Fagopyrum tataricum showed a small sequence difference, indicating that its genetic relationship is relatively close. The above results provide a theoretical basis for the study on the relationship between the genetic diversity and evolution of genus Buckwheat.