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利用1D-SDS-PAGE分离了261份青藏高原农家青稞的淀粉颗粒结合蛋白,旨在为青藏高原青稞淀粉品质改良和淀粉颗粒结合蛋白机制研究提供依据和基础信息。在分子量45～100kD区域共有21种多态性蛋白条带和78种组合带型,其中2、3、5、10、11为新条带。利用PCR技术克隆了236份农家青稞GBSSI基因5′前导序列,出现1000bp和800bp的多态性片段,且以前者为主,其频率为80.1%。在8份农家青稞及4份引进的低直链淀粉材料的GBSSI基因5′前导序列中共检测到32个多态性位点,包括9个InDels和23个SNPs。GBSSI基因5′前导序列中出现了特有的序列差异,如未出现600bp类型(约400bp的特异缺失),而该缺失被认为是低直连淀粉大麦形成的原因;材料yf127、yf70、011Z1396和09Z586出现了特异点突变。因此认为,青藏高原农家青稞品种的淀粉颗粒结合蛋白具有丰富的多态性和独特性。低直链淀粉青稞GBSSI基因序列的新特征表明可能存在新的低直链淀粉形成机制。
The starch particle-binding proteins of 261 Tibetan barley cultivars from Qinghai-Tibet Plateau were separated by 1D-SDS-PAGE, which provided the basis and basic information for the study on the starch quality improvement and the mechanism of starch granule-binding protein in Qinghai-Tibet Plateau. There were 21 polymorphic bands and 78 bands in the region of molecular weight 45 ~ 100kD, of which 2, 3, 5, 10 and 11 were new bands. The 5 ’leader sequence of GBSSI gene was cloned by PCR and the polymorphic fragments of 1000bp and 800bp were obtained. The former was predominant and the frequency was 80.1%. A total of 32 polymorphic sites, including 9 InDels and 23 SNPs, were detected in the 5 ’leader sequence of GBSSI gene in 8 farmland barley and 4 introduced low amylose materials. The unique sequence divergence occurred in the 5 ’leader sequence of GBSSI gene, such as the absence of a 600bp type (about 400bp specific deletion), which is believed to be responsible for the formation of low direct starch barley. Materials yf127, yf70, 011Z1396 and 09Z586 A specific point mutation occurred. Therefore, it is considered that there are abundant polymorphisms and unique features of Starch Granule-Binding Protein in farmland barley cultivars in Qinghai-Tibet Plateau. The new characterization of the low amylose barley GBSSI gene sequence suggests that there may be a new mechanism of low amylose formation.