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作物学报 ›› 2012, Vol. 38 ›› Issue (07): 1148-1154.doi: 10.3724/SP.J.1006.2012.01148

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

青藏高原青稞农家品种淀粉颗粒结合蛋白组成及GBSSI基因5′前导序列的多态性

王春萍1,2,**，高丽鹃1,2,**，潘志芬1,*，尼玛扎西3，唐亚伟3，曾兴权3，李俏1,2，邓光兵1,2，龙海1，余懋群1,*   

  1. 1中国科学院成都生物研究所，四川成都 610041；2中国科学院研究生院，北京 100049； 3西藏农牧科学院 / 农业部藏区青稞生物学与遗传育种重点实验室，西藏拉萨 850032
  • 收稿日期:2011-12-05 修回日期:2012-04-16 出版日期:2012-07-12 网络出版日期:2012-05-11
  • 通讯作者: 潘志芬, E-mail: panzf@cib.ac.cn, Tel: 028-85226136; 余懋群, E-mail: yumq@cib.ac.cn, Tel: 028-85229053
  • 基金资助:

    本研究由国家自然科学基金项目(31101150), 国家科技支撑计划项目(2012BAD03B01), 国家重点基础研究计划(973计划)前期研究专项(2012CB723006), 国家科技基础性工作专项规划(2006FY110700)和中国科学院西部之光人才培养计划项目资助。

Polymorphism of Starch Granule-Associated Proteins and 5′ Leader Sequence of GBSSI Gene in Indigenous Naked Barley (Hordeum vulgare L.) from Qinghai-Tibetan Plateau in China

WANG Chun-Ping1,2,**,GAO Li-Juan1,2,**,PAN Zhi-Fen1,*,NIMA Zha-Xi3,TANG Ya-Wei3,ZENG Xing-Quan3,LI Qiao1,2,DENG Guang-Bing1,2,LONG Hai1,YU Mao-Qun1,*   

  1. 1 Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China; 2 Graduate School of Chinese Academy of Sciences, Beijing 100039, China; 3 Tibet Academy of Agriculture and Animal Sciences / Key Laboratory of Biology and Genetic Breeding of Tibetan Naked Barley, Ministry of Agriculture, Lhasa 850032, China
  • Received:2011-12-05 Revised:2012-04-16 Published:2012-07-12 Published online:2012-05-11
  • Contact: 潘志芬, E-mail: panzf@cib.ac.cn, Tel: 028-85226136; 余懋群, E-mail: yumq@cib.ac.cn, Tel: 028-85229053

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摘要： 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45~100 kD区域共有20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1 000 bp和800 bp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InDel和23个SNP。GBSSI基因5′前导序列中出现了特有的序列差异，如未出现600 bp类型(约400 bp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，可能存在新的形成机制。

关键词: 农家品种, 青稞, 淀粉颗粒结合蛋白, GBSSI基因

Abstract: Starch granule-associated proteins (SGAPs) are minor components tightly bound with starch granules, most of which are believed to be starch biosynthetic enzymes. We separated SGAPs from 261 indigenous naked barley accessions in Qinghai-Tibetan Plateau of China using 1D-SDS-PAGE technique. A total of 20 polymorphic banding patterns with molecular weights of 45–100 kD and 78 SGAPs patterns were detected, of which bands 2, 3, 5, 10, and 11 were novel protein bands. The 5′ leader sequences of GBSSI gene were cloned using 236 accessions of indigenous naked barley. Two types of length polymorphisms (1 000 bp and 800 bp) were found with frequencies of 80.1% and 19.9%, respectively. According to sequencing results, there were 32 polymorphic sites in the 5′ leader sequences of GBSSI gene, including nine InDels and 23 SNPs. Specific mutations were found in Qinghai-Tibetan Plateau accessions. For example, the 400 bp deletion reported corresponding to low amylose content in barley was absent in this study; and special SNPs were detected in accessions yf127, yf70, 011Z1396, and 09Z586. These results imply that the SGAPs of the Qinghai-Tibetan Plateau indigenous naked barley are abundant and polymorphic. The novel sequence characteristics in the GBSSI5′ leader sequence of the low amylose accessions suggest a different mechanism for amylose biosynthesis in Qinghai-Tibetan Plateau indigenous naked barley.

Key words: Indigenous variety, Naked barley, Starch granule-associated proteins (SGAPs), GBSSI gene

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