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西藏青稞品种喜玛拉雅10号干旱胁迫的SSH文库构建及干旱诱导表达基因分析
  • ISSN号:1009-1041
  • 期刊名称:《麦类作物学报》
  • 分类:S512.3[农业科学—作物学] S330[农业科学—作物遗传育种;农业科学—农艺学]
  • 作者机构:[1]西藏自治区青稞种质改良和牦牛繁育重点实验室,西藏拉萨850002, [2]西藏自治区农牧科学院,西藏拉萨850002, [3]中国农业大学农业生物技术重点实验室,北京100086
  • 相关基金:国家“973”计划前期研究专项(2011CBlll512;2012CB723006);中国农业大学农业生物技术重点实验室开放课题(2011SKI.AB064);国家自然科学基金项目(31160283);国家科技支撑计划项目(2012BAD03801).
中文摘要:

为了研究西藏青稞抗旱的分子机制,以西藏耐旱青稞(Hordeum vulgare L.var.nudum HK.f.)品种喜玛拉雅10号为材料,利用抑制性差减杂交技术(SSH)构建青稞苗期叶片干旱胁迫诱导基因表达的正向差减文库。挑取600个阳性克隆进行PCR验证,并对验证后的单克隆进行测序分析,共获得420个有效序列。去除冗余序列和嵌合序列后,获得220条高质量EST序列,其中183条singlets,37条contigs。BlastN分析表明,其中189个EST序列可以在GenBank的unigene中找到同源序列,31个未能找到unigene同源序列;BlastX分析表明,62个EST序列unigene与未知功能蛋白或假定蛋白有较高的相似性;158条EST序列与已知功能蛋白有较高的同源性。获得的EST序列多数不仅与植物的非生物胁迫相关,也与植物的生物胁迫反应相关,说明植物在胁迫反应中有明显的共同机制。其中与大麦表达序列高度同源的EST占58%;这些基因涉及参与信号转导和转录调节的基因(11.96%),编码保护、防御和胁迫耐受蛋白的基因(9.78%),跨膜转运相关基因(5.43%),光合作用基因(6.52%),参与结构和功能代谢合成途径中的一些酶(30.43%)等代谢过程。实验结果表明,青稞在干旱复水条件下可诱导一系列特异基因的表达,如参与抗旱反应相关酶和蛋白基因锌指蛋白、衰老相关蛋白、CAS、细胞色素P450、热激蛋白、胁迫诱导蛋白,以及LEA蛋白、脱水蛋白、P5CS等保护蛋白基因。用KOBAS系统将56个EST定位到32个Pathways中,初步分析发现,谷氨酸盐代谢途径(Glutamate metabolism)、精氨酸和脯氨酸代谢途径(Arginine and proline metabolism)、泛素蛋白介导的蛋白质水解代谢(Ubiquitin mediated proteolysis)途径可能与青稞抗旱相关性较大。

英文摘要:

A forward cDNA-SSH library was established by suppression subtractive hybridization u- sing seedling leaf of Himalaya 10, a drought-tolerant Tibet barley (Hordeum vulgate I.. var. nudum HK. f. ), among which 600 positive clones were selected for sequencing, after detection by PCR for each clone, each single clone was sequenced. Totally 420 available sequences were assembled and 220 uniESTs, in which 37 contigs and 183 singlets were obtained by cluster analyses of the ESTs. The re- sults of BlastN showed that 189 ESTs had homologous sequences in unigenes database of GenBank, while the other 31 ESTs had no protein homologous. The BlastX results indicated that 158 ESTs had significant homology with proteins encoded by unigenes and 62 were unknown proteins and putative proteins. Most of those genes were not only related to abiotic stresses, but also related to biotic stres- ses in plants, which suggesting the cross-interactions in response to different stresses in plants, and 58 % of the ESTs were homologous with barley ESTs. These UniGenes involved in signal transduction and transcriptional regulation (11.96%), encoding protection, defense and stress protein (9. 78%), transport related (5.43%), photosynthesis genes (6.52%), and enzymes for structure and function of metabolic synthesis pathway and other metabolic processes (30.43G). The experimental results in- :ticated that the barley can induce a series of specific expressed genes in the drought-rehydration condi- Lions, not only some known enzymes and protein genes participated in drought response, such as zinc protein, senescence associated protein, CAS, cytochrome P450, heat shock proteins and stress pro- eins, were found this study, but also some protective protein genes, such as LEA protein, dehydra- ed protein, PSCS. KOBAS mapped 56 ESTs of the 220 unigSTs to 32 KEGG pathways. This study uggested that the drought tolerance of Tibet barley was closely related with glutamate metabolism, rginine and proline metabolism and Ubiquitin medi

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期刊信息
  • 《麦类作物学报》
  • 北大核心期刊(2011版)
  • 主管单位:中华人民共和国教育部
  • 主办单位:西北农林科技大学 中国作物学会 国家小麦工程技术研究中心
  • 主编:孙其信
  • 地址:陕西省杨凌邰城路3号《麦类作物学报》编辑部
  • 邮编:712100
  • 邮箱:mlzwxb@163.com
  • 电话:029-87082032
  • 国际标准刊号:ISSN:1009-1041
  • 国内统一刊号:ISSN:61-1359/S
  • 邮发代号:52-66
  • 获奖情况:
  • 从2003年起被收录为国家科技部“中国科技核心期刊”,2002年获第三届全国优秀农业期刊一等奖,1999年获陕西省科技期刊一等奖,1995年获陕西省优秀期刊二等奖
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,美国剑桥科学文摘,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:13981