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从耐盐水稻品种 Pokkali 的 cDNA 文库中进行功能筛选,鉴定出甘露糖-1-磷酸鸟苷转移酶基因(OsMPG1)是盐胁迫响应的关键成员。

Functional screening of cDNA library from a salt tolerant rice genotype Pokkali identifies mannose-1-phosphate guanyl transferase gene (OsMPG1) as a key member of salinity stress response.

机构信息

Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India.

出版信息

Plant Mol Biol. 2012 Aug;79(6):555-68. doi: 10.1007/s11103-012-9928-8. Epub 2012 May 29.

Abstract

Salinity, one of the most deleterious stresses, affects growth and overall yield of crop plants. To identify new "candidate genes" having potential role in salinity tolerance, we have carried out 'functional screening' of a cDNA library (made from a salt tolerant rice-Pokkali). Based on this screening, we identified a cDNA clone that was allowing yeast cells to grow in the presence of 1.2 M NaCl. Sequencing and BLAST search identified it as mannose-1-phosphate guanyl transferase (OsMPG1) gene from rice. Analysis of rice genome sequence database indicated the presence of 3 additional genes for MPG. Out of four, three MPG genes viz. OsMPG1, 3 and 4 were able to functionally complement yeast MPG mutant -YDL055C. We have carried out detailed transcript profiling of all members of MPG family by qRT-PCR using two contrasting rice genotypes (IR64 and Pokkali) under different abiotic stresses (salinity, drought, oxidative stress, heat stress, cold or UV light). These MPG genes showed differential expression under various abiotic stresses with two genes (OsMPG1 and 3) showing high induction in response to multiple stresses. Analysis of rice microarray data indicated higher expression levels for OsMPG1 in specific tissues such as roots, leaves, shoot apical meristem and different stages of panicle and seed development, thereby indicating its developmental regulation. Functional validation of OsMPG1 carried out by overexpression in the transgenic tobacco revealed its involvement in enhancing salinity stress tolerance.

摘要

盐度是最具危害性的胁迫之一,会影响作物的生长和整体产量。为了鉴定具有耐盐潜力的新“候选基因”,我们对来自耐盐水稻-Pokkali 的 cDNA 文库进行了“功能筛选”。基于该筛选,我们鉴定了一个 cDNA 克隆,该克隆使酵母细胞能够在 1.2 M NaCl 存在的情况下生长。测序和 BLAST 搜索将其鉴定为来自水稻的甘露糖-1-磷酸鸟苷转移酶 (OsMPG1) 基因。对水稻基因组序列数据库的分析表明,存在另外 3 个 MPG 基因。在这 4 个基因中,3 个 MPG 基因(OsMPG1、3 和 4)能够在酵母 MPG 突变体 -YDL055C 中功能性互补。我们使用两种不同的水稻基因型(IR64 和 Pokkali)在不同的非生物胁迫(盐度、干旱、氧化应激、热应激、冷或紫外光)下通过 qRT-PCR 对 MPG 家族的所有成员进行了详细的转录谱分析。这些 MPG 基因在各种非生物胁迫下表现出不同的表达模式,其中 2 个基因(OsMPG1 和 3)对多种胁迫表现出高诱导。对水稻微阵列数据的分析表明,OsMPG1 在特定组织(如根、叶、茎尖分生组织以及颖花和种子发育的不同阶段)中的表达水平较高,表明其发育调控。通过在转基因烟草中过表达 OsMPG1 进行的功能验证表明,它参与增强耐盐胁迫。

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