Cloning and characterization of three genes encoding Qb-SNARE proteins in rice
Cloning and characterization of three genes encoding Qb-SNARE proteins in rice
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DOI：
10.1007/s00438-007-0313-2
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发表时间：
2008-03-01
影响因子：
3.1
通讯作者：
Zhang, Hong-Sheng
中科院分区：
文献类型：
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作者：
Bao, Yong-Mei;Wang, Jian-Fei;Zhang, Hong-Sheng
Qb-SNARE蛋白属于SNARE（可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体）的超家族，并且作为真核细胞中囊泡运输机制的重要组分起作用。本文报道了从水稻中分离到的三个新的植物SNARE基因，分别命名为OsNPSN 11、OsNPSN 12和OsNPSN 13。它们在其整个编码区上具有约70%的核苷酸同一性，并且每个基因中具有相似的基因组结构，具有10个外显子和9个内含子。推导的氨基酸序列的多重比对表明OsNPSNs蛋白与拟南芥AtNPSNs同源，在C端区域含有Qb-SNARE结构域和跨膜结构域。半定量RT-PCR分析表明，OsNPSNs在根、茎、叶、幼穗和开花穗中均存在差异表达。OsNPSNs的表达在H2O2处理的水稻幼苗中被显著激活，而在NaCl和PEG 6000胁迫下被下调。在洋葱表皮细胞中的瞬时表达方法显示OsNPSNs定位于质膜。OsNPSNs转化的酵母细胞在含有不同浓度H2O2的固体或液体选择性培养基上的生长速率比空载体转化体更好，但对NaCl和甘露醇胁迫更敏感。35S：OsNPSNll转基因烟草也表现出比非转基因烟草更强的H2O2耐受性以及对NaCl和甘露醇的敏感性。这些结果表明OsNPSNs可能参与植物和酵母对非生物胁迫响应的信号转导的不同方面。
Qb-SNARE proteins belong to the superfamily of SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) and function as important components of the vesicle trafficking machinery in eukaryotic cells. Here, we report three novel plant SNARE (NPSN) genes isolated from rice and named OsNPSN11, OsNPSN12 and OsNPSN13. They have about 70% nucleotide identity over their entire coding regions and similar genomic organization with ten exons and nine introns in each gene. Multiple alignment of deduced amino acid sequences indicate that the OsNPSNs proteins are homologous to AtNPSNs from Arabidopsis, containing a Qb-SNARE domain and a membrane-spanning domain in the C-terminal region. Semi-quantitative RT-PCR assays showed that the OsNPSNs were ubiquitously and differentially expressed in roots, culms, leaves, immature spikes and flowering spikes. The expression of OsNPSNs was significantly activated in rice seedlings treated with H2O2, but down-regulated under NaCl and PEG6000 stresses. Transient expression method in onion epidermal cells revealed that OsNPSNs were located in the plasma membrane. Transformed yeast cells with OsNPSNs had better growth rates than empty-vector transformants when cultured on either solid or liquid selective media containing various concentrations of H2O2, but more sensitive to NaCl and mannitol stresses. The 35S:OsNPSN11 transgenic tobacco also showed more tolerance to H2O2 and sensitivity to NaCl and mannitol than non-transgenic tobacco. These results indicate that OsNPSNs may be involved in different aspects of the signal transduction in plant and yeast responses to abiotic stresses.