Agarwal Pradeep K, Agarwal Parinita, Jain Parul, Jha Bhavanath, Reddy M K, Sopory S K
International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Road, New Delhi, 110 067, India.
Plant Cell Rep. 2008 Jan;27(1):105-15. doi: 10.1007/s00299-007-0446-0. Epub 2007 Sep 27.
The Rab GTPases are important components of endocytic network in plant cells. Endocytosis participates in the cell's reaction to extracellular stimuli by desensitizing, down-regulating or recycling receptors and membrane proteins. Rab7 is a small GTP-binding protein involved in intracellular vesicle trafficking from late endosome to the vacuole. We have isolated Rab7 cDNA from Pennisetum glaucum, a relatively drought-stress tolerant food grain crop grown commonly in India, during cDNA-subtractive hybridization of dehydration-stress treated plants. The PgRab7 ORF, encoding 207 aminoacids, was over-expressed in E. coli. The recombinant PgRab7 protein showed GTP-binding and GTPase activity. Transcript expression of PgRab7 gene was differentially up-regulated by different environmental stimuli such as cold, dehydration and NaCl and also by a plant hormone IAA. Overexpression of PgRab7 gene enhanced tolerance to NaCl and mannitol in transgenic tobacco. Transgenic plants also had increased alkaline phosphatase (ALP) activity. These results show that PgRab7 is a potential candidate gene for developing both salinity and dehydration tolerance in planta.
Rab GTP酶是植物细胞内吞网络的重要组成部分。内吞作用通过使受体和膜蛋白脱敏、下调或循环利用来参与细胞对细胞外刺激的反应。Rab7是一种小GTP结合蛋白,参与从晚期内体到液泡的细胞内囊泡运输。在对脱水胁迫处理的植物进行cDNA消减杂交过程中,我们从狼尾草(一种在印度普遍种植的相对耐旱的粮食作物)中分离出了Rab7 cDNA。编码207个氨基酸的PgRab7开放阅读框在大肠杆菌中过表达。重组PgRab7蛋白表现出GTP结合和GTP酶活性。PgRab7基因的转录表达受到不同环境刺激(如寒冷、脱水和NaCl)以及植物激素IAA的差异上调。PgRab7基因的过表达增强了转基因烟草对NaCl和甘露醇的耐受性。转基因植物的碱性磷酸酶(ALP)活性也有所增加。这些结果表明,PgRab7是在植物中培育耐盐性和耐旱性的潜在候选基因。