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Early effects of salinity on water transport in Arabidopsis roots. Molecular and cellular features of aquaporin expression

机译:盐度对拟南芥根系水分传输的早期影响。水通道蛋白表达的分子和细胞特征

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Aquaporins facilitate the uptake of soil water and mediate the regulation of root hydraulic conductivity (Lp(r)) in response to a large variety of environmental stresses. Here, we use Arabidopsis (Arabidopsis thaliana) plants to dissect the effects of salt on both Lp(r) and aquaporin expression and investigate possible molecular and cellular mechanisms of aquaporin regulation in plant roots under stress. Treatment of plants by 100 mM NaCl was perceived as an osmotic stimulus and induced a rapid (half-time, 45 min) and significant (70%) decrease in Lp(r), which was maintained for at least 24 h. Macroarray experiments with genespecific tags were performed to investigate the expression of all 35 genes of the Arabidopsis aquaporin family. Transcripts from 20 individual aquaporin genes, most of which encoded members of the plasma membrane intrinsic protein (PIP) and tonoplast intrinsic protein (TIP) subfamilies, were detected in nontreated roots. All PIP and TIP aquaporin transcripts with a strong expression signal showed a 60% to 75% decrease in their abundance between 2 and 4 h following exposure to salt. The use of antipeptide antibodies that cross-reacted with isoforms of specific aquaporin subclasses revealed that the abundance of PIP1s decreased by 40% as early as 30 min after salt exposure, whereas PIP2 and TIP1 homologs showed a 20% to 40% decrease in abundance after 6 h of treatment. Expression in transgenic plants of aquaporins fused to the green fluorescent protein revealed that the subcellular localization of TIP2;1 and PIP1 and PIP2 homologs was unchanged after 45 min of exposure to salt, whereas a TIP1; 1-green fluorescent protein fusion was relocalized into intracellular spherical structures tentatively identified as intravacuolar invaginations. The appearance of intracellular structures containing PIP1 and PIP2 homologs was occasionally observed after 2 h of salt treatment. In conclusion, this work shows that exposure of roots to salt induces changes in aquaporin expression at multiple levels. These changes include a coordinated transcriptional down-regulation and subcellular relocalization of both PIPs and TIPs. These mechanisms may act in concert to regulate root water transport, mostly in the long term (>= 6 h).
机译:水通道蛋白可促进土壤水的吸收并响应各种环境胁迫,调节根系水力传导率(Lp(r))。在这里,我们使用拟南芥(Arabidopsis thaliana)植物解剖盐对Lp(r)和水通道蛋白表达的影响,并研究胁迫下植物根中水通道蛋白调节的分子和细胞机制。用100 mM NaCl处理植物被认为是一种渗透刺激,并导致Lp(r)迅速(半小时,45分钟)显着下降(70%),并保持至少24 h。进行具有基因特异性标签的宏阵列实验以研究拟南芥水通道蛋白家族的所有35个基因的表达。在未处理的根中检测到来自20个水通道蛋白基因的转录本,其中大多数编码质膜内在蛋白(PIP)和液泡膜内在蛋白(TIP)亚家族的成员。所有具有强表达信号的PIP和TIP水通道蛋白转录物在暴露于盐后2至4小时内,其丰度降低60%至75%。与特定水通道蛋白亚型的亚型交叉反应的抗肽抗体的使用表明,PIP1的丰度最早在盐接触后30分钟下降了40%,而PIP2和TIP1的同系物在盐接触后的丰度下降了20%至40%。治疗6小时。与绿色荧光蛋白融合的水通道蛋白在转基因植物中的表达表明,暴露于盐45分钟后,TIP2; 1和PIP1和PIP2同源物的亚细胞定位没有变化,而TIP1; 1-绿色荧光蛋白融合物重新定位到暂时被鉴定为真空内侵的细胞内球形结构。盐处理2小时后,偶尔会观察到含有PIP1和PIP2同源物的细胞内结构。总之,这项工作表明,根部暴露于盐会在多个水平上诱导水通道蛋白表达的变化。这些变化包括PIP和TIP的协调转录下调和亚细胞再定位。这些机制可能共同作用以调节根系水的运输,主要是长期(> = 6 h)。

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