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EEB：西北農(nóng)林科技大學(xué)丨MsPIP2; 2提升轉(zhuǎn)基因擬南芥耐鹽性
期刊：Environmental and Experimental Botany（EEB）
主題：MsPIP2; 2提升轉(zhuǎn)基因擬南芥耐鹽性
標(biāo)題：MsPIP2; 2, a novel aquaporin gene from Medicago sativa, confers salt tolerance in transgenic Arabidopsis
影響因子：3.712
檢測(cè)指標(biāo)： Na+、K+流速
檢測(cè)部位：伸長(zhǎng)區(qū)，距離根尖550-600μm
Na+、K+流實(shí)驗(yàn)處理方法：
7日齡擬南芥，150mM NaCl處理24小時(shí)
Na+、K+流實(shí)驗(yàn)測(cè)試液成份：
文獻(xiàn)無(wú)
推薦測(cè)試液成份：
- Na+：0.5mMNaCl ，0.3mM MES，pH6.0
- K+：0.1mMKCl ，0.3mM MES，pH6.0
通訊作者：西北農(nóng)林科技大學(xué)楊培志
英文摘要
Aquaporins (AQPs) are channel proteins that facilitate water transport across cell membranes and play important roles in many biological processes. However, most AQP functions are still poorly understood in the plant kingdom. Here, MsPIP2;2 was isolated and identified from alfalfa (Medicago sativa). MsPIP2;2 was localized to the plasma membrane, and its expression was induced by salt and abscisic acid (ABA) treatment. Overexpression of MsPIP2;2 in Arabidopsis increased the seed germination rate, seedling root length, survival rate, proline content and antioxidant defence activity and decreased cell membrane damage and reactive oxygen species (ROS) accumulation compared to those in WT under salt stress.
The salt tolerance of MsPIP2;2 was affected by Ca2+ and pH in transgenic Arabidopsis plants. MsPIP2;2-overexpressing plants maintained a better K+/Na+ ratio and higher Ca2+ content under salt stress. The higher K+/Na+ maintenance in transgenic plants was mainly achieved by increasing Na+ efflux and K+ retention in roots via regulating the expression of the related ion channel genes. Stress-responsive genes, including P5CS1, RD29A, DREB2 and KIN2, were upregulated in transgenic plants under salt stress.
These results suggest that MsPIP2;2 confers salt tolerance by regulating antioxidant defence system-mediated ROS scavenging, K/Na ion homeostasis and stress-responsive gene expression in plants.
中文摘要（谷歌機(jī)翻）
水通道蛋白（AQPs）是促進(jìn)水跨細(xì)胞膜轉(zhuǎn)運(yùn)的通道蛋白，在許多生物過(guò)程中起重要作用。然而，大多數(shù)AQP功能在植物界仍然知之甚少。在此，從紫花苜蓿（Medicago sativa）中分離并鑒定了MsPIP2; 2。MsPIP2; 2定位于質(zhì)膜，其表達(dá)由鹽和脫落酸（ABA）處理誘導(dǎo)。與鹽脅迫下WT相比，擬南芥中MsPIP2; 2的過(guò)量表達(dá)提高了種子萌發(fā)率，幼苗根長(zhǎng)，存活率，脯氨酸含量和抗氧化防御活性，降低了細(xì)胞膜損傷和活性氧（ROS）積累。
MsPIP2; 2的耐鹽性受轉(zhuǎn)基因擬南芥植物中Ca2+和pH的影響。MsPIP2; 2過(guò)表達(dá)植物在鹽脅迫下保持更好的K+ / Na+比率和更高的Ca2+含量。轉(zhuǎn)基因植物中較高的K+ / Na+維持率主要通過(guò)調(diào)節(jié)相關(guān)離子通道基因的表達(dá)來(lái)增加根中的Na+流出和K+保留來(lái)實(shí)現(xiàn)。應(yīng)激反應(yīng)基因，包括P5CS1，RD29A，DREB2和KIN2，在鹽脅迫下在轉(zhuǎn)基因植物中上調(diào)。
這些結(jié)果表明，MsPIP2; 2通過(guò)調(diào)節(jié)抗氧化防御系統(tǒng)介導(dǎo)的ROS清除，K / Na離子穩(wěn)態(tài)和植物中的應(yīng)激反應(yīng)基因表達(dá)來(lái)賦予耐鹽性。
Fig. 9. The net ?ux of Na+ and K+ at the root tip elongation zones of 7-day-old Arabidopsis seedlings. (A) The net Na+ ?ux under normal conditions (0 mM NaCl). (B) The net Na+ ?ux after 150 mM NaCl stress. (C) The mean Na+ ?ux. (D) The net K+ ?ux under normal conditions (0 mM NaCl). (E) The net K+ ?ux after 150 mM NaCl stress. (F) The mean K+ ?ux. Bars represent the mean ± SE (n = 3). Asterisks and double asterisks above the bars indicate signi?cant di?erences between the transgenic lines and the WT under the same growth conditions: *, P < 0.05; **, P < 0.01.