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Genes ：北京林業(yè)大學(xué)丨鹽脅迫下鉀轉(zhuǎn)運(yùn)體LrKUP8抑制K+外排
NMT是基因功能的活體檢測(cè)技術(shù)，已被103位諾貝爾獎(jiǎng)得主所在單位，及北大、清華、中科院使用。
期刊：Genes
主題：鹽脅迫下鉀轉(zhuǎn)運(yùn)體LrKUP8抑制K+外排
標(biāo)題：Potassium Transporter LrKUP8 Is Essential for K+Preservation in Lycium ruthenicum, A Salt-Resistant Desert Shrub
影響因子：3.331
檢測(cè)指標(biāo)：K+流速
檢測(cè)樣品：黒枸杞愈傷組織
K+流實(shí)驗(yàn)處理方法：
有/無(wú)300 mM NaCl培養(yǎng)12h
K+流實(shí)驗(yàn)測(cè)試液成份：
0.5 mM KCl, 0.5 mM NaCl, 0.1mM MgCl2, 0.2 mM CaCl2, 2.5% sucrose，pH 5.7
作者：北京林業(yè)大學(xué)陳金煥、戴逢斌
英文摘要
Salt stress is a major constraint for many crops and trees. A wild species of Goji named Lycium ruthenicum is an important economic halophyte in China and has an extremely high tolerance to salinity. L. ruthenicum grows in saline soil and is known as a potash-rich species.
However, its salt adaptation strategies and ion balance mechanism remains poorly understood. Potassium (K+) is one of the essential macronutrients for plant growth and development. In this study, a putative salt stress-responsive gene encoding a HAK (high-affinity K+)/KUP (K+ uptake)/KT (K+ transporter) transporter was cloned and designated as LrKUP8.
This gene belongs to the cluster II group of the KT/HAK/KUP family. The expression of LrKUP8 was strongly induced under high NaCl concentrations. The OE-LrKUP8 calli grew significantly better than the vector control calli under salt stress conditions. Further estimation by ion content and micro-electrode ion flux indicated a relative weaker K+ efflux in the OE-LrKUP8 calli than in the control.
Thus, a key gene involved in K+ uptake under salt condition was functionally characterized using a newly established L. ruthenicum callus transformation system. The importance of K+ regulation in L. ruthenicum under salt tolerance was highlighted.
中文摘要（谷歌機(jī)翻）
鹽脅迫是許多農(nóng)作物和樹(shù)木的主要限制因素。枸杞的一種野生種，稱為枸杞，是中國(guó)重要的經(jīng)濟(jì)鹽生植物，對(duì)鹽分的耐受性*。ruthenicum生長(zhǎng)在鹽漬土壤中，被稱為富含鉀肥的物種。
然而，其鹽適應(yīng)策略和離子平衡機(jī)制仍然知之甚少。鉀（K+）是植物生長(zhǎng)和發(fā)育*的大量營(yíng)養(yǎng)素之一。在這項(xiàng)研究中，克隆了編碼HAK（高親和力K+）/ KUP（K+攝?。? KT（K+轉(zhuǎn)運(yùn)蛋白）轉(zhuǎn)運(yùn)蛋白的假定鹽脅迫響應(yīng)基因，并將其命名為L(zhǎng)rKUP8。
該基因?qū)儆贙T / HAK / KUP家族的II類(lèi)。在高NaCl濃度下強(qiáng)烈誘導(dǎo)LrKUP8的表達(dá)。在鹽脅迫條件下，OE-LrKUP8愈傷組織的生長(zhǎng)明顯好于載體對(duì)照的愈傷組織。通過(guò)離子含量和微電極離子通量的進(jìn)一步估計(jì)表明，與對(duì)照相比，OE-LrKUP8愈傷組織中的K+外排相對(duì)較弱。
因此，使用新建立的黑麥草愈傷組織轉(zhuǎn)化系統(tǒng)對(duì)在鹽條件下參與鉀離子吸收的關(guān)鍵基因進(jìn)行了功能鑒定。強(qiáng)調(diào)了在耐鹽性條件下釕中的K+調(diào)節(jié)的重要性。
結(jié)果表明：LrKUP8抑制了愈傷組織中K+外排。不存在NaCl脅迫時(shí)，兩種愈傷組織的K+外排速率較小，且無(wú)顯著差異。經(jīng)鹽脅迫后，K+外排速率遠(yuǎn)高于無(wú)脅迫組；對(duì)照組（pBI121）的K+外排速率明顯高于過(guò)表達(dá)組（OE-LrKUP8）。過(guò)表達(dá)組由于其更強(qiáng)的保K+能力，因而維持高的K+/Na+比。保持的K+/Na+比是植物耐鹽性的關(guān)鍵特征之一，證明LrKUP8的保K+能力促進(jìn)了黒枸杞在鹽脅迫下的生長(zhǎng)發(fā)育。