大车前（Plantago major L. "Giant Turkish."）体外优化再生体系的建立及Na+/H+逆向转运蛋白基因SeNHX1 和TtNHX1 的功能鉴定

大车前（Plantago major L. "Giant Turkish."）不仅有很高的药用价值，在生态学研究方面也是重要模式植物。大车前的组织培养工作，目前报道很少。对其组织培养体系的建立，为筛选大车前耐盐突变体和基因转化建立高效的体外再生系统和实验平台体系。通过愈伤组织诱导和直接不定芽再生途径， 建立了大车前（Plantago major L. "Giant Turkish."）的快速高效再生系统。叶片外植体在含有1.0 mg/L NAA的MS培养基中培养3周后，形成愈伤组织，愈伤组织在含4.0 mg/L 6-BA的MS培养基中成功再生，得到完整植株。种子外植体在含0.2 mg/L IAA和1.0 mg/L TDZ的MS培养基中培养4周后产生大量的丛生芽，对9株再生植株进行RAPD检测表明，部分植株在DNA水平上发生了变异。 植物抵御盐胁迫的一个重要机制是在液泡中积累Na+，从而使细胞质内Na+保持在较低水平，并且降低细胞渗透势。Na+运输到液泡是由液泡Na+/H+逆向转运蛋白完成的。本实验室已从盐生植物盐角草（Salicornia europaea）和番杏（Tetragonia tetragonioides）中分别克隆得到SeNHX1和TtNHX1基因。本文研究了SeNHX1和TtNHX1基因在酵母突变体里的作用。TtNHX1和SeNHX1蛋白在缺陷型酵母菌株里的表达能够提高这些菌株对NaCl、LiCl和潮霉素的抗性，提高到与野生型相当的抗性水平。说明TtNHX1和SeNHX1有着与酵母ScNHX1相似的细胞定位和作用机制，是ScNHX1的功能类似蛋白。

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null RAE2008

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