荷花NnNHX1基因耐盐性在转化烟草中的验证

园艺学报 ›› 2012, Vol. 39 ›› Issue (2): 323-332.

荷花NnNHX1基因耐盐性在转化烟草中的验证

郭会敏，顾春笋，陈发棣，徐迎春，刘兆磊^*

南京农业大学园艺学院，南京 210095

出版日期:2012-02-25 发布日期:2012-02-25
通讯作者: 刘兆磊 E-mail:lzl@njau.edu.cn

Salt Tolerance Verification of Lotus NnNHX1 in Transformed Tobacco

GUO Hui-min，GU Chun-sun，CHEN Fa-di，XU Ying-chun，and LIU Zhao-lei^*

College of Horticulture，Nanjing Agricultural University，Nanjing 210095，China

Online:2012-02-25 Published:2012-02-25
Contact: LIU Zhao-lei E-mail:lzl@njau.edu.cn

摘要/Abstract

摘要： 为研究荷花液泡膜型Na⁺/H⁺逆向转运蛋白基因NnNHX1的耐盐性，构建了植物表达载体pCAMBIA1301-220-NnNHX1，并通过农杆菌介导的叶盘转化法将NnNHX1基因转入烟草。利用荧光定量PCR分析NnNHX1在转基因烟草中的表达情况。对转基因烟草进行盐胁迫试验，并测定其在盐胁迫条件下叶绿素、脯氨酸和丙二醛含量、相对电导率、SOD和POD活性等相关生理指标及后代种子的发芽率。结果显示，在获得的7个转NnNHX1基因烟草株系中，T-N1、T-N2和T-N10等3个株系NnNHX1表达量较高。盐胁迫下，转基因烟草组培苗和盆栽苗的生长状况都好于对照植株，在叶绿素含量、脯氨酸含量、细胞质膜的完整性（相对电导率和丙二醛含量）和抗氧化酶（SOD和POD）活性等方面都好于对照，并且后代种子的发芽率也明显高于对照。表明荷花NnNHX1基因能够在烟草中正常翻译和表达，NnNHX1的过量表达提高了转基因烟草的耐盐性。

关键词: 荷花, NnNHX1基因, 转基因, 烟草, 耐盐性

Abstract: To study the salt tolerance of tobacco plants transformed with lotus vacuolar Na⁺/H⁺ antiporter gene NnNHX1，a plant expression vector pCAMBIA1301-220-NnNHX1 was constructed and then was transformed into wild Nicotiana tabacum by Agrobacterium-mediated leaf disc transformation. The expression of NnNHX1 in transgenic tobacco was detected by quantitative PCR analysis. Salt stress test on transgenic tobacco and then the chlorophyll，proline and MDA content，relative conductivity，SOD，POD and other physiological indexes and the generation rate of transgenic tobacco seed were measured in salt stress condition. The results showed that among the seven transgenic tobacco strains，the expression of NnNHX 1 in T-N1，T-N2 and T-N10 strains was higher. Under salt stress，the growth conditions of both tissue culture seedlings and potted seedlings of transgenic tobacco were better than control tobacco. The chlorophyll and proline content and cell membrane integrity（relative conductivity and MDA）and antioxidant enzyme（SOD and POD）activity in transgenic tobacco were better than that of control tobacco，and the transgenic tobacco seed germination was significantly higher than control. Indicated that lotus NnNHX1 gene could normaly translated and expressed in tobacco，and over expression of NnNHX1 enhanced salt tolerance of transgenic tobacco.

Key words: lotus, NnNHX1 gene, transgenic, tobacco, salt tolerance

中图分类号:

S 682.32

郭会敏, 顾春笋, 陈发棣, 徐迎春, 刘兆磊. 荷花NnNHX1基因耐盐性在转化烟草中的验证[J]. 园艺学报, 2012, 39(2): 323-332.

GUO Hui-Min, GU Chun-Sun, CHEN Fa-Di, XU Ying-Chun, LIU Zhao-Lei. Salt Tolerance Verification of Lotus NnNHX1 in Transformed Tobacco[J]. ACTA HORTICULTURAE SINICA, 2012, 39(2): 323-332.

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