AtNHX1基因转化番茄及其耐盐碱性分析  

Introduction of AtNHX1 Gene into Tomato and Salt and Alkali Tolerance Analysis in Transgenic Tomato

作　　者：王东明[1] 崔继哲[1] 杨淳淋[1] 

机构地区：[1]哈尔滨师范大学生命科学与技术学院,哈尔滨150025

出　　处：《中国农学通报》2009年第22期52-56,共5页Chinese Agricultural Science Bulletin

基　　金：黑龙江省科技攻关计划项目"Na+/H+反向转运蛋白基因克隆及AtNHX1对番茄的遗传转化研究"(GA06B103-7)

摘　　要：高盐是限制作物生长发育和产量最严重的非生物胁迫之一,提高作物的耐盐性已成为一个日益紧迫的研究课题。将含有AtNHX1基因的表达载体pFYC-AtNHX1和含有bar基因的质粒pCAMBIA3300等量混合,采用花粉管通道法共转化番茄。经PPT抗性筛选后,PCR扩增获得12株导入了AtNHX1基因的番茄植株,Southern杂交分析表明该基因已经整合到番茄基因组中,通过RT-PCR检测到了AtNHX1基因的表达。在150 mmol/L NaCl处理下,转基因植株T1代对NaCl耐性显著增强。对转AtNHX1基因番茄T2代进行80 mmol/L碱性盐NaHCO(3pH 8.25)胁迫,以此评价转AtNHX1基因番茄对碳酸盐的耐受性。叶片相对电导率检测结果表明,AtNHX1基因的导入确实提高了番茄对碱性盐NaHCO3的耐受性。培育并栽种转AtNHX1基因的耐盐碱番茄将会降低盐渍化环境对番茄生长发育的影响。High salinity is one of the major abiotic stresses which constraints plant development, growth and productivity, improving the salt tolerance of plants has been an important project. The expression vector pFYC-AtNHX1containing AtNHX1 and pCAMBIA3300 containing bar gene mixed in equal concentrations were co-transformed into tomato varieties via pollen-tube pathway. 12 AtNHX1 positive plants were selected by spraying 300 mg/L Basra on transgenic tomato leaves and by PCR assay of specific AtNHX1 for PPT resistant plants. The presence and integration of the AtNHX1 in transgenic tomato genome were confirmed by Southern blotting. Expression of the AtNHX1 in transgenic positive plants was determined by RT-PCR .The salt tolerance of transgenic plants were enhanced when exposed to 150 mmol/L NaC1. The results demonstrated that AtN- HX1 gene could improve the salt tolerance of tomato effectively. In order to investigate whether the AtNHX1 gene transformation can enhance the NaHCO3 tolerance of the T2 transgenic plants, relative conductivity in leaves of the transgenic plant and control were estimated when they were cultured under the 80 mmol/L Nail- CO3 （pH 8.25） stress. The results indicated that the intergration ofAtNHX1 gene enhanced the resistance to the NaHCO3 stress of the transgenic tomato. Breeding salt and alkali tolerant transgenic AtNHX1 tomato cultivars is one approach to reducing the deleterious effects of saline environment on tomato.

关 键 词：ATNHX1基因 转基因番茄 耐盐性 NAHCO3胁迫 相对电导率 

分 类 号：Q945[生物学—植物学]