转TaNHX2大豆的耐盐性分析  

Salt Tolerance Analysis of Ta NHX2 Over-Expression Transgenic Soybean

作　　者：林抗雪[1] 刘修杰 孙石[1] 陈莉[1] 韩天富[1] 侯文胜[1] 

机构地区：[1]中国农业科学院作物科学研究所/国家植物转基因技术研究中心/农业部北京大豆生物学重点实验室,北京100081

出　　处：《中国农业科学》2015年第20期3998-4007,共10页Scientia Agricultura Sinica

基　　金：科技部国家转基因生物新品种培育重大科技专项(2014ZX08010-004);中国农业科学院科技创新工程

摘　　要：【目的】在盐胁迫条件下,通过对转Ta NHX2(小麦Na+/H+转运蛋白编码基因)大豆的盐害表型、光合作用强度以及产量相关农艺性状的考察,评估其生产应用潜力,以期获得具有一定应用价值的耐盐大豆新种质。【方法】以实验室前期获得并初步鉴定具有一定耐盐性的转Ta NHX2大豆T4代株系为材料,在主茎第二片复叶完全展开时(V3期)进行草丁膦抗性、PCR和RT-PCR检测。选取阳性植株,在主茎第三片复叶完全展开时(V4期)进行Na Cl胁迫处理,处理期间观察记录盐害表型;待植株长至初荚期(R3期)测定其光合作用参数;最后,分单株收获已生长至完熟期(R8期)的大豆植株,考察其株高、节数、单株荚数、单株粒数和单株粒重。其中,昌平春播试验点设置0、150和200 mmol·L-1 3个Na Cl浓度,北圃场夏播试验点设置0和200 mmol·L-1 2个Na Cl浓度,盐害表型观察以及光合作用参数测定只在北圃场试验点进行。【结果】分子鉴定表明外源Ta NHX2在转基因后代中成功转录表达,遗传稳定。在无盐胁迫条件下,转基因与受体植株长势相当,叶片大小相似,光合作用强度相近。而在200 mmol·L-1 Na Cl溶液胁迫处理下,转基因与受体植株均出现矮化、叶片变小、光合作用强度降低的现象,但与受体植株相比,转基因大豆株系C12、C21和C19的矮化程度较低,叶片较大,光合作用相关参数数值较大,其中株系C12和C21与受体的净光合速率(Pn)差异具有显著性。另外,无盐胁迫条件下,转基因株系与受体品种自贡冬豆各产量相关农艺性状数值相近,而在昌平试验点设置的150和200 mmol·L-1 Na Cl溶液胁迫下,转基因株系各农艺性状数值均高于受体对照,其中150 mmol·L-1 Na Cl胁迫下,C12、C21和C19的单株粒重,C19的单株荚数与受体差异显著。200 mmol·L-1 Na Cl胁迫处理时,株系C12、C21和C19的单株荚数、单株粒数和单株粒重,C12和C19的株高均与受体对照【Objective】 In order to obtain valuable salt-tolerance soybean germplasms, the productive potential of Ta NHX2（wheat Na^＋/H^＋ transporter code gene）over-expression transgenic soybeans was studied through observing their salt induced phenotype, photosynthetic intensity and agronomic traits under salt stress.【Method】The T4 generation of Ta NHX2 over-expression transgenic soybeans, which the transgenic authenticity confirmed by Glufosinate-resistance assay, PCR and RT-PCR detections at V3 stage were chosen as materials. The positive individuals were then treated with NaCl solution at V4 stage, during the treatment phase, the salt induced phenotype of transgenic and wild-type plants was observed. Furthermore, the photosynthetic intensity of the soybeans at R3 stage was measured. Finally, the plant height, node number on main stem, pod number per plant, seed number per plant and seed weight per plant of soybean plants which harvested at R8 stage were measured. Two concentrations of NaCl solution（0 and 200 mmol·L^-1） were applied in Beipuchang research site, while three concentrations were applied（0, 150 and 200 mmol·L^-1） in Changping, Beijing. The observation of salt induced phenotype and measurement of photosynthetic intensity were only carried out at Beipuchang research site. 【Result】 The successful expression of foreign gene Ta NHX2 was confirmed by molecular identification, its inheritance was stable. Transgenic soybean and wild-type plants had similar growth vigor, leaf size and photosynthetic intensity under normal growing condition. While both the transgenic soybean and wild-type plants became shorter under 200 mmol·L^-1 NaCl treatment, their leaves became smaller and their photosynthetic intensity decreased. When compared with wild-type, the transgenic soybean lines（C12, C21 and C19） had lower dwarf degree, larger leaf size and photosynthetic intensity, the photosynthetic rate（Pn） of lines C12 and C21 significantly larger than wild-type. In addition, the yield related ag

关 键 词：转Ta NHX2大豆 盐胁迫 产量表现 生产应用 

分 类 号：S565.1[农业科学—作物学]