转TaNHX2大豆的耐盐性分析

doi:10.3864/j.issn.0578-1752.2015.20.002

摘要/Abstract

摘要： 【目的】在盐胁迫条件下，通过对转TaNHX2(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TaNHX2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NaCl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NaCl浓度，北圃场夏播试验点设置0和200 mmol·L^-1 2个NaCl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TaNHX2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^-1NaCl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 NaCl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 NaCl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 NaCl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品种存在显著性差异。在北圃场试验点设置的200 mmol·L^-1NaCl胁迫处理下，转基因株系C12、C21和C19的株高、C19的单株粒数和单株粒重与受体对照品种具有显著性差异。【结论】在盐胁迫条件下，与受体对照品种相比，转TaNHX2大豆株系C12、C21和C19盐害程度较低，能维持较强的光合作用和一定的产量，其中株系C19在2个试验点的产量表现均较佳，具有较大的育种应用价值。

关键词: 转TaNHX2大豆, 盐胁迫, 产量表现, 生产应用

Abstract: 【Objective】 In order to obtain valuable salt-tolerance soybean germplasms, the productive potential of TaNHX2（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 TaNHX2 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 TaNHX2 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^-1NaCl 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 agronomic trait value of transgenic soybean lines and wild-type were comparable under normal condition, while under 150 mmol·L^-1 and 200 mmol·L^-1NaCl stress, the agronomic trait values of transgenic soybean lines were larger than wild-type. Under 150 mmol·L^-1NaCl stress (Changping), seed weight per plant of lines C12, C21 and C19, pod number per plant of line C19 had a significant difference with wild-type. Under 200 mmol·L^-1NaCl stress (Changping), pod number per plant, seed number per plant and seed weight per plant of lines C12, C21 and C19, plant height of lines C12 and C19 had a significant difference with wild-type. At Beipuchang research site, plant height of lines C12, C21 and C19, seed number per plant and seed weight per plant of line C19 were significantly larger than wild-type. 【Conclusion】Compared with wild-type, transgenic soybean lines (C12, C21 and C19) had lower salt damage degree, kept a higher photosynthesis and certain yield under salt stress, line C19 had an excellent yield potential both at two research sites, therefore, it is practical valuable for breeding and production.

Key words: TaNHX2 over-expression transgenic soybean, salt stress, yield potential, application in production

林抗雪，刘修杰，孙石，陈莉，韩天富，侯文胜. 转TaNHX2大豆的耐盐性分析[J]. 中国农业科学, 2015, 48(20): 3998-4007.

LIN Kang-xue, LIU Xiu-jie, SUN Shi, CHEN Li, HAN Tian-fu, HOU Wen-sheng. Salt Tolerance Analysis of TaNHX2 Over-Expression Transgenic Soybean[J]. Scientia Agricultura Sinica, 2015, 48(20): 3998-4007.

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