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通过异位表达小麦反向转运蛋白基因(TaNHX2)提高转基因向日葵植株的耐盐性和生长性能。

Improved salinity tolerance and growth performance in transgenic sunflower plants via ectopic expression of a wheat antiporter gene (TaNHX2).

机构信息

Aegis Agro Chemical India Pvt Ltd, Plot No: B 11/1, Industrial Developmental Area, Uppal, Hyderabad, Telangana, 500039, India.

Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, Telangana, 500049, India.

出版信息

Mol Biol Rep. 2019 Dec;46(6):5941-5953. doi: 10.1007/s11033-019-05028-7. Epub 2019 Aug 10.

Abstract

Sunflower (Helianthus annuus. L) is one of the principal oil seed crops affected by the salinity stress, which limits the oil content and crop yield of sunflower plants. The acclimatization of plants to abiotic stresses such as salinity tolerance is mainly mediated by the vacuolar Na/H antiporters (NHX) by tagging Na into vacuoles from the cytosol. We show here that the over-expression of wheat TaNHX2 gene in transgenic sunflower conferred improved salinity stress tolerance and growth performance. Transgenic sunflower plants were produced by infecting the embryonic axis ex-plants with Agrobacterium tumefaciens strain EHA105 containing a pBin438-TaNHX2 binary vector that carried a wheat antiporter (TaNHX2) gene under the control of a double CaMV 35S promoter with NPT II gene as a selectable marker. PCR analysis of T and T transgenic plants confirmed the integration of TaNHX2 in sunflower genome. Stable integration and expression of TaNHX2 in sunflower genome was further verified by Southern hybridization and semi-quantitative RT-PCR analyses. As compared to the non-transformed plants, TaNHX2 expressing transgenic plants showed better growth performance and accumulated higher Na, K contents in leaves and roots under salt stress (200 mM NaCl). Transgenic sunflower plants displayed improved protection against cell damage exhibiting stable relative water content, chlorophyll content, increased proline accumulation and improved reactive oxygen species (ROS) scavenging because of higher activities of the antioxidant enzymes like superoxide dismutase and ascorbate peroxidase, along with decreased production of hydrogen peroxide, free oxygen radical and malondialdehyde (MDA) under salt stress (200 mM NaCl). Taken together, our findings suggest that TaNHX2 expression in sunflower plants contributed towards improving growth performance under sodium chloride stress.

摘要

向日葵(Helianthus annuus. L)是受盐胁迫影响的主要油料作物之一,盐胁迫限制了向日葵植物的含油量和产量。植物对非生物胁迫(如耐盐性)的适应主要是通过液泡中的 Na/H 反向转运蛋白(NHX)来介导的,该蛋白将 Na 从细胞质标记到液泡中。我们在这里表明,在转基因向日葵中过表达小麦 TaNHX2 基因赋予了其提高的耐盐性和生长性能。通过用含有 pBin438-TaNHX2 双元载体的根癌农杆菌菌株 EHA105 感染胚胎轴外植体,生产转基因向日葵植物,该载体在双 CaMV 35S 启动子的控制下携带小麦反向转运蛋白(TaNHX2)基因,带有 NPT II 基因作为可选择标记。T1 和 T2 转基因植物的 PCR 分析证实 TaNHX2 已整合到向日葵基因组中。通过 Southern 杂交和半定量 RT-PCR 分析进一步证实了 TaNHX2 在向日葵基因组中的稳定整合和表达。与非转化植物相比,在盐胁迫(200 mM NaCl)下,TaNHX2 表达的转基因植物表现出更好的生长性能,叶片和根部积累了更高的 Na、K 含量。由于抗氧化酶如超氧化物歧化酶和抗坏血酸过氧化物酶的活性较高,同时由于过氧化氢、游离氧自由基和丙二醛(MDA)的产生减少,转基因向日葵植物表现出对细胞损伤的改善保护,表现为稳定的相对含水量、叶绿素含量、脯氨酸积累增加和活性氧(ROS)清除增强。综上所述,我们的研究结果表明,TaNHX2 在向日葵植物中的表达有助于提高其在氯化钠胁迫下的生长性能。

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