转甜瓜CmSAMDC基因拟南芥的获得及其耐盐性研究
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陕西省科技创新与攻关项目(2016NY032);


Regeneration of Transgenic Arabidopsis thaliana Overexpressing Melon CmSAMDC Gene and Its Salt Tolerance
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    摘要:

    该研究以哥伦比亚生态型野生拟南芥为材料,将甜瓜CmSAMDC基因构建到植物双元表达载体pCAMBIA1304上,采用农杆菌介导法转入拟南芥,在含有50 mg/L潮霉素(Hyg)MS固体培养基上筛选转基因后代,并利用T3代转基因幼苗进行耐盐性分析。结果显示:(1)成功构建了植物超表达载体35S∷CmSAMDC,并经农杆菌介导法转化拟南芥,潮霉素抗性筛选后获得了转CmSAMDC基因拟南芥T3代植株。(2)转CmSAMDC基因拟南芥T3代幼苗在含100、150、200 mmol/L NaCl培养基中,侧根长势比野生型植株更为健壮;在200 mmol/L NaCl浇灌处理后,转CmSAMDC基因T3代植株仍能维持正常生长,而野生型植株的生长明显受到抑制;在400 mmol/L NaCl浇灌处理后16 d,野生型植株逐渐死亡,而转基因植株仍能继续存活;对盐胁迫后植株的脂质过氧化程度(MDA)测定显示,野生型植株MDA水平较转基因植株上升更为明显。研究表明,过表达甜瓜CmSAMDC基因增强了转基因拟南芥的耐盐性。

    Abstract:

    In this study, we used wild type of Arabidopsis thaliana as tested materials, and the CmSAMDC gene from melon was built into the pCAMBIA1304 binary vector, then the vector was transformed into A. thaliana by using Agrobacteriummediated method. The transgenic plants overexpressing CmSAMDC were obtained by screened on MS medium containing 50 mg/L hygromycin, and then the salt tolerance test was performed by T3 transgenic seedlings. The results showed that: (1) the plant overexpression vector 35S∷CmSAMDC was successfully constructed and finally the T3 transgenic A. thaliana plants were obtained after transformed by Agrobacterium and resistance screened by hygromycin. (2) The lateral roots of the transgenic T3 lines were stronger than that of wildtype under 100, 150 and 200 mmol/L NaCl treatment. Meanwhile, the transgenic T3 lines grew up normally under 200 mmol/L NaCl treatment by irrigating roots, while the wild type plants were significantly inhibited under the same condition. In addition, the T3 transgenic seedlings were still alive under 400 mmol/L NaCl treatment by irrigating roots 16 days later whereas the wildtype plants gradually died. Furthermore, the level of MDA was higher in wildtype plants than that of transgenic plants under the same salt stress. The results showed that overexpressing of CmSAMDC gene could obviously enhance the salt tolerance of transgenic A. thaliana.

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刘长命,张 显,王永琦.转甜瓜CmSAMDC基因拟南芥的获得及其耐盐性研究[J].西北植物学报,2018,38(7):1179-1186

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  • 在线发布日期: 2018-08-17
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