| 盐生植物盐爪爪液泡膜钠氢反向运输载体基因(KfNHX1)遗传转化拟南芥的耐盐性鉴定 |
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| 引用本文： | 银芳柳,毛晓菲,曾幼玲.盐生植物盐爪爪液泡膜钠氢反向运输载体基因(KfNHX1)遗传转化拟南芥的耐盐性鉴定[J].新疆农业科学,2021,58(3):565-572. |
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| 作者姓名： | 银芳柳 毛晓菲 曾幼玲 |
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| 作者单位： | 新疆大学生命科学与技术学院/新疆生物资源基因工程重点实验室,乌鲁木齐 830046 |
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| 基金项目： | 国家自然科学基金 (31160186) |
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| 摘 要： | 【目的】 研究盐爪爪液泡膜Na+/H+反向运输载体KfNHX1 (AY825250) 基因的耐盐功能,为耐盐育种提供候选基因。【方法】 采用农杆菌介导花序浸染的方法,将KfNHX1转入拟南芥中,结合基因组PCR和RT-PCR方法鉴定符合3∶1分离比的转基因株系;利用在盐胁迫下的萌发率、根长和表型分析,结合原子吸收分光光度计法测定叶片的Na+、K+含量,推断其耐盐性。【结果】 对抗生素筛选符合3∶1的转基因纯合株系进行基因组PCR和RT-PCR分析,证实KfNHX1基因在拟南芥基因组中整合和表达。盐胁迫下转基因株系的拟南芥种子的萌发率和根长明显高于野生型。200 mM NaCl胁迫处理15 d的拟南芥成苗,相较野生型叶片萎黄和死亡,转基因植株的生长表型较好,且积累了较高的Na+和K+。外源ABA的处理下,转基因植株的发芽率和生长表型也好于野生型。【结论】 盐爪爪(Kalidium foliatum)是一种藜科(Chenopodiaceae)盐生灌木,对盐的耐受性很强。液泡膜Na+/H+反向运输体(NHX)是在离子稳态中起重要作用的膜蛋白,通过调节胞间离子的跨膜转运来维持细胞内离子和pH平衡。盐生植物盐爪爪KfNHX1能够提高转基因拟南芥的耐盐性,具有提高植物耐盐性的潜力。
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| 关 键 词： | 盐生植物 盐爪爪 KfNHX1 耐盐性 拟南芥 遗传转化 |
| 收稿时间： | 2020-10-15 |
Salt-Tolerant Identification of Genetic Transformation in Arabidopsis with the KfNHX1 Gene from the Halophyte Kalidium foliatum |
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| YIN Fangliu,MAO Xiaofei,ZENG Youling.Salt-Tolerant Identification of Genetic Transformation in Arabidopsis with the KfNHX1 Gene from the Halophyte Kalidium foliatum[J].Xinjiang Agricultural Sciences,2021,58(3):565-572. |
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| Authors: | YIN Fangliu MAO Xiaofei ZENG Youling |
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| Institution: | Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China |
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| Abstract: | 【Objective】 The halophyte Kalidium foliatum is a very salt-tolerant shrub belonging to Chenopodiaceae with strong resistance to salinity.Vacuolar Na+/H+ antiporter (NHX) is a membrane protein that plays an important role in ionic homeostasis.It maintains intracellular ion and pH balance by regulating the transmembrane transport of ions between cells.To study the salt tolerance function of the vacuolar Na+/H+ antiporter gene (KfNHX1, AY825250) from K.foliatum, to provide candidate genes for salt tolerance breeding.【Methods】 The KfNHX1 gene was transferred into Arabidopsis thaliana by the method of Agrobacterium tumefaciens mediated inflorescence transformation; combined with genomic PCR and RT-PCR methods to identify transgene lines with 3:1 segregation ratio; utilize the germination rate, root length and phenotype analysis under salt stress; combine the content of Na+ and K+ in leaves with an atomic absorption spectrophotometer to infer the salt tolerance.【Results】 Genomic PCR and RT-PCR analysis were performed on the homozygous transgenic plants by antibiotic screening of 3∶1 to confirm that the KfNHX1 gene was stably integrated into the genome and expressed in Arabidopsis.Further salt tolerant physiological function data show that the transgenic Arabidopsis had higher germination rate and root length than the wild-type under salt stress.The WT and transgenic plants were stressed with 200 mM NaCl for 15 days, the transgenic plants grew well and accumulated high Na+ and K+ contents in the leaves, but the WT plants were chlorotic and dying.Besides, the transgenic plants have higher germination rate and growth phenotype under exogenous ABA applyment.【Conclusion】 The results showed that the K.foliatum KfNHX1 can enhance the salt tolerance of transgenic Arabidopsis and has the potential to improvement of plant salt tolerance. |
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| Keywords: | halophyte Kalidium foliatum KfNHX1 salt tolerance Arabidopsis thaliana genetic transformation |
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