木薯MeNRT2.1基因的克隆及表达分析

发布时间：2018-05-30 18:26

本文选题：木薯 + 硝态氮　；参考：《热带作物学报》2016年01期

【摘要】：木薯具有产量高、抗贫瘠等特点,为了了解其耐贫瘠的作用机理,提高木薯在贫瘠土壤中对氮素的利用率,以培养20 d的"华南5号"木薯组培苗为实验材料,采用同源克隆和RT-PCR技术,获得一个高亲和硝态氮转运蛋白(NRT2)基因,命名为Me NRT2.1,该基因含有1 593 bp的开放阅读框架,编码530个氨基酸。生物信息学分析结果表明,木薯Me NRT2.1与苜蓿、拟南芥、可可、杨树等物种的NRT2.1同源性高,其中与可可树Tc NRT2.1的亲缘关系最近,氨基酸相似性达到90%。实时荧光定量PCR检测结果表明,Me NRT2.1在木薯组培苗的根中表达,并且在NO_3~-浓度为0.2 mmol/L时,其相对表达量较高,NO_3~-浓度为10 mmol/L时其相对表达量较低,NO_3~-浓度为0时几乎不表达;Me NRT2.1在茎、叶中也几乎不表达,即该基因具有诱导型组织特异性表达模式。原生质体瞬时表达发现Me NRT2.1定位在细胞膜上。此研究为进一步通过NRT2基因提高木薯的抗逆性奠定了基础。
[Abstract]:Cassava has the characteristics of high yield and resistance to barren. In order to understand its mechanism of tolerance to barren, and to improve the utilization rate of nitrogen in barren soil, the tissue culture seedling of "South China No.5" cassava was cultured for 20 days as the experimental material. By using homologous cloning and RT-PCR techniques, a highly compatible nitrate-nitrogen transporter (NRT2) gene was obtained, named me NRT2.1, which contains an open reading frame of 1 593bp and encodes 530 amino acids. Bioinformatics analysis showed that me NRT2.1 had high NRT2.1 homology with alfalfa, Arabidopsis thaliana, cocoa, poplar and so on. The results of real-time fluorescence quantitative PCR showed that me NRT2.1 was expressed in the roots of cassava plantlets in tissue culture, and the relative expression of me NRT2.1 was relatively high when the concentration of NO3- was 0.2 mmol/L. When the concentration was 10 mmol/L, the relative expression of me NRT2.1 was almost non-expressed in stems. There was almost no expression in leaves, that is, the gene had inducible tissue specific expression pattern. It was found that me NRT2.1 was located on the cell membrane by transient expression in protoplasts. This study laid a foundation for further enhancing the stress resistance of cassava by NRT2 gene.
【作者单位】：海南大学农学院;中国热带农业科学院热带生物技术研究所农业部热带作物生物学与遗传资源利用重点实验室;
【基金】：国际科技合作与交流项目(No.2013DFA32020) 海南省重大科技项目子课题热带生物种质与基因资源研究(No.ZDZX2013023-1) 国家高技术研究发展计划重点项目(No.2012AA101204)
【分类号】：S533

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