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木薯MeNRT2.5基因的克隆及表达分析

任宁,  陈秀珍,  夏幽泉,  白雪杨,  江行玉,  周扬

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文章导航 > 热带生物学报  > 2019 >  10(2): 111-118
任宁, 陈秀珍, 夏幽泉, 白雪杨, 江行玉, 周扬. 木薯MeNRT2.5基因的克隆及表达分析[J]. 热带生物学报, 2019, 10(2): 111-118. doi: 10.15886/j.cnki.rdswxb.2019.02.003
引用本文: 任宁, 陈秀珍, 夏幽泉, 白雪杨, 江行玉, 周扬. 木薯MeNRT2.5基因的克隆及表达分析[J]. 热带生物学报, 2019, 10(2): 111-118. doi: 10.15886/j.cnki.rdswxb.2019.02.003
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REN Ning, CHEN Xiuzhen, XIA Youquan, BAI Xueyang, JIANG Xingyu, ZHOU Yang. Cloning and Expression Analysis of MeNRT2.5 Gene in Cassava[J]. Journal of Tropical Biology, 2019, 10(2): 111-118. doi: 10.15886/j.cnki.rdswxb.2019.02.003
Citation: REN Ning, CHEN Xiuzhen, XIA Youquan, BAI Xueyang, JIANG Xingyu, ZHOU Yang. Cloning and Expression Analysis of MeNRT2.5 Gene in Cassava[J]. Journal of Tropical Biology, 2019, 10(2): 111-118. doi: 10.15886/j.cnki.rdswxb.2019.02.003
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木薯MeNRT2.5基因的克隆及表达分析

doi: 10.15886/j.cnki.rdswxb.2019.02.003

  • 海南大学 热带农林学院/海南省热带生物资源可持续利用重点实验室, 海口 570228

基金项目: 

海南省重大科技计划项目(HNGDhs201502)

国家自然科学基金(31660253)

海南大学科研启动项目(KYQD(ZR)1845)

海南大学科研团队项目(hdkytg201706)

详细信息
    第一作者:

    任宁(1993-),女,海南大学热带农林学院2016级硕士研究生,E-mail:renning_hu@163.com

    通信作者:

    周扬(1988-),男,博士,讲师.研究方向:植物抗逆分子生物学,E-mail:zhouyang@hainanu.edu.cn

  • 中图分类号: Q786

Cloning and Expression Analysis of MeNRT2.5 Gene in Cassava

  • Institute of Tropical Agriculture and Forestry, Hainan University/Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresources, Haikou, Hainan 570228, China

  • 摘要: 木薯具有高产、耐旱、耐贫瘠等特点,为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30 d龄的"华南8号"组培苗为实验材料,采用同源克隆技术获得1个高亲和性硝酸根转运蛋白基因NRT2。生物信息学分析结果表明,该基因开放阅读框全长1 479 bp,编码492个氨基酸,命名为MeNRT2.5。MeNRT2.5蛋白有10个跨膜区域,与橡胶树、油桐树、可可树等物种的NRT2.5蛋白具有较高的同源性,其氨基酸序列相似性分别为94%,89.84%,87.40%。半质量RT-PCR结果表明,MeNRT2.5基因在根、茎、叶、花等各个器官均有表达,在成熟木薯植株的根中和组培苗的叶中表达量较高。实时荧光定量RT-PCR分析表明,MeNRT2.5在低浓度NO3-(0.3 mmol·L-1)处理后,其在根中的表达量在6 h时达到峰值;高浓度NO3-(3 mmol·L-1)处理后,其表达量在根茎叶中均没有明显变化,即该基因的表达被高浓度NO3-所抑制。该研究结果为进一步分析MeNRT2.5在木薯中的功能验证奠定理论基础。
    Abstract: Cassava（Manihot esculenta Crantz） is characteristic of high yield, drought tolerance and poor soil tolerance. To probe into the molecular basis for tolerance of poor soils and enhance the utilization rate of nitrogen in cassava, two years old mature plants and 1 month-old tissue cultured plants of cassava cultivar "Huanan 8" were collected and cultured for 30 days in a medium as the experimental material, from which a high-affinity nitrate transporter gene NRT2 was isolated by using homologous cloning technique. Sequence analysis revealed that the open reading frame of this gene called MeNRT2.5 is 1 479 bp and encodes 492 amino acids. Bioinformatics analyses showed that the MeNRT2.5 protein had 10 transmembrane regions, and had a a high homology with the NRT2.5 protein of Hevea brasiliensis, Jatropha curcas and,Theobroma cacao, with its amino acid sequences being 94%, 89.84% and 87.4% in similarity, respectively. Semi-Quantitative real-time PCR analysis showed that the MeNRT2.5 gene was expressed in root, stem, leaf, flower and other organs, with its expression being higher in the roots of mature cassava plants and in the leaves of tissue cultured plants. Quantitative real-time PCR analysis showed that transcript of the MeNRT2.5 in root had a peak expression at 6 h under the treatment with low concentration NO3-(0.3 mmol·L-1) but its expression did not change significantly in the root, stem and leaf after the treatment with high concentration of NO3-(3 mmol·L-1), indicating the transcript of the MeNRT2.5 gene is inhibited by high concentrations of NO3-. All these results provide a theoretical foundation for further analysis of the functional verification of the MeNRT2.5 gene in cassava.
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    • 收稿日期:  2018-03-16
    • 修回日期:  2019-03-19

    木薯MeNRT2.5基因的克隆及表达分析

    doi: 10.15886/j.cnki.rdswxb.2019.02.003
    • 海南大学 热带农林学院/海南省热带生物资源可持续利用重点实验室, 海口 570228
      • 基金项目:

      海南省重大科技计划项目(HNGDhs201502)

      国家自然科学基金(31660253)

      海南大学科研启动项目(KYQD(ZR)1845)

      海南大学科研团队项目(hdkytg201706)

      作者简介:

      任宁(1993-),女,海南大学热带农林学院2016级硕士研究生,E-mail:renning_hu@163.com

      通讯作者: 周扬(1988-),男,博士,讲师.研究方向:植物抗逆分子生物学,E-mail:zhouyang@hainanu.edu.cn
    • 收稿日期: 2018-03-16
    • 修回日期: 2019-03-19

    • 中图分类号: Q786

    摘要: 木薯具有高产、耐旱、耐贫瘠等特点,为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30 d龄的"华南8号"组培苗为实验材料,采用同源克隆技术获得1个高亲和性硝酸根转运蛋白基因NRT2。生物信息学分析结果表明,该基因开放阅读框全长1 479 bp,编码492个氨基酸,命名为MeNRT2.5。MeNRT2.5蛋白有10个跨膜区域,与橡胶树、油桐树、可可树等物种的NRT2.5蛋白具有较高的同源性,其氨基酸序列相似性分别为94%,89.84%,87.40%。半质量RT-PCR结果表明,MeNRT2.5基因在根、茎、叶、花等各个器官均有表达,在成熟木薯植株的根中和组培苗的叶中表达量较高。实时荧光定量RT-PCR分析表明,MeNRT2.5在低浓度NO3-(0.3 mmol·L-1)处理后,其在根中的表达量在6 h时达到峰值;高浓度NO3-(3 mmol·L-1)处理后,其表达量在根茎叶中均没有明显变化,即该基因的表达被高浓度NO3-所抑制。该研究结果为进一步分析MeNRT2.5在木薯中的功能验证奠定理论基础。

    English Abstract

    任宁, 陈秀珍, 夏幽泉, 白雪杨, 江行玉, 周扬. 木薯MeNRT2.5基因的克隆及表达分析[J]. 热带生物学报, 2019, 10(2): 111-118. doi: 10.15886/j.cnki.rdswxb.2019.02.003
    引用本文: 任宁, 陈秀珍, 夏幽泉, 白雪杨, 江行玉, 周扬. 木薯MeNRT2.5基因的克隆及表达分析[J]. 热带生物学报, 2019, 10(2): 111-118. doi: 10.15886/j.cnki.rdswxb.2019.02.003
    shu
    REN Ning, CHEN Xiuzhen, XIA Youquan, BAI Xueyang, JIANG Xingyu, ZHOU Yang. Cloning and Expression Analysis of MeNRT2.5 Gene in Cassava[J]. Journal of Tropical Biology, 2019, 10(2): 111-118. doi: 10.15886/j.cnki.rdswxb.2019.02.003
    Citation: REN Ning, CHEN Xiuzhen, XIA Youquan, BAI Xueyang, JIANG Xingyu, ZHOU Yang. Cloning and Expression Analysis of MeNRT2.5 Gene in Cassava[J]. Journal of Tropical Biology, 2019, 10(2): 111-118. doi: 10.15886/j.cnki.rdswxb.2019.02.003
    shu

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