甘蔗钾转运蛋白基因SsHAK2的克隆及表达特性分析

发布时间：2018-05-26 23:01

本文选题：甘蔗 + 钾转运蛋白　；参考：《农业生物技术学报》2017年03期

【摘要】：甘蔗(Saccharum species hybrid)是最重要的糖料作物,由于其生育周期长,生物量大,对钾的需求量也大。南方是我国甘蔗的主要种植区域,由于土壤本身特性,有效钾含量低,限制了甘蔗的生产。因此,培育耐低钾甘蔗品种是提高甘蔗钾吸收效率的有效途径之一。本研究以甘蔗品种新台糖22号为材料进行低钾胁迫处理,利用RT-PCR技术从其根系中克隆得到钾转运蛋白基因,命名为SsHAK2(Gen Bank登录号:KM98738)。该基因全长为2 798 bp,包含一个完整的2 352 bp的ORF,编码784个氨基酸,相对分子量为87.602 k D,等电点为8.85,预测其为碱性蛋白。SsHAK2苷酸序列包含了12个跨膜结构域(S1~S12),有80%的概率定位在细胞质膜上。同时,该基因还包含3个保守结构域,分别为钾转运蛋白结构域和氨基酸转运蛋白结构域。SsHAK2基因与玉米(Zea mays)、大麦(Hordeum vulgare)、水稻(Oryza sativa)等其他作物中HAK基因具有高度的同源性,一致的核苷酸比例在52%~95%。q PCR分析结果表明,在低钾、干旱和盐胁迫下,SsHAK2的表达都发生改变。在低钾胁迫条件下处理96 h,该基因相对表达量最大,约达到对照的1.70倍;在盐胁迫条件下处理48 h,该基因的相对表达量出现显著上调,96 h相对表达量最高,约为对照的4.37倍。在干旱胁迫12 h,该基因的相对表达量达到最高,约为对照的4.07倍。干旱胁迫处理24 h,该基因表达量约为对照的1.69倍。但是,在干旱胁迫24到48 h过程中表达由下调转为上调。干旱胁迫48 h后,该基因的表达迅速下调,96 h时该基因表达量最低,约为对照表达量的1/4。q PCR分析结果表明该基因在低钾、干旱和盐胁迫下发挥重要的调控作用。本研究结果为进一步研究甘蔗钾吸收分子机制提供基础。
[Abstract]:Saccharum species hybrid is the most important sugar crop. Because of its long growing period and large biomass, the demand for potassium is also large. The south is the main planting area of sugarcane in China. Because of the characteristic of soil and the low content of available potassium, the production of sugarcane is restricted. Therefore, the cultivation of low potassium tolerant sugarcane varieties is one of the effective ways to improve the potassium absorption efficiency of sugarcane. In this study, the sugarcane cultivar Xintaitang 22 was used for low potassium stress. Potassium transporter gene was cloned from its root system by RT-PCR technique and named as SsHAK2(Gen Bank accession number: KM98738. The gene is 2 798 BP in length and contains a complete 2 352 BP ORF encoding 784 amino acids. The relative molecular weight is 87.602 KD and the isoelectric point is 8.85. It is predicted that the nucleotide sequence of basic protein SsHAK2 contains 12 transmembrane domain S1S12, and the probability is 80% located on the cytoplasmic membrane. At the same time, the gene also contains three conserved domains, namely, potassium transporter domain and amino acid transporter domain. The SsHAK2 gene has high homology with other crops, such as Zea mayssos, Hordeum vulgarea, Oryza sativa, etc. The results of PCR analysis showed that the expression of SsHAK2 changed under low potassium, drought and salt stress. Under the condition of low potassium stress for 96 h, the relative expression of the gene reached 1.70 times of that of the control, and the relative expression of the gene was significantly up-regulated at 96 h after 48 h of salt stress, about 4.37 times of that of the control. At 12 h of drought stress, the relative expression of the gene reached the highest level, about 4.07 times of that of the control. Under drought stress for 24 h, the expression of the gene was 1.69 times as much as that of the control. However, the expression changed from down-regulation to upregulation during 24 to 48 h of drought stress. After 48 h of drought stress, the expression of this gene was the lowest at 96 h after drought stress. The results of 1 / 4.q PCR analysis showed that the gene played an important role in regulation under low potassium, drought and salt stress. The results provide a basis for further study on the molecular mechanism of potassium absorption in sugarcane.
【作者单位】：广东省生物工程研究所(广州甘蔗糖业研究所)广东省甘蔗改良与生物炼制重点实验室;
【基金】：国家甘蔗产业技术体系(CARS-20-1-4) 广东省科技计划项目(No.2014B070705002;No.2014A040401033;No.2014B090907006) 广东省科学院科研平台环境与能力建设专项资金项目(No.2016GDASPT-0306) 亚热带农业生物资源保护与利用国家重点实验室开放课题
【分类号】：S566.1

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