过表达OsMPK17激酶蛋白质增强了水稻的耐旱性  

Overexpression of OsMPK17 protein enhances drought tolerance of rice

作　　者：马金姣 兰金苹[1,2] 张彤 陈悦 郭亚璐 刘玉晴[1] 燕高伟 魏健 窦世娟 杨明[1] 李莉云[1] 刘国振[1] MA Jin-Jiao;LAN Jin-Ping;ZHANG Tong;CHEN Yue;GUO Ya-Lu;LIU Yu-Qing;YAN Gao-Wei;WEI Jian;DOU Shi-Juan;YANG Ming;LI Li-Yun;LIU Guo-Zhen(College of Life Sciences,Hebei Agricultural University,Baoding 071001,Hebei,China;Research Center for Life Sciences,Hebei North University,Zhangjiakou 075000,Hebei,China;Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen 518116,Guangdong,China)

机构地区：[1]河北农业大学生命科学学院,河北保定071001 [2]河北北方学院生命科学研究中心,河北张家口075000 [3]中国农业科学院农业基因组研究所,广东深圳518116

出　　处：《作物学报》2020年第1期20-30,共11页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31171528)资助~~

摘　　要：促分裂原活化蛋白质激酶(mitogen-activated protein kinase, MAPK)在真核生物中高度保守,在水稻逆境应答反应中也发挥着重要作用。本研究表达纯化了水稻OsMPK17蛋白质,制备了特异性抗体,对多种非生物逆境胁迫下的蛋白质样品进行免疫印迹分析,发现OsMPK17蛋白质在干旱胁迫下诱导表达,提示该蛋白质在干旱胁迫应答中发挥作用。对脱落酸和茉莉酸甲酯处理的离体叶片蛋白质分析发现,OsMPK17蛋白质表达丰度下降,提示该蛋白质的功能发挥可能受激素调控。为此,构建了过表达OsMPK17蛋白质的载体,转化水稻后筛选获得了OsMPK17蛋白质过表达的纯合株系。田间种植鉴定结果表明,转基因株系的株高变矮、穗长变短、结实率降低。种子萌发期拟旱(PEG-6000)处理条件下,过表达OsMPK17株系的种子长势明显比野生型好,根长与芽长均显著大于野生型。幼苗期失水试验表明,转基因植株的失水率低于野生型。在土培干旱胁迫后恢复浇水的试验中,过表达OsMPK17蛋白质的转基因水稻生长也好于野生型。综上,过表达OsMPK17蛋白质提高了水稻的耐旱性。本研究增进了对水稻OsMPK17基因功能的了解。Mitogen-activated protein kinase(MAPK) highly conserved in eukaryotes plays important roles in stress responses in plant. In this study, full-length Os MPK17 gene was cloned and fusion protein was expressed. The purified protein was used as immunogen to generate monoclonal antibody. Western blot(WB) analyses were carried out for protein samples isolated from tissues under different abiotic stresses. The expression of Os MPK17 was induced by drought stress, suggesting that the OsMPK17 protein may play a role in drought stress response. Proteins isolated from leaves treated with abscisic acid(ABA) or methylene jasminate acid(MeJA) demonstrated a decrease of OsMPK17 protein abundance, suggesting that hormones may be involved in the function of the protein. The overexpression vector of OsMPK17 protein was established and transformed into TP309 via Agrobacteria-mediated protocol. Homozygous transgenic lines for overexpression of OsMPK17 protein were obtained. In the field planting experiment, the plant height and the spike length of transgenic lines shortened and the seed setting rate decreased. At seed germination stage, under the condition of PEG-6000 treatment, the seeds of overexpressed Os MPK17 protein lines grew better and the length of root and shoot was significantly longer than those of the wild type. At seedling stage, transgenic plants showed lower water loss rate when exposed in the air. The transgenic rice with overexpressed OsMPK17 protein grew better than the wild type in the experiment with soil drought stress and re-watering then. In conclusion, the overexpressed OsMPK17 protein enhances drought tolerance of rice. This study enhances the understanding for the function of OsMPK17 protein.

关 键 词：水稻 MAPK蛋白质 免疫印迹 逆境胁迫 过表达 基于抗体的蛋白质组学 

分 类 号：S51[农业科学—作物学]