核桃JrNPR1基因的克隆与抗病响应潜力

病程相关基因非表达子NPR1（Nonexpressor of pathogenesis-related gene 1）基因，在植物抗病过程中起核心调控作用。核桃（Juglans regia）是我国乡村振兴的重要经济油料树种，其生长和产量严重受病虫害制约。为探索核桃抗病生理机制，筛选抗病基因，以‘香玲’核桃为试材，克隆获得JrNPR1基因，对其基本生物信息和病害响应进行分析，预测JrNPR1响应病害的功能。结果显示：JrNPR1基因开放读码框（ORF）长1 782 bp，包含593个氨基酸，理论等电点为6.40；与杨梅（Morella rubra）、蓖麻（Ricinus communis）等同源蛋白进行多序列比对，发现均有NPR1-like-C保守结构域，且JrNPR1与杨梅和欧洲栓皮栎（Quercus suber）等的同源蛋白具有较近的进化关系。其上游1 233 bp启动子中含有多种与植物抗病相关的顺式作用元件，如，WRKY71OS。在胶孢炭疽菌（Colletotrichum gloeosporioides）、矩圆黑盘孢菌（Melanconium oblongum）、黄单胞菌（Xanthomonas campestris）等病原处理下，JrNPR1被显著诱导，且其表达在水杨酸（Salicylic acid，SA）存在下高出单一病原处理的1.31~178.89倍。表明JrNPR1基因可能是抵抗核桃炭疽病、枝枯病、细菌性黑斑病的重要基因，且涉及SA信号通路。

Cloning and Disease Resistance Response Potential of Walnut JrNPR1 Gene

Nonexpressor of pathogenesis-related gene 1(NPR1)played a key role in regulating plant disease resistance.Walnut(Juglans regia)was an important economic oil tree for rural revitalization in China,and its growth and yield were severely restricted by pests and diseases.In order to explore the physiological mechanism of walnut disease resistance and screen disease resistance genes,the JrNPR1 gene was cloned from‘Xiangling’walnut,and its basic biological information and disease response were analyzed to predict the function of JrNPR1.The results showed that the ORF of JrNPR1 gene was 1782 bp,encoded 593 amino acids,and the theoretical isoelectric point was 6.40.Multi-sequence comparison with Morella rubra,Ricinus communis and other homologous proteins showed that they all had NPR1-like-C conserved domains,and JrNPR1 had a relatively close evolution with M.rubra and Quercus suber.The upstream 1233 bp promoter contained a variety of cis-acting elements related to plant disease resistance,such as WRKY71 OS.Under treatment with pathogens such as Colletotrichum gloeosporioides,Melanconium oblongum,and Xanthomonas campestris,JrNPR1 was significantly induced,and its expression was up-regulated by salicylic acid(SA).The expression level of JrNPR1 under CgTJ+SA,MoZK+SA,XcHB+SA was 1.31~178.89 fold of that under single stress of CgTJ,MoZK,XcHB,respectively.These results suggested that JrNPR1 might be an important disease resistance gene against walnut anthracnose,branch blight and bacterial black spot,and might be involved in the SA signal pathway.