文心兰NPR1基因的克隆与诱导抗性过程中的表达分析  

NPR1 Cloning and Expression during Development of Induced Systemic Resistance in Oncidium hybridum

作　　者：郑世仲 王培育 张春柳 江胜滔 江金兰 颜沛沛 叶炜 赖钟雄[4] ZHENG Shi-zhong;WANG Pei-yu;ZHANG Chun-liu;JIANG Sheng-tao;JIANG Jin-lan;YAN Pei-pei;YE Wei;LAI Zhong-xiong(College of Life Sciences,Ningde Normal University,Ningde,Fujian 352100,China;Sanming Academy of Agricultural Sciences,Sanming,Fujian 365059,China;Ningde Agricultural Bureau,Ningde,Fujian 352100,China;Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]宁德师范学院生命科学学院,福建宁德352100 [2]福建省三明市农业科学研究院,福建三明365059 [3]福建省宁德市农业局,福建宁德352100 [4]福建农林大学园艺植物生物工程研究所,福建福州350002

出　　处：《福建农业学报》2020年第2期140-149,共10页Fujian Journal of Agricultural Sciences

基　　金：福建省林业厅科技项目(闽林科[2018]26号);福建省自然科学基金面上项目(2014J01382)。

摘　　要：【目的】探明文心兰NPR1基因在诱导抗性过程中的生理作用。【方法】利用RACE技术克隆文心兰NPR1基因全长,并进行相关生物信息学分析和遗传进化树的构建;以印度梨形孢共培养、水杨酸(SA)和茉莉酸甲酯(MeJA)处理文心兰,分析菊欧文氏杆菌导致的软腐病抗性影响及NPR1基因的表达特性变化。【结果】克隆得到的2条基因序列长度分别为1804 bp和1902 bp,编码555、556个氨基酸,均为NPR1类型,分别命名为OgNPR1-1和OgNPR1-2;文心兰两个NPR1基因在未接菌的情况下均呈现低表达量,在接种共生菌印度梨形孢后轻微上调表达,在接种菊欧文氏杆菌后显著上调表达,而在印度梨形孢共生菌的存在下,接种菊欧文氏杆菌使两个NPR1基因表达更为活跃;同时,相比印度梨形孢共生菌处理,两个NPR1基因均在外源SA、MeJA处理下接种菊欧文氏杆菌表现为更显著上调表达;此外,共生印度梨形孢可显著提高文心兰对菊欧文氏杆菌导致软腐病的抗性,限制软腐病病症在非接病叶片的扩展。【结论】在共生印度梨形孢的文心兰植株中,印度梨形孢显著提高了文心兰NPR1基因响应菊欧文氏杆菌的表达水平,虽然上调水平不及SA和MeJA激素处理的水平,却使文心兰植株表现出更强的对抗软腐病的能力。【Objective】Physiological role of NPR1 gene plays in the development of induced systemic resistance(ISR)in Oncidium hybridum was studied.【Method】NPR1 of the orchid was cloned using RACE technique,and bioinformatics and phylogenenetic analyses on the gene conducted.After co-cultured with Piriformospora indica or treated with salicylic acid(SA)or methyl jasmonate(MeJA)to induce ISR,the orchids were analyzed for the treatment effect against soft rot disease caused by Erwinia chrysanthemi.And,expressions of NPR1 during the treatments were investigated.【Result】Two fulllength cDNA sequences were cloned.They were of a same type,one named OgNPR1-1 with a length of 1804 bp encoding 555 amino acids,and the other,OgNPR1-21902 bp encoding 556 amino acids.Their expressions were slightly upregulated by the inoculation of symbiotic P.indica.However,as both P.indica and E.chrysanthemi were present in the orchids,the expressions were significantly upregulated.It indicated that,in the presence of P.indica,the orchid plants became significantly more resistant to E.chrysanthemi inhibiting spread of the soft rot disease.On the other hand,the symbiosis of P.indica did not upregulated the expressions of the two genes more than the treatment by exogenous SA or MeJA on the orchids.【Conclusion】The expression of NPR1 in the orchids inoculated with the symbiotic P.indica was significantly upregulated in the presence of E.chrysanthemi.Although the increased expression by P.indica was lower than what induced by exogenous SA or MeJA hormone treatment,nonetheless,the orchids benefitted from the symbiosis and became more resistance to the soft rot disease.

关 键 词：文心兰 NPR1 诱导性抗性 印度梨形孢 菊欧文氏杆菌 

分 类 号：S682.31[农业科学—观赏园艺]