转GhB301基因烟草的防御酶活性及抗病相关基因表达分析  

Defense Enzyme Activities and Expression Analysis of the Disease-related Genes in Transgenic GhB301 Tobacco

作　　者：刘戈辉 朱金成 郭文婷[1] 张鹏飞[1] 张薇[1,2] LIU Gehui;ZHU Jincheng;GUO Wenting;ZHANG Pengfei;ZHANG Wei(College of Agronomy,Shihezi University,Shihezi,Xinjiang 832003,China;Key Laboratory of Oasis Ecological Agriculture,Xinjiang Production and Construction Corps,Shihezi,Xinjiang 832003,China)

机构地区：[1]石河子大学农学院,新疆石河子832003 [2]新疆生产建设兵团绿洲生态农业重点实验室,新疆石河子832003

出　　处：《西北植物学报》2019年第11期2011-2018,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31560407)

摘　　要：为了明确棉花ERF-B3亚族转录因子基因GhB301在烟草异位表达后(抗枯萎病中)的功能,该研究以过表达GhB301基因烟草和野生型烟草为材料,采用枯萎病菌孢子悬浮液接菌方法,分析病原菌侵染前后防御酶活性变化以及防卫相关基因的表达变化与抗病性的关系。结果显示:(1)棉花枯萎病菌处理15 d后,2个转基因株系烟草叶片黄化程度与野生型相比较轻。(2)棉花枯萎病菌处理后,过表达GhB301转基因烟草和野生型烟草叶片过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)的活性较未接菌对照显著提高,并且酶活峰值出现均早于野生型材料;转基因材料叶片的POD、PAL、PPO活性均在处理3 d后达到峰值,而野生型材料叶片的POD、PAL活性在处理5 d后才达到峰值。(3)接种棉花枯萎病菌后活性氧相关基因、乙烯(ET)/茉莉酸(JA)途径相关基因、病程相关基因的表达量在转基因株系OE1和OE2中均受到明显影响。研究推测,GhB301在烟草中的异位表达激活了防卫相关基因的表达,提高了防御酶的活性,从而增强了烟草对枯萎病菌的抗性。The objectives of this study are to determine the function of the cotton ERF-B3 subgroup transcription factor gene GhB301 after ectopic expression of tobacco(in resistance to Fusarium wilt).The over-expression of the GhB301 transgenic tobacco and wild-type tobacco treated with Fusarium wilt spore suspension,were used for analysis of the relationship between the defense enzyme activities and the expression of defense-related genes and disease resistance before and after infection.The results showed that:(1)after treatment with Fusarium wilt for 15 days,the degree of yellowing of tobacco leaves in two transgenic lines was lighter than that in wild type;(2)Compared with non-inoculated controls,the over-expression of the GhB301 transgenic tobacco and wild-type tobacco leaves showed a drastic increase of the enzyme activities of the oxide enzyme(POD),phenylalanine ammonia lyase(PAL)and polyphenol oxidase(PPO),and the peak of enzyme activities appeared earlier than that of wild type material.POD,PAL and PPO activities in the leaves of transgenic materials peaked at 3 d,while the POD and PAL activity in leaves of wild-type materials reached a peak at 5 d;(3)The expression level of reactive oxygen-related genes,ethylene(ET)/jasmonic acid(JA)pathway-related genes,and pathogenesis-related genes were affected in both the transgenic materials OE1 and OE2 after inoculation with Fusarium wilt.It indicated that the ectopic expression of the GhB301 in tobacco activated the expression of defense-related genes and increased the activities of the defense enzymes,thus enhancing the resistance of tobacco to Fusarium wilt.

关 键 词：烟草 GhB301基因 枯萎病 防御酶活性 防卫相关基因 

分 类 号：Q786[生物学—分子生物学] Q945.78