摘要
【目的】本文研究了转hpal_(Xoo)基因棉花对棉花炭疽病的抗性水平与其应用潜力。【方法】采用棉花炭疽菌(Colletotrichum gossypii)侵染转hpal_(xoo)基因的棉花材料和普通棉花,将棉花炭疽菌接种于转基因棉花(出发品种为陆地棉品种854)和普通棉花(854)的叶片上,分别于接种后0、12、24、48、72 h取样,进行叶片H_2O_2含量和H_2O_2DAB定位检测,过氧化物酶(POD)和苯丙氨酸解氨酶(PAL)活性测定及对棉花抗性相关基因的表达情况进行分析。【结果】在接种棉花炭疽菌后48 h,转hpal_(xoo)基因棉花的H_2O_2含量及其定位检测可显示出峰值,随后又下降到和0 h相当的水平;POD与PAL活性也显示出类似的变化趋势。然而普通棉花叶片在接种后24 h,H_2O_2含量及其定位检测可显示出峰值;POD酶活性只有在72 h才略微上升。抗性相关基因荧光定量检测显示,在接种后24 h转hpal_(xoo)基因棉花中防卫基因hsr203J表达高于普通棉,而在72 h转基因与普通棉花两者几乎都没有表达;另外,在接种后0～48 h转基因植株中NPR1基因的表达一直低于普通棉花,但其表达量比普通棉花增长速度快,在72 h两者的表达量相同。在接种后0～12 h转基因植株中PR-1b基因就有微弱的表达,在24 h表达量最大,后续逐渐减弱,而普通棉花一直处于低表达。在受到棉花炭疽菌侵染时,hpal_(xoo)基因能够诱导寄主产生高水平活性氧及相关基因表达,说明hpal_(xoo)编码的Harpin_(Xoo)表达赋予转基因棉花对棉花炭疽菌的抗性。【结论】转hpal_(xoo)基因棉花为某些防卫基因超水平表达提供了分子基础。
【Objective】This paper aimed to study the resistance level of transgenic hpalXoocotton to Colletotrichum gossypii and find out its potential applications. 【Method】Colletotrichum gossypii was inoculated on the leaves of transgenic hpalXoocotton( departure variety is 854)and ordinary cotton( 854),and the concentration of H2O2 and accumulation of H2O2 in the leaves by DAB staining after the inoculation at 0,12,24,48,72 hours were detected,additionally the activity of the relative enzyme( peroxidase and phenylalnineammonialyase) were measured,and analysis the expression of cotton resistance related gene at different inoculated time as stated. 【Result】The H2 O2 content of transgenic hpalXoocotton leaves reach the highest level at 48 hours after inoculated,and subsequently declined to the level of 0 hours. Besides,the activity of peroxidase and phenylalnine ammonialyase also showed a similar trend. However,the H2 O2 content of ordinary cotton leaves was the highest at 24 hours after inoculated,and the POD activity was rising slightly at 72 hours. For the expression of the resistancerelated genes,hsr203 J gene in transgenic hpalXoocotton leaves was evidently higher express than that in ordinary cotton at 24 hours after in-oculated,but both of them shown very low expression at 72 hours; The expression of NPR1 gene of transgenic hpalXoo cotton leaves kept less than that of ordinary cotton all through at0-48 hours after inoculated,but the expression of transgenic hpalXoocotton leaves was growing faster than that of ordinary cotton and reached the same expression at 72 hours. There was weak the expression of PR-1 b gene in transgenic hpalXoocotton leaves at 0-12 hours after inoculated and reached the highest level at 24 hours,however,ordinary cotton kept low expression through the time after inoculated. hpalxoogene could induce thehost to produce high levels of reactive oxygen species and also induce the expression of defence gene,which means that hpalXoogene enhanced the cotton resistance to Colletotrichum gossy
作者
刘文波
何其光
蔡加挺
靳鹏飞
林春花
邬国良
缪卫国
LIU Wen-bo;HE Qi-guang;CAI Jia-ting;JIN Peng-fei;LIN Chun-hua;WU Guo-liang;MIAO Wei-guo(Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource/Institute of Tropical Agriculture and Forestry,Hainan University,Hainan Haikou 570228,China)
出处
《西南农业学报》
CSCD
北大核心
2018年第12期2478-2485,共8页
Southwest China Journal of Agricultural Sciences
基金
南繁区生物安全监测预警及控制关键技术研究与示范(201403075)
海南省自然科学基金(20153131)
海南大学青年教师基金(hdkyxj201708)
国家自然科学基金(31360029
31160359)
