稻瘟病菌蛋白激酶MoCbk1的功能研究  

Function Analysis of Protein Kinases MoCbk1 in Magnaporthe oryzae

作　　者：陈培蕊 张冬梅[2,3] 张连虎[2,3] 陈小强 林婕[1] 陈曦 王宗华[2,3] 

机构地区：[1]福建农林大学生命科学学院,福州350002 [2]福建农林大学生物农药与化学生物学教育部重点实验室,福州350002 [3]福建农林大学植物保护学院,福州350002

出　　处：《分子植物育种》2016年第3期529-535,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31301630)资助

摘　　要：真核生物的丝氨酸/苏氨酸蛋白激酶,在细胞形态建成、胞内信号转导和胚胎发育等过程中,发挥着重要作用,并已经在酵母和一些真菌中得到证实。前期研究发现,在丝状真菌稻瘟病菌(Magnaporthe oryzae)中存在一个丝氨酸/苏氨酸蛋白激酶Kin1,能够调节各种细胞进程,且在许多致病真菌中非常保守。为了更好地研究Kin1蛋白激酶在稻瘟病菌中的功能,我们用生物信息学方法,预测获得了一个与Sc Kin1互作的蛋白Sc Cbk1激酶,在稻瘟病菌数据库中搜索得到了4个与酿酒酵母Cbk1同源性相对较高的蛋白,并与Sc Cbk1和Hs Ndr一起进行了结构域分析,以及常见真菌同源物的系统进化分析。比对结果显示,稻瘟病菌基因MGG_09519和MGG_05376为2个Mo Cbk1。并运用遗传学方法对2个Mo Cbk1进行功能分析。Mo Cbk1a(MGG_09519)基因敲除后,敲除突变体相比野生型菌株和异位整合突变体,生长速率减慢,对水稻大麦等致病力减弱;敲除突变体产孢量略有增加,孢子萌发和附着胞的形成却没有变化。然而,经过多次实验,却无法得到Mo Cbk1b(MGG_05376)的敲除体,敲除体可能是真菌生存所必须的。综上,在稻瘟病菌基因组中,有2个Mo Cbk1基因,它们可能参与稻瘟病菌的营养生长和对水稻的致病过程的调控。In eukaryotes,serine/threonine protein kinase plays an important role in cell growth and morphogenesis,intracellular signal transduction and embryonic development,and it is verified in yeast and a number of other fungi.Previously study indicated that there was a Serine/Threonine kinase Kin1 in Magnaporthe oryzae which could regulate various cellular processes and also was very conserved in a range of fungi.In order to further study the function of Mokin1 in Magnaporthe oryzae,we predicted one of the Sc Kin1-interacing protein Sc Cbk1 by using bioinformatic analysis.By searching Saccharomyces cerevisiae homologs in the rice blast genome database,we further found four high identitical genes and then analysed their conserved domains with Sc Cbk1 in S.cerevisiae and Hs Ndr in Homo sapiens.And at the same time,we constructed the phylogenetic analysis among a range of fungi.The results of alignment indicated that MGG_09519 and MGG_05376 were two Mo Cbk1 proteins.Then they were used for functional analysis by genetic approach.ΔMo Cbk1a(MGG_09519) showed lower growth rate and pathogenicity to rice and barley compared to wild type and ectopic strain.The mutants showed higher conidiation but no changes in conidial germination,and appressorium formation.However,Mo Cbk1b(MGG_05376) may be essential for the fungal survival,and we could not get knockout of Mo Cbk1b(MGG_05376) after several experiments.In summary,there are two Mo Cbk1 genes(Mo Cbk1 a and Mo Cbk1b) in M.oryzae,and may be involved in regulation of vegetative growth and pathogenicity to the rice plant.

关 键 词：稻瘟病菌 蛋白激酶 MoCbk1 功能 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]