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1、水稻细菌性条斑病菌GX01菌株关键胞外蛋白酶基因的鉴定黄萍I,刘伟2,卢香浓I,蒋屮伟I,蒋蓉I,玉延华1,何勇强1(1.广西大学生命科学与技术学院，南宁530004：2.通化师范学院生命科学学院，吉林通化134002)摘要：胞外蛋门酶是许多病原菌的巫要的致病因子z—,起降解寄主细胞组分或免疫相关蛋白的作用。水稻细菌性条斑病菌GX01菌株基因组屮注释了5个胞外蛋白酚基因，H前，尚不确定哪一个属于该菌的关键胞外蛋白醮基因。采用基于同源整合突变技术，构建了这5个基因的整合突变体，平板法检测结果显示，XOC1545基因的突变体NK1545对脱脂牛奶不产生
2、水解圈。针刺接种结果农明，NK1545対水稻口木晴品种的致病力极显若低于野生菌株，而其它应试基因突变体的表型与野生型菌株差别不显著。带有全长XOC1545基【人1的PLAFR3质粒能够互补突变体的表型，表明XOC1545基因是该菌株关键的胞外蛋白酶基因。该基因预测的编码产物为M35家族金加蛋白酶，与水稻细菌性条斑病菌的毒力和关。关键词：植物病理学；水稻细菌性条斑病菌；胞外蛋白酶；基因；毒力中图分类号：S432.1Identificationofthekeygenewhichisresponsibleforthemajorextracellularp
3、roteaseactivityofXanthomonasoryzaepv.oryzicolastrainGX01HUANGPingi,LIUWei2,LUXiangnongi,JIANGZhongweii,JiangRongi,YUYanhuai,HEYongqiangi(1.CollegeofLifeScienceandTechnology,GuangxiUniversity,NanNing530004;2.CollegeofLifeScience,TonghuaNormalUniversity,JiLinTongHua134002)Abstra
4、ct:Extracellularproteaseisoneoftheimportantvimlencefactorsformanypathogens・Itplaysimportantrolesindigestionsofthehostcellwallandmembranecomponentsandimmuneresponseproteins.TherearefivegeneswereannotatedtoencodeextracellularproteaseinthegenomeoftherecentlysequencedXanthomonasor
5、yzaepv.oiyzicola(Xoc)GX01strain.Itisunknownforwhichputativeextracellularproteasegeneplaystheactualrolestoberesponsiblefortheextracellularproteaseactivities・AnintegrationmutantofeachgenewasconstructedbyusinghomologousintegrationwithasuicideplasmidpK18mob.OnlythemutantNK1545,the
6、non-polarmutantofXOC1545ofXocGX01,showeddramaticallyreducedextracellularproteaseactivitydisplayedontheagarplatetestandthesignificantlyreducedinvirulenceonthehostplantricecultivarsNipponbarecomparedtothewildtypestrainandthemutantsaffectedinotherputativeextracellularprotease-enc
7、odinggenes・ThephenotypesofmutantNK1545canbefullyrestoredbycomplementationbyintroductionofapLAFR3carryingtheintactCDSsequenceofXOC1545.ThisstudydemonstratedthattheXOC1545isthekeyextracellularproteasegeneofXoc.XOC1545ispredictedtoencodeanM35familypeptidase(deuterolysinmetallopro
8、teas)whichisresponsibleforthemajorextracellularproteaseactivi
