水稻类病变突变体及抗病性的研究进展

植物病理学报ACTA PHYTOPATHOLOGICA SINICA㊀41(1):1-9(2011)收稿日期:2009-05-04;修回日期:2010-09-23基金项目:国家自然科学基金资助项目(30771329;30800677);国家转基因重大专项(2008ZX 08009-003);浙江师范大学创新团队项目通讯作者:马伯军,教授,主要从事植物抗病分子遗传学研究;E-mail:mbj @第一作者:陈析丰(1980-),男,浙江温州人,硕士,主要从事植物抗病分子遗传学研究㊂췍췍췍췍췍췍췍췍췍췍췍췍췍췍췍췍专题评述水稻类病变突变体及抗病性的研究进展陈析丰,金杨,马伯军*(浙江师范大学化学与生命科学学院,金华321004)摘要:植物类病变突变体是一类在没有病原物侵染情况下就能自发产生坏死斑的突变体,这类突变往往导致植株的抗病增强和防御相关基因的组成性表达㊂水稻中已报道了将近200个来源不同的类病变突变体,截至2009年5月底52个水稻类病变突变体已被鉴定和命名,其中6个控制类病变性状的基因被克隆,它们分别编码不同的蛋白,包括热激蛋白转录因子㊁E3泛素连接酶㊁酰基转移酶㊁质膜蛋白激酶㊁锌指蛋白和锚蛋白㊂尽管这些蛋白不是直接与植物抗病途径相关,但是在41个已鉴定的水稻类病变突变体中,绝大多数提高了对白叶枯病或稻瘟病的抗性,表明这些类病变基因的突变激活了植株的防御系统,并且不同的类病变基因可能参与了不同的抗病信号传导途径㊂深入研究水稻类病变突变体对作物抗病的分子机理研究和栽培品种的遗传改良都具有重要的意义㊂关键词:水稻;类病变突变体;基因鉴定;抗病性Progress on the studies of rice lesion mimics and their resistant mechanism to the pathogens㊀CHEN Xi-feng ,JIN Yang ,MA Bo-jun㊀(College of Chemistry and Life Sciences ,Zhejiang NormalUniversity ,Jinhua 321004,China )Abstract :Lesion mimic mutants are characterized as the formation of necrotic lesions in the absence of pathogens ,and such genetic defects often result in enhanced resistance to pathogen infection and constitutive expression of defense response genes.About 200rice lesion mimics were found by different strategies ,52of which had been identified by the end of May ,2009.Six genes had been cloned from these identified mutants and encoded distinct proteins including heat stress transcription factor ,E3ubiquitin ligase ,acyltransferase ,cy-toplasmic protein kinase ,zinc finger protein and ankyrin repeats protein.Although none of these proteins ap-peared to be directly involved in plant defense pathways ,most of the 41lesion mimics exhibited the resistance to Xanthomonas campestris pv.oryzae or Magnaporthe grisea and constitutive expression of typical defense-response marker genes such as callose ,autofluorescence martials ,phytoalexin ,reactive oxygen specie ,and pathogen-related genes.It was indicated that these lesion mimic mutations activated the defense system in plants ,but they probably functioned in distinct signaling pathways.Thus ,extensive studies of rice lesion mi-mics will promote the better understanding of molecular mechanisms on the crop resistance to pathogens and genetically improve resistance breeding in crops.Key words :rice ;lesion mimics ;gene identification ;pathogen resistance中图分类号:S432.21㊀㊀㊀文献标识码:A㊀㊀㊀文章编号:0412-0914(2011)01-0001-09㊀㊀植物类病变突变体(lesion mimic mutant )是指一类在没有逆境㊁损伤或病害情况下就能自发形成㊀植物病理学报41卷类似病原物侵染的坏死斑(lesion)的突变体[1]㊂第一个植物类病变突变体是从玉米中被发现的[2],近年来在拟南芥[3]㊁大麦[4]㊁水稻[5]㊁玉米[6]㊁花生[7]和大豆[8]等其他植物中也发现了大量这类突变体㊂由于植物类病变突变体都能自发地表现出坏死的表型,这使学者们推测它们可能具有一些共同的机制[9]㊂近年来发现类病变坏死的表型与植物防卫过程中的过敏反应(hypersensitive response,HR)非常相似,而且许多类病变突变体对某些植物病原体也表现出一定的抗性[3,4,10]㊂HR 是植物受非亲和性病原菌感染后,侵染部位细胞迅速死亡,从而限制病原菌增殖的一种植物的特异性防卫反应,属于细胞程序性死亡(programmed cell death,PCD)[11]㊂因此,类病变突变体成为了研究植物抗病性机制和PCD途径的理想材料,受到了诸多学者的广泛关注㊂目前,一些控制类病变性状的基因已经被克隆,如拟南芥lsd1基因编码1个含3个锌指结构域的类似转录因子蛋白[12],大麦mlo基因编码1个60kD的跨膜蛋白[4],玉米lls1基因的表达产物与芳香烃需氧降解有关的ARH(aromatic ring-hy-droxylating)双加氧酶相似[6]㊂尽管这些已克隆的类病变基因没有同源性,但是这些突变体均不同程度地提高了植株对病原菌的抗性,表现出一些发生抗病反应时的组织化学和分子特征,甚至启动了系统获得抗性(systematic acquired resistance,SAR)相关基因的表达[4,6,12]㊂这表明引发植物类病变坏死的原因可能是多样的,植物抗病信号转导网络也是非常复杂的,只有更多地鉴定和研究这些类病变突变体,才能全面解析植物类病变坏死及其抗病的机制㊂水稻既是单子叶模式植物,又是世界上最重要的粮食作物之一,所以近年来国内外对水稻的类病变突变体的研究相对比较多,也积累了许多重要的研究成果,但是目前还未见到有对水稻类病变进行综述性的报道㊂本文综述了国内外水稻类病变突变体的最新研究进展,主要介绍水稻类病变突变体的发掘与命名㊁突变基因的定位与克隆㊁突变体的抗病性及其机理研究等,对作物抗病机制及分子育种具有重要参考价值㊂1㊀水稻类病变突变体的发掘1.1㊀突变体的来源20世纪70年代,Sekiguchi等[13]在水稻中首次发现了类病变突变体,取名为sekiguchi lesion(sl),后来又相继报道了许多类似的水稻突变体㊂这些突变体除了少数来源于自然变异,绝大部分由物理诱变㊁化学诱变㊁转基因等方法得到㊂近年来,水稻类病变突变体的数目不断增加,到目前为止报道了将近有200个,如Iwata等[14]发现了9个类病变,其中2个自发突变㊁5个γ射线诱变和2个化学剂MNU(N-methyl-N-nitrosourea)诱变;Taka-hashi等[15]从MNU诱变库中筛选到93个类病变; Wu等[16]分别利用γ射线㊁快中子(fast neutron)和化学剂Diepoxybutane得到了70个类病变;Wang 等[17]筛选到18个γ射线诱变的类病变;由T-DNA和Tos17逆转座子插入引起的类病变突变也各报道了1个[18,19]㊂这些研究表明水稻中类病变突变的发生概率相当高,Wu等[16]曾报道突变率达到0.6%,如此高的诱发率说明许多突变都能导致植物细胞死亡,这可能就是植物类病变来源广泛的重要原因㊂1.2㊀突变体的命名尽管水稻中报道了大量的类病变突变体,但是大部分突变没有进行基因的等位分析和正式命名㊂国际谷物基因组数据库Gramene(http://www. /)目前只注册了32个水稻类病变基因㊂根据国内外的研究报道,截止2009年5月一共有52个水稻类病变基因被鉴定和命名(表1)㊂水稻类病变基因的命名主要有两种方式,Gramene 上统一以lrd(lesion resembling disease)命名,如Wang等[17]将18个筛选到的类病变命名为lrd27 ~lrd44(只鉴定分析了lrd32㊁lrd39㊁lrd40和lrd42),不过Gramene注册的lrd基因又有各自不同的名称,所以另一种命名方式是根据类病变发生时间㊁颜色㊁形状或导致植株抗病的特点,如spl (spotted leaf)㊁bl(brown leaf spot)㊁cdr(cell death and resistance to the blast fungus)㊁blm(blast lesion mimic)㊁ysl(yellow leaf spot)㊁fgl(faded green2㊀㊀1期㊀㊀陈析丰,等:水稻类病变突变体及抗病性的研究进展Table1㊀Summary of identified rice lesion mimic mutants up to May2009Gene symbol Source Original variety Resistance Inherence Chr.PutativefunctionReferencespl1Spontaneous Banshinriki-byogata M Recessive12-[14,16,20-22] spl2Spontaneous Katsumonbyo X Recessive2-[14,16,20-22] spl3γ-ray Norin8X Recessive3-[14,16,20-22] spl4γ-ray Norin8M,X Recessive6-[14,16,20-22] spl5γ-ray Norin8M,X Recessive7-[14,16,20-22] spl6MNU Kinmaze None Recessive1-[14,16,20-22]spl7γ-ray Norin8M,X Recessive5Heat stresstranscription factor[14,16,20-23]spl8MNU Kinmaze None Recessive5-[14,20,21] spl9γ-ray Norin8M Recessive7-[14,16,21,22,24] spl10MNU Kinmaze M,X Recessive10-[20,21,25] spl11EMS IR68M,X Recessive12E3ubiquitin ligase[4,16,22,26] Spl12MNU Hinohikari M,X Dominant--[21]spl12(t)Hybrid AsominoriˑIR24-Recessive2-[27]Spl13MNU Hinohikari M,X Dominant--[21]Spl14MNU Taichung65M,X Recessive--[21]Spl15MNU Koshihikari M,X Dominant--[21]spl16Diepoxybutane IR64None Recessive--[16]spl17Diepoxybutane IR64M,X Recessive--[16]Spl18T-DNA Nipponbare M,X Dominant10Acyltransferase[18]spl19Diepoxybutane IR64X Recessive--[16]spl20Fast neutron IR64M Recessive--[16]spl21Diepoxybutane IR64X Recessive--[16]spl22Diepoxybutane IR64X Recessive--[16]spl23Diepoxybutane IR64X Recessive--[16]Spl24Diepoxybutane IR64X Dominant--[16]spl25Diepoxybutane IR64X Recessive--[16]Spl26Diepoxybutane IR64M,X Dominant--[16]Spl27Diepoxybutane IR64X Dominant--[16]bl1Spontaneous Norin8-Recessive2-[28]bl2Spontaneous Norin8-Recessive6-[29]bl3Hybrid E-48ˑA-31-Recessive6-[29]bl4β-ray Norin8-Recessive3-[30]bl5β-ray Norin8-Recessive--[30]bl6β-ray Norin8-Recessive--[30]cdr1MNU Kinmaze M Recessive--[15]cdr2MNU Kinmaze M Recessive--[15]Cdr3MNU Kinmaze M Dominant--[15]sl Hybrid Asahi M Recessive--[13,31-34]3㊀植物病理学报41卷Contiued Table1Genesymbol Source Original variety Resistance Inherence Chr.Putativefunction Referenceyslβ-ray Norin8-Recessive--[30] fgl Spontaneous Houki Asahi-Recessive10-[35] znγ-ray AC581-3-Recessive6-[36] lrdγ-ray Norin8M Recessive--[37] lrd32γ-ray Shuangkezao X Recessive--[17,38] lrd39γ-ray Zhonghua11X Recessive--[17,38] lrd40γ-ray Zhonghua11X Recessive--[17,38] lrd42γ-ray Zhonghua11X Recessive--[17,38] blm MNU Hwacheong M Recessive--[39,40] lmiγ-ray Zhongxian3037-Recessive8-[41,42] lmm1EMS Katy M,T Recessive--[43] attm1Tos17Nipponbare M,X Recessive5Cytoplasmic protein kinase[19] oslsd1RNAi Nipponbare M Recessive8Zinc finger protein[44] OsNPR1Overexpression LG X Dominant1Ankyrin repeats protein[45,46] M:Magnaporthe grisea;X:Xanthomonas campestris pv.oryzae;T:Thanatephorus cucumeris(Frank)Donk; None:No resistance to the pathogens tested.The symbol - means the item not determined.leaf)㊁zn(zebra necrosis)㊁lmi(lesion mimic initia-tion)等㊂其中,以spl命名的突变体最多,已经有28个(spl1~Spl12㊁spl12(t)㊁Spl13~Spl27)㊂2㊀水稻类病变基因的克隆水稻类病变的表型受单个显性或隐性基因控制,在52个水稻类病变中,9个显性遗传,43个隐性遗传㊂通过形态学标记㊁DNA分子标记和反向遗传学技术,22个类病变基因已被定位在水稻12条染色体上(表1)㊂其中,6个基因spl7[23]㊁spl11[26]㊁Spl18[18]㊁attm1[19]㊁oslsd1[44]和Os-NPR1[45]已经被克隆,它们各自编码不同功能的蛋白,参与不同的代谢途径,表明了导致植物细胞坏死的途径非常复杂和多样㊂此外,还有3个基因被精细定位,lmi基因被定位在8号染色体距标记C4135-9的0.08cM处[41],spl1基因被定位在12号染色体70kb的区间内[47],本课题组也利用DNA分子标记技术将spl5基因定位在了7号染色体80kb的区间内[48],目前正在对候选基因进行图位克隆和功能验证㊂2.1㊀spl7和spl11基因spl7和spl11基因都是用图位克隆的方法得到的㊂spl7基因编码1个热激转录因子HSF(heat stress transcription factor),在热胁迫下HSF能激活热激蛋白Hsp(heat shock protein)基因的表达,诱导产生热激应答反应[23];spl11编码1个具有E3泛素连接酶活性的蛋白,含有1个U-box和1个armadillo(ARM)重复保守结构域,前者在酵母中被认为与泛素化有关,而后者则在哺乳动物中被证明参与蛋白间的互作,推测SPL11蛋白可能是通过泛素化系统对植物的PCD和防卫反应起负调控作用[26]㊂这2个基因的突变都是由于基因编码区中单个碱基的替换造成的,spl7是保守结构域中氨基酸发生了替换,,而spl11是无义突变使蛋白的翻译提前终止[23,26]㊂2.2㊀Spl18和attm1基因Spl18是T-DNA激活标签(T-DNA activation tagging)插入突变,该T-DNA能显著性增强插入位点周围基因的表达[18]㊂Mori等[18]利用TAIL-PCR技术分析插入位点,发现Spl18基因编码1个酰基转移酶(acyltransferase),命名为OsAT1㊂野生型水稻中OsAT1基因表达水平很低,但是在Spl18突变体中其表达量非常高,用35S启动子过量表达该基因也能形成明显的类病变表型[18]㊂attm14㊀㊀1期㊀㊀陈析丰,等:水稻类病变突变体及抗病性的研究进展(Tos17triggered mutation1)是逆转座子Tos17插入突变,突变植株产生了一系列的防卫应答,并增强了对稻瘟病的抗性[19]㊂Takahashi等[19]通过TAIL-PCR技术检测到attm1突变位点发生在Os-Pti1a基因的第3个外显子中,该基因与西红柿抗病基因Pti1(Pro interacting1)高度同源,编码1个质膜蛋白激酶(cytoplasmic protein kinase),过量表达OsPti1a基因会降低植株的抗病能力,推测OsP-ti1a可能在水稻的防卫系统中起负调控的作用[19]㊂2.3㊀oslsd1和OsNPR1基因拟南芥类病变基因LSD1(lesions simulating disease resistance)编码1个锌指蛋白,在植物防卫反应和细胞程序性死亡信号途径中起负调控作用[12]㊂Wang等[44]通过反向遗传学的方法,在水稻中克隆了1个与LSD1同源的基因,命名为OsLSD1㊂在水稻中反义抑制OsLSD1基因的表达能引起类病变表型,过量表达该基因能促进转基因水稻愈伤组织的分化和增加植株中叶绿素B的含量,在烟草中过量表达OsLSD1基因能增强植株对1种真菌毒素(Fumonisins B1)的抗性,推测OsLSD1可能是植物PCD和愈伤组织分化的双功能调控因子[44]㊂拟南芥类病变基因AtNPR1(non-expresser of pathogenesis-related genes)是SAR抗病途径中的1个关键调节因子,它编码1个含双向核定位序列(bipartite nuclear localization sequence)和锚定蛋白(ankyrin)重复结构域的新蛋白[49]㊂Chern等[45]和Yuan等[46]分别在水稻中也克隆到了AtNPR1的同源基因 OsNPR1,过量表达Os-NPR1能使水稻植株表现出类病变坏死斑和增强对白叶枯病的抗性,RNAi抑制OsNPR1则使水稻对白叶枯病更加敏感㊂3㊀水稻类病变突变体的抗病性3.1㊀对水稻病原菌的抗性目前研究表明尽管不是所有的类病变基因都是植物防卫信号途径中所必须的,但是许多类病变突变体表现出了广谱抗病性,如拟南芥37个类病变突变体中,绝大部分提高了对1个或多个病原菌的抗性[50]㊂水稻类病变突变体也同样存在普遍抗病的现象,经抗病性鉴定的41个突变体中,38个对水稻病原菌增强了抗性,其中24个抗稻瘟病(Magnaporthe grisea),27个抗白叶枯病(Xan-thomonas campestris pv.oryzae),1个抗纹枯病[Thanatephorus cucumeris(Frank)Donk](表1)㊂这些用来鉴定的病原菌生理小种来自中国㊁日本㊁菲律宾㊁韩国㊁葡萄牙㊁哥伦比亚㊁美国等不同地区,水稻类病变突变体对这些生理小种的抗性存在显著的差异,如spl21㊁spl22㊁Spl24㊁cdr1~Cdr3等只对1个小种有抗性,spl1~spl3㊁spl10㊁Spl13~ Spl15㊁blm等则对多个小种有抗性,而spl4㊁spl5㊁spl7㊁spl11㊁Spl12等具有广谱的抗性[15,16,21,22]㊂3.2㊀抗病相关的组织和生理特征在类病变出现时,许多突变植株都伴有与病原菌侵染相关物质的组成性表达,包括酚类物质(au-tofluorescent phenolic compound)积累㊁胼胝质(cal-lose)沉积㊁植保素(phytoalexin)水平上升和活性氧中间代谢物(reactive oxygen intermediate,ROI;主要指H2O2和O-㊃2)产生等,这些物质具有抵抗或抑制病原菌的作用,尤其是ROI能直接杀死病原菌并启动HR[50]㊂水稻类病变中也普遍存在这样的现象,如spl5㊁Spl12~Spl15㊁cdr1~Cdr3㊁blm等叶片中有明显的胼胝质或酚类物质沉积[15,39,51]; Spl18㊁cdr1~Cdr3㊁blm等在类病变形成后,植株中2种植保素Sakuranetin和Momilactone A的含量迅速上升[15,18,39];lrd40㊁cdr1~Cdr3㊁spl2㊁spl7㊁spl11等在类病变坏死斑的周围出现H2O2或O-㊃2的过量积累[15,38,52]㊂3.3㊀抗病相关基因和蛋白表达特征一般情况下,植物在受到病原物侵染时,一些与HR㊁SAR相关的抗病基因才会被诱导表达,而在水稻类病变中这些基因呈组成性表达㊂如受抗稻瘟病诱导表达的基因PR1(pathogenesis-related gene)和PBZ1(probenazole-inducible gene)在cdr1 ~Cdr3㊁spl1~spl7㊁spl9㊁spl11等中都有明显的表达,受大麦白粉病菌(powdery mildew fungus)诱导表达的草酸氧化酶(oxalate oxidase)基因HvOxOa 和HvOxOLP在spl1~spl7㊁spl9㊁spl11等中也有不同程度的表达,此外在spl3中还检测到过氧化物酶(Peroxidases)基因POX22.3和POX8.1的表达,这2个基因参与H2O2的代谢并受白叶枯病诱5㊀植物病理学报41卷导表达[15,22]㊂5个水稻类病变blm㊁cdr1㊁cdr2㊁spl1和spl6的蛋白组学研究也表明这些突变体中许多蛋白的表达量增加,其中包括防御相关的蛋白PBZ1㊁PR5㊁PR10㊁COMT(caffeic acid3-O-methyl-transferase)等,ROS代谢相关的酶CAT(catalase)㊁APX(ascorbate peroxidae)㊁SOD(superoxide dis-mutase)㊁GST(glutathione S-transferase)等,以及细胞衰老和死亡相关的蛋白PHB1(prohibi-tin)[40,53~56]㊂4㊀水稻类病变基因所涉及的信号转导途径由于类病变突变体往往表现出细胞死亡的调控异常㊁对病原菌的抗性增强以及防御系统的组成性表达,因此这类突变基因很可能参与了植物细胞中一些基础的㊁重要的信号传导途径,如活性氧的信号途径㊁水杨酸(salicylic acid,SA)信号途径㊁茉莉酸(jasmonate acid,JA)信号途径㊁乙烯信号途径㊁R基因(resistance gene)介导的抗病信号途径等[50]㊂氧迸发(oxidative burst)即ROI急剧增加是植物细胞防卫应答中最早期的信号之一,许多类病变突变体中都有H2O2和O-㊃2的积累[50],拟南芥㊁水稻类病变已有的研究和我们对spl5测定分析表明,这些突变体中ROI降解途径基本没有受到破坏,ROI的积累可能是其生物合成途径中的某些关键酶(如NADPH氧化酶)的活性失调所引起[15,54,57]㊂SA被认为是诱导SAR的必需信号分子,施加外源的SA及其类似物都能启动SAR[58]㊂拟南芥类病变基因AtNPR1是SA介导SAR途径中1个重要的调控因子,水稻中也存在类似的信号途径,过量表达NPR1基因能使植株产生SAR,但是它们的调节方式有所不同,水稻中内源SA水平较高,使过量表达的OsNPR1蛋白处于激活状态,而拟南芥中内源SA水平较低,过量表达的At-NPR1蛋白保持失活状态,只有施加化学诱导物或病原菌侵染才被激活[45,49]㊂JA-乙烯是植物发育和抗病信号网络中重要的介导因子,拟南芥类病变突变体cpr5㊁cpr22㊁ssi1和hrl1中的1个激活JA-乙烯途径的标记基因 PDF1.2基因呈组成性表达[59],而突变体cet组成性表达thionin基因,该基因的表达受JA 途径的直接调控[60]㊂此外,一些拟南芥类病变基因(EDS1㊁PAD4㊁NDR1)还参与R基因抗病途径的调控,这些基因属于R基因的2个下游途径[61]㊂虽然,在水稻中还没有报道类病变基因参与JA-乙烯途径和R基因介导的抗病途径,但是这些途径在单子叶和双子叶植物中相对比较保守,随着类病变基因研究的不断深入,很有可能通过这些途径来揭示它们的功能㊂5㊀水稻类病变突变体的应用前景由于类病变突变体普遍具有抗病性的特点,因此除了研究基因突变的原因及其信号途径,如何在作物育种和生产实践中合理利用和开发这一遗传资源是一个值得考虑的问题㊂目前,植物类病变的研究还处于理论性的阶段,主要原因可能是类病变往往会伴随细胞坏死㊁植株矮小㊁产量减少等一些不良的农艺性状,限制了它们在生产实践中的直接应用㊂尽管如此,有些具有非小种专一抗性的类病变还是引起了育种工作者们的重视,大麦类病变突变体mlo就是一个典型的实例㊂该突变体对几乎所有已知的白粉病(Erysiphe graminis f.sp.Hor-dei)的生理小种都具有持久抗性[4],自1979年第一次商业化推广带有mlo基因的大麦品种以来,一直保持高效的抗性,现在在欧洲许多国家已经开始大面积种植[62]㊂水稻中虽然发现了许多抗病基因,其中一些基因已经运用到抗病育种中去,如Xa21㊁xa5㊁Xa7㊁xa13㊁Pi-z t㊁Pi-k s㊁Pi-sh等[63,64],但是这些基因存在小种专一化和抗性不持久等问题,而且病原菌也在不断进化,这给水稻的抗病育种带来了很多困难,急需寻找一些具有广谱㊁持久抗性的遗传资源㊂类病变突变体的研究为水稻的抗病育种开启了一条新的道路,这些突变体不仅大部分提高了抗病性,而且有些还激活了植株的整个防御系统,表现出广谱的抗病性,将可能成为作物抗病分子育种的宝贵材料㊂参考文献[1]㊀Johal G S,Hulbert S H,Briggs S P.Disease lesionmimics of maize:a model for cell death in plants[J].BioEssays,1995,17(8):685-692.[2]㊀Hoisington D A,Neuffer M G,Virginia Walbot.Di-sease lesion mimics in maize:I.Effect of geneticbackground,temperature,developmental age,andwounding on necrotic spot formation with Les1[J].Dev.Biol.,1982,93:381-388.6㊀㊀1期㊀㊀陈析丰,等:水稻类病变突变体及抗病性的研究进展[3]㊀Dietrich R A,Delaney T P,Uknes S J.et al.Arabi-dopsis mutants simulating disease resistance response[J].Cell,1994,77(4):565-577.[4]㊀Buschges R,Hollricher K,Panstruga R.et al.Thebarley MLO gene:a novel control element of plantpathogen resistance[J].Cell,1997,88(5):695-705.[5]㊀Singh K,Multani D S,Khush G S.A new spottedleaf mutant in rice[J].Rice Genet.News.,1995,12:192-193.[6]㊀Gray J,Close P S,Briggs S P.et al.A novel sup-pressor of cell death in plants encoded by the Lls1geneof maize[J].Cell,1997,89(1):25-31. 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