玉米矮花叶病毒病的理化性质是什么?

2025-03-28 09:51:00
推荐回答(1个)
回答1:

病毒名称:甘蔗花叶病毒Sugarcane mosaic virus(SCMV)和白草花叶病毒Pennisetum mosaic virus,)(PenMV)。

分类地位:马铃薯Y病毒属Potyvirus(SCMV),马铃薯Y病毒科Potyviridae。在国际病毒分类委员会(ICTV)中的编码为00.057.0.01.062(SCMV)。

病毒提纯:每300g切碎的组织,加300ml水,水内含0.3%抗坏血酸、0.01mol/L二乙基二硫代胺基甲酸钠和0.3%2-巯基乙醇,搅成匀浆。每次用300ml的抽提液,连续抽提3次100g的叶子,每次都取出残渣而保留提出液。经粗布过滤后加等量氯仿与滤液匀浆。低速离心后再经高速离心,将高速离心沉淀重新悬浮于0.1mol/L硼酸盐、0.01mol/L乙二胺四醋酸(EDTA)、pH8.2的溶液中。收集沉淀并使其重新悬浮的溶液用量尽可能少;因为如果再作第二次的高速离心,侵染性就会丧失90%以上。经第二次低速离心后,将上清液悬浮于含10%~40%蔗糖梯度的0.1mol/L硼酸盐、0.01mol/L乙二胺四醋酸(EDTA),pH8.2液上,以90000g离心2h,病毒即形成一个光散射区带(Bond和Pirone,1971)。

用一个极为相似的方法,即将密度梯度用的蔗糖溶解于0.01mol/L柠檬酸钠溶液中,来提纯玉米矮花叶病毒的A、B毒株以及甘蔗花叶病毒的毒株(Snazelle et al.,1971),制剂可再经蔗糖梯度区带电泳,或者经蔗糖梯度平衡离心而进一步纯化。

病毒理化特性:

①病毒粒子:SCMV线状病毒,(750~770)nm*13nm,沉降系数148~176S,A260/A280为1.22。PenMV病毒粒子长375~750nm,多数集中在550~650nm之间,宽13~14nm。

②核酸:SCMV核酸含量5%,基因组为正单链RNA,长为9.6kb。

③蛋白:SCMV蛋白含量95%,分子质量约为36.4ku。外壳蛋白的氨基酸组成:ala23,arg14,asx26,cys1,glx37,gly18,his7,ile7,leu14,lys22,met11,phe8,pro8,ser15,thr19,trp4,tyr9,val13(Von Baumgarten&Ford,1981)。

株系:石银鹿等(1986)从白草(Pennisetum flaccidum 或P.centrasiaticum)花叶病株上分离到MDMV的一个新株系,并将其命名为MDMV-G(PenMV);该株系的TDP为52~58℃,DEP为10-4,LIV为3d。病毒粒子长375~750nm,多数集中在550~650nm,宽13~14nm。有风轮状内含体。与MDMV-B(SCMV)抗血清呈阳性反应,但寄主范围略有不同,此分离物可侵染狼尾草Pennisetum alopecuroides,在二色蜀黍品种Atlas上表现为花叶,而MDMV-B(SCMV)不侵染狼尾草,在Atlas上为红色条斑。朱福成等(1986)也从白草及其他禾本科植物上分离到一种类似的病毒(株系),将其暂定为白草花叶病毒(FPMV)。范在丰等从显花叶症的白草、玉米等植物上分离、鉴定了一种新的病毒,并将其命名为“Pennisetum mosaic virus(PenMV)”(Fan et al.,2003b;2004),已经得到国际上的初步承认(Adams et al.,2005),其中文名称仍沿用“白草花叶病毒”。根据近年国际上病毒分类的变化以及对我国各地玉米矮花叶病的鉴定研究,我国以前鉴定的MDMV-B应为甘蔗花叶病毒(SCMV,见下文)(Chen et al.,2002;Fan et al.,2003a);而MDMV-G与FPMV应为白草花叶病毒(PenMV)。

其他:对侵染禾本科植物的蚜虫传播的马铃薯Y病毒属成员的分类研究(Shukla et al.,1989)表明,以前的甘蔗花叶病毒(SCMV)的株系(Pirone,1972)实际上包括4种不同的病毒。其中之一为玉米矮花叶病毒(美国株系A、D、E和F)。其他三种病毒分别为石茅高粱花叶病毒(以前澳大利亚甘蔗花叶病毒JG株系、美国玉米矮花叶病毒O株系和Kansas1株系)、甘蔗花叶病毒(美国玉米矮花叶病毒B株系、甘蔗花叶病毒株系A、B、D和E,以及澳大利亚甘蔗花叶病毒株系SC,BC与Sabi)和高粱花叶病毒(SrMV)(美国甘蔗花叶病毒株系H、I和M)。这4种病毒在一些寄主上诱发相似的症状,寄主范围通常局限于禾本科,并且有一些共同的蚜虫介体。然而,它们在血清学上不相关或只有微弱的血清学关系。以前有关这些病毒及其株系间的血清学关系的报道是由于应用了针对在马铃薯Y病毒属成员外壳蛋白中保守的核心区域中的抗原决定簇的多克隆抗体(抗血清)。这些抗体可识别马铃薯Y病毒属的多数成员(Shukla et al.,1988)。这些病毒在其外壳蛋白的氨基酸组成和氨基酸序列差异很大(Gough&Shukla,1981;Von Baumgarten&Ford,1981;Shukla et al.,1987;Jilka,1990),并且可通过利用高效液相层析法对外壳蛋白的降解物很容易地加以区分(Shukla et al.,1988)。也可以根据一些高粱自交系在受到侵染后的反应对这4种病毒加以区分(Teakle&Grylls,1973;Persley et al.,1985;Giorda et al.,1986;Shukla&Teakle,1989;Teakle et al.,1989;Tosic et al.,1990)。MDMV在其系统侵染的高粱品种Rio的叶片上诱发花叶与坏死斑点,而其他3种病毒只诱发花叶或无症侵染。石茅高粱花叶病毒(Johnsongrass mosaic virus)在二色蜀黍自交系OKY8与SA8735上诱发坏死红条,而其他3种病毒只诱发花叶症状。SCMV的株系在Atlas的新叶上诱发坏死,而SrMV诱发典型的红叶症状,其他两种病毒只诱发花叶症状。

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