从云南蝴蝶兰上检测到番茄斑萎病毒属病毒

 应用电镜观察、DAS-ELISA以及RT-PCR检测,从症状表现黄化、环斑的云南蝴蝶兰(Phalaenopsis amabilis)病样中分离得到的一个病毒分离物Tospo-Pha。该分离物粒子近球形、具包膜、直径约90nm,与番茄斑萎病毒属(Tospovirus)的西瓜银色斑驳病毒(Watermelon silver mosaic virus,WSMoV)/花生芽坏死病毒(Groundnut bud necrosis virus,GBNV)复合抗血清呈强阳性反应,分子检测发现该分离物SRNA5'末端序列与CaCV大岩桐分离物(CaCV-Gloxinia)同源性最高(91.0%),在系统进化树中与CaCV聚于同一分支。上述结果表明,从云南蝴蝶兰中分离到的Tospo-Pha属于Tospovirus病毒。     

花生病毒病发生和防治研究进展

花生病毒病是世界性的花生重要病害。在世界范围内，经济上重要的、侵染花生的病毒有９种，包括主要发生在美国的花生斑驳病毒（PMV）和番茄斑萎病毒（TSWV），在印度和南亚国家花生上发生的花生芽枯病毒（PBNV）和印度花生丛矮病毒（IPCV），在非洲国家发生的花生丛簇病毒（GRV）和花生丛矮病毒（PSV），以及我国花生上发生的条纹病毒（PStV）、黄瓜花叶病毒（CMV）和花生矮化病毒（PSV）。在过去的十余年内，国外对花生病毒的研究迅速深入到分子生物学的水平，不仅加深了对病毒自身的认识，也提高了病毒检测、病害诊断的技术…     

逆转录环介导等温扩增技术检测花生条纹病毒

花生条纹病毒病(Peanut stripe virus,PStV)是我国花生上流行最广[1]、田间发病率最高的一种花生病毒病,严重影响花生的产量和品质.由于PStV存在不同症状类型株系,在中国占优势的轻斑驳株系,引起花生症状较轻,并且与其他花生病毒病症状类似,不易分辨,目前主要利用PCR方法对PStV进行检测[2],但该方法检测时间长,并且需要专业仪器,较难普及.     

西花蓟马传播病毒病的研究进展

西花蓟马［Frankliniella occidentalis (Pergande)］是一种世界性的重要农业害虫,目前在69个国家和地区已有报道。西花蓟马能以持久性的方式传播番茄斑萎病毒属(Tospovirus)的病毒,所传播病毒造成的经济损失远远大于其本身所造成的损失。因此,许多学者对西花蓟马及其传播的番茄斑萎病毒属病毒进行了大量研究。本文主要综述了近年来西花蓟马传播病毒的种类、番茄斑萎病毒属病毒的结构以及西花蓟马的传毒机制等方面的研究进展。     

利用RNAi 介导的抗病性获得抗2 种花生病毒的转基因烟草

 分别以花生条纹病毒红安株系(PStV-Hongan)外壳蛋白基因(CP),花生矮化病毒轻型株系(PSV-Mi)和花生黄瓜花叶病毒(CMV-CA)复制酶基因2a为模板,通过PCR 方法分别得到PStV-CP 5′ 端,PSV-Mi 和CMV-CA 2a3′ 端150 bp 的片段,3 种片段混合物为模板经PCR 拼接得到450 bp 的片段,此拼接片段通过Gateway 系统重组至植物表达载体pK7GW1WG2,得到含反向重复拼接片段的植物表达载体pK450。冻融法导入根癌农杆菌(Agrobacterium tumefaciens)菌株GV3101 后,叶盘法转化本生烟(Nicotiana benthamiana),经PCR 检测,获得转基因植株。对T1 代转基因植株分别人工接种3 种病毒,66. 7% 的植株表现对PStV 免疫,9% 的植株表现对CMV-CA 的恢复抗性,全部植株对PSV 感病。siRNA 的Northern blot 结果表明,所有转基因烟草植株都含有病毒特异siRNA,siRNA 含量随接种后时间延长而衰减。     

番茄斑萎病毒(TSWV)广州分离物生物学特性研究

 1984年南方花生病害调查,发现由番茄斑萎病毒(TSWV)引起的新病害。     

番茄斑萎病毒RT-RPA检测方法的建立

为了快速、准确地鉴定番茄斑萎病毒,根据GenBank中韩国的番茄斑萎病毒株系(序列号:KC261961)ss RNA-S基因组编码保守外壳蛋白基因序列设计特异性引物TSWV 5F/5R,建立了番茄斑萎病毒RT-RPA检测方法。特异性检测结果表明,仅番茄斑萎病毒样品扩增出215 bp目标条带,而其他4种病毒样品无扩增出目标条带。灵敏度检测结果表明番茄斑萎病毒RPA检测方法可以达到10-5 cDNA稀释度。     

基于TaqMan MGB探针的花生矮化病毒检测研究

花生矮化病毒(Peanut stunt virus,PSV)是我国进境检疫性有害生物。本研究根据该病毒不同分离株外壳蛋白基因(coat protein,CP)的保守序列,设计了特异性引物与TaqMan MGB荧光探针,建立了PSV的实时荧光RT-PCR检测方法。方法特异性研究表明,针对PSV的2个不同株系PSV-E和PSV-W,均能够得到典型扩增曲线,Ct值分别为20.10和21.22;而对于黄瓜花叶病毒、番茄不孕病毒、马铃薯Y病毒、菜豆荚斑驳病毒以及烟草环斑病毒等其他毒株则没有典型扩增曲线,也无Ct值。灵敏度比较发现,该方法比普通RT-PCR检测方法的灵敏度提高100倍,具有快速、灵敏和高特异性的优点,适合对PSV的检测。     

云南辣椒正番茄斑萎病毒属病毒的分子鉴定

辣椒是云南省主要经济作物之一,近年来病毒病尤其是正番茄斑萎病毒属病毒发病严重,影响了辣椒产量和品质。利用RT-PCR技术对从云南辣椒主产区采集的疑似感染正番茄斑萎病毒属病毒的25份辣椒样品进行分子鉴定,结果显示,12份样品检测出正番茄斑萎病毒属病毒,检出率为48.0%,其中6份是番茄斑萎病毒tomato spotted wilt orthotospovirus (TSWV),检出率为24.0%;5份是番茄环纹斑点病毒tomato zonate spot orthotospovirus (TZSV),检出率为20.0%;有1份是TSWV和TZSV复合侵染,检出率为4.0%,这是在云南辣椒生产上首次发现TSWV和TZSV的复合侵染。通过鉴定,初步了解正番茄斑萎病毒属病毒在云南辣椒生产中的发生情况和种类,为制定云南地区该属病毒的防治策略提供理论依据。     

自然侵染菜豆的花生矮化病毒研究

 对从北京郊区菜豆皱缩花叶病株上获得的一个病毒分离物进行了系统研究。根据该分离物的寄主范围、传播特性、病毒颗粒形态、血清学性质和理化特性等,确定为花生矮化病毒(Peanut Stunt Virus,PSV),并称为PSV菜豆分离物,即PSV Bean Isolate,简称PSV-B。这是国内从菜豆上分离PSV的首次报道。在测定的5科28种植物中,PSV-B侵染5科21种。PSV-B致死温度60~65℃,稀释限点10^-4~10^-5,存活期限6 d以上。PSV-B能被豆蚜(Aphis craccivora Koch)以非持久性方式传播。提纯病毒颗粒球形,直径30 nm。制备的PSV-B抗血清,采用琼脂双扩散法测定效价为1 128。在血清学关系上,P SV-B与已报道的中国PSV-Mi株系相近,与美国PSV-E株系差异明显。PSV-B外壳蛋白亚基分子量为25 000道尔顿;含有4个RNA片段。     

Commonwealth Agricultural Bureaux First Commonwealth Helminthological Meeting

Abstract

Fourteen of the 62 taxonomically characterized viruses known to infect peanuts naturally or under experimental conditions are potyviruses (Sreenivasulu et al. 1991). Peanut stripe potyvirus (PStV) was first noted in 1982 when virus‐like leaf symptoms were observed on peanut in plots in Georgia (USA) that seemed different from those of the endemic peanut mottle potyvirus (Demski and Lovell, 1985). Demski et al. (1984a,b,c) reported it to be a new virus disease of groundnut (Arachis hypogaea L.) characterized by dark green stripes and discontinuous banding along the lateral veins of young leaves and an oakleaf pattern on the older leaves. The virus was detected in germplasm received from China (Demski et al. 1984a,b,c; Demski and Lovell, 1985). PStV is now known to occur in most of the south‐east Asian countries growing groundnut including India (Prasada Rao et al., 1988a). The virus is reported to be seedborne and transmitted by species of aphids. It has assumed considerable quarantine importance throughout the world. Much scientific information has been generated and published on various aspects of PStV, but the information is scattered. This paper reviews research on peanut stripe disease and considers future priorities and control strategies.     

刺槐—花生矮化病毒的初侵染源

 1988~1991年调查河北、河南、山东和北京地区刺槐花叶病树率为4~87.5%,一般30%左右。蚜传试验说明:豆蚜(Aphis craccivora)能将花生矮化病毒(PSV)从刺槐传播到花生。在田间经常观察到PSV由刺槐花叶病树林向花生传播,形成病害发生梯度。1990~1991两年河南开封田间系统观察,5月中旬至6月上、中旬,蚜虫在刺槐上形成繁殖高峰并产生有翅蚜虫向花生地迁飞,随后花生地内出现PSV病株。1990和1991两年河北滦县、迁安病区调查,未发现PSV种传苗,开封PSV感染病株种子,PSV种传率0.025%。PSV引起花生病害一般年份发病很轻。与种传相比,刺槐作为病害初侵染源,对PSV在花生上流行起更为重要的作用。     

感染花生条纹病毒(PStV)后花生生理生化性状变化的研究

 研究表明,花生接种条纹病毒发病后叶片的光合强度、叶绿素含量及超氧化物歧化酶(SOD)活性明显下降,而过氧化物酶(POD)活性却显著提高。高感花生品种白沙1016上述指标的变化幅度较大,发病植株叶片中POD同功酶出现了4条健株中没有的新带,迁移率分别为0.67、0.73、0.80和0.89,而原有的迁移率为0.38的1条谱带变弱消失,发病后SOD同功酶谱带数没有显著变化。     

Nucleotide sequence and symptom modulating analysis of a Peanut stunt virus-associated satellite RNA from Poland: high level of sequence identity with the American PSV satellites

Two peanut stunt virus isolates from Poland (PSV-Ag and PSV-P) have been studied. The isolates produce similar systemic symptoms onNicotiana tabacum plants but the symptoms onN. benthamiana, Pisum sativum andDatura stramonium plants are much stronger for the PSV-P isolate. Analysis of the RNA extracted from purified virions by gel electrophoresis and RT-PCR amplification allowed the detection of a satellite RNA in the PSV-P isolate. The nucleotide sequence of this European PSV satellite was determined and found to have a high degree of identity with the sequences of the four American PSV satellites previously studied, which were found to have either no effect or ameliorate the PSV symptoms in tobacco plants (Collmer et al., 1985; Naidu et al., 1991). The possible role of the European PSV satellite in the modulation of viral symptoms has been studied but no effect was observed when the purified satellite was used with the PSV-Ag isolate as helper virus on any of the three hosts cited above.     

芝麻上花生条纹病毒的发生规律

ＥＬＩＳＡ和生长法检测结果表明,芝麻种不种传ＰＳｔＶ,感染ＰＳｔＶ的花生是芝麻黄花叶病毒病的主要初侵染源。桃蚜、豆蚜和大事蚜能传播芝麻上ＰＳｔＶ,传毒率分别为３７％、１９．３％和１３．８％,而经为０。黄花叶病害流行程度肥芝麻生育期蚜虫发生量互作作用影响。芝麻黄花叶病害年度间流行程度差异大,发病率与６月下旬７月上旬平均气温、降雨量及雨日在。芝麻苗期至蕾期为感病生育期,进入开花后期,芝麻对ＰＳｔＶ 表     

Characterisation of black locust isolates of <Emphasis Type="Italic">Peanut stunt virus</Emphasis> (PSV) from the Pannon ecoregion show the frequent occurrence of the fourth taxonomic PSV subgroup

Biological and molecular characterisation of ten Peanut stunt virus (PSV) isolates from Robinia pseudoacacia was carried out. The host range of these isolates was similar to that of the previously described PSV strains in most cases, but on Pisum sativum and Lens culinaris latent infection was induced. Variability in systemic symptoms was observed only on Nicotiana glutinosa. The partial RNA3 sequences were determined, including the carboxyl terminal region of the movement protein gene, the intergenic region, the entire coat protein gene and the 3′ untranslated region. Nucleotide sequence comparison of the coat protein genes showed 77.6–84.2% identity with most of the known PSV strains and 96.3–98.0% identity with PSV-Rp the typical member of subgroup IV. Phylogenetic analysis indicated the presence of the ancient homologous recombination in all of the examined black locust isolates and all the isolates were members of the fourth PSV subgroup. These results showed that the isolates of the fourth subgroup are widely distributed in black locust in this region.