福建口岸首次从日本进境水仙种球上截获4种病毒

<正>2016年3月,福建出入境检验检疫局技术中心采用血清学ELISA、电镜观察及分子生物学RT-PCR方法从一批旅检截获的日本进境水仙种球上检出水仙普通潜隐病毒(Narcissus common latent virus,NCLV)、水仙退化病毒(Narcissus degeneration virus,NDV)、水仙迟季黄化病毒(Narcissus late season     

福建口岸从英国进境水仙种球上截获南芥菜花叶病毒和水仙潜隐病毒

2011年11月,福建出入境检验检疫局技术中心对一批英国进境水仙种球进行检疫,采用ELISA初筛,RT-PCR及序列测定确认的方法,从中同时检出南芥菜花叶病毒(Arabis mosaic virus,ArMV)和水仙潜隐病     

江苏省扬州市水仙病毒检测鉴定与序列分析

水仙属于石蒜科水仙属,是重要的观赏花卉之一。2016年-2017年,在江苏省扬州市采集的水仙上发现典型的花叶、条斑等病毒病症状。利用已报道的水仙病毒通用引物对疑似病样进行RT-PCR检测,结果表明,扬州采集到的3份病样上均能扩增到与预期大小相同的DNA片段。经克隆测序和序列分析发现,所得序列包含3种不同的类型,分别与水仙普通潜隐病毒Narcissus common latent virus(NCLV)、水仙退化病毒Narcissus degeneration virus(NDV)和水仙黄条病毒Narcissus yellow stripe virus(NYSV)有93%、98%和99%的相似性,表明江苏水仙受到了NCLV、NDV、NYSV 3种病毒侵染。这是江苏水仙上第一次发现NCLV、NDV、NYSV。在系统树中,NCLV可分为两个组,本研究所得NCLV-YZ与上海分离物位于同一组中;NDV也可分为两个组,本研究所得NDV与其他中国分离物均位于第二组;而NYSV具有很高的多样性,可分为4个组,本研究所得NYSV与其他部分中国分离物和日本分离物位于第一组。NCLV和NDV分组具有清晰的地理差异性,而NYSV受到地理隔离的影响不明显。     

侵染唐菖蒲和百合三种病毒的多重实时荧光RTPCR检测方法

南芥菜花叶病毒Arabis mosaic virus(ArMV)、百合无症病毒Lily symptomless virus(LSV)和菜豆黄花叶病毒Bean yellow mosaic virus(BYMV)是侵染百合和唐菖蒲的主要病原物,受这三种病毒侵染的唐菖蒲和百合植株长势减弱、切花质量降低、种球产量下降.     

从进境水仙上检测出南芥菜花叶病毒

利用DAS-ELISA对从荷兰进口的6个品种的水仙种球进行检测发现,品种为Recurvus的水仙种球对ArMV的多抗血清呈阳性反应达85%.免疫吸附电镜观察到在感病水仙种球中存在直径约30nm的球状病毒粒子.根据ArMV外壳蛋白(CP)基因的保守序列设计引物,RT-PCR能从感病样品中扩增到预期580bp特异条带,序列测定分析表明此条带的序列为ArMV部分CP基因,在系统关系树上与ArMV的其它分离物形成一簇、亲缘关系很近,表明从进境水仙上检测到了ArMV.     

从葎草中检出复合侵染的多种病毒

采用抗原直接包被酶联免疫吸附测定法（ELISA）对采自重庆近郊的34个葎草样品进行了主要病毒种类的检测。其中马铃薯Y病毒(Potato virus Y, PVY)的侵染最普遍,其阳性检出率达44.12%;马铃薯X病毒(Potato virus X, PVX)的阳性检出率最低,仅为26.47%,其余5种病毒,烟草花叶病毒(Tobacco mosaic virus, TMV)、黄瓜花叶病毒(Cucumber mosaic virus, CMV)、番茄花叶病毒(Tomato mosaic virus, ToMV)、芜菁花叶病毒(Turnip mosaic virus, TuMV)及蚕豆萎蔫病毒2号(Broad bean wilt virus 2, BBWV-2)的阳性检出率均为35.29%。葎草样品受多种病毒的复合侵染现象非常严重,15个阳性样品中病毒复合侵染率为80%,其中75%的样品检测到7种病毒复合侵染。     

进境荷兰郁金香种苗中南芥菜花叶病毒的鉴定

为了防止南芥菜花叶病毒(Arabis mosaic virus,ArMV)传入我国,采用血清学、分子生物学、电镜观察及生物学接种等方法对从荷兰进口的郁金香种苗进行了ArMV检测。结果表明:ArMV血清学反应为强阳性;RT-PCR反应扩增出370bp的特异性目标条带;RT-PCR产物与已报道的3个ArMV部分外壳蛋白基因的核苷酸序列同源性为91.00%～93.24%,氨基酸序列同源性为99%～100%;免疫电镜观察到叶片病汁液中含有直径约30nm的球状病毒粒体;该病毒在黄花烟、白肋烟、矮牵牛和昆诺藜等鉴别寄主上引起坏死和褪绿等典型症状,经DAS-ELISA检测,这些接种寄主均呈ArMV血清学阳性反应。故将该病毒鉴定为南芥菜花叶病毒。     

侵染冬瓜的病毒病原种类鉴定

本研究在我国主要冬瓜产区采集具有典型病毒病症状的病叶材料105份,根据葫芦科作物上常见的5种病毒病原的CP基因设计特异性引物,对105份待检冬瓜材料进行RT-PCR检测。检测结果表明:5对特异引物可分别在105份待检材料的95份中检测到小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)、西瓜花叶病毒(Watermelon mosaic virus,WMV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)、番木瓜环斑病毒(Papaya ringspot virus,PRSV)4种病毒,未检测到南瓜花叶病毒(Squash mosaic virus,SqMV);并且发现不同的冬瓜主产区致病的病毒种类有较大差异;同时还发现,在这些待检样品中4种病毒复合侵染现象较普遍,其中以PRSV与WMV组合最常见,占复合侵染现象的31.25%;未发现有4种病毒复合侵染。     

中山局从进镜花卉种球中检出3种病毒

2006年10～12月期间，中山检疫局技术中心植检实验室对从荷兰进境的2000多头百合种球和400多头剑兰种球进行检疫，利用RT—PCR分子检测方法，从样品中检出带有百合无症病毒和百合斑驳病毒（Lily mottle virus）复合感染的百合种球1株；在剑兰种球中检出2株带有菜豆黄色花叶病毒（Bean yellow mosaic virus），这是中山局首次检出这3种植物病毒，另外菜豆黄色花叶病毒也是广东辖区内首次检出的病毒。     

High prevalence of viruses in table grape from Spain detected by real-time RT-PCR

Table grapes from one of the most important growing area in Spain (Vinalopó, Alicante) protected by the Designation of Origin “Vinalopó bagged table grape”, were surveyed and analysed to determine the prevalence of the five viruses included in the Spanish certification program: Arabis mosaic virus (ArMV), Grapevine fanleaf virus (GFLV), Grapevine fleck virus (GFkV), Grapevine leafroll associated virus-1 (GLRaV-1) and Grapevine leafroll associated virus-3 (GLRaV-3). Ninety five sampling points were selected and the position of grapevine plants georeferenced. Samples were collected in two different vegetative periods and analyses were performed by ELISA and real-time RT-PCR. Purified RNA and immobilized viral targets from plant extracts on nylon membranes were used in parallel assays as templates for PCR assays. In order to analyse these five viral species by real-time RT-PCR, new specific primers and TaqMan probes were designed for detection of ArMV and GFkV. Real time RT-PCR from purified RNA was more sensitive than spot version and ELISA tests. The most prevalent virus was GFLV (95.8%) followed by GLRaV-3 (94.7%), GLRaV-1 (66.3%) and GFkV (65.3%). ArMV was not detected in any sample. The high level of viral infections and the presence of mixed infections suggest that initial infected plant material and uncontrolled traffic of propagation material have played an important role in the spread of viruses.     

同步检测为害百合种球3种检疫性病毒的多重RT-PCR方法

 The multiplex RT-PCR approach was developed for simultaneous detections of Arabis mosaic virus(ArMV),Strawberry latent ringspot virus(SLRSV)and Lily symptomless virus(LSV)from the imported lily bulbs.The results indicated that good specificity and sensitivity for simultaneous detection were obtained.The ultimate of RNA detection with three viruses mixture was 305 pg.The ultimate of RNA detection with ArMV,SLRSV and LSV were 156.0,13.4 pg and 1.12 ng respectively.This approach has potential to be used in quarantine of imports and exports.     

Molecular epidemiology of cassava mosaic disease in Madagascar

Cassava is the staple food for hundreds of millions of people in Africa but its cultivation is seriously constrained by cassava mosaic disease (CMD) in Madagascar, and in Africa in general. This study identified the cassava mosaic geminiviruses (CMGs) involved in CMD in Madagascar and their associated epidemiological characteristics from countrywide surveys. Molecular characterization of CMGs in Madagascar revealed an unprecedented diversity and co‐occurrence of six viruses: African cassava mosaic virus (ACMV), East African cassava mosaic Cameroon virus (EACMCV), East African cassava mosaic Kenya virus (EACMKV), East African cassava mosaic virus (EACMV), South African cassava mosaic virus (SACMV) and the recently described Cassava mosaic Madagascar virus (CMMGV). Distinct geographical distributions were observed for the six viruses. While ACMV was more prevalent in the central highlands, EACMV and EACMKV were prevalent in lowlands and coastal regions. Both EACMCV and SACMV occurred in almost all the localities visited. PCR diagnosis revealed that mixed infection (up to four co‐infected viruses) occurred in 21% of the samples and were associated with higher symptom severity scores. Pairwise comparisons of virus associations showed that EACMCV was found in mixed infections more often than expected while ACMV and SACMV were mostly found in single infections. A greater abundance of whiteflies was observed in lowland and coastal areas. Nevertheless, infected cuttings remain the primary source of CMD propagation (95%) in Madagascar.     

High incidence of <Emphasis Type="Italic">Pelargonium line pattern virus</Emphasis> infecting asymptomatic <Emphasis Type="Italic">Pelargonium</Emphasis> spp. in Spain

Geraniums (Pelargonium spp.) are traditional ornamental plants largely cultivated in Europe and northern America. Vegetative propagation makes them prone to viral infections, which have detrimental effects on crop production and quality. Asymptomatic samples collected in Spain were tested for a range of viruses using ELISA. The tobamovirus, Tobacco mosaic virus (TMV), the cucumovirus, Cucumber mosaic virus (CMV), and several viruses in the family Tombusviridae, namely, Pelargonium line pattern virus (PLPV), Pelargonium flower break virus (PFBV), and Pelargonium leaf curl virus (PLCV), were detected either singly or in combination in 59.2% of 800 samples. PLPV and PFBV infections were confirmed by dot-blot hybridisation. The most relevant viral infection found on Spanish asymptomatic geraniums was by Pelargonium line pattern virus (PLPV). Symptoms did not develop for 3 years on most of the PLPV infected geranium plants under greenhouse conditions.     

Multiple virus infections of lablab [<Emphasis Type="Italic">Lablab purpureus</Emphasis> (L.) Sweet] in Nigeria

Leaf samples of Lablab purpureus collected from two agroecological zones of Nigeria—the northern guinea savanna zone (NGSZ) and the derived savanna zone (DSZ)—were infected with viruses when serologically indexed against available antisera. Approximately 31.1 and 81.1% of the leaf samples collected from the NGSZ and DSZ, respectively, were infected. Seven viruses were found: Bean common mosaic virus (BCMV), Cowpea aphid-borne mosaic virus (CABMV), Cucumber mosaic virus (CMV), Cowpea mottle virus (CPMoV), Cowpea severe mosaic virus (CPSMV), Southern bean mosaic virus (SBMV) and Tobacco mosaic virus (TMV) were detected from samples collected from NGSZ, while CMV, CPMoV, Cowpea mosaic virus (CPMV) and CPSMV were detected from samples from DSZ.     

Evaluating insecticides for the control of narcissus flies under field conditions in Israel

In Israel, narcissus bulb flies (Diptera: Syrphidae) are serious pests of cultivated flower bulbs of the families Liliaceae and Amaryllidaceae. The large narcissus fly(Merodon eques) is the major pest, whereas the small narcissus fly (a new species in the genusEumerus, yet to be described) is only a secondary pest. The large narcissus fly is also considered a quarantine pest by the U.S.A. authorities. Narcissus bulbs,Narcissus tazetta (var. ‘Ziva’), were planted in an experimental field at Bizzaron during November 1995 and harvested during June 1996. Currently aldicarb (Temik) is recommended for the control of narcissus fly larvae. We compared the control efficacy of imidacloprid (Confidor) and isazofos (Miral) with that of aldicarb. These latter insecticides were applied to the soil in February, in April, or on both dates. The mean level of damaged bulbs in the untreated plots was 32%. Two applications of aldicarb, one in February and one in April, reduced the damage to the lowest level of 0.5%. A single application of aldicarb in February, and two applications of imidacloprid—one in February and one in April—reduced the damage to 5-10%. Treatments with imidacloprid in February or in April, reduced the damage to 12-13%. Neither one application of aldicarb in April, nor any of the treatments with isazofos, was effective. In all treatments, larvae of the large narcissus fly were found in only approximately one-third of the damaged bulbs. The level of infestation with the small narcissus fly in the untreated bulbs was only approximately 2%. The effects of the insecticide treatments on the small narcissus fly were similar to those recorded for the large narcissus fly .     

Phytosanitary status of grapevine in Montenegro

During 2006 and 2007, a survey on the incidence and distribution of fourteen grapevine viruses was carried out in the Skadar Lake basin, one of the two main grapevine‐growing areas of Montenegro. In total 165 samples were collected from four red (‘Vranac’, ‘Krato?ija’, ‘Merlot’ and ‘Cardinal’), two white (‘Chardonnay’ and ‘Rkaciteli’) and a few unknown grapevine varieties in the vicinity of Podgorica and Bar. The phytosanitary status of the collected samples was analysed by DAS‐ELISA and the presence of Grapevine fanleaf virus (GFLV), Grapevine leafroll‐associated virus 1 (GLRaV‐1), Grapevine leafroll‐associated virus 2 (GLRaV‐2) and Grapevine leafroll‐associated virus 3 (GLRaV‐3) was confirmed in some of them. The most frequently found virus in assayed samples was GLRaV‐3 (54.5%), followed by GFLV (23%), GLRaV‐1 (20%) and GLRaV‐2 (0.6%). These serological analyses showed the absence of Grapevine leafroll‐associated virus 6 (GLRaV‐6), Grapevine leafroll‐associated virus 7 (GLRaV‐7), Raspberry bushy dwarf virus (RBDV), Strawberry latent ringspot virus (SLRSV), Tomato ringspot virus (ToRSV), Raspberry ringspot virus (RpRSV), Arabis mosaic virus (ArMV), Tobacco ringspot virus (TRSV), Tomato black ring virus (TBRV) and Cherry leaf roll virus (CLRV) from all tested samples.