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我国部分地区樱桃病毒病害初步调查和病原检测 总被引：2，自引：0，他引：2
对山东泰安、辽宁大连和北京的樱桃病毒病发生情况进行调查,发现8个果园/栽培区均有病毒病发生,主要症状为叶片皱缩、畸形、卷叶、花叶、植株矮缩等。采集20份样品,利用12种病毒的引物进行RT-PCR检测。结果表明,在样品中扩增出与樱桃病毒A(Cherry virus A,CVA)、李属坏死环斑病毒(Prunus necrotic ringspot virus,PNRSV),李矮缩病毒(Prune dwarf virus,PDV)、李树皮坏死与茎痘伴随病毒(Plum bark necrosis stem pitting-associated virus,PBNSPaV)、樱桃绿环斑驳病毒(Cherry green ring mottle virus,CGRMV)、樱桃小果病毒-1(Little cherry virus-1,LChV-1)预期大小一致的目的片段;序列分析表明,与GenBank中注册所测的病毒核苷酸序列均具有较高的一致性。其中,大连、泰安和北京样品均检测到CVA;大连和北京样品中检测到PNRSV和PDV;北京样品中检测到PBNSPaV;大连苗木样品枝条中检测到CGRMV和LChV-1。这是在我国樱桃上首次检测到LChV-1。 相似文献
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侵染肥城桃的病毒和类病毒的分子检测与鉴定 总被引：1，自引：0，他引：1
为明确山东肥城桃种植区桃树上主要存在的病毒和类病毒及其发生情况,采集具有花叶、斑驳和皱缩典型症状的肥城桃样品,提取叶片总RNA后,分别选用桃树上已报道的啤酒花矮化类病毒Hopstuntviroid(HSVd)、桃潜隐花叶类病毒Peach latent mosaic viroid(PLMVd)、苹果褪绿叶斑病毒Apple chlorotic leaf spot virus(ACLSV)、樱桃锉叶病毒Cherry rasp leaf virus(CRLV)、桃花叶病毒Peach mosaic virus(PMV)、李属坏死环斑病毒Prunus necrotic ringspot virus(PNRSV)、李痘病毒Plum pox virus(PPV)、李矮缩病毒Prunus dwarf virus(PDV)、樱桃绿环斑驳病毒Cherry green ring mottle virus(CGRMV)、杏假褪绿叶斑病毒Apricot pseudo-chlorotic leaf spot virus(APCLSV)、李树皮坏死茎纹孔伴随病毒Plum bark necrosis stem pitting-associated virus(PBNSPaV)和小樱桃病毒1号Little cherry virus 1(LchV1)的特异性引物进行RT-PCR检测。PCR结果显示仅HSVd、PLMVd、ACLSV、PNRSV和PBNSPaV的扩增产物中得到了预期大小的目的片段,将目的片段克隆测序后,经NCBI BLAST比对发现,山东肥城桃分离物HSVd、PLMVd、ACLSV、PNRSV和PBNSPaV与GenBank已报道分离物序列一致性均达90%以上。表明山东肥城桃已感染HSVd、PLMVd 2种类病毒和ACLSV、PNRSV、PBNSPaV 3种病毒。 相似文献
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A. Bouani M. Al Rwahnih N. Abou Ghanem-Sabanadzovic I. Alami M. Zemzami A. Myrta V. Savino 《EPPO Bulletin》2004,34(3):399-402
Field surveys were carried out in the main stone-fruit growing areas of Morocco to evaluate the sanitary status of commercial orchards, varietal collections and nurseries. The presence of virus and virus-like diseases was checked by ELISA, sap transmission to herbaceous hosts, testing on woody indicators and molecular hybridization (dot-blot and tissue-printing). 1211 samples (382 almond, 339 peach, 291 plum, 150 apricot and 49 cherry) were tested by ELISA for the presence of Prunus necrotic ring spot virus (PNRSV), Prune dwarf virus (PDV), Apple chlorotic leaf spot virus (ACLSV), Apple mosaic virus (ApMV) and Plum pox virus (PPV). The overall average of virus infection rate was 16.4%, whereas that of single species was: 22.6% for almond, 17.8% for plum, 15% for peach, 10.2% for cherry, and 2.7% for apricot. The following viruses were detected: PNRSV, PDV, ACLSV and ApMV. 565 samples were tested by dot-blot and tissue-printing hybridization for the presence of Peach latent mosaic viroid (PLMVd) and Hop stunt viroid (HSVd). 48 samples were infected, 41 by PLMVd and 7 by HSVd. In addition, nested-PCR tests identified Plum bark necrosis and stem-pitting associated virus (PBNSPaV) in a few almond trees affected by stem pitting. 相似文献
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Simultaneous detection and identification of eight stone fruit viruses by one-step RT-PCR 总被引：12，自引：0，他引：12
A sensitive and reliable one step RT-PCR reaction with an internal control has been developed to detect and differentiate eight important viruses that affect stone fruit tress: Apple mosaic virus (ApMV), Prunus necrotic ringspot virus (PNRSV), Prune dwarf virus (PDV), American plum line pattern virus (APLPV), Plum pox virus (PPV), Apple chlorotic leaf spot virus (ACLSV), Apricot latent virus (ApLV) and Plum bark necrosis stem pitting associated virus (PBNSPaV). In addition, we investigated the detection limit and the efficiency of three different nucleic acid extraction methods that avoid the use of organic solvents, for both multiplex RT-PCR and dot-blot hybridisation assays. The primer cocktail was used to analyse 38 stone fruits originating from nine different countries and six species. A large number of virus combinations was detected and up to three different viruses were observed in five samples. A decrease in sensitivity was observed when the primer cocktail contained more than five different pair primers. However, comparative analyses showed that the multiplex RT-PCR containing the eight virus pair primers was even more sensitive than the ELISA or molecular hybridisation assays. The use of the multiplex RT-PCR technology in routine diagnosis of stone fruit tree viruses is discussed. 相似文献
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Occurrence of Citrus psorosis virus in Campania, southern Italy 总被引：1，自引：0，他引：1
D. Alioto A. Troisi A. Peluso G. Quatrano V. Masenga R.G. Milne 《European journal of plant pathology / European Foundation for Plant Pathology》2000,106(8):795-799
Citrus psorosis virus (CPsV), genus Ophiovirus, is associated with a severe disease of citrus worldwide. Double antibody sandwich (DAS) ELISA using a polyclonal antiserum, and triple antibody sandwich (TAS) ELISAs, employing the IgG monoclonal antibody (mab) 13C5, and the IgM mab 2A3, were used to detect CPsV in orchards of different citrus varieties in Campania, southern Italy. TAS ELISA with 13C5 detected all the infections detected by DAS ELISA. Overall, 14% of trees younger than 15 years were positive, but only 1% of older trees, suggesting that infected propagating material has been increasingly used in recent years, in the absence of certification. Highest infection rates were in younger trees of sweet orange (22.8%) and clementine (18.6%). CPsV could easily be detected at all seasons of the year tested (June–January); these and earlier results indicate that TAS ELISA using 13C5 is a sensitive, broad-spectrum and reliable diagnostic method useful for routine tests and certification programmes. Of 44 field isolates responding strongly to DAS ELISA and 13C5-TAS ELISA, mab 2A3 gave similar results with 29 isolates, but gave low values with the others, thus providing a degree of differentiation among isolates. To confirm that the ELISA tests were indeed detecting CPsV, samples of 42 ELISA-positive plants were analysed by ISEM in a blind test, and in 38 of these, characteristic virus particles were clearly seen. Although CPsV was frequently and consistently detected in the area sampled, bark scaling symptoms were not seen: possible reasons for this are discussed. 相似文献
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Liming Lin Ruhui Li Margarita Bateman Raymond Mock Gary Kinard 《European journal of plant pathology / European Foundation for Plant Pathology》2013,137(4):797-804
Asian prunus viruses (APV 1, APV 2 and APV 3), Plum bark necrosis stem pitting associated virus (PBNSPaV) and Peach latent mosaic viroid (PLMVd) are pathogens that infect Prunus species. A single-tube multiplex, TaqMan real-time RT-PCR assay was developed for the simultaneous detection and identification of these pathogens. The protocol includes amplification and detection of a fluorogenic cytochrome oxidase gene (COX) as an internal control. The results of the multiplex TaqMan RT-PCR assay correlated with those from conventional RT-PCR, with a 10-fold increase in sensitivity in the multiplex real-time format. The efficiency and accuracy of the assay was evaluated by testing stone fruit trees from positive control collections and several orchard locations. Several mixed infections of target pathogens were detected in peach orchard samples. This assay is simple, rapid and cost-effective and can be used by quarantine and certification programs where numerous stone fruit trees need to be tested for these pathogens. 相似文献
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Surveys were carried out in vineyards in the main grapevine-growing areas of Palestine (Hebron, Bethlehem, Gaza, Jerusalem, Ramallah, Jenin, Jericho and Nablus) to assess the presence and incidence of virus and virus-like diseases. Leafroll symptoms were observed in Bethlehem, Ramallah and Jerusalem in native and imported cultivars, with higher rates in the red-fruited Shami, Beitoni and Smari. Rugose-wood symptoms were also observed in local and foreign cultivars, especially on grafted vines with a high incidence in Bethlehem. Fanleaf symptoms were rarely observed, while phytoplasma-induced symptoms were observed in Jenin, Jericho and Bethlehem on cvs Biadi, Superior Seedless and Beitoni. ELISA tests showed that 463 out of 566 (82%) tested vines were infected by at least one virus. GVA was the prevailing virus (66.1%), followed by GLRaV-1 (45.6%), GLRaV-3 (21.7%), GFkV (15.7%) and GLRaV-2 (8.3%). GVB and GFLV were also detected to a lesser extent, their incidence ranging between 3.7 and 1.2%, whereas GLRaV-7 was detected in a single vine of cv. Sultanina of foreign origin. Vineyards in the Bethlehem area were particularly badly damaged (97.5%), and some local cultivars were totally (Jandali, Marrawi and Shoyoukhi) or heavily infected (Zaini, Biadi and Shami). ELISA testing of 69 young rootstock mother plants showed a relatively high incidence of virus infection (20.3%). Vein necrosis and vein mosaic diseases were also ascertained on graft-inoculated 110R and Vitis riparia indicator plants, whereas no viruses other than GFLV were mechanically transmitted from about 200 vines onto inoculated herbaceous hosts. 相似文献
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河南甜樱桃病毒病害调查及病原检测 总被引：1，自引：0，他引：1
在河南省郑州市、巩义市、荥阳市、新郑市选择具有代表性的甜樱桃生产园对病毒病发生情况进行调查,采集表现为疑似病毒病症状的样本65份,利用7种病毒引物进行RT-PCR检测。5种病毒检测结果呈阳性,分别是李属坏死环斑病毒(Prunus necrotic ringspot virus,PNRSV)、李矮缩病毒(Prune dwarf virus,PDV)、樱桃绿环斑驳病毒(Cherry green ring mottle virus,CGRMV)、樱桃坏死锈斑病毒(Cherry necrotic rusty mottle virus,CNRMV)及樱桃病毒A(Cherry virus A,CVA);序列分析结果表明,5种病毒扩增片段与GenBank中注册的相应病毒核苷酸序列均具有较高的一致性;样本病毒检出率为100%,其中13份样本为单独侵染,其余52份样本均为多病毒复合侵染,占比高达80%,复合侵染比例随着侵染病毒种类的增多逐渐降低;病毒侵染组合与叶片表型症状无明显对应关系。 相似文献
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Surveys were carried out in commercial vineyards in the main grapevine-growing areas of south-eastern (Adiyaman, Diyarbakir, Merdin, Sanliurfa, Elazig) and central (Nevşehir) Anatolia (Turkey) to assess the presence and incidence of virus and virus-like diseases. Typical fanleaf symptoms were observed in most of the surveyed areas, but they were particularly frequent in Elazig in cvs Kirmizi, Agin, Sirfoni and Kohnu. Leafroll symptoms were present in most vineyards in Adiyaman, Sanliurfa and Elazig, primarily in the red-fruited cvs Antep Karasi, Humusi, Kohnu and Siyah Kabarcik, and in Nevşheir. Rugose wood symptoms were common in Adiyaman, where vineyards were established with grafted planting material, but not in any of the self-rooted Cappadocian cultivars. Phytoplasma-like symptoms were sometimes observed in Elazig and Nevşheir. Biological (sap inoculation to herbaceous plants and graft transmission to woody indicators) and serological (ELISA) assays were used for virus detection and identification. A total of 55.3% of ELISA-tested vines (296 out of 535) were infected by one (11.4%) or more (43.9%) viruses. GVA 1 was the most widespread virus (42.4%), followed by GLRaV-1 (38.5%), GFLV (10.7%) and GFkV (7.1%). Surprisingly low (2.4%) was the infection rate by GLRaV-3, and even lower (<1%) that of the other viruses tested, i.e. GLRaV-2, GLRaV-6, GVB and ArMV. GLRaV-7 was not detected. The occurrence of vein mosaic and vein necrosis was ascertained by testing on woody indicators. A putative nepovirus was isolated from a single vine of cv. Kizlar Tahtasi, the identification and characterization of which is still under way. 相似文献
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The purpose of this study was a comprehensive survey on the occurrence and distribution of the viruses affecting almond nursery trees in western Anatolia (Turkey). Three provinces (Aydýn, Balýkesir and Ýzmir) which are well known for their state and commercial nursery plantations of almond (mixed with other stone fruits) were visited and 19 300 and 19 000 almond nursery trees were surveyed in 1992 and 1993, and 56 and 40 leaf samples collected, respectively. The main viral symptoms were chlorotic and necrotic spots, calico, mosaic, narrowing and deformation of the leaves; gummosis, weak growth, dwarfing and short internodes of the trees. DAS-ELISA was used to detect prunus necrotic ringspot ilarvirus (PNRSV), prune dwarf ilarvirus (PDV), raspberry ringspot nepovirus (RRV), peach rosette mosaic nepovirus (PRMV) and tomato ringspot nepovirus (ToRSV) in almond nursery trees in west Anatolia. PNRSV was found to be the most widespread according to ELISA. PDV was the second most important virus, and some samples were infected with both ilarviruses. Fewer samples were found infected with PRMV, RRV and only a few samples were infected with ToRSV. 相似文献
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A study was carried out to clarify the role of the fungus Chalara fraxinea in decline of Fraxinus excelsior , which is observed on a large scale in central and northern Europe with high incidence of tree mortality. The aims of this work were: (i) to check for the presence of C. fraxinea in various tissues of declining F. excelsior by agar culture isolations and by direct analysis of plant tissues using molecular techniques (DNA extraction, ITS-PCR, cloning, ITS sequencing and T-RFLP); (ii) to study fungal communities inhabiting tissues with symptoms; and (iii) to test the pathogenicity of C. fraxinea to F. excelsior . Chalara fraxinea was isolated from 93% of stem cankers, 91% of necrotic leaf stalks, 27–28% of bark wounds and 30% of visually healthy leaf stalks. Molecular analyses of necrotic leaves, leaf stalks and bark revealed the presence of 25 different fungal taxa, 14 of which were detected in all three types of tissue sample. Chalara fraxinea was the second most common species (61% of samples), and only Cryptococcus foliicola occurred more often (70%). All eight of the tested C. fraxinea isolates induced necroses in bark and cambium on each of 86 inoculated trees, and all controls remained healthy. Average length of necroses caused by different C. fraxinea strains varied from 4·2 to 8·9 cm, but the differences were statistically insignificant. Instead, differences in resistance of individual trees to C. fraxinea were observed. 相似文献
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番茄斑萎病毒(tomato spotted wilt virus, TSWV)和烟草花叶病毒(tobacco mosaic virus, TMV)是2种重要的植物病原病毒, 对多种经济作物的产量和品质均造成严重影响。2021年-2022年, 在云南省丽江市烟草种植区不同烟区采集叶片黄化、皱缩以及无症状的青蒿Artemisia caruifolia样品共计14份, 利用免疫金标速测卡和RT-PCR对其病原病毒进行检测。利用免疫金标速测卡检测结果显示, 在所检样品中有9份样品检测出TSWV, 检出率为64.28%, 有3份样品检测出TMV, 检出率为21.43%, 2种病毒复合侵染的检出率同样为21.43%;利用RT-PCR对复合侵染的3份样品进行分子检测, 结果显示, 在3份复合侵染青蒿样品中获得3条TSWV N基因序列、3条TMV cp基因序列和2条TMV RdRp部分序列。TSWV青蒿分离物与分离自云南的TSWV-2分离物相似性最高, 为99.6%;TMV青蒿分离物与分离自辽宁的TMV-Shenyang分离物和分离自云南的TMV-Yongren-1相似性最高, 均大于99.4%。这是首次发现TSWV和TMV 2种不同属病毒复合侵染青蒿。 相似文献
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Variability among Spanish citrus tristeza virus isolates revealed by double-stranded RNA analysis 总被引：2，自引：2，他引：2
A survey for citrus tristeza virus (CTV) strains, based on double-stranded RNA (dsRNA) analysis, was carried out in five locations on the eastern citrus-growing area of Spain. CTV was recovered from 137 trees of different ages, citrus species and varieties, sampled in 53 orchards. The best months for dsRNA recovery were April, May, September, October, and November, and the highest dsRNA yield was obtained from sweet orange cultivars. Sixteen dsRNA profiles differing by the number and/or position of subgenomic bands were detected. One of these profiles was detected in more than half the trees analysed. Maximum diversity of dsRNA patterns was found in the location with the oldest citrus orchards and the highest CTV incidence (Alzira-Carcaixent). In many instances, several dsRNA profiles were detected in neighbouring trees of the same orchard, notably in Alzira-Carcaixent, where 70% of the plots sampled contained more than one profile. The possible causes for the diversity of CTV isolates found in this specific area are discussed. 相似文献
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Plum pox virus detection in dormant plum trees by PCR and ELISA 总被引：1，自引：0，他引：1
An immunocapture polymerase chain reaction (IC-PCR) protocol and ELISA were compared for their effectiveness in detecting plum pox virus (PPV) in dormant plum material. Although the IC-PCR was about one thousand times more sensitive than ELISA, PPV was detected by ELISA in 71–80% of bark samples collected in December, January and March 1996/97 from pot-grown rootstock trees inoculated with PPV the previous March, compared with 85–86% detection in the same samples by IC-PCR. In similar samples from one-year-old shoots taken from infected branches of orchard trees, 66–81% were positive by ELISA compared with 81–87% by IC-PCR. With bulked samples taken from the fibrous roots of the pot-grown trees, PPV was detected in 92–100% of samples by IC-PCR in winter compared with only 38–65% by ELISA. These results were confirmed in samples from the roots and shoots of the same trees in 1997/98. Three samples per shoot would have been sufficient to detect PPV by ELISA in 87 of the 88 infected shoots tested during the two winters. However, infected shoots are irregularly distributed in diseased trees and PCR assays of root samples offer the potential for improving the reliability of identifying trees infected with PPV. 相似文献
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Bassanezi RB Bergamin Filho A Amorim L Gimenes-Fernandes N Gottwald TR Bové JM 《Phytopathology》2003,93(4):502-512
ABSTRACT Citrus sudden death (CSD), a new disease of unknown etiology that affects sweet orange grafted on Rangpur lime, was visually monitored for 14 months in 41 groves in Brazil. Ordinary runs analysis of CSD-symptomatic trees indicated a departure from randomness of symptomatic trees status among immediately adjacent trees mainly within rows. The binomial index of dispersion (D) and the intraclass correlation (k) for various quadrat sizes suggested aggregation of CSD-symptomatic trees for almost all plots within the quadrat sizes tested. Estimated parameters of the binary form of Taylor's power law provided an overall measure of aggregation of CSD-symptomatic trees for all quadrat sizes tested. Aggregation in each plot was dependent on disease incidence. Spatial autocorrelation analysis of proximity patterns suggested that aggregation often existed among quadrats of various sizes up to three lag distances; however, significant lag positions discontinuous from main proximity patterns were rare, indicating a lack of spatial association among discrete foci. Some asymmetry was also detected for some spatial autocorrelation proximity patterns, indicating that within-row versus across-row distributions are not necessarily equivalent. These results were interpreted to mean that the cause of the disease was most likely biotic and its dissemination was common within a local area of influence that extended to approximately six trees in all directions, including adjacent trees. Where asymmetry was indicated, this area of influence was somewhat elliptical. Longer-distance patterns were not detected within the confines of the plot sizes tested. Annual rates of CSD progress based on the Gompertz model ranged from 0.37 to 2.02. Numerous similarities were found between the spatial patterns of CSD and Citrus tristeza virus (CTV) described in the literature, both in the presence of the aphid vector, Toxoptera citricida. CSD differs from CTV in that symptoms occur in sweet orange grafted on Rangpur lime. Based on the symptoms of CSD and on its spatial and temporal patterns, our hypothesis is that CSD may be caused by a similar but undescribed pathogen such as a virus and probably vectored by insects such as aphids by similar spatial processes to those affecting CTV. 相似文献
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我国发现由甘薯褪绿矮化病毒和甘薯羽状斑驳病毒协生共侵染引起的甘薯病毒病害 总被引：12，自引：0，他引：12
甘薯病毒病害(Sweet potato virus disease,SPVD)是由毛形病毒属(Crinivirus)的甘薯褪绿矮化病毒(Sweet potato chlorotic stunt virus,SPCSV)和马铃薯Y病毒属(Potyvirus)的甘薯羽状斑驳病毒(Sweet potato feathery mottle virus,SPFMV)协生共侵染甘薯引起的病毒病害[1]. 相似文献
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中国甘薯病毒的血清学检测 总被引：21，自引：2，他引：21
作者用4种甘薯病毒抗体(IgG),3种血清学方法(DAS-ELISA、Dot-blot-ELISA和ISEM)对北京,江苏、四川、山东四省(市)的253份甘薯病毒病样品进行了检测。结果表明:上述地区甘薯中普遍存在甘薯羽状斑驳病毒(SPFMV)和甘薯潜隐病毒(SPLV),尚难确定是否存在甘薯轻斑驳病毒(SPMMV)和甘薯花叶菜花叶状病毒(Sweet Potato Caulimo-like Virus,SPCLV)。21%的显症样品同上述4种病毒的抗血清不产生反应,显示我国甘薯上尚存在其它病毒。用Dot blot-ELISA和ISEM检测甘薯病毒比用DAS-ELISA灵敏准确。 相似文献