Closterovirus infection and mealybug exposure are necessary for the development of mealybug wilt of pineapple disease

ABSTRACT The roles of Pineapple mealybug wilt-associated viruses (PMWaVs) and mealybug (Dysmicoccus spp.) feeding in the etiology of mealybug wilt of pineapple (MWP) were evaluated. Container-grown pineapple (Ananas comosus) plants from five commercially grown Hawaiian proprietary selections and a field study utilizing a randomized complete block design were used to test four treatments for induction of MWP: PMWaV-1-free and PMWaV-1-infected plants maintained mealybug-free, and PMWaV-1-free and PMWaV-1-infected plants that received monthly applications of nonviruliferous mealybugs. A second PMWaV, PMWaV-2, was identified in some of the test plants during the course of these studies and was shown to be an integral factor in MWP etiology. Typical MWP symptoms developed only in plants infected with PMWaV-2 and exposed to mealybugs. MWP did not develop in PMWaV-1-free or PMWaV-1-infected plants that were exposed to mealybugs, or in mealy-bug-free plants infected with PMWaV-1, PMWaV-2, or both viruses. Plants from all five Hawaiian proprietary selections developed MWP when PMWaV-2 infected plants were exposed to mealybug feeding. A PMWaV-2-specific monoclonal antibody was produced that decorated the particles in immunosorbent electron microscopy and detected the virus in tissue blot immunoassays. PMWaV-2 was acquired and transmitted by pink and gray pineapple mealybugs (Dysmicoccus spp.) to pineapple plants, and these plants subsequently developed MWP symptoms while sustaining mealybug populations.     

Detection of pineapple closterovirus in pineapple plants and mealybugs using monoclonal antibodies

Monoclonal antibodies (MAbs) were produced to the pineapple closterovirus (PCV) in Hawaii. These antibodies were shown to be specific for PCV by decoration of the virus particles in immunosorbent electron microscopy (ISEM) and indirect enzyme-linked immunosorbent assay (ELISA). Several methods of ELISA were compared. An indirect DAS ELISA using a polyclonal antibody to trap virus particles followed by reaction with monoclonal antibody was shown to be the method of choice for detecting PCV in pineapple plants. Pineapple root tissue was found to be most suitable for detecting PCV in crude samples by indirect ELISA. PCV was detected in symptomatic and asymptomatic pineapple plants collected from Oahu and Maui, and pineapple collections in the USDA/ARS National Clonal Germplasm Repository, but was not detected from pineapple plants grown from seed. At least two serotypes of PCV were detected. In addition. PCV was detected from mealybugs collected from wilted pineapple plants, but not from mealybugs of the same species collected from a colony reared on squash. The role of PCV in mealybug wilt of pineapple is being investigated.     

New Mealybug Species Vectoring Grapevine Leafroll-Associated Viruses-1 and -3 (GLRaV-1 and -3)

Many grape viruses, such as filamentous Grapevine leafroll-associated viruses in the Closteroviridae family, are spread primarily through infected propagating material. However, there is increasing evidence that leafroll disease are spread in the field by insect vectors, namely mealybugs and other scale insects. This study was carried out in the northern wine-growing regions of France where Grapevine leafroll-associated virus-1 and -3 (GLRaV-1 and -3) are the most widespread grape Ampelovirus species. The vineyards were inspected for presence of mealybug and scale insects and grapes infected by GLRaV-1 and -3. Mealybugs, Heliococcus bohemicus, Phenacoccus aceris (Pseudococcidae) and the soft scale insect Parthenolecanium corni (Coccidae), were capable of a transmission efficiency of 14%, 23% and 29% respectively. GLRaV-1 and -3 infections that resulted from virus transmission were confirmed with DAS-ELISA using polyclonal antibodies. This is the first report of GLRaV-1 and -3 transmission by mealybug and coccid species in France, and the first report of the ability of H. bohemicus and Phenacoccus aceris to transmit these viruses to grapevines. The relevance of these findings with regards to maintenance of virus-free grapevine stocks and to control leafroll spread in commercial vineyards is discussed.     

Maize fine streak virus, a New Leafhopper-Transmitted Rhabdovirus

ABSTRACT A previously uncharacterized virus was isolated from fall-planted sweet corn (Zea mays L., Syngenta GSS 0966) leaves showing fine chlorotic streaks. Symptomatic plants were negative in enzyme-linked immunosorbent assay against many maize viruses, but reacted weakly with antisera to Sorghum stunt mosaic virus suggesting a distant relationship between the viruses. The virus was readily transmitted by vascular puncture inoculation (VPI), but not by leaf-rub inoculation. Symptoms on maize included dwarfing and fine chlorotic streaks along intermediate and small veins that developed 12 to 17 days post-VPI. The isolated virus was bacilliform (231 +/- 5 nm long and 71 +/- 2 nm wide), with a knobby surface, and obvious helical structure typical of rhabdovirus morphology. Nucleorhabdovirus virions were observed by transmission electron microscopy of infected maize leaf tissue sections. Proteins unique to infected plants were observed in extracts of infected leaves, and the isolated virion contained three proteins with molecular masses 82 +/- 2, 50 +/- 3, and 32 +/- 2 kDa. Preliminary sequence analysis indicated the virus had similarity to members of the family Rhabdoviridae. The virus was transmitted by Graminella nigrifrons under persistent conditions. The data indicate the virus, provisionally designated Maize fine streak virus, is a new species in the genus Nucleorhabdovirus.     

Solanum nigrum: an indigenous weed reservoir for a tomato yellow leaf curl geminivirus in southern Spain

A tomato yellow leaf curl geminivirus (TYLCV-AL), was first identified in tomato plants in Almeria, southern Spain in 1992. This virus is transmitted by the tobacco whitefly, Bemisia tabaci (Gennadius), and is presently infecting tomato crops throughout the south eastern region of Spain. Solanum nigrum, collected from a field in south east Spain and exhibiting leaf curl symptoms, was squash blotted onto nylon membrane and gave a positive signal when hybridised to a TYLCV-Is DNA probe. Laboratory tests showed B. tabaci to transmit TYLCV-AL from infected tomato plants to S. nigrum seedlings. The virus could then be acquired by B. tabaci and transmitted back from infected S. nigrum plants to tomato, inducing typical TYLCV symptoms. These results indicate the importance of S. nigrum as a weed host/reservoir for a TYLCV and its possible role in the spread of this virus within Europe.     

Ceratothripoides claratris, a New Vector of a Capsicum chlorosis virus Isolate Infecting Tomato in Thailand

ABSTRACT Ceratothripoides claratris, the predominant thrips species on tomato in Thailand, was tested for vector competence and efficiency to transmit Capsicum chlorosis virus (CaCV) (isolate AIT) to tomato. The efficiency of adult-stage transmission was influenced by the larval stage at which virus was acquired. Adult C. claratris showed 69% transmission efficiency after acquiring the virus as freshly emerged (<1 h) first-instar larvae. However, when just molted (<1 h) second-instar larvae acquired the virus, the percentage of adult transmitters significantly decreased (48%). Transmission efficiency of up to 47% was detected with second-instar larvae of C. claratris which had acquired the virus as freshly emerged first-instar larvae. Transmission efficiency did not significantly differ between adult males and females, irrespective of the larval stage at which the virus was acquired. Highest transmission efficiency for CaCV was recorded in adult C. claratris derived from second-instar larvae collected from infected tomato plants in a greenhouse. Lowest transmission efficiency was observed in adults directly collected from infected tomato plants in the greenhouse. The spread of CaCV on tomato plants in greenhouses showed a close association with thrips infestations.     

Comparative whitefly transmission of Tomato chlorosis virus and Tomato infectious chlorosis virus from single or mixed infections

Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV) are two criniviruses that are emerging worldwide, and induce similar yellowing diseases in tomato crops. While TICV is transmitted only by Trialeurodes vaporariorum , ToCV is transmitted by three whitefly species in two genera Trialeurodes vaporariorum , T. abutilonea and Bemisia tabaci . The efficiency of transmission by T. vaporariorum from plants infected by one virus or by both was compared, and the probability of virus transmission by a single whitefly was derived from group testing experiments. The estimated transmission probabilities ranged from 0·01 to 0·13, and were not significantly different between ToCV and TICV, or between single and mixed infections. Experiments using B. tabaci as a vector and source plants infected by TICV and ToCV did not reveal any functional trans-complementation for transmission of TICV by ToCV, suggesting that if this phenomenon occurs in nature, it is at a very low frequency. Possible reasons why TICV did not establish in southern France while ToCV is now endemic are discussed.     

The question of seed transmissibility of Plum pox virus

For many years, Plum pox virus (PPV) was considered to be transmissible by seed, increasing the fear of long-distance spread of the disease. In the late 1970s, it was claimed on the basis of biological transmission of the virus to herbaceous indicator plants and the development of serological diagnosis based on polyclonal antibodies, that PPV was seed-transmitted, with a different infection rate according to the plant species and part of the seed which was tested. In the 1990s, PPV was characterized into four different types, and specific monoclonal antibodies were produced for them. These new and more sensitive diagnostic techniques, together with RT-PCR with different sets of specific primers, were used to approach once again the problem of PPV transmission through seeds. The virus was detected in seed coats and cotyledons, but embryonic tissue and seedlings obtained from germinated seeds never showed symptoms, and gave negative results for PPV with both ELISA and PCR assays. No PPV isolate is currently recognized to be seed transmitted, so vertical transmission of PPV from infected mother plants to their progeny does not occur. Hypothetically, the only possibility of seed transmission would arise from a mutation in the helper component of the virus, associated with high susceptibility of the infected Prunus cultivar.     

The role of the mealybug Phenacoccus aceris in the spread of Grapevine leafroll-associated virus −1 (GLRaV-1) in two French vineyards

Spread patterns of a Grapevine leafroll-associated virus 1 (GLRaV-1) epidemic and a mealybug infestation survey over 10 year were recorded in two Burgundy French vineyards to investigate the relation between them. The temporal evolution of leafroll spread at both study sites was compared on disease incidence data with logistic regression models. We first tested if the spatial distribution of the disease and the mealybug were aggregated using permutation methods, then we tested the independence between the two spatial patterns by randomly shifting one pattern. In Bonzon, an increase from 5 % to 86 % of leafroll prevalence was observed over an 8-year time span, whereas leafroll prevalence remained stable around 5 % in Marsannay-la-Côte during the same period. In Bonzon, the disease spread rapidly from older neighbouring vineyards in four main patches while no spread of the disease was recorded from infected vines in Marsannay-la-Côte. The mealybug Phenacoccus aceris was recorded on 74 % of vines in Bonzon throughout the study and only 6 % of vines in Marsannay-la-Côte. In the latter location, the disease was not associated with the presence of the mealybug, so that it may have arisen from infected plant material escaping the sanitary inspection. In Bonzon, the significant statistical correlation between the mealybug distribution and diseased plants suggests that P. aceris was responsible for the rapid spread of GLRaV-1 in the vineyard. This is the first report of GLRaV-1 natural spread in Europe.     

The transmission by nematodes of tobraviruses is not determined exclusively by the virus coat protein

The coat protein gene of the nematode non-transmissible, SP5 isolate of pea early-browning tobravius was replaced with that of the highly nematode transmissible, PPK20 isolate of tobacco rattle tobravirus. Plants were infected with the recombinant virus when mechanically inoculated and the virus invaded the plants systemically. However, although the PPK20 isolate of TRV was transmitted by nematodes from these plants, the recombinant virus was not transmitted. Therefore, the virus coat protein is not the exclusive determinant of nematode transmission.     

香石竹脉斑驳病毒的研究——Ⅰ.病毒的生物学、病理学变化及血清学

 香石竹脉斑驳病毒的寄主范围比较狭窄,只能侵染石竹科、藜科、苋科等几个科中的少数几种植物。昆诺藜和美国石竹是该病毒理想的鉴别寄主和繁殖寄主。病毒粒体为微弯曲的线条状,长790nm,宽12nm,能为国外提供的香石竹脉斑驳病毒抗血清均一包被。该病毒能被桃蚜以非持久性的方式传播,传播效率可达58%。本文还对病组织的超薄切片、病毒侵染组织中的双链RNA、病毒的体外抗性进行了研究。     

Pfaffia mosaic virus: a new potyvirus found infecting Pfaffia glomerata in Brazil

In Brazil plants of Pfaffia glomerata with mosaic symptoms were found to be infected with a previously undescribed potyvirus, Pfaffia mosaic virus (PfMV). Virus particles were long and flexuous, c.  10 × 700–800 nm, and cylindrical inclusions typical of potyviruses were present in cells of infected tissue. Partial host-range studies revealed that in addition to P. glomerata , PfMV infected only Chenopodium amaranticolor and Chenopodium quinoa . It was efficiently transmitted by the aphids Aphis gossypii and Myzus persicae . Polyclonal antiserum produced against the PfMV coat protein (CP) reacted with Potato virus Y (PVY), but not with four other potyviruses in PTA-ELISA. The similarity of the nucleotide sequence of the PfMV coat-protein gene ( CP ) varied from 7 to 76% when compared with other members of the family Potyviridae . Similarity of the 3' NTR sequence varied from 4 to 23%. In both cases the highest similarity was with PVY. These data indicate that PfMV is a new species in the genus Potyvirus .     

Acquisition and subsequent transmission of tobacco rattle virus isolates by Paratrichodorus and Trichodorus nematode species

Isolates of the PRN serotype of tobacco rattle virus (TRV) were transmitted with different efficiencies by the nematode vectorParatrichodorus pachydermus. Virus isolates which belonged to other serotypes were not acquired and/or transmitted by this vector, nor were PRN serotype isolates which had been obtained from naturally infected potato plants and maintained by mechanical transmission in the glasshouse for several years. PRN serotype TRV isolates from the Netherlands or from Scotland were equally well transmitted by initially virus-freeP. pachydermus populations from either country. Allowing a naturally viruliferous nematode population access for 3 weeks to uninfected or TRV-infected roots resulted in an increased proportion of the trichodorid population transmitting TRV.     

Characterization of an isolate of beet mosaic virus from South Kazakhstan

A filamentous virus isolated from a sugar-beet plant showing systemic mosaic collected in South Kazakhstan was identified as an isolate of beet mosaic virus (BMV-K). BMV-K was transmitted by the green peach aphid Myzus persicae in a non-persistent manner, and by sap inoculation to 11 out of 19 species from seven families tested. The virus could not be transmitted to Nicotiana tabacum, N. debneyi, N. glutinosa and N. clevelandii, cither mechanically or with M. persicae. The thermal inactivation point of BMV-K in sugar-beet sap was 55-60 C, dilution end point 1:1000 and longevity in vitro 2 days at 20 C. A purification procedure produced 1-5-3 mg of purified virus from 100 g of infected Stellaria media plants. Purified virus contained a single protein species of molecular weight 34 700 Da. In ELISA tests, BMV-K reacted positively with BMV-specifc antisera obtained from Japan. Germany and Portugal. By competitive DAS- ELISA, the virus isolate was shown to be closely serologically related to all the three isolates of BMV, and very distantly related to bean yellow mosaic and soy bean mosaic viruses.     

新菠萝灰粉蚧在中国目前及未来的潜在地理分布研究

新菠萝灰粉蚧(Dysmicoccus neobrevipes(Beardsley)),属同翅目(Homoptera)粉蚧科(Pseudococci-dae),被列为我国进境植物检疫性有害生物。该虫主要分布在美洲地区,特别是墨西哥到巴西之间,另外欧洲、非洲和亚洲也有少量分布。新菠萝灰粉蚧于1998年在我国首次发现,随后在广东省的部分地区大量暴发,造成严重经济损失。本研究利用CLIMEX 3.0与ArcGIS 9.3结合的方法研究该虫在我国目前以及未来的潜在地理分布。研究结果发现当前气候条件下,新菠萝灰粉蚧在我国的适生范围主要分布在18.3°N～27.3°N,包括海南、广东、广西、云南、贵州、台湾、香港、澳门、福建、湖南、江西、四川、重庆、浙江、西藏等省市,适生区面积占全国总面积的13.03%。其中整个海南、广东和广西的大部分地区、云南南部、福建东南部和台湾西南部高度适生。基于我国未来气候变化的预测结果显示,2020年,适生区向北扩展,面积将增加至全国总面积的18.97%;到2050年,适生区北界将向北移至32.8°N,上海、江苏和安徽南部均将成为适生区。鉴于新菠萝灰粉蚧在我国的潜在地理分布研究结果,应进一步加强针对该虫的检疫、根除措施,完善疫情监测体系,预防其新的进入和扩散。     

Transmission by aphids of potato spindle tuber viroid encapsidated by potato leafroll luteovirus particles

Potato spindle tuber viroid (PSTVd) was transmitted by Myzus persicae to Physalis floridana from P. floridana plants that also were infected with potato leafroll luteovirus (PLRV), whereas it was not transmitted by aphids from plants infected with PSTVd alone. Dot-blot hybridisation was used to detect PSTVd. The results indicate that PLRV can assist PSTVd in its transmission by M. persicae. Doubly infected, aphid-inoculated P. floridana plants from the previous experiment were used as the source plants in aphid transmission tests to the tomato cv. Rutgers, P. floridana and Datura stramonium. PSTVd was detected in 17 of 30 plants of tomato. The viroid was not detected by dot-blotting in any plant of P. floridana and D. stramonium in this experiment, but it was recovered from some plants by sap inoculation of the Rutgers plants. Treatment with RNase A of PLRV preparations purified from doubly infected plants indicated that PSTVd was encapsidated by PLRV particles.     

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。