Whiteflies on tomato crops in Portugal*

In Portugal during the 1960/1980s, there was intensive development of vegetable crop production, in particular protected crops, of which tomato was the most important. The main producing regions now are Ribatejo e Oeste, Alentejo and Algarve. Tomato presents extensive phytosanitary problems, being host to a wide range of pests, including the whiteflies Trialeurodes vaporariorum, in protected crops, and Bemisia tabaci, in protected and field crops. Portugal has applied for the status of an EU ‘protected zone’ for this latter pest. As B. tabaci is an important vector of a large number of viruses, including tomato yellow leaf curl viruses (TYLCV), a monitoring programme of the tomato crop was implemented in Portugal. Preliminary data are presented concerning B. tabaci in the three main tomato‐growing regions of the country.     

<Emphasis Type="Italic">Bemisia tabaci</Emphasis> Biotype Q is present in Costa Rica

Whiteflies are an insect group that comprises multiple species and biotypes, capable of affecting crops by phloem feeding, virus transmission and promotion of fungal colonization. The distribution of these pests is worldwide. In Costa Rica, a country located in the tropics, the most problematic whiteflies are Bemisia tabaci biotype B and Trialeurodes vaporariorum. In September 2009, two greenhouses in the Alfaro Ruiz region, northwest of the country’s capital, San Jose, were surveyed as part of a larger effort to determine the occurrence of species and races of whiteflies in this agronomically important region. In addition, the insect samples were analyzed to determine the presence of Tomato chlorosis virus (ToCV), a yield-affecting crinivirus transmitted by whiteflies. The results revealed the presence of the Q biotype of B. tabaci, and important invasive species, as well as the expected T. vaporariorum. Viral detection assays identified potentially viruliferous individuals for Tomato chlorosis virus. These results identified a new pest capable of harbouring plant viruses has been identified, as well as a viral agent (ToCV) in a region where it was not reported, and which might cause significant yield losses.     

烟粉虱传播的番茄褪绿病毒和番茄黄化曲叶病毒对不同番茄品种的复合侵染

为明确烟粉虱传播的番茄褪绿病毒(Tomato chlorosis virus,ToCV)与番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)对不同番茄品种的复合侵染情况,于2015年11月在山东省寿光市温室内采集13个番茄品种共390份疑似发病植株叶片,对不同番茄品种的TYLCV抗性和2种病毒的复合侵染以及温室内发病番茄植株上烟粉虱成虫的带毒率进行检测。结果表明,采集的13个番茄品种经分子标记检测鉴定均为TYLCV杂合抗性;不同番茄品种ToCV与TYLCV的复合侵染率存在明显差异,大果番茄粉宴和贝瑞上复合侵染率最高可达73.3%,而樱桃番茄八喜上未检测到这2种病毒的复合侵染。此外,在发病番茄植株上采集的烟粉虱成虫体内可检测到2种病毒,其中烟粉虱ToCV带毒率为90.7%,TYLCV带毒率为80.0%,同时检测到ToCV与TYLCV的概率为71.3%。表明ToCV和TYLCV的复合侵染在山东省番茄生产中普遍发生,烟粉虱可同时携带这2种病毒并广泛传播。     

Potato, an experimental and natural host of the crinivirus Tomato chlorosis virus

Tomato chlorosis virus (ToCV, genus Crinivirus, family Closteroviridae) causes yellowing of tomatoes in many countries worldwide. Symptoms of ToCV infections in tomatoes include inter-veinal yellow chlorotic areas that develop first on lower leaves and then advance towards the upper part of the plant. ToCV is transmitted in nature by the whiteflies Bemisia tabaci, Trialeurodes vaporariorum, and Trialeurodes abutilonea in a semi-persistent manner. In the summer of 2006, a few potato (Solanum tuberosum) volunteer plants heavily infested with the whitefly B. tabaci were found growing within a pepper crop in the province of Málaga, southern Spain. Leaf samples from volunteer plants were tested for the presence of ToCV by molecular hybridization and RT-PCR, and were shown to be infected. Furthermore, potato plants were readily infected by ToCV after experimental transmission using B. tabaci biotype Q as vector. ToCV was also detected in the tubers from infected plants that subsequently produced infected plants. Potato also served as virus source for tomato infection via B. tabaci transmission.     

Tomato yellow leaf curl virus accumulates in vesicle-like structures in descending and ascending midgut epithelial cells of the vector whitefly, Bemisia tabaci, but not in those of nonvector whitefly Trialeurodes vaporariorum

Begomoviruses are transmitted by a single species of vector insect, the whitefly Bemisia tabaci, in a circulative manner. However, the mechanisms of this strict vector specificity have not been clarified. By immunoelectron microscopy, we showed that a begomovirus, Tomato yellow leaf curl virus (TYLCV), can enter midgut epithelial cells of the vector whitefly B. tabaci but not those of a nonvector whitefly, Trialeurodes vaporariorum, belonging to the same family. In midgut epithelial cells of viruliferous B. tabaci, the virus was localized in vesicle-like structures, suggesting endocytosis as an entry mechanism. These structures were also observed in midgut cells of nonviruliferous B. tabaci that had fed on healthy plants and in those of the nonvector T. vaporariorum that had fed on virus-infected plants. Vesicles containing TYLCV particles were observed most frequently in cells in the anterior part of the descending midgut, suggesting that this is the major entry site. These results clearly demonstrated that the virus-containing vector and nonvector whiteflies differ in the cellular localization of the virus and strongly suggest that a critical step in determining the vector insect specificity of begomoviruses is the entry of the viruses into midgut epithelial cells.     

Lack of synergistic effects in tomato plants co-infected with tomato severe rugose virus and tomato chlorosis virus

The begomovirus Tomato severe rugose virus (ToSRV) and the crinivirus Tomato chlorosis virus (ToCV), in single and co-infections, are very common in tomato crops in Brazil. Both viruses are transmitted by the whitefly Bemisia tabaciMEAM1 (biotype B). The objective of this study was to analyse the interaction between ToSRV and ToCV in tomato plants of cultivars Santa Clara and Kada. Plants at 15, 30 and 45 days after emergence were inoculated with 30 viruliferous B. tabaci per plant. The following treatments were compared: plants inoculated with ToSRV, ToCV, ToSRV + ToCV, and healthy (control). The interaction between these viruses was analysed by measuring the virus titre by qPCR and the fresh and dry weights of the aerial parts of the tomato plants. Based on two independent assays, no significant effects for co-infection of ToSRV and ToCV on virus titres and plant development were observed compared to single infections. The dry weight of tomato plants of both cultivars infected with ToSRV, ToCV, or co-infected did not differ significantly. However, the dry weight of Santa Clara tomato plants infected with ToSRV, ToCV and ToSRV + ToCV showed mean reductions of 21.5%, 25.5% and 32%, respectively, compared to healthy plants, and mean reductions for Kada were 31.7%, 37.5% and 38%, respectively.     

Note:Bemisia tabaci biotype B associated with tomato yellow leaf curl disease epidemics on rhodes Island, Greece

In 2006 an outbreak of tomato yellow leaf curl disease occurred in tomato crops on Rhodes Island, Greece. Diseased plants were found to be infested with the B biotype of theBemisia tabaci (Gennadius) complex and greenhouse and open-field-grown tomato crops were infected withTomato yellow leaf curl virus (TYLCV) introduced from the Middle East. This is the first report of TYLCV and the B biotype ofB. tabaci on Rhodes Island in Greece. http://www.phytoparasitica.org posting Dec. 11, 2007.     

Arabidopsis thaliana,an experimental host for tomato yellow leaf curl disease‐associated begomoviruses by agroinoculation and whitefly transmission

Tomato yellow leaf curl disease is one of the most devastating viral diseases affecting tomato crops worldwide. This disease is caused by several begomoviruses (genus Begomovirus, family Geminiviridae), such as Tomato yellow leaf curl virus (TYLCV), that are transmitted in nature by the whitefly vector Bemisia tabaci. An efficient control of this vector‐transmitted disease requires a thorough knowledge of the plant–virus–vector triple interaction. The possibility of using Arabidopsis thaliana as an experimental host would provide the opportunity to use a wide variety of genetic resources and tools to understand interactions that are not feasible in agronomically important hosts. In this study, it is demonstrated that isolates of two strains (Israel, IL and Mild, Mld) of TYLCV can replicate and systemically infect A. thaliana ecotype Columbia plants either by Agrobacterium tumefaciens‐mediated inoculation or through the natural vector Bemisia tabaci. The virus can also be acquired from A. thaliana‐infected plants by B. tabaci and transmitted to either A. thaliana or tomato plants. Therefore, A. thaliana is a suitable host for TYLCV–insect vector–plant host interaction studies. Interestingly, an isolate of the Spain (ES) strain of a related begomovirus, Tomato yellow leaf curl Sardinia virus (TYLCSV‐ES), is unable to infect this ecotype of A. thaliana efficiently. Using infectious chimeric viral clones between TYLCV‐Mld and TYLCSV‐ES, candidate viral factors involved in an efficient infection of A. thaliana were identified.     

Control of the Bemisia tabaci/Tomato yellow leaf curl virus complex on protected tomato crops in Algarve (Portugal)*

The whitefly Bemisia tabaci has been a serious pest in protected tomato crops since 1995 in the south of Portugal (Algarve), causing severe losses mainly resulting from Tomato yellow leaf curl virus (TYLCV), first reported in the autumn/winter season. In order to manage and control the B. tabaci/TYLCV complex, experimental field trials were carried out between 1997 and 2000. Several control methods were tested, such as the application of white screen nets to windows and doors, chemical treatments against B. tabaci and the use of cucumber as a trap crop. The results show that the percentage of plants with TYLCV symptoms mainly reflects B. tabaci infestation level in the first 6 weeks. Screen net protection was the control method that clearly provided a low incidence of plants with TYLCV symptoms in the autumn/winter season. Additionally, insecticide treatments, made weekly in the first 6 weeks, may give improved protection. In the winter/spring season, the B. tabaci/TYLCV complex is a minor problem because of the low populations of B. tabaci and the climatic conditions. Information about the work in progress and results was given to growers through visits to the experimental fields and oral presentations.     

繁育寄主植物对浅黄恩蚜小蜂生防潜能的影响

为明确以雪莲果Smallanthus sonchifolius为寄主植物繁育的浅黄恩蚜小蜂Encarsia sophia的生防潜能,测定雪莲果繁育的浅黄恩蚜小蜂个体大小以及其对烟粉虱Bemisia tabaci MED隐种和温室白粉虱Trialeurodes vaporariorum的致死能力,并解析其寄生2种粉虱若虫后的子代发育情况。结果表明,雪莲果繁育的浅黄恩蚜小蜂雌雄蜂体长、头宽及后足胫节长度均显著高于番茄繁育的浅黄恩蚜小蜂。雪莲果繁育的浅黄恩蚜小蜂对烟粉虱和温室白粉虱的平均致死数量分别为24.7头和25.0头,显著高于番茄繁育的21.4头和21.0头。相对于番茄,雪莲果繁育的浅黄恩蚜小蜂寄生烟粉虱和温室白粉虱若虫后其子代发育时间更短,平均分别为13.2 d和12.5 d;而且子代羽化率也显著高于番茄繁育的子代羽化率,分别为84.1%和86.9%。表明与番茄相比,雪莲果为寄主植物繁育的浅黄恩蚜小蜂对烟粉虱和温室白粉虱具有更强的生防潜能。     

Virus transmission studies and diagnostic detection of four begomovirus isolates in selected crop hosts and in Bemisia tabaci biotype B*

Virus transmission studies were conducted under glasshouse conditions using the vector Bemisia tabaci biotype B to determine how effectively isolates of the begomoviruses Tomato yellow leaf curl virus (TYLCV) and Tomato leaf curl Bangalore virus (ToLCBV) could be transmitted to phaseolus bean, capsicum and tomato test plants, the latter host used as a positive control for transmission. Diagnostic detection of viruses in these host crops and vector was also evaluated. Polymerase chain reaction (PCR) detection of TYLCV in bean cv. Wade and capsicum cv. Bellboy was achieved 4 weeks after fumigation in asymptomatic plants. Detection of TYLCV in tomato controls was achieved 2 weeks after fumigation with improved frequency of detection at 4 weeks. PCR was found to be a more sensitive method than triple‐antibody sandwich enzyme‐linked immunosorbent assay (TAS‐ELISA) for the detection of TYLCV isolates in all hosts. ToLCBV was detected by PCR and TAS‐ELISA in bean. TYLCV was also detected by PCR in the vector, with a novel internal positive control. This work was carried out to facilitate the development of a diagnostic protocol for the begomoviruses causing tomato yellow leaf curl under the EU SMT programme project –‘Diagnostic protocols for organisms harmful to plants’ (DIAGPRO).     

Methods of controlling Tomato yellow leaf curl virus (TYLCV) and its vector Bemisia tabaci in the Maltese Islands*

Bemisia tabaci and Tomato yellow leaf curl virus (TYLCV) were first observed in Malta in the early 1990s and caused serious damage to glasshouse and outdoor tomato crops. Chemical, physical and biological control methods have been developed, but the effective method has been the use of virus‐tolerant cultivars.     

Simultaneous detection of six RNA plant viruses affecting tomato crops using a single digoxigenin-labelled polyprobe

A polyprobe for the simultaneous detection by non-isotopic molecular hybridisation has been developed to detect any of the following six viruses causing important economic losses in tomato crops: Tomato spotted wilt virus, Tomato mosaic virus, Pepino mosaic virus, Cucumber mosaic virus, Potato Y virus and Parietaria mottle virus. The polyprobe detected all six viruses with similar sensitivity to that obtained using individual riboprobes. In addition, we evaluated the possible use of the tissue-printing as a sample preparation technique applied to routine diagnosis of tomato plants with the polyprobe. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

First report of the Q biotype ofBemisia tabaci in Japan by mitochondrial cytochrome oxidase I sequence analysis

The recent upsurgence ofBemisia tabaci (Genn.) as an important insect pest and vector ofTomato yellow leaf curl virus (TYLCV) is directly linked to serious damage to tomato crops grown throughout Japan. The molecular genetic identification and phylogenetic relationships of 12B. tabaci populations collected from representative locations in Japan were determined based on the mitochondrial cytochrome oxidase I (mtCOI) sequence. Phylogenetic analysis of the whitefly mtCOI sequence indicated that both the invasive B and Q biotypes now occur in Japan. The Q biotype was found at four locations: Mihara in Hiroshima, Nishigoshi in Kumamoto, Miyanojo and Okuchi in Kagoshima prefectures; the remaining eight collections were identified as the B biotype. This is the first report of the introduction of Q biotype in Japan. http://www.phytoparasitica.org posting July 21, 2006.     

Tomato chlorosis virus in pepper: prevalence in commercial crops in southeastern Spain and symptomatology under experimental conditions

Tomato chlorosis virus (ToCV), a member of the genus Crinivirus (family Closteroviridae), has been present in Spain since at least 1997, causing annual epidemics of yellowing in protected tomato crops. In 1999, sweet pepper plants exhibiting stunting and symptoms of interveinal yellowing and mild upward curling in the leaves, were found to be infected with ToCV in a greenhouse heavily infested with the whitefly Bemisia tabaci in the province of Almería, southeastern Spain. This study investigated the prevalence of ToCV in tomato and pepper crops in the major growing areas of southeastern Spain (Murcia, Almería and Málaga provinces) over a 3‐year period. In addition, an experimental system was developed for ToCV inoculation using B. tabaci as a vector, which allowed analysis of susceptibility of different pepper cultivars to the virus. The disease syndrome and yield losses induced by ToCV in pepper were also studied under experimental conditions, confirming severe yield reduction in infected plants.     

Bioassay of insecticides on mortality of <Emphasis Type="Italic">Bemisia tabaci</Emphasis> biotype B and transmission of <Emphasis Type="Italic">Tomato severe rugose virus</Emphasis> (ToSRV) on tomatoes

Tomato severe rugose virus (ToSRV) is a serious and prevalent begomovirus that causes severe mosaic and yield loss of tomato plants in Brazil. The virus is transmitted by Bemisia tabaci bitotype B (MEAM1) in a persistent circulative manner. This study evaluated the efficacy of cyantraniliprole foliar spray, cyantraniliprole root drenching, spiromesifen, thiamethoxam, and cartap on the mortality of Bemisia tabaci byotipe B and simulated primary and secondary transmission of ToSRV. None of the insecticides were effective in controlling primary transmission of ToSRV by B. tabaci. Cyantraniliprole root drenching, cartap, and cyantraniliprole foliar spray were effective in controlling secondary transmission of the virus, and infections were reduced by 94.5, 89.6, and 81%, respectively, compared to each of their controls. Thiamethoxam and spiromesifen did not provide effective control of secondary infection. Elimination of external sources of inoculum before starting new plantings and rational use of insecticides to reduce secondary infection may contribute to better disease management of tomato crops.     

日光温室番茄褪绿病毒病的发生规律及其与Q型烟粉虱种群动态关系

为有效控制日光温室番茄褪绿病毒病,于2014—2015年通过RT-PCR检测方法研究了济南市日光温室番茄褪绿病毒(Tomato chlorosis virus,ToCV)的发生规律、其与Q型烟粉虱Bemisia tabaci种群动态的关系及防虫网对该病毒病的防控效果。结果表明,春季日光温室番茄植株上Q型烟粉虱成虫数量呈增长趋势,5月下旬最高达到0.10头/叶,秋季日光温室番茄植株上Q型烟粉虱成虫数量9月上旬达最高7.42头/叶,后逐渐下降;日光温室Q型烟粉虱带毒率随着定植时间的延长而逐渐上升,之后维持相对稳定状态,即春季为20.00%~24.14%,秋季为30.00%~40.00%。日光温室ToCV发生与Q型烟粉虱成虫数量和带毒率密切相关,春季番茄最高发病率为12.00%;秋季番茄植株最高发病率为93.02%。番茄育苗和生长期用100目防虫网隔离可显著降低番茄植株带毒率。因此,秋季是日光温室ToCV防控关键期,覆盖防虫网阻隔烟粉虱可有效防治ToCV,推荐在日光温室使用。     

Comparative spatial dispersal ofTomato yellow leaf curl virus vectored by B and Q biotypes ofBemisia tabaci in Tomato glasshouses

The spatial dispersal patterns ofTomato yellow leaf curl virus (TYLCV) disease vectored by the B and Q biotypes of the whiteflyBemisia tabaci in tomato glasshouses were compared. Tomato plants were arranged in glasshouses and TYLCV-infected plants were placed in the center of each plot. Adult whiteflies of each biotype were released onto TYLCV-infected plants and the insects were then freely dispersed in the glasshouses under high or moderate temperature conditions. The abundance and spatial distribution of dispersed whiteflies did not differ between the B and Q biotypes in tomato glasshouses. The disease incidence and dispersion of TYLCV as a result of short-distance movement of the whiteflies were also similar between the two biotypes, although on several investigation dates there was a tendency for the disease incidence caused by the B biotype to be slightly greater than that caused by the Q biotype. These results demonstrated that the aspects of spatial spread of TYLCV vectored by the B and Q biotypes ofB. tabaci in tomato glasshouses are similar. http://www.phytoparasitica.org posting Dec. 11, 2007.     

Pyriproxyfen,a novel insect growth regulator for controlling whiteflies: Mechanisms and resistance management

Pyriproxyfen, a novel juvenile hormone mimic, is a potent suppressor of embryogenesis and adult formation of the sweetpotato whitefly, Bemisia tabaci (Gennadius), and the greenhouse whitefly, Trialeurodes vaporariorum (Westwood). Dipping of cotton or tomato seedlings infested with 0 to 1-day-old eggs in 0.1 mg litre^?1 resulted in over 90% suppression of egg hatch of both B. tabaci and T. vaporariorum. Older eggs were affected to a lesser extent. Exposure of whitefly females to cotton or tomato seedlings treated with pyriproxyfen resulted in oviposition of non-viable eggs. The LC90 values for egg viability of B. tabaci and T. vaporariorum exposed to treated plants were 0.05 and 0.2 mg litre^?1, respectively. Treatment of whitefly larvae with 0.04–5 mg litre^?1 resulted in normal development until the pupal stage; however, adult emergence was totally suppressed. Second instars of B. tabaci exposed to 5 mg litre^?1 pyriproxyfen, excreted honeydew at a level similar to the control level until the fourth instar (pupation), after which a strong reduction was observed. Inhibition of egg-hatch on the lower surface of cotton leaves was observed when their upper surface was treated with 1–25 mg litre^?1, indicating a pronounced translaminar effect. These findings indicate that pyriproxyfen is an efficient control agent of both B. tabaci and T. vaporariorum. The compound has been used successfully for controlling whiteflies in Israeli cotton fields since 1991. Adults of B. tabaci collected from a rose greenhouse and from adjacent cotton fields were monitored during 1991–1993 for their susceptibility to pyriproxyfen. A high level of resistance was recorded in whiteflies collected from a greenhouse after three successive applications of pyriproxyfen. Based on LC50 values, the resistance ratio for egg-hatch suppression was 554-fold and, for adult emergence failure, 10-fold. However, a single treatment of pyriproxyfen in cotton fields during the summer season (according to an insecticide resistance management (IRM) strategy) did not alter appreciably the susceptibility of B. tabaci to this compound. In order to prevent development of resistance, an attempt should be made to restrict its use to one treatment per crop season applied during the peak activity of the pest. Pyriproxyfen can be alternated with other novel compounds such as buprofezin and diafenthiuron for controlling whiteflies in cotton, vegetables and ornamentals as part of integrated pest management (IPM) and IRM strategies. In pyriproxyfen- or buprofezin-resistant strains of B. tabaci or T. vaporariorum, no appreciable cross-resistance was observed among pyriproxyfen, buprofezin and diafenthiuron.     

Host Range Studies for Tomato chlorosis virus, and Cucumber vein yellowing virus Transmitted by Bemisia tabaci (Gennadius)

The Bemisia tabaci (Gennadius) biotype B transmitted host range of Tomato chlorosis virus (ToCV), genus Crinivirus, Family Closteroviridae, and Cucumber vein yellowing virus (CVYV), genus Ipomovirus, Family Potyviridae, was studied. New experimental hosts were identified for each of these viruses. Seventeen species in eight plant families were assessed as potential hosts for ToCV. Infection in asymptomatic Anthriscus cereifolium (chervil) test plants by ToCV was confirmed by using a Real-Time PCR assay designed for ToCV. The presence of readily transmissible, infectious ToCV virions in A. cereifolium was confirmed by re-isolation of the virus via whitefly-transmission from A. cereifolium to Lycopersicon esculentum and A. cereifolium. This is the first report of the experimental transmission of ToCV by B. tabaci to a species within the Umbelliferae. All other hosts assessed for the presence of ToCV were found to be uninfected. Ten species in five families were assessed as potential hosts for CVYV. The CVYV host range identified included some important crops and common weeds, such as L. esculentum, Nicotiana tabacum, A. cereifolium, Datura stramonium, Nicotiana benthamiana, Nicotiana clevlandii and Cucumis sativus. Symptoms were present on D. stramonium, N. benthamiana and C. sativus control plants. The presence of infectious whitefly transmitted CVYV virions was confirmed solely for D. stramonium and N. tabacum, following re-isolation of the virus via B. tabaci transmission from all infected species to C. sativus. This is the␣first report of experimental CVYV transmission by B. tabaci to non-cucurbitaceous crop and weed hosts belonging to the Solanaceae or Umbelliferae.