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.     

Occurrence and Diagnosis of Tomato chlorosis virus in Portugal

Tomato chlorosis virus (ToCV), a new whitefly-transmitted and phloem-limited Crinivirus infecting tomatoes in Europe, is reported for the first time in Portugal. Tomato plants with symptoms of interveinal chlorosis, collected during autumn 1998 and summer and autumn 1999 in Algarve, southern Portugal, were positive in RT-PCR assays using ToCV-specific primers. The amplified 439bp fragment was sequenced and showed 99% homology with the ToCV sequence in the GenBank database. A digoxigenin–DNA probe was produced and tested in dot-blot with total RNAs extracted from tomato samples. Both the RT-PCR and dot-blot hybridisation procedures enabled rapid and reliable detection of ToCV from field samples.     

Viruses affecting tomato crops in Serbia

In a two-year survey (2011–2012), 3220 samples were collected and analyzed in order to determine the presence and distribution of viruses in tomato crops at 56 localities of 18 districts in Serbia. Out of 12 viruses tested, Cucumber mosaic virus (CMV), Potato virus Y (PVY), Alfalfa mosaic virus (AMV), Tomato spotted wilt virus (TSWV), Tomato mosaic virus (ToMV) and Tobacco mosaic virus (TMV) were detected in 42.1, 40, 11, 8.6, 2.3 and 1.3% of the total tested samples, respectively. The results revealed that CMV was prevalent in 2011 and PVY in 2012. CMV and PVY, apart from being predominant, were also the most widespread viruses. In general, single infections were the most frequent type of infection. Additionally, the most common mixed infections were double infections and the most prevalent combination was CMV and PVY. In 2011, the incidence of diseases and the percentage of all infection types were significantly higher than in 2012. Furthermore, in 2011, regardless of total single infections being prevalent compared to mixed infections, two prevailing viruses were commonly detected in mixed infections. The additional molecular testing of ELISA-negative samples using virus specific primers did not reveal the presence of Pepino mosaic virus (PepMV), Tomato yellow leaf curl virus (TYLC), Tomato infections chlorosis virus (TICV) and Tomato chlorosis virus (ToCV).     

<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.     

<Emphasis Type="Italic">Physalis ixocarpa</Emphasis> and <Emphasis Type="Italic">P. peruviana</Emphasis>, new natural hosts of <Emphasis Type="Italic">Tomato chlorosis virus</Emphasis>

Tomato chlorosis virus causes yellow leaf disorder epidemics in many countries worldwide. Plants of Physalis ixocarpa showing abnormal interveinal yellowing and plants of Physalis peruviana showing mild yellowing collected in the vicinity of tomato crops in Portugal were found naturally infected with ToCV. Physalis ixocarpa and P. peruviana were tested for susceptibility to ToCV by inoculation with Bemisia tabaci, Q biotype. Results confirmed that ToCV is readily transmissible to both species. The infection was expressed in P. ixocarpa by conspicuous interveinal yellow areas on leaves that developed into red or brown necrotic flecks, while P. peruviana test plants remained asymptomatic. Infected plants of both P. ixocarpa and P. peruviana served as ToCV sources for tomato infection via B. tabaci transmission. This is the first report of P. ixocarpa and P. peruviana as natural hosts of ToCV.     

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。     

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.     

Serological and molecular detection of Tomato chlorosis virus and Tomato infectious chlorosis virus in tomato

Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV) are two criniviruses inducing similar yellowing symptoms in tomato. An approximately 4 kb central region of the genomic RNA2 of French ToCV and TICV isolates was sequenced. TICV, for which no other sequences were available, appeared as a distant species in the genus, being close only to LIYV ( Lettuce infectious yellows virus ) for some, but not all, proteins. ToCV has more than 98% nucleotide identity with isolates from the US and Spain, and sequencing the CP gene of several isolates collected in different regions in southern France during 2 years suggested a unique origin. Polyclonal antisera were produced using capsid proteins of both viruses expressed in Escherichia coli . DAS-ELISA assays were developed for routine diagnosis and conditions for preparing samples for an optimized detection were determined. No cross-reactions were observed. However, some false-negative results, corresponding to samples giving ELISA readings close to the detection limit were regularly detected, particularly for ToCV (approximately 5% of the samples). A triplex RT-PCR assay was thus developed, which allowed detection of both viruses in a one-step protocol. An internal PCR control was included, which in addition showed that it could be used as a control for the entire RT-PCR procedure. Finally, combining DAS-ELISA in a first round, and triplex RT-PCR for doubtful samples, appeared the best way to achieve a reliable diagnosis of these viruses.     

Ultrastructural aspects of tomato leaves infected by Tomato torrado virus (ToTV) and co‐infected by other viruses

Optical and electron microscopy studies were carried out to investigate the cytopathology induced in tomato leaves infected by Tomato torrado virus (ToTV), a new picorna‐like virus associated with the ‘Torrado’ disease. Infected leaves, showing typical Torrado disease symptoms were surveyed in commercial greenhouses in the main tomato production areas of Spain. The effect of the co‐infection of ToTV with other viruses which commonly infect tomato crops was also studied. Ultra‐thin sections of ToTV‐infected tomato leaves did not show a strong cellular alteration. However, crystalline arrays of isometric virus‐like particles (VLPs) of 20–30 nm in the inclusion bodies were observed in phloem parenchyma cells of the infected tissues. Tissues co‐infected by ToTV and either Tomato chlorosis virus (ToCV) or Pepino mosaic virus (PepMV) presented more severe cellular alterations. The most deleterious consequences for tomato cells were found in triple infections of ToTV, PepMV and Tomato spotted wilt virus (TSWV), where characteristic cell wall overgrowth was distinguishable, together with a large amount of necrotic cells.     

北京地区茄子感染番茄褪绿病毒的分子鉴定

为明确北京地区茄子感染番茄褪绿病毒(Tomato chlorosis virus,To CV)的情况,于2015年3—6月收集了10份疑似感染To CV的设施茄子样品,通过PCR分子鉴定法进行检测,并进一步对扩增阳性样品To CV的CP基因采用邻接法进行了系统进化分析。结果表明,10个检测样品中有4个扩增得到约463 bp的特异条带,经测序与Gen Bank中To CV序列相似性达到99.0%以上,表明这4个样品均携带有该病毒,检出率为40%;To CV茄子分离物的CP基因序列大小为774 bp,Gen Bank登录号为KT751008,与To CV北京番茄分离物(KC311375)同源性最高,达到99.9%,确认该序列为To CV片段;系统进化树显示,To CV茄子分离物与To CV日本分离物(AB513443)处于同一分支,具有密切的亲缘关系,表明不同国家To CV分离物之间的亲缘关系与地理距离具有一定的相关性。     

A DNA probe mix for the multiplex detection of ten artichoke viruses

Artichoke Italian latent virus (AILV), Artichoke latent virus (ArLV), Artichoke mottled crinkle virus (AMCV), Bean yellow mosaic virus (BYMV), Cucumber mosaic virus (CMV), Pelargonium zonate spot virus (PZSV), Tomato infectious chlorosis virus (TICV), Tobacco mosaic virus (TMV), Tomato spotted wilt virus (TSWV) and Turnip mosaic virus (TuMV) are damaging to artichoke. We have developed a protocol enabling the simultaneous detection of these artichoke viruses by non-isotopic dot blot hybridisation with DNA probes. The probe mix detected all viruses with high specificity and identical to that obtained using individual probes. The approach is proposed for the routine assessment of phytosanitary status for certified nursery production of globe artichoke.     

Tomato chlorosis virus: a new whitefly-transmitted, Phloem-limited, bipartite closterovirus of tomato

ABSTRACT Tomato chlorosis virus (ToCV) is the second whitefly-transmitted, phloem-limited, bipartite closterovirus described infecting tomato. ToCV is distinct from tomato infectious chlorosis virus (TICV), based on lack of serological and nucleic acid cross-reactions and differences in vector specificity. TICV is transmitted only by the greenhouse whitefly (Trialeurodes vaporariorum), whereas ToCV is transmitted by the greenhouse whitefly, the banded-wing whitefly (T. abutilonea), and Bemisia tabaci biotypes A and B (B. argentifolii). Double-stranded (ds) RNA analyses of ToCV show two prominent dsRNAs of approximately 7,800 and 8,200 bp, with several small dsRNAs. Digoxigenin-11-UTP-labeled riboprobes derived from cDNA clones representing portions of RNAs 1 and 2 were used in Northern blot hybridizations to detect two large nonhomologous dsRNAs and a subset of smaller dsRNAs. These probes were used in dot blot hybridizations to detect ToCV in infected tomato. Inclusion bodies and cytoplasmic vesicles were consistently observed in phloem tissues of ToCV-infected Nicotiana clevelandii. Computer-assisted sequence analysis showed significant homology between ToCV clones that hybridize specifically with RNAs 1 and 2 and the lettuce infectious yellows virus methyltransferase of RNA 1 and the HSP70 heat shock protein homolog of RNA 2, respectively. Thus, ToCV is another member of the growing subgroup of bipartite closteroviruses transmitted by whiteflies.     

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.     

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。     

Occurrence of Tomato infectious chlorosis virus (TICV) in Jordan

A new disease on tomato ( Lycopersicon esculentum L.) caused by Tomato infectious chlorosis virus (TICV) has been detected for the first time in Jordan. Disease symptoms consisted of interveinal yellowing areas in older leaves followed by generalized yellowing. Using specific primers, Tomato infectious chlorosis virus was detected in symptomatic plants by RT-PCR. The amplified fragment (416 bp) was cloned and sequenced. Results of sequence analysis showed that the Jordanian isolate of TICV shared high nucleotide similarity (> 98%) with two other isolates from Japan and France. The distribution of TICV has been investigated in four regions in the Jordan Valley by non-radioactive dot blot hybridization. Data from the study showed high incidence of the disease in all surveyed regions. In addition, the expected size of the coat protein gene of TICV could be amplified from two symptomatic weeds species, Chenopodium album and Chenopodium murale , indicating that these weeds are natural hosts for the virus.     

Screening tomato genotypes for resistance and tolerance to Tomato chlorosis virus

Tomato chlorosis virus (ToCV) is an emerging crinivirus in Brazil that causes an economically important disease in tomato (Solanum lycopersicum) and other solanaceous species. ToCV is transmitted predominantly by the whitefly Bemisia tabaci Middle East‐Asia Minor 1 (MEAM1, formerly biotype B), in a semipersistent manner. As all cultivated tomato varieties and hybrids are susceptible to this crinivirus, the main alternatives for the control of the disease are the use of healthy seedlings for transplanting and the chemical control of the insect vector. The objective of this work was to evaluate the responses of tomato genotypes to infection with this crinivirus and their tolerance to the disease in order to support the development of other alternatives for disease control. Resistance to infection was evaluated by ToCV inoculation with viruliferous B. tabaciMEAM1 followed by virus detection by RT‐PCR and RT‐qPCR. To measure tolerance to the disease, plant development and fruit yield of ToCV‐infected and healthy plants were compared. Among 56 genotypes, only the lineage IAC‐CN‐RT (S. lycopersicum ‘Angela Gigante’ × S. peruvianum ‘LA 444‐1’) was highly resistant to infection with ToCV. Tolerance to the disease over two trials with different genotypes showed variable results. The effect of ToCV on plant development varied from 2.9% to 71.9% reduction, while yield loss varied from 0.2% to 51.8%. The highly ToCV‐resistant lineage IAC‐CN‐RT, which is also resistant to a Spanish isolate of ToCV, might be useful for tomato breeding programmes.     

Situation of the whiteflies Bemisia tabaci and Trialeurodes vaporariorum in protected tomato crops in Algarve (Portugal)*

Protected tomato is the most important horticultural crop in the Algarve (south of Portugal). However, the growing area has decreased by almost 48% since 1995, mainly as a result of the pests Bemisia tabaci and Trialeurodes vaporariorum and the epidemics of Tomato yellow leaf curl virus (TYLCV), a Begomovirus transmitted by B. tabaci. Both whiteflies are vectors of economically important viruses. Recently, Tomato chlorosis virus (ToCV), a member of the genus Crinivirus, transmitted by both B. tabaci and T. vaporariorum, was reported infecting tomato crops in Algarve. A study was carried out to evaluate the dynamics of whitefly populations on tomato crops in Algarve. Population counts of B. tabaci were high in the first months of autumn, then decreased until January, when numbers of T. vaporariorum became higher. Counts of B. tabaci then increased again.