Control of the Bemisia tabaci/Tomato yellow leaf curl virus complex in tomato nurseries in Portugal*

According to EU and Portuguese law, inspection of horticultural nurseries is an important task for the Phytosanitary Inspection Services of the NPPO, which requires the introduction of some rules: official registration of producers and production of plant material of EC quality. Mainly as a result of the introduction of Tomato yellow leaf curl virus (TYLCV) in 1995, tight control of registered nurseries was implemented in Algarve in order to ensure the production of young horticultural plants of good varietal and phytosanitary quality. A survey system was established in order to contain pests by trying to eradicate them in nurseries. Nurseries were obliged to adopt new technologies, including for example the compulsory use of screen nets, the improvement of the existing glasshouses and the implementation of strict rules for access of people to these places. The Administration (MADRP/DRAALG) has provided financial support in order to have new nursery glasshouses (without increasing the area) suitable for the control of Bemisia tabaci and other pests which may be disease vectors. It should be stressed that about 50% of nursery production in Algarve is used in other regions of Portugal, considered as EU Protected Zones for B. tabaci.     

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.     

Effect of Host Plant Resistance to Tomato yellow leaf curl virus (TYLCV) on Virus Acquisition and Transmission by Its Whitefly Vector

ABSTRACT The effect that Tomato yellow leaf curl virus (TYLCV)-infected resistant tomato plants may have on virus epidemiology was studied. Four tomato genotypes that exhibit different levels of viral resistance, ranging from fully susceptible to highly resistant, served as TYLCV-infected source plants. Viral acquisition and transmission rates by white-flies following feeding on the different source plants were evaluated. TYLCV transmission rate by whiteflies that had fed on infected source plants 21 days postinoculation (DPI), shortly after the appearance of TYLCV symptoms, was negatively correlated with the level of resistance displayed by the source plant. Therefore, the higher the resistance, the lower the transmission rate. In addition, TYLCV DNA accumulation was shown to be lower in the resistant source plants compared with the susceptible plants. Whitefly survival rate, following feeding on source plants 21 DPI, was similar for all the cultivars tested. Significant differences in whitefly survival were found, however, following feeding on the infected source plants at 35 DPI; here, whitefly survival rate increased with higher levels of resistance displayed by the source plant. At 35 DPI, the susceptible plants had developed severe TYLCV disease symptoms, and transmission rates from these plants were the lowest, presumably due to the poor condition of these plants. Transmission rates from source plants displaying a medium level of resistance level were highest, with rates declining following feeding on source plants displaying higher levels of TYLCV resistance. TYLCV DNA accumulation in whiteflies following feeding on infected source plants at both 21 and 35 DPI was directly correlated with viral DNA accumulation in source plants. Results show that, in essence, the higher the resistance expressed, the less suitable the plant was as a viral source. Consequently, following acquisition from a highly resistant plant, TYLCV transmission by whiteflies will be less efficient.     

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.     

Whitefly resistance traits derived from the wild tomato Solanum pimpinellifolium affect the preference and feeding behavior of Bemisia tabaci and reduce the spread of Tomato yellow leaf curl virus

Breeding of tomato genotypes that limit whitefly (Bemisia tabaci) access and feeding might reduce the spread of Tomato yellow leaf curl virus (TYLCV), a begomovirus (genus Begomovirus, family Geminiviridae) that is the causal agent of tomato yellow leaf curl disease. TYLCV is restricted to the phloem and is transmitted in a persistent manner by B. tabaci. The tomato breeding line ABL 14-8 was developed by introgressing type IV leaf glandular trichomes and secretion of acylsucroses from the wild tomato Solanum pimpinellifolium accession TO-937 into the genetic background of the whitefly- and virus-susceptible tomato cultivar Moneymaker. Results of preference bioassays with ABL 14-8 versus Moneymaker indicated that presence of type IV glandular trichomes and the production of acylsucrose deterred the landing and settling of B. tabaci on ABL 14-8. Moreover, electrical penetration graph studies indicated that B. tabaci adults spent more time in nonprobing activities and showed a reduced ability to start probing. Such behavior resulted in a reduced ability to reach the phloem. The superficial type of resistance observed in ABL 14-8 against B. tabaci probing significantly reduced primary and secondary spread of TYLCV.     

Experimental transmission of the mild strain of Tomato yellow leaf curl virus (TYLCV) to Amaranthus dubius by Bemisia tabaci

Tomato yellow leaf curl virus (TYLCV) is a monopartite begomovirus from the Old World. The mild strain of this virus (TYLCV-Mld) was described for South America in Venezuela in 2007. Due to the potential risks of establishment of this virus in the field, six common weeds were evaluated for susceptibility to an isolate of TYLCV-Mld by using adults of Bemisia tabaci (Hemiptera: Aleyrodidae) to mediate viral transmission. In this work, detection based on PCR amplification with TYLCV specific primers showed Amaranthus dubius (Amaranthaceae) as the only infected weed. In A. dubius, viral symptoms were observed from 11.0?±?1.3?days post-inoculation and the transmission rate of TYLCV-Mld to this plant species was 83.3%. The successful back-transmission of TYLCV-Mld from A. dubius to tomato plants was demonstrated.     

Characterisation of a Quantitative Resistance to Vector Transmission of Tomato yellow leaf curl virus in Lycopersicon pimpinellifolium

Two wild genotypes from the same species Lycopersicon pimpinellifolium, WVA106 (susceptible) and INRA-Hirsute (so-called ‘resistant’), were compared with respect to their reaction to Tomato yellow leaf curl virus isolate Réunion (TYLCV-Mld[RE]), using both whitefly-mediated inoculation and graft inoculation. Disease incidence and symptom severity were scored. Presence and quantification of viral DNA were assessed by dot blot hybridisation. Upon insect inoculation, accession INRA-Hirsute showed a moderate resistance against TYLCV that was overcome by a high inoculation pressure obtained by increasing the cumulative number of inoculative whiteflies. Temporal analyses of the disease progress in relation to this criterion exhibited that the protection was quantitative, mainly reducing the TYLCV-Mld[RE] incidence by at maximum 50% at low inoculation pressure. When graft inoculated, the final TYLCV-Mld[RE] disease incidence was 100% in both susceptible and resistant genotypes with severe symptoms, suggesting a reduction of virus transmission by a vector resistance as a possible mechanism. Implications of using such type of resistance in breeding programmes are discussed.     

Tomato yellow leaf curl Sardinia virus and its vector Bemisia tabaci in Sicilia (Italy): present status and control possibilities*

Tomato yellow leaf curl Sardinia begomovirus (TYLCSV) appeared in Sicilia (IT) in 1988, creating great threats to agriculture and causing huge losses, especially in south‐eastern areas of the island, where protected tomato cultivation is widespread. Towards the mid‐1990s, a reduction occurred in the virus epidemics, probably due to new approaches which have been applied to rational control of its vector, the whitefly Bemisia tabaci. More recently, TYLCSV has increased again, creating great concern among local tomato producers and stimulating new research. Besides studies on natural enemies of the vector, aiming to investigate their role and distribution, the main current research lines in Sicilia concern the possibility of reducing both whitefly activity, using photoselective plastics as covers, and virus damage, by growing tolerant tomato genotypes.     

Detection of Tomato yellow leaf curl Sardinia virus in Tunisia

Surveys were undertaken for tomato yellow leaf curl viruses in the main Tunisian tomato-growing areas in fields and plastic houses. Symptoms included yellowing, leaf curling and stunting. Collected samples were submitted to molecular analysis using two approaches: (1) hybridization tests with two DNA probes corresponding to an intergenic region derived from a cloned dimer of an Egyptian full-length TYLCV and to the coat region of a Tunisian TYLCV isolate; (2) PCR amplification coupled to RFLP allowing both identification and clustering of Tunisian isolates. Only Tomato yellow leaf curl Sardinia virus was detected in Tunisia.     

河北省番茄黄化曲叶病毒病的分子鉴定初报

双生病毒是一类具有孪生颗粒形态的植物单链病毒,目前双生病毒病害已在多个国家和地区的作物上造成严重危害。近年来,我国多省报道作物上有这类病毒的发生,且有逐年加重和扩散的趋势。根据危害番茄的双生病毒DNA A序列保守区设计简并引物,对2009年4月采自河北省魏县表现叶片黄化、曲叶症状的4个番茄样品进行PCR检测,均为双生病毒阳性,对样品的扩增片断进行了克隆测序,经序列比对分析,与番茄黄化曲叶病毒（Tomato leaf curl virus,TYLCV）山东分离物（FJ646611.1）序列相似性为99.25%~99.55%,说明河北番茄黄化曲叶病由TYLCV引起。     

Current situation of Tomato yellow leaf curl virus in Portugal

The name Tomato yellow leaf curl virus (TYLCV) has been applied to a group of virus species of the genus Begomovirus in the family Geminiviridae that cause a similar tomato disease worldwide. In 1995, TYLCV was first reported in Algarve (southern Portugal) as responsible for an epidemic outbreak of a severe tomato disease. Molecular data have shown that this Portuguese TYLCV isolate was distinct from those previously reported in Europe, as it belonged to the TYLCV-Israel species ¹ . Since then, TYLCV epidemics have occurred annually, being a limiting factor mainly for autumn/winter glasshouse tomato crops. In 1998, TYLCV was also found associated with the emergence of a novel disease of Phaseolus vulgaris in Algarve. The affected bean plants were severely stunted and gave no marketable yield. However, the disease occurs only sporadically, even in conditions of high TYLCV infection pressure. Recently, Tomato chlorosis virus (ToCV), a whitefly-transmitted bipartite closterovirus (genus Crinivirus , family Closteroviridae ), was found associated with an unusual tomato yellow leaf syndrome, in single or mixed infection with TYLCV. The impact of this new pathosystem on tomato production has yet to be determined. Surveys are in progress in mixed cropping systems infested with whiteflies. So far, TYLCV and ToCV diseases are limited to the Algarve region.     

侵染广西番茄的番茄黄化曲叶病毒的分子鉴定

番茄黄化曲叶病毒病是番茄生产上的毁灭性病害,严重影响番茄产量及品质。2019年从广西百色市采集疑似番茄黄化曲叶病叶片样品,采用滚环扩增(RCA)及基因克隆等方法,获得了4个烟粉虱传双生病毒的全基因序列,4个分离物的全长序列分别为2 776、2 781、2 752、2 781 bp,均编码6个开放阅读框;核苷酸相似性比较发现,4个分离物彼此间的相似性均在90%以上,与已报道的番茄黄化曲叶病毒Tomato yellow leaf curl virus (TYLCV)各分离物间的相似性也在91%以上;系统进化分析表明,TYLCV广西分离物BS-2和BS-4与TYLCV-Hunan、BS-1和BS-3与TYLCV-YN6553等分离物处于独立的小分支,说明TYLCV广西分离物与TYLCV-Hunan、TYLCV-YN6553具有较近的亲缘关系。本研究首次报道TYLCV在广西发生。     

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.     

Tomato yellow leaf curl virus: host range and virus-vector relationships

The Jordanian isolate of tomato yellow leaf curl virus has a narrow host range restricted to a few solanaceous plants. Severe symptoms developed on tomatoes and Datura stramonium , whereas Nicotiana glutinosa and N. tabacum cvs Samsun and Havana 423 were infected without showing symptoms. The whitefly Bemisa tabaci is an efficient vector; a single whitefly was able to transmit the virus. The minimum acquisition and inoculation feeding periods were 60 and 30 min, respectively, and the latent period was 20–24 h. The virus was retained by Bemisia tabaci for 11 days. The host range and virus vector relationships of the Jordanian isolate are reported for the first time.     

北京地区番茄黄化曲叶病毒病的鉴定及防治对策

番茄黄化曲叶病毒病是一种由烟粉虱传播的病毒病,给番茄生产造成严重威胁。2009年在北京郊区调查时发现部分保护地种植的番茄植株表现典型黄化曲叶症状。通过提取典型症状样品总DNA利用粉虱传双生病毒检测简并引物PA/PB,进行PCR扩增到541bp的特异条带。通过测序和核苷酸序列比对表明该序列与番茄黄化曲叶病毒序列相似性最高为99%。分子检测结果表明北京郊区部分保护地种植的番茄已被烟粉虱传播的番茄黄化曲叶病毒侵染危害。     

A simple, efficient agroinoculation soaking procedure for Tomato yellow leaf curl virus

Tomato yellow leaf curl virus (TYLCV, genus Begomovirus, family Geminiviridae), poses a serious threat to tomato crops in tropical and subtropical regions. We developed a simple agroinoculation method with an infectious clone of TYLCV. Dipping of excised sections of susceptible tomato shoots in an Agrobacterium suspension successfully introduced the replicating virus with high efficiency. An additional vacuum treatment for 5 min ensured uniform infection without escapes, allowing evaluation of differences in TYLCV resistance among tomato cultivars. The method can be used in laboratory experiments for virological studies and in breeding programs for resistant cultivars.     

番茄褪绿病毒与番茄黄化曲叶病毒复合侵染对番茄褪绿病毒传播的影响

番茄褪绿病毒Tomato chlorosis virus(ToCV)是一种由烟粉虱Bemisia tabaci传播的正义单链RNA病毒,在田间常与番茄黄化曲叶病毒Tomato yellow leaf curl virus(TYLCV)复合侵染而造成番茄生产上重大的经济损失。为了明确ToCV与TYLCV的复合侵染对烟粉虱传播ToCV所造成的影响,本文采用RT-PCR以及qRT-PCR检测了复合侵染的番茄对烟粉虱获取和传播ToCV的影响。研究表明,烟粉虱取食复合侵染的番茄后对ToCV的传播效率显著提高,仅25头烟粉虱的传毒率即可达到100%,ToCV在烟粉虱以及番茄体内的累积量均显著提高。说明这种复合侵染促进了烟粉虱对ToCV的传播,在田间应当及时防控烟粉虱,警惕病毒与烟粉虱的蔓延。