Tomato Yellow Leaf Curl Virus Infection in a Monocotyledonous Weed (Eleusine indica)

Tomato yellow leaf curl virus (TYLCV) is one of the most important plant viruses belonging to the genus Begomovirus of the family Geminiviridae. To identify natural weed hosts that could act as reservoirs of TYLCV, 100 samples were collected at a TYLCV-affected tomato farm in Iksan from 2013 to 2014. The sample weeds were identified as belonging to 40 species from 18 families. TYLCV was detected in 57 samples belonging to 28 species through polymerase chain reaction using root samples including five species (Eleusine indica, Digitaria ciliaris, Echinochloa crus-galli, Panicum dichotomiflorum, and Setaria faberi) from the family Poaceae. Whitefly Bemisia tabaci-mediated TYLCV transmission from TYLCV-infected E. indica plants to healthy tomatoes was confirmed, and inoculated tomatoes showed typical symptoms, such as leaf curling and yellowing. In addition, TYLCV was detected in leaf and root samples of E. indica plants inoculated by both whitefly-mediated transmission using TYLCV-viruliferous whitefly and agro-inoculation using a TYLCV infectious clone. The majority of mastreviruses infect monocotyledonous plants, but there have also been reports of mastreviruses that can infect dicotyledonous plants, such as the chickpea chlorotic dwarf virus. No exception was reported among begomoviruses known as infecting dicots only. This is the first report of TYLCV as a member of the genus Begomovirus infecting monocotyledonous plants.     

番茄抗黄化曲叶病毒育种研究进展

本文分别对近年番茄抗黄化曲叶病毒的传统育种、分子辅助育种、基因工程育种进展进行了综述.栽培番茄均不抗番茄黄化曲叶病毒,所以传统育种采用从野生近缘种中筛选抗性材料,以其为亲本与栽培番茄进行杂交来获得抗性;野生近缘种中的抗性位点Ty-1、Ty-2和Ty-3及一些QTLs先后被定位,也筛选出了可鉴定Ty-1基因的SSR-47标记及鉴定Ty-3的SCAR标记;通过转基因技术获得抗性是研究热点之一,目前转入番茄后表现出抗性的序列有TYLCV病毒的CP基因、尺EP基因的部分序列或反义序列、不编码的保守序列以及源于白粉虱的GroEL基因.同时讨论了今后的主要发展方向.     

Within-Host Dynamics of the Emergence of Tomato Yellow Leaf Curl Virus Recombinants

Tomato yellow leaf curl virus (TYLCV) is a highly damaging begomovirus native to the Middle East. TYLCV has recently spread worldwide, recombining with other begomoviruses. Recent analysis of mixed infections between TYLCV and Tomato leaf curl Comoros begomovirus (ToLCKMV) has shown that, although natural selection preserves certain co-evolved intra-genomic interactions, numerous and diverse recombinants are produced at 120 days post-inoculation (dpi), and recombinant populations from different tomato plants are very divergent. Here, we investigate the population dynamics that lead to such patterns in tomato plants co-infected with TYLCV and ToLCKMV either by agro-inoculation or using the natural whitefly vector Bemisia tabaci. We monitored the frequency of parental and recombinant genotypes independently in 35 plants between 18 and 330 dpi and identified 177 recombinants isolated at different times. Recombinants were detected from 18 dpi and their frequency increased over time to reach about 50% at 150 dpi regardless of the inoculation method. The distribution of breakpoints detected on 96 fully sequenced recombinants was consistent with a continuous generation of new recombinants as well as random and deterministic effects in their maintenance. A severe population bottleneck of around 10 genomes was estimated during early systemic infection–a phenomenon that could account partially for the heterogeneity in recombinant patterns observed among plants. The detection of the same recombinant genome in six of the thirteen plants analysed beyond 30 dpi supported the influence of selection on observed recombination patterns. Moreover, a highly virulent recombinant genotype dominating virus populations within one plant has, apparently, the potential to be maintained in the natural population according to its infectivity, within-host accumulation, and transmission efficiency - all of which were similar or intermediate to those of the parent genotypes. Our results anticipate the outcomes of natural encounters between TYLCV and ToLCKMV.     

上海地区番茄黄化曲叶病毒病的鉴定及嫁接接种法研究

番茄黄化曲叶病毒(tomato yellow leafcurl virus,TYLCV)是一种由烟粉虱(Bemisia tabaci)和嫁接传播的双生病毒,在热带、亚热带地区给番茄生产造成严重威胁.根据番茄黄化曲叶病毒的保守序列设计一对引物,运用PCR技术从上海地区的感病番茄中扩增出一条575bp的特异带,而健康植株无此带.测序表明该序列与番茄黄化曲叶病毒具有极高的同源性(97%～99%).将健康接穗嫁接到感染番茄黄化曲叶病毒的番茄砧木上,间隔15 d和30 d,分别提取接穗的DNA,并用PCR法检测病毒,发现嫁接15 d后在部分接穗中检测到TYLCV病毒,嫁接30 d后在所有的接穗中均检测到病毒,因此,嫁接法可以作为番茄黄化曲叶病毒病的接种鉴定方法.     

番茄黄化曲叶病毒对番茄叶片光合特性和叶绿体超微结构的影响

以番茄为试验材料,研究番茄黄化曲叶病毒(TYLCV)侵染对植株叶片叶绿素含量、净光合速率、气孔导度、胞间CO2浓度和叶绿体超微结构的影响.结果表明:TYLCV侵染番茄后,叶片叶绿素a、b以及总叶绿素含量分别下降50.2%、24.19%和43.84%,叶片净光合速率和气孔导度分别下降43.28%、27.07%,胞间CO2浓度增加13.04%.与健康叶片相比,叶绿体变形,叶绿体基质片层大部分消解,基粒结构消失,叶绿体外膜和内膜剥离,质壁分离和细胞膜内陷,细胞器消解.研究表明,TYLCV侵染破坏了番茄叶片的叶绿体结构,严重影响番茄叶片的光合作用.     

Reservoir Weed Hosts of Tomato Yellow Leaf Curl Begomovirus from Tanzania

A survey was initiated to detect tomato yellow leaf curl virus (TYLCV) and identify its reservoir weed hosts in six regions (Arusha, Morogoro, Dodoma, Iringa, Kilimanjaro and Dar es Salaam) in Tanzania. Three farms were randomly selected in each region. Assessment of TYLCV incidence was done by relating the number of infected tomato plants to the total number of plants assessed along a diagonal in five quadrants measuring 4m ‐ 4m in size (one at each corner of the farm and one at the centre). Disease severity was scored on a scale of 0 to 4 (where 0 = no symptoms and 4 = very severe symptoms). Within and outside each farm, weeds showing TYLCV-like symptoms were collected and either squash-blotted, dot-blotted or both on nylon membranes. The membranes were hybridized with DIG-labelled probe synthesized for the detection of TYLCV from Sardinia (TYLCV-Sar) following standard protocols. Selected plant species were experimentally inoculated with screenhouse cultures of TYLCV representative isolates from the six regions using Bemisia tabaci to determine their host status. Results indicated that TYLCV incidence and severity were significantly higher (P = 0.05) in Dodoma region than the rest of the regions. In Iringa region, the incidence and severity of TYLCV were the lowest of all regions. TYLCV was detected in 12 of the 17 dot-blotted samples and in all the 21 squashed samples using the non-radioactively labelled riboprobes. Similarly, five plant species (Capsicum annuum, Datura stramonium, Lycopersicon esculentum, Nicotiana glutionsa and N. tabacum) tested in the screenhouse were infected by the six TYLCV isolates used. It is recommended that weeds within and outside tomato farms be removed to eliminate or reduce sources of virus inoculum. The dot and squash blot techniques are convenient for field detection of the virus, and are especially useful for the detection of early and latent infections so that management strategies can be initiated and implemented.     

Tomato Yellow Leaf Curl Virus DNA Forms in the Viral Capside, in Infected Plants and in the Insect Vector

Tomato yellow leaf curl virus (TYLCV) DNA was used as a probe to identify and analyze virus-related DNAs in the viral capside, in infected tomato plants and in the virus vector, the whitefly. In addition to the single-stranded viral genomic DNA, double-stranded virus-related DNA molecules were detected in infected plants. Not all of the virus-related DNA forms are present simultaneously in the infected plant. The double-stranded molecules, which are probably the replicative form of the viral genome, have been purified from an infected tomato plant. In the viruliferous whitefly, only the single-stranded unit-size viral genome was detected.     

Tomato Yellow Leaf Curl Virus Benefits Population Growth of the Q Biotype of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae)

Plant viruses can directly influence their insect vectors, and indirectly through their shared host plant, altering their behavior and performance in a mutualistic or rather antagonistic manner. One of the most studied begomovirus, Tomato yellow leaf curl virus (TYLCV), may also facilitate the expansion of its vector, the whitefly Bemisia tabaci (Gennadius). Considering the likely expansion of the disease and its major vector, we studied the direct and the indirect effects of a Mediterranean isolate of this virus (TYLCV-IL) on the biological performance of the Q biotype of B. tabaci. The following parameters were examined: development time and viability of nymphs, sex ratio, fecundity, and fertility and longevity. The results varied from positive to neutral depending on the parameter and the effect studied. TYLCV accelerated nymphal developmental and increased male longevity of B. tabaci when viruliferous insects developed on TYLCV-immune eggplants (direct effects). An indirect, positive effect of TYLCV-infected plants was observed on fecundity of B. tabaci, which laid more eggs on virus-infected than on noninfected tomato plants. Our results show that TYLCV enhances the population increase of its whitefly vector and that there is a high risk of rapid expansion of both the virus and its vector—the MED species of B. tabaci—into new areas when both agents interact together.     

Molecular Variability in the Replicase Gene of Viruses Causing Tomato Leaf Curl Disease in India

Tomato Leaf Curl Virus (ToLCV)-infected samples collected from four different ecological zones of India: Northern zone, NewDelhi (IARI) [ToLCNDV-IARI]; Central zone, Jabalpur [ToLCV-Jb], Raipur [ToLCV-Rp] and Southern zone, Dharwad [ToLCV-Dh] were maintained by whitefly transmission into tomato cv Pusa Early Dwarf. Replicase gene coding for replication initiator protein (Rep) was amplified by PCR using gene specific primers, yielding an amplicon of 1086 bp for all the isolates. Sequence analysis of the rep amplicons of all the Indian isolates of ToLCV causing the disease clearly shows the presence oftwo subgroups irrespective of their geographical evolution. Isolates belonging to subgroup I comprising of tomato leaf curl virus-New Delhi (ToLCV-NDe) having bipartite genome are conserved within themselves and showed 94–95% nucleotide sequence homology. Second subgroup comprising of tomato leaf curl virus-Bangalore (ToLCBV) having monopartite genome and conserved among themselves, but showed only 73-75% homology with the subgroup I. In phylogenetic tree analysis, ToLCV-NDe appeared in a different cluster than ToLCV-Ban. All the four isolates used in this study fall in ToLCV-NDe group based on their replicase gene sequences. The specificity determinant located at N terminal suggests ToLCV-NDe and ToLCV-Rp as mild isolates.     

Infectivity of Tomato Leaf Curl Hainan Virus in Tomato and Tobacco in China

Tomato leaf curl Hainan virus (ToLCHnV) was previously reported as a distinct begomovirus infecting tomato in Hainan, China. To investigate the infectivity of ToLCHnV, an infectious clone of ToLCHnV‐[CN: HaNHK7] was constructed and agro‐inoculated into Solanum lycopersicum, Nicotiana benthamiana, Nicotiana glutinosa, Petunia hybrida, Cucumis sativus, Solanum melongena and Capsicum annuum plants; it induced severe leaf curling and crinkling symptoms in these plant species except C. sativus, S. melongena and C. annuum. The induced symptoms were compared with those induced by Papaya leaf curl China virus.     

Papaya Leaf Curl Guangdong Virus and Ageratum Yellow Vein Virus Associated with Leaf Curl Disease of Tobacco in China

Two samples (YC7, YC27) of Nicotiana tabacum showing leaf curling, vein swelling and enations on undersides of leaves were collected in the Fujian Province of China in 2007. Virus isolates YC7‐1 and YC7‐2 (associated with betasatellite, YC7‐2β) were detected in both samples. The complete DNA‐A sequence of YC7‐1 (FJ869907) comprised 2741 nucleotides (nt). The complete DNA‐A (FJ869908) and betasatellite (FJ869909) sequence of YC7‐2 consisted of 2754 and 1344 nt, respectively. YC7‐1 had the highest nucleotide sequence identity (97.3%) with Papaya leaf curl Guangdong virus (PaLCuGuV‐[CN:Gd2:02], AJ558122). YC7‐2 had the highest sequence identity (90.1%) with Ageratum yellow vein virus (AYVV‐TW[TW:Tai:99], AF307861) and its betasatellite (96.5%) with Ageratum yellow vein betasatellite (AYVB‐[TW:CHu:02], AJ542495). These indicate that YC7‐1 and YC7‐2 are isolates of PaLCuGuV and AYVV, respectively. Symptoms including leaf curling, vein swelling and enations on undersides of leaves were observed in N. tabacum and N. glutinosa when infected by whiteflies with sample YC7 as the viral source under greenhouse conditions. PCR results showed that these infected plants contained both YC7‐1 and YC7‐2/YC7‐2β. To our knowledge, this is the first report of PaLCuGuV and AYVV/AYVB co‐infecting N. tabacum in China.     

Cytological Evidence of the Association of a Geminivirus with the Tomato Yellow Leaf Curl Disease in Tunisia

Samples of tomato plants from Tunisia naturally or artificially infected with the yellow leaf curl disease were examined in thin sections with the electron microscope. Cytological changes were compared with those previously observed in similarly affected tomato plants from Israel. Evidence was obtained that also in Tunisia a non-mechanically transmissible geminivirus is associated with the disease.