Engineering Dual Begomovirus- Bemisia tabaci Resistance in Plants
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In the fall of 2008,the cowpea growing in a greenhouse in Shanghai suburban suffered a disease characterized by chlorotic and yellowing leaves.The complete nucleotide sequence of a tomato yellow leaf curl virus was amplified from DNA extracted from diseased plants.The sequence comparisons and phylogenetic analysis revealed that the pathogen was Tomato yellow leaf curl virus-Israel(TYLCV-IL),a member of the genus Begomovirus.The genome consisted of circular ssDNA with 2781 nucleotides,which has the typical characteristic of Old World monopartite begomoviruses with six ORFs corresponding to V2 and V1(Capsid Protein gene)on the viral strand,and C3,C2,C1 and C4 on the complementary strand.This is the first report that a natural Tomato yellow leaf curl virus infected V.sinensis.
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Tomato yellow leaf curl disease (TYLCD) is one of the most devastating viral diseases affecting tomato crops in tropical, subtropical and temperate regions of the world. Here, we focus on the interactions through recombination between the different begomovirus species causing TYLCD, provide an overview of the interactions with the cellular genes involved in viral replication, and highlight recent progress on the relationships between these viruses and their vector, the whitefly Bemisia tabaci.The tomato yellow leaf curl virus-like viruses (TYLCVs) are a complex of begomoviruses (family Geminiviridae, genus Begomovirus) including 10 accepted species: Tomato yellow leaf curl Axarquia virus (TYLCAxV), Tomato yellow leaf curl China virus (TYLCCNV), Tomato yellow leaf curl Guangdong virus (TYLCGuV), Tomato yellow leaf curl Indonesia virus (TYLCIDV), Tomato yellow leaf curl Kanchanaburi virus (TYLVKaV), Tomato yellow leaf curl Malaga virus (TYLCMalV), Tomato yellow leaf curl Mali virus (TYLCMLV), Tomato yellow leaf curl Sardinia virus (TYLCSV), Tomato yellow leaf curl Thailand virus (TYLCTHV), Tomato yellow leaf curl Vietnam virus (TYLCVNV) and Tomato yellow leaf curl virus(TYLCV). We follow the species demarcation criteria of the International Committee on Taxonomy of Viruses (ICTV), the most important of which is an 89% nucleotide identity threshold between full-length DNA-A component nucleotide sequences for begomovirus species. Strains of a species are defined by a 93% nucleotide identity threshold.The primary host of TYLCVs is tomato (Solanum lycopersicum), but they can also naturally infect other crops [common bean (Phaseolus vulgaris), sweet pepper (Capsicum annuum), chilli pepper (C. chinense) and tobacco (Nicotiana tabacum)], a number of ornamentals [petunia (Petuniaxhybrida) and lisianthus (Eustoma grandiflora)], as well as common weeds (Solanum nigrum and Datura stramonium). TYLCVs also infect the experimental host Nicotiana benthamiana.Infected tomato plants are stunted or dwarfed, with leaflets rolled upwards and inwards; young leaves are slightly chlorotic; in recently infected plants, fruits might not be produced or, if produced, are small and unmarketable. In common bean, some TYLCVs produce the bean leaf crumple disease, with thickening, epinasty, crumpling, blade reduction and upward curling of leaves, as well as abnormal shoot proliferation and internode reduction; the very small leaves result in a bushy appearance.
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The current epidemic of cotton leaf curl disease (CLCuD) in Pakistan started in 1988 with the natural host range limited to a few plant species in the family Malvaceae. However, we have observed expansion in the host range of the virus, and several non-Malvaceous plants were found to be infected with the virus. Characteristic symptoms of CLCuD such as leaf curl and enations have been observed on radish plants, primarily in kitchen gardens. However, in 1999, levels of infection of 10 to 90% were observed both in commercial fields and kitchen gardens in the Punjab province of Pakistan. Both symptomatic and nonsymptomatic samples were collected from five different locations. Total DNA was isolated, dot-blotted on nylon membrane, and a full-length clone corresponding to DNA A of cotton leaf curl virus was labeled with 32P dCTP and used as a probe for the detection of a begomovirus. Strong signals were observed in symptomatic plants while no signals were observed in nonsymptomatic plants. Infection with a begomovirus was further confirmed by polymerase chain reaction (PCR) using degenerate primers for DNA A (1). Primers specific for the two distinct begomoviruses associated with CLCuD were also used in PCR reactions (2), and products of the expected size were obtained from all symptomatic samples, confirming infection with begomoviruses similar to those associated with CLCuD. A full-length probe of a nanovirus-like molecule associated with cotton leaf disease (3), called DNA 1 was labeled with 32P dCTP and detected the virus only in symptomatic plants. Similarly, primers specific for DNA 1 (3) amplified a product of expected size when used in PCR. On the basis of symptomatology and the detection of specific viral components associated with the disease, we confirmed that radish plants are infected with Cotton leaf curl virus (CLCuV). Since radish is a short duration crop, infection of CLCuV in radish may not serve as a direct source of infection for the next cotton crop. However, it is a potential threat to tomato crops which overlap with radish in the Punjab province. The detection of CLCuD in radish is another example of the mobilization of begomoviruses to previously unknown hosts. References: (1) M. R. Rojas et al. Plant Dis. 77:340, 1993. (2) S. Mansoor et al. Pak. J. Bot. 31:115, 1999. (3) Mansoor et al. Virology 259:190, 1999.
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TYLCV transmission to different tomato accessions (Solanum lycopersicon L.) mediated by Bemisia tabaci (Gennadius) During December 2006- February 2007 transmission of TYLCV to tomato potted plants mediated by Bemisia tabaci (Biotype B) was evaluated in insect proof cages. Tomato plants of Rio Orinoco hybrid (ROH), Rio Grande hybrid (RGH), Cid hybrid (CH) and Rio Grande variety (RGV) were exposed for 48 hours in individual cages to whiteflies raised either on viral infected tomato plants or healthy cotton plants. Thereafter, plants were observed daily for symptoms detection. Presence of Begomovirus in source plants was determined by PCR amplifying the 550 and 851 bp fragments of the central region of the gene coding for the viral coat protein. The sequences of the amplified fragments have 99% and 96% identity respectively with a TYLCV isolate. For experimental plants only presence of Begomovirus was determined amplifying by PCR the 550 bp fragment. ROH, RGH and RGV showed symptoms 11.3±1.4, 11.2±1.5 and 11.2+1.4 days post exposure period to the whiteflies, respectively. No symptoms were observed on CH tomato plants despite detection of Begomovirus by PCR. Symptomatic plants reached 100% for RGH and RGV, as well as 95% for ROH. These results suggest specific resistance of CH tomato plants to TYLCV infection. Additional key words: Resistant cultivars, vector, Begomovirus, Geminiviridae
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Abstract Background Many plant viruses are vector-borne and depend on arthropods for transmission between host plants. Begomoviruses, the largest, most damaging and emerging group of plant viruses, infect hundreds of plant species, and new virus species of the group are discovered each year. Begomoviruses are transmitted by members of the whitefly Bemisia tabaci species complex in a persistent-circulative manner. Tomato yellow leaf curl virus (TYLCV) is one of the most devastating begomoviruses worldwide and causes major losses in tomato crops, as well as in many agriculturally important plant species. Different B. tabaci populations vary in their virus transmission abilities; however, the causes for these variations are attributed among others to genetic differences among vector populations, as well as to differences in the bacterial symbionts housed within B. tabaci. Results Here, we performed discovery proteomic analyses in 9 whitefly populations from both Middle East Asia Minor I (MEAM1, formerly known as B biotype) and Mediterranean (MED, formerly known as Q biotype) species. We analysed our proteomic results on the basis of the different TYLCV transmission abilities of the various populations included in the study. The results provide the first comprehensive list of candidate insect and bacterial symbiont (mainly Rickettsia) proteins associated with virus transmission. Conclusions Our data demonstrate that the proteomic signatures of better vector populations differ considerably when compared with less efficient vector populations in the 2 whitefly species tested in this study. While MEAM1 efficient vector populations have a more lenient immune system, the Q efficient vector populations have higher abundance of proteins possibly implicated in virus passage through cells. Both species show a strong link of the facultative symbiont Rickettsia to virus transmission.
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Abstract This study investigated the role of vector acquisition and transmission on the propagation of single and co-infections of tomato yellow leaf curl virus (TYLCV,) and tomato mottle virus (ToMoV) (Family: Geminiviridae , Genus: Begomovirus ) by the whitefly vector Bemisia tabaci MEAM1 (Gennadius) in tomato. The aim of this research was to determine if how viruses are co-acquired and co-transmitted changes the probability of acquisition, transmission and new host infections. Whiteflies acquired virus by feeding on singly infected plants, co-infected plants, or by sequential feeding on singly infected plants. Viral titers were also quantified by qPCR in vector cohorts, in artificial diet, and plants after exposure to viruliferous vectors. Differences in transmission, infection status of plants, and titers of TYLCV and ToMoV were observed among treatments. All vector cohorts acquired both viruses, but co-acquisition/co-inoculation generally reduced transmission of both viruses as single and mixed infections. Co-inoculation of viruses by the vector also altered virus accumulation in plants regardless of whether one or both viruses were propagated in new hosts. These findings highlight the complex nature of vector-virus-plant interactions that influence the spread and replication of viruses as single and co-infections.
Geminiviridae
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Geminiviridae
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Plant-pathogenic begomoviruses have a complex association with their whitefly vector and aspects concerning virus genetic activity (genome replication and gene transcription) within the insect remain highly controversial. Virus transcript abundance was assessed by quantifying selected gene transcripts of Tomato mottle virus (ToMoV, a New World bipartite begomovirus) and Tomato yellow leaf curl virus (TYLCV, an Old World monopartite begomovirus) in whiteflies (Bemisia tabaci biotype B) after feeding on virus-infected tomato plants and after subsequent transfer to cotton, a plant that is immune to the selected begomoviruses. Real-time RT-PCR was performed using specific primers for three ToMoV genes (AV1, BC1 and BV1) and three TYLCV genes (V1, V2 and C3). The ToMoV gene transcripts rapidly became undetectable in whiteflies following transfer from tomato to cotton, probably because degradation was not accompanied by new synthesis. On the other hand, TYLCV transcripts increased after transfer of whiteflies to cotton, indicating active TYLCV transcription. Interestingly, the difference observed in ToMoV and TYLCV transcripts in the vector parallel observations on the different biological effects of these viruses on whiteflies, i.e. TYLCV, but not ToMoV, reduces whitefly fitness.
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Tomato yellow leaf curl virus (TYLCV) (Begomovirus, Geminiviridae) is the type member and representative of the complex of viruses associated with the tomato yellow leaf curl disease (TYLCD) with ssDNA genome, a plant-infecting group of viruses that have single or double genomic components enveloped by an icosahedral coat protein. These viruses infect tomatoes and other vegetable and ornamental crops and cause severe losses estimated by billions of dollars each year. Begomoviruses are exclusively transmitted by the whitefly Bemisia tabaci in a persistent circulative manner. First epidemics of TYLCV were reported in Israel in the early 1960s and later on the causative agent was identified as TYLCV. Epidemics were often associated with the presence of whiteflies. Since then, extensive research in many laboratories in the world was conducted to better understand the interactions between TYLCV, the tomato plant and its only vector B. tabaci. These studies resulted in hundreds of research papers and reviews, a...
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Leaf curl
Geminiviridae
Ornamental plant
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