A New Begomovirus Species Causing Tomato Leaf Curl Disease in Patna, India

During December of 2007, a severe leaf curl disease of tomato (ToLCD) occurred in tomato-growing areas in the Patna District of Bihar, India. Viral DNA was isolated from symptomatic tomato plants (2) and begomovirus association was confirmed by PCR using DNA-A degenerate primers (3). Isolated viral DNA was restricted with KpnI and full-length genome was cloned in pUC18. DNA-β was amplified using degenerate primers (1) and cloned in pTZ57RT. Partial tandem repeats of viral genome and DNA-β could infect Nicotiana benthamiana and tomato through Agrobacterium-mediated inoculation. Infected test plants exhibited typical symptoms characteristic of ToLCD. Full-length viral genome (GenBank Accession No. EU862323) consists of 2,752 nt and showed the highest identity (85.8%) with Tomato leaf curl Laos virus-[Laos] (GenBank Accession No. AF195782). The satellite DNA-β component (GenBank Accession No. EU862324) consists of 1,349 nt and showed the highest identity (75.8%) with Tomato leaf curl Joydebpur betasatellite (GenBank Accession No. AJ966244). On the basis of the ICTV species demarcation criteria of 89% of DNA-A sequence identity, the present isolate was considered as a new begomovirus species and named Tomato leaf curl Patna virus (ToLCPaV). Since the isolated DNA-β satellite shares less than 78% identity, it is considered a new species of betasatellite and the name, Tomato leaf curl Patna betasatellite (ToLCPaB) is proposed. These results show that severe ToLCD in Patna is caused by a newly identified species of begomovirus and betasatellite. References: (1) R. W. Briddon et al. Mol. Biotechnol 20:315, 2002. (2) S. Chakraborty et al. Phytopathology 93:1485, 2003. (3) S. D. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996.

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A New Begomovirus Species Causing Tomato Leaf Curl Disease in Varanasi, India

Plant Disease ◽

10.1094/pdis.2003.87.3.313a ◽

2003 ◽

Vol 87 (3) ◽

pp. 313-313 ◽

Cited By ~ 7

Author(s):

S. Chakraborty ◽

P. K. Pandey ◽

M. K. Banerjee ◽

G. Kalloo ◽

C. M. Fauquet

Keyword(s):

Infectious Clone ◽

Uttar Pradesh ◽

Tomato Leaf ◽

Full Length ◽

Degenerate Primers ◽

Tomato Leaf Curl Disease ◽

Leaf Curling ◽

Tomato Leaf Curl ◽

Leaf Curl Disease ◽

Leaf Curl

In November 2001, a leaf curl disease of tomato, manifested by yellowing of leaf lamina, upward leaf curling, leaf distortion, shrinking of leaf surface, and stunted plant growth was observed in tomato-growing areas in the Varanasi and Mirzapur districts of eastern Uttar Pradesh, India, which caused yield losses up to 100%. The causal agent was infective to tomato cv. Punjab Chuhara by whiteflies and grafting. Inoculated plants developed symptoms observed in naturally infected tomatoes. Viral DNA was isolated from artificially inoculated tomato plants using 1% CTAB (2) followed by a concentration of supercoiled DNA by alkaline denaturation (1). A geminivirus was confirmed by polymerase chain reaction using DNA-A degenerate primers (3), and a 550-bp amplified product was obtained from artificially and naturally infected plants. Full-length viral genomes of DNA-A and DNA-B were cloned in plasmid pUC18 at HindIII and XbaI sites, respectively. Partial tandem dimers of the viral clones were infective to Nicotiana benthamiana and tomato cv. Organ Spring through particle bombardment. Infected N. benthamiana plants exhibited downward and upward leaf curling, big veins, leaf puckering with interveinal chlorosis, and stunting. On tomato, symptoms were the same as those seen on naturally infected plants. Cloned DNA also infected Capsicum annuum cv. California Wonder (upward leaf curling and stunting) and tobacco cv. Xanthi (leaf curling and crinkling) but failed to infect Phaseolus vulgaris, okra, cotton, and N. glutinosa. The Varanasi isolate was sap transmissible (0.1 M potassium phosphate buffer, pH 7.0) from the bombarded plants to N. benthamiana and tomato cv. Organ Spring. DNA-A alone infected N. benthamiana (upward leaf curling and big veins) and tomato cv. Organ Spring (mild leaf curl), but symptoms were delayed and milder. Full-length genome sequencing revealed DNA-A (AY190290) contained 2,757 nt and DNA-B (AY190291) contained 2,688 nt. DNA-A of the Varanasi isolate shares 98.4% identity with a DNA-A sequence (AF449999) obtained from a tomato showing leaf curl symptoms from the same region and 97.1% identity with an isolate from Gujarat (900 km from Varanasi). All three sequences represent isolates of the same species, herein called Tomato leaf curl Gujarat virus, based on the priority of submission of the DNA sequence for the Gujarat region (ToLCGV; AF 413671). All isolates noted were obtained from GenBank. However, except for the DNA-A sequence, no other information is available for these ToLCGV isolates. DNA-A of the ToLCGV-Varanasi isolate shares 66.8% identity with Tomato leaf curl New Delhi virus, severe strain (ToLCNdV-Svr) (U15015), and 84.1% with Tomato leaf curl Karnataka virus (U38239). No DNA-B has been reported for these two ToLCGV isolates, and no infectious clone proving the etiology of the disease has been constructed, except for ToLCGV-Varanasi. DNA-B of ToLCGV-Varanasi shares 79.2% homology with ToLCNdV-Svr and 84.1% with ToLCNdV-Luc (X89653). These results suggest that the isolate from Varanasi belongs to ToLCGV, a previously undescribed geminivirus species causing a devastating tomato leaf curl disease in Gujarat and Uttar Pradesh. References: (1) H. C. Birnboim and J. Doly. Nucleic Acids Res. 7:1513, 1979. (2) K. M. Srivastava et al. J. Virol. Methods 51:297, 1995. (3) S. D. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996.

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Evidence for the Association of a Bipartite Geminivirus with Tomato Leaf Curl Disease in Pakistan

Plant Disease ◽

10.1094/pdis.1997.81.8.958c ◽

1997 ◽

Vol 81 (8) ◽

pp. 958-958 ◽

Cited By ~ 11

Author(s):

S. Mansoor ◽

S. H. Khan ◽

M. Saeed ◽

A. Bashir ◽

Y. Zafar ◽

...

Keyword(s):

Tomato Leaf ◽

Cotton Leaf ◽

Degenerate Primers ◽

Tomato Plants ◽

Tomato Leaf Curl Disease ◽

Eastern Hemisphere ◽

Tomato Leaf Curl ◽

Leaf Curl Virus ◽

Leaf Curl Disease ◽

Leaf Curl

Tomato leaf curl disease is the most important constraint on tomato production in Pakistan, where it is found throughout the country. The disease, which occurs in high incidence in Punjab and Sindh provinces, causes 30 to 40% yield losses in the spring crop and uneconomically high losses when grown as an autumn crop. The symptoms of the disease include upward or downward leaf curling, vein thickening, and stunting of the plant. The disease is transmitted by Bemisia tabaci whiteflies (non-B, biotype K) and is suspected to be caused by a geminivirus. For the detection of geminivirus, total DNA was extracted from infected plants, fractionated in an agarose gel, transferred to a nylon membrane, and Southern blotted. A full-length clone of DNA-A of cotton leaf curl virus from Pakistan (S. Mansoor, I. Bedford, M. S. Pinner, A. Bashir, R. Briddon, J. Stanley, Y. Zafar, K. A. Malik, and P. G. Markham, unpublished) was labeled with [32P]dCTP by the oligo-labeling method and hybridized at medium stringency. Geminivirus DNA forms that are normally found in infected plants were detected in plants with tomato leaf curl disease but not in healthy plants. To further confirm the presence of a whiteflytransmitted geminivirus, universal primers for dicot-infecting geminiviruses (1) were used in polymerase chain reaction (PCR) and a product of expected size (approximately 2.7 kb) was detected. The 2.7-kb PCR-amplified DNA from diseased tomato plants was labeled with [32P]dCTP and used as probe in Southern hybridization. This probe also detected geminivirus DNA forms at medium stringency. Both monopartite and bipartite geminiviruses transmitted by whiteflies have been reported to cause leaf curl symptoms on tomato from the Eastern hemisphere. Degenerate primers (PBLv2040 and PCRc1), which amplify B component DNA, were used to determine if tomato leaf curl was monopartite or bipartite (2). A product of expected size (0.65 kb) was amplified, suggesting this virus to be bipartite. DNA-B PCR product obtained from diseased tomato plants was hybridized as described above and detected geminivirus DNA forms at medium stringency. Samples of diseased tomato plants were collected from tomato fields throughout Punjab. DNA-A was detected in all 20 samples whereas DNA B was detected in 17 samples when hybridized by dot blot method at medium stringency. Our data show that tomato leaf curl virus from Pakistan is a bipartite geminivirus. This is the first evidence for a bipartite geminivirus in tomato plants from Pakistan. References: (1) R. W. Briddon and P. G. Markham. Mol. Biotechnol. 1:202, 1993. (2) M. R. Rojas et al. Plant Dis. 77:340, 1993.

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First Report and Molecular Characterization of DNA A of Three Distinct Begomoviruses Associated with Tomato Leaf Curl Disease in Ghana

Plant Disease ◽

10.1094/pdis-92-11-1585b ◽

2008 ◽

Vol 92 (11) ◽

pp. 1585-1585 ◽

Cited By ~ 14

Author(s):

M. K. Osei ◽

R. Akromah ◽

S. L. Shih ◽

L. M. Lee ◽

S. K. Green

Keyword(s):

Molecular Characterization ◽

Tomato Leaf ◽

Begomovirus Species ◽

Sequence Comparisons ◽

Tomato Leaf Curl Disease ◽

Sequence Identity ◽

Tomato Leaf Curl ◽

Leaf Curl Disease ◽

Leaf Curl

Tomato leaf curl disease is reported to be widespread in Ghana and to cause severe yield losses (4). So far, the causal agent has not been identified. Thirty-three tomato (Solanum lycopersicum L.) samples with symptoms such as curling, yellowing, small leaves, and stunting were collected from the Ashanti Region, the main tomato-production area in Ghana, including three samples from Akumandan in the autumn of 2007 and 30 samples from Kumasi in the spring of 2008. The observed leaf curl disease incidence in the farmer's field in Kumasi was approximately 75%. Viral DNAs were extracted from the 33 samples and tested for the presence of begomoviral DNA-A, DNA-B, and associated satellite DNA by PCR with previously described primers (1,3). The expected 1.4-kb DNA-A begomovirus fragment was obtained from one of the samples from Akumadan and from 25 samples from Kumasi. DNA-B and DNA-beta were not detected by PCR. The 1.4-kb PCR products from all positive samples were cloned and sequenced. Sequence comparison by MegAlign software (DNASTAR, Inc., Madison, WI) showed three distinct virus groups. One isolate from each group was selected and specific primers were designed to complete the DNA-A sequence. The DNA-As of GH5-3 (group 1), GOTB2-2 (group 2), and GHK2 (group 3) isolates consisted of 2,803 (GenBank Accession No. EU350585), 2,794 (GenBank Accession No. EU847739), and 2,792 nt (GenBank Accession No. EU847740) respectively. All contain the geminiviral conserved nonanucleotide sequence TAATATTAC in the intergenic region and the six predicted open reading frames (ORFs V1, V2, C1, C2, C3, and C4). BLASTn analysis was conducted with geminivirus sequences available in the GenBank database at National Center for Biotechnology Information (Bethesda, MD). Further sequence comparisons were performed by Clustal V algorithm of MegAlign software with the representative isolates of begomovirus species reported by Fauquet et al (2) and the sequences that showed high scores in BLASTn search. The DNA-A sequence of isolate GHK2 from Kumasi showed highest sequence identity (96.5%) with Tomato yellow leaf curl Mali virus (TYLCMLV; GenBank Accesssion No. AY502934). The DNA-A sequence of GH5-3 and GOTB2-2 isolates had 87.5% sequence identity with each other. Both had highest sequence identities of 76.7 and 77.6%, respectively, with Tomato leaf curl Antsiranana virus, Madagascar (GenBank Accession No. AM701764). They constitute two distinct begomovirus species based on DNA-A sequence comparisons and the International Committee on Taxonomy of Viruses proposed species demarcation of 89% sequence identity. The names Tomato leaf curl Ghana virus for isolate GH5-3 and Tomato leaf curl Kumasi virus for isolate BOTB2-2 are proposed, respectively. To our knowledge, this is the first report of molecular characterization of begomoviruses associated with tomato leaf curl disease in Ghana and of the presence of three distinct tomato begomoviruses. This presence should be considered for recommending or developing stable begomovirus resistant tomato cultivars for Ghana. References: (1) R. W. Briddon et al. Mol. Biotechnol. 20:315, 2002. (2) C. M. Fauquet et al. Arch. Virol. 153:783, 2008. (3) S. K. Green et al. Plant Dis. 85:1286, 2001. (4) D. Horna et al., eds. Online publication. Int. Food Policy Res. Inst. PBS Policy Brief 2, 2007.

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A New Begomovirus Species in Association with Betasatellite Causing Tomato Leaf Curl Disease in Gandhinagar, India

Plant Disease ◽

10.1094/pdis-07-13-0719-pdn ◽

2014 ◽

Vol 98 (3) ◽

pp. 428-428 ◽

Cited By ~ 4

Author(s):

S. Rathore ◽

B. S. Bhatt ◽

B. K. Yadav ◽

R. K. Kale ◽

A. K. Singh

Keyword(s):

New Species ◽

Tomato Leaf ◽

Begomovirus Species ◽

Leaf Rolling ◽

Tomato Leaf Curl Disease ◽

Total Dna ◽

Tomato Leaf Curl ◽

Leaf Curl Disease ◽

A New Species ◽

Leaf Curl

In December 2012, tomato leaf curl disease (ToLCD) (2) was observed in tomato-growing areas of Gandhinagar District of Gujarat, a state in northwestern India. Incidence of ToLCD was estimated to be between 40 and 70% depending on the cultivars used. Infected plants exhibited symptoms consisting of leaf rolling, leaf curling, and yellowing typical of begomoviruses. Total DNA was isolated from a single affected tomato plant (2). Begomovirus infection in this sample was established by amplification of the expected-size 550-bp DNA fragment from this extract by PCR with degenerate DNA-A primers (3). Rolling circle amplification (RCA) using ϕ29 DNA polymerase was carried out on the total DNA, followed by digestion with Bam HI. An amplicon of ~2.8 kb was gel-eluted and cloned into Bam HI linearized pBluescript II KS(+). Restriction enzyme digestion of plasmid DNA from the resulting clones indicated the presence of one type of molecule. Using PCR and universal betasatellite primers, the expected 1.3-kb fragment was amplified from the DNA extract (1). An amplicon of ~1.3 kb was gel-eluted and cloned into pTZ57RT vector. Sequence analysis revealed that DNA-A (GenBank Accession No. KC952005) is composed of 2,753 nt and showed the highest identity (87.8%) with Tomato leaf curl Kerala virus[India:Kerala:2008] (GenBank Accession No. EU910141). An analysis for recombination showed this begomovirus DNA likely to have originated by recombination between Tomato leaf curl Kerala virus and Tomato leaf curl Karnataka virus. The satellite DNA-β (GenBank Accession No. KC952006) is composed of 1,365 nt and showed the highest identity (75.6%) with Tomato leaf curl betasatellite[India:Ludhiana:2004] (ToLCB-[IN:Lud:04]) (GenBank Accession No. AY765255). On the basis of DNA-A sequence analysis, the ICTV species demarcation criteria of 89% DNA-A sequence identity, and genome organization, the present isolate was considered as a new begomovirus species and named Tomato leaf curl Gandhinagar virus (ToLCGNV). The betasatellite shares less than 78% identity with (ToLCB-[IN:Lud:04]), it is considered a new species of betasatellite and the name, Tomato leaf curl Gandhinagar betasatellite (ToLCGNB) is proposed. Multimeric clones of the begomovirus and betasatellite DNAs were generated in a binary vector and these plasmids transformed into Agrobacterium tumefaciens. Nicotiana benthamiana and tomato plants agroinoculated with the cloned begomovirus DNA developed leaf curl symptoms, whereas plants co-agroinoculated with the cloned begomovirus and betasatellites developed more severe symptoms, including leaf rolling, leaf curling, and yellowing. The symptoms induced by the begomovirus and betasatellite DNAs were indistinguishable from those observed in the field. Thus, ToLCGNV is a new monopartite begomovirus which, in association with a new species of betasatellite, causes ToLCD in Gandhinagar, India. The presence of ToLCGNV needs to be considered, along with the already reported begomoviruses infecting tomatoes in this state, e.g., Tomato leaf curl Gujarat virus (2), in studies aimed to developing tomato cultivars with stable resistance to these tomato-infecting begomoviruses in India. References: (1) R. W. Briddon et al. Mol. Biotechnol. 20:315, 2002. (2) C. Reddy et al. Arch Virol. 150:845, 2005. (3) S. D. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996.

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A new begomovirus species and betasatellite causing severe tomato leaf curl disease in Ranchi, India

New Disease Reports ◽

10.5197/j.2044-0588.2011.023.011 ◽

2011 ◽

Vol 23 ◽

pp. 11 ◽

Cited By ~ 5

Author(s):

Punam Kumari ◽

Achuit K Singh ◽

Brotati Chattopadhyay ◽

Supriya Chakraborty

Keyword(s):

Tomato Leaf ◽

Begomovirus Species ◽

Tomato Leaf Curl Disease ◽

Tomato Leaf Curl ◽

Leaf Curl Disease ◽

Leaf Curl

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Potato apical leaf curl disease: current status and perspectives on a disease caused by tomato leaf curl New Delhi virus

Journal of Plant Diseases and Protection ◽

10.1007/s41348-021-00463-w ◽

2021 ◽

Author(s):

Ravinder Kumar ◽

Rahul Kumar Tiwari ◽

Arjunan Jeevalatha ◽

Sundaresha Siddappa ◽

Mohd. Abas Shah ◽

...

Keyword(s):

Tomato Leaf ◽

Current Status ◽

New Delhi ◽

Apical Leaf ◽

Tomato Leaf Curl ◽

Leaf Curl Disease ◽

Leaf Curl

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Diversity analysis of Tomato leaf curl New Delhi virus-[potato], causing apical leaf curl disease of potato in India

Phytoparasitica ◽

10.1007/s12600-017-0563-4 ◽

2017 ◽

Vol 45 (1) ◽

pp. 33-43 ◽

Cited By ~ 5

Author(s):

Arjunan Jeevalatha ◽

Swarup Kumar Chakrabarti ◽

Sanjeev Sharma ◽

Vinay Sagar ◽

Kamlesh Malik ◽

...

Keyword(s):

Tomato Leaf ◽

Diversity Analysis ◽

New Delhi ◽

Apical Leaf ◽

Tomato Leaf Curl ◽

Leaf Curl Disease ◽

Leaf Curl

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Molecular Characterization of a Begomovirus, α-Satellite, and β-Satellite Associated with Leaf Curl Disease of Parthenium hysterophorus in India

Plant Disease ◽

10.1094/pdis-09-15-0982-re ◽

2016 ◽

Vol 100 (11) ◽

pp. 2299-2305 ◽

Cited By ~ 9

Author(s):

Susheel Kumar ◽

Ashish Srivastava ◽

Meraj Jaidi ◽

Puneet Singh Chauhan ◽

S. K. Raj

Keyword(s):

Satellite Dna ◽

Rolling Circle Amplification ◽

Tomato Leaf ◽

Parthenium Hysterophorus ◽

Alternate Host ◽

Invasive Weed ◽

Tomato Leaf Curl Disease ◽

Tomato Leaf Curl ◽

Leaf Curl Disease ◽

Leaf Curl

Parthenium hysterophorus plants exhibiting severe leaf curl and stunting symptoms were observed near agriculture fields in Lucknow, India. The association of a begomovirus, β-satellite, and α-satellite with these symptoms of a Parthenium disease was investigated by sequence analyses of virus and satellite DNA amplified by rolling circle amplification and polymerase chain reaction. The highest sequence identities and closest phylogenetic relationships for the begomovirus, β-satellite, and α-satellite detected in P. hysterophorus plants were to Tomato leaf curl virus (ToLCV), papaya leaf curl β-satellite (PaLCuB), and Ageratum yellow vein India α-satellite (AYVIA), respectively. These findings identified the virus and satellites infecting the Parthenium sp. as ToLCV, PaLCuB, and AYVIA, respectively. P. hysterophorus and tomato seedlings infected with cloned ToLCV, PaLCuB, and AYVIA by agroinoculation developed leaf curl symptoms, whereas plants infected with ToLCV alone or with ToLCV and AYVIA developed mild yellowing. The results show that this complex infects and causes disease in P. hysterophorus and tomato. P. hysterophorus is an invasive weed commonly found around agricultural fields and along roadsides in India. These results indicate that P. hysterophorus plants infected with ToLCV and associated satellite DNA act as an alternate host (reservoir), and that could lead to increased incidence of tomato leaf curl disease.

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