The begomovirus species Melon chlorotic leaf curl virus is composed of two highly divergent strains that differ in their genetic and biological properties

Plant Disease ◽  
2021 ◽  
Author(s):  
Minor R Maliano ◽  
Tomas Melgarejo ◽  
Maria J. Rojas ◽  
Natalia Barboza ◽  
Robert Gilbertson
Keyword(s):  
Costa Rica ◽  
Transmitted Disease ◽  
Biological Properties ◽  
Begomovirus Species ◽  
Full Length ◽  
Bipartite Begomovirus ◽  
Species Demarcation Threshold ◽  
Chlorotic Leaf ◽  
Leaf Curl Virus ◽  
Leaf Curl

Since the early 1990s, squash production in Costa Rica has been affected by a whitefly-transmitted disease characterized by stunting and yellow mottling of leaves. The squash yellow mottle disease (SYMoD) was shown to be associated with a bipartite begomovirus, originally named squash yellow mild mottle virus (SYMMoV). It was subsequently established that SYMMoV is a strain of melon chlorotic leaf curl virus (MCLCuV), a bipartite begomovirus that causes a chlorotic leaf curl disease of melons in Guatemala. In the present study, the complete sequences of the DNA-A and DNA-B components of a new isolate of the strain MCLCuV-Costa Rica (MCLCuV-CR) were determined. Comparisons of full-length DNA-A sequences revealed 97% identity with a previously characterized isolate of MCLCuV-CR, and identities of 90 to 91% with those of isolates of the strain MCLCuV-Guatemala (MCLCuV-GT), which is below or at the current begomovirus species demarcation threshold of 91%. A more extensive analysis of the MCLCuV-CR and -GT sequences revealed substantial divergence in both components and different histories of recombination for the DNA-A components. The cloned full-length DNA-A and DNA-B components of this new MCLCuV-CR isolate were infectious and induced SYMoD in a range of squashes and in pumpkin, thereby fulfilling Koch’s postulates for this disease. However, in contrast to MCLCuV-GT, MCLCuV-CR induced mild symptoms in watermelon and no symptoms in melon and cucumber. Taken together, our results indicate that MCLCuV-CR and -GT have substantially diverged, genetically and biologically, and have evolved to cause distinct diseases of different cucurbit crops. Taxonomically, these viruses are at the strain/species boundary, but retain the designation as strains of Melon chlorotic leaf curl virus under current ICTV guidelines.

scholarly journals First Report of Tomato yellow leaf curl virus Infecting Tomato, Tomatillo, and Peppers in Guatemala

Plant Disease ◽  
2010 ◽  
Vol 94 (4) ◽  
pp. 482-482 ◽  
Author(s):  
R. Salati ◽  
M. Shorey ◽  
A. Briggs ◽  
J. Calderon ◽  
M. R. Rojas ◽  
...  
Keyword(s):  
Blot Hybridization ◽  
Tomato Yellow Leaf ◽  
Full Length ◽  
Bipartite Begomovirus ◽  
Yellow Leaf ◽  
Sequence Analyses ◽  
Degenerate Primers ◽  
Begomovirus Infection ◽  
Leaf Curl Virus ◽  
Leaf Curl

In Guatemala and other Central American countries, whitefly-transmitted geminiviruses (begomoviruses) cause economically important diseases of tomato (Solanum lycopersicum) and pepper (Capsicum annuum). Disease symptoms include stunted and distorted growth and leaf curling, crumpling, light green to yellow mosaic, purpling, and vein swelling. In Guatemala, at least eight bipartite begomovirus species infect tomato or peppers (1), but their role and relative importance is unclear. As part of an Integrated Pest Management strategy to manage these diseases, surveys for begomovirus symptoms in pepper and tomato have been conducted in the Salama Valley, Sanarate, and other locations since 2003, and begomoviruses were identified by squash blot hybridization, PCR and DNA sequencing. Beginning in 2006, a new type of symptom, stunted upright growth and upcurled leaves with yellowing of the margins and interveinal areas, was observed in tomato and tomatillo plants in the Salama Valley and Sanarate. These symptoms were similar to those induced by the exotic monopartite begomovirus Tomato yellow leaf curl virus (TYLCV). Evidence that TYLCV caused these symptoms came from positive results in high stringency squash blot hybridization tests with a TYLCV probe, and amplification of the expected size of ~0.3- and 2.8-kb fragments in PCR tests with TYLCV capsid protein (CP) gene and full-length component primer pairs, respectively (3). Sequence analyses of PCR-amplified CP fragments and portions of full-length fragments revealed 97 to 99% identity with isolates of TYLCV-Israel (TYLCV-IL). The complete nucleotide sequence of an isolate from the Salama Valley (GenBank Accession No. GU355941) was >99% identical to those of TYLCV-IL isolates from the Dominican Republic, Florida, and Cuba and ~97% identical to those of isolates from Mexico and California. Thus, this TYLCV-IL isolate (TYLCV-IL[GT:06]) was probably introduced from the Caribbean Region. To further characterize begomoviruses in the Salama Valley, leaf samples were collected from 44 and 118 tomato plants showing symptoms of begomovirus infection in March 2006 and 2007, respectively, and from 106 symptomatic pepper plants in March 2007. Begomovirus infection was confirmed in 42 of 44 and 93 of 118 of the tomato samples and 100 of 106 of the pepper samples based on PCR amplification of the expected size of ~0.6- and 1.1-kb DNA fragments with the begomovirus degenerate primers pairs AV494/AC1048 and PAL1v1978/PAR1c496, respectively (2,4). Sequence analyses of cloned PCR-amplified fragments revealed that 3 of the 44 and 16 of the 118 tomato samples collected in 2006 and 2007, respectively, and 9 of the 106 pepper samples were infected with TYLCV based on >97% identity with TYLCV-IL. In all samples, TYLCV was present in mixed infections with other begomoviruses. The introduction of TYLCV adds to the already high level of genetic complexity of bipartite begomovirus infection of tomatoes and peppers in Guatemala and will undoubtedly complicate disease management efforts. References: (1) M. K. Nakhla et al. Acta Hortic. 695:277, 2005. (2) M. R. Rojas et al. Plant Dis. 77:340, 1993. (3) R. Salati et al. Phytopathology 92:487, 2002. (4) S. D. Wyatt and J. Brown. Phytopathology 86:1288, 1996.

scholarly journals Emergence of a New Cucurbit-Infecting Begomovirus Species Capable of Forming Viable Reassortants with Related Viruses in the Squash leaf curl virus Cluster

2002 ◽  
Vol 92 (7) ◽  
pp. 734-742 ◽  
Author(s):  
J. K. Brown ◽  
A. M. Idris ◽  
C. Alteri ◽  
Drake C. Stenger
Keyword(s):  
Nucleotide Sequence ◽  
Host Range ◽  
Nucleotide Sequence Identity ◽  
Begomovirus Species ◽  
Progeny Virus ◽  
Bipartite Begomovirus ◽  
Sequence Identity ◽  
Squash Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

Cucurbit leaf curl virus (CuLCV), a whitefly-transmitted geminivirus previously partially characterized from the southwestern United States and northern Mexico, was identified as a distinct bipartite begomovirus species. This virus has near sequence identity with the previously partially characterized Cucurbit leaf crumple virus from California. Experimental and natural host range studies indicated that CuLCV has a relatively broad host range within the family Cucurbitaceae and also infects bean and tobacco. The genome of an Arizona isolate, designated CuLCV-AZ, was cloned and completely sequenced. Cloned CuLCV-AZ DNA A and B components were infectious by biolistic inoculation to pumpkin and progeny virus was transmissible by the whitefly vector, Bemisia tabaci, thereby completing Koch's postulates. CuLCV-AZ DNA A shared highest nucleotide sequence identity with Squash leaf curl virus-R (SLCV-R), SLCV-E, and Bean calico mosaic virus (BCaMV) at 84, 83, and 80%, respectively. The CuLCV DNA B component shared highest nucleotide sequence identity with BCaMV, SLCV-R, and SLCV-E at 71, 70, and 68%, respectively. The cis-acting begomovirus replication specificity element, GGTGTCCTGGTG, in the CuLCV-AZ origin of replication is identical to that of SLCV-R, SLCV-E, and BCaMV, suggesting that reassortants among components of CuLCV-AZ and these begomoviruses may be possible. Reassortment experiments in pumpkin demonstrated that both reassortants of CuLCV-AZ and SLCV-E A and B components were viable. However, for CuLCV-AZ and SLCV-R, only one reassortant (SLCV-R DNA A/CuLCV-AZ DNA B) was viable on pumpkin, even though the cognate component pairs of both viruses infect pumpkin. These results demonstrate that reassortment among sympatric begomovirus species infecting cucurbits are possible, and that, if generated in nature, could result in begomoviruses bearing distinct biological properties.

scholarly journals Widespread Occurrence of Cotton leaf curl virus on Radish in Pakistan

Plant Disease ◽  
2000 ◽  
Vol 84 (7) ◽  
pp. 809-809 ◽  
Author(s):  
S. Mansoor ◽  
S. Mukhtar ◽  
M. Hussain ◽  
I. Amin ◽  
Y. Zafar ◽  
...  
Keyword(s):  
Host Range ◽  
Full Length ◽  
Cotton Leaf ◽  
Degenerate Primers ◽  
Nylon Membrane ◽  
Punjab Province ◽  
Cotton Leaf Curl Virus ◽  
Kitchen Gardens ◽  
Leaf Curl Virus ◽  
Leaf Curl

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.

scholarly journals Cucurbit leaf curl virus, a New Whitefly Transmitted Geminivirus in Arizona, Texas, and Mexico

Plant Disease ◽  
2000 ◽  
Vol 84 (7) ◽  
pp. 809-809 ◽  
Author(s):  
J. K. Brown ◽  
A. M. Idris ◽  
M. W. Olsen ◽  
M. E. Miller ◽  
T. Isakeit ◽  
...  
Keyword(s):  
Binding Site ◽  
Field Isolate ◽  
Bipartite Begomovirus ◽  
Degenerate Primers ◽  
Field Isolates ◽  
Squash Leaf Curl Virus ◽  
Cp Gene ◽  
Honeydew Melon ◽  
Leaf Curl Virus ◽  
Leaf Curl

In 1998 to 1999, geminivirus-like symptoms were observed in whitefly-infested pumpkin, honeydew melon, and muskmelon in Arizona and Texas and in Coahuilla, Mexico (MX), respectively. Plants exhibited leaf curl and/or mottling, reminiscent of symptoms caused by Squash leaf curl virus (SLCV-WAZ) described from Arizona in 1981 (2). The isolate from Arizona pumpkin fields was experimentally transmitted to pumpkin seedlings by the “B type” of Bemisia tabaci (Genn.), and symptoms were indistinguishable from those observed in infected fields. Samples from AZ, MX, and TX were assessed for begomovirus presence by polymerase chain reaction (PCR) using degenerate primers that amplify a contiguous fragment containing the viral coat protein (Cp) gene and common region (CR) of the A component (CR-A) (~2,100 bp) and a fragment containing the CR of the B component (CR-B) (~1,100 bp). One to four isolates from each location were examined by PCR using both primer pairs, and at least three amplicons per isolate were cloned and their sequences determined. Alignment of viral Cp nucleotide (nt) sequences revealed that AZ [AF256199], MX, and TX field isolates shared 98.7 to 100% sequence identity, but were only 84.5 to 85.6% identical to the Cp gene of SLCV-extended (SLCV-E) [M38183] and SLCV-restricted (SLCV-R) (S. G. Lazarowitz, unpublished), respectively, suggesting a new, previously undescribed begomoviral species (3). Further, the Cp nt sequence of the three field isolates was 6 nt shorter than SLCV-E, SLCV-WAZ [AF256203], and SLCV-R Cp sequences. The CR-A [AF256200] and CR-B [AF256201] sequences (179 nt, each) of field isolates, including the theoretical Rep binding element, GGTGT, were 100% identical. Although the Rep binding site is identical among field isolates, SLCV-E, SLCV-R, and SLCV-WAZ, the field isolate CR sequence shared only 64.2, 67.5, and 66.9% overall identity with CR-A SLCV-E, SLCV-R [M63155], and SLCV-WAZ [AF256202], respectively. Prior to 1998 to 1999, SLCV-WAZ was the only New World begomovirus of cucurbits known to infect both melon (Cucumis) and pumpkin (Cucurbita) (1). Therefore, SLCV was initially suspected as the causal agent. However, here we provide evidence for a new, previously undescribed bipartite begomovirus of cucurbits in AZ, MX, and TX that is herein provisionally designated Cucurbit leaf curl virus (CuLCV). Prediction of its closest begomovirus relatives by Cp nt sequence and Rep binding site comparisons suggest that CuLCV is a new member of the SLCV lineage, also containing Bean calico mosaic virus, Cabbage leaf curl virus, SLCV-E, and Texas pepper virus-TAM. References: (1) J. K. Brown and M. R. Nelson. Phytopathology 74:1136, 1984. (2) J. K. Brown and M. R. Nelson. Ann. Appl. Biol. 115:243, 1986. (3) M. A. Mayo and C. R. Pringle. J. Gen. Virol. 97:649, 1998.

scholarly journals Squash Leaf Curl Virus: A New World Bipartite Begomovirus Threatening Squash Production in Oman

Plant Disease ◽  
2020 ◽  
Vol 104 (9) ◽  
pp. 2533-2533
Author(s):  
M. S. Shahid ◽  
H. Al-Sulaimani ◽  
A. M. Al-Sadi
Keyword(s):  
New World ◽  
Bipartite Begomovirus ◽  
Squash Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

scholarly journals Melon chlorotic leaf curl virus, a New Begomovirus Associated with Bemisia tabaci Infestations in Guatemala

Plant Disease ◽  
2001 ◽  
Vol 85 (9) ◽  
pp. 1027-1027 ◽  
Author(s):  
J. K. Brown ◽  
A. M. Idris ◽  
D. Rogan ◽  
M. H. Hussein ◽  
M. Palmieri
Keyword(s):  
Nucleotide Sequence ◽  
Bemisia Tabaci ◽  
Nucleotide Sequences ◽  
Bipartite Begomoviruses ◽  
Degenerate Primers ◽  
The Core ◽  
Cp Gene ◽  
Chlorotic Leaf ◽  
Leaf Curl Virus ◽  
Leaf Curl

In 2000, geminivirus-like symptoms were widespread in muskmelon (Cucumis melo L.) fields (70 to 80% incidence) in Zacapa Valley, Guatemala. Muskmelon fields were infested with the whitefly Bemisia tabaci (Genn.), and plants exhibited patchy foliar chlorosis, leaf curling, and reduced fruit set, which is reminiscent of symptoms caused by certain whitefly-transmitted geminiviruses. Quarantine restrictions prevented experimental transmission experiments from being carried out with the whitefly vector or biolistic inoculation. Leaves collected from six symptomatic plants were assessed for the presence of begomovirus DNA by polymerase chain reaction (PCR) with the use of degenerate primers that amplify the core region of the coat protein (CP) gene of most begomoviruses (1). PCR products of the expected size (approximately 576 bp) were obtained from all three melon samples. The core CP amplicons were cloned, and their nucleotide sequences were compared. Nucleotide sequences of core CP fragments shared 99.7% identity, suggesting the presence of a single begomovirus in all assayed symptomatic melon plants. Two additional pairs of degenerate primers were used to obtain contiguous viral fragments containing the CP gene, the common region of the A component (CR-A; approximately 2,100 bp), and a fragment containing the CR of the B component (CR-B; approximately 1,100 bp), respectively (2). At least three amplicons obtained with each primer pair were cloned and their nucleotide sequence was determined. Virus-specific PCR primers were then designed within the CP open reading frame and used to obtain fragments that overlapped with the 2,100-bp fragment to yield an apparent full-length A component of 2,662 nucleotides (accession no. AF325497). CR-A and CR-B (accession no. AF325498) sequences (161 nucleotides) shared 98.1% identity and contained an identical directly repeated, replication-associated protein (REP) binding site: GGTGT CCT GGTGT. Nucleotide sequence alignment, with CLUSTAL W, of the melon virus A-component with that of other well-studied begomoviruses revealed that its closest relatives were members of the Squash leaf curl virus (SLCV) group. The melon virus from Guatemala shared its greatest sequence identity, 83.1%, with SLCV extended (SLCV-E) (accession no. M38183), indicating that it is a new, previously unidentified begomovirus species, herein referred to as Melon chlorotic leaf curl virus (MCLCV). The next closest relatives of MCLCV were SLCV restricted (SLCV-R; 78.6%) (S. G. Lazarowitz, unpublished) Cucurbit leaf curl virus-Arizona (CuLCV-AZ; accession no. AF256199; 74.1%) (3), Cabbage leaf curl virus (CaLCV; 72.0%), Bean calico mosaic virus (BCMoV; 71.7%), and Texas pepper virus-Tamaulipas (71.4%). Additionally, the theoretical REP binding element, GGTGT, is 100% identical among MCLCV and BCMoV, CaLCV, CuLCV-AZ, SLCV-E, and SLCV-R. On the basis of shared nucleotide sequence identities with other begomoviruses described to date and the presence of B. tabaci in melon fields, it is likely that MCLCV also is whitefly-transmitted. Collectively, CP and CR sequences suggest that MCLCV is a new species of the SLCV lineage that contains other bipartite begomoviruses indigenous to Central America, Mexico, and the U.S. Sunbelt states. References: (1) S. D. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996. (2) A. M. Idris and J. K Brown. Phytopathology 88:648, 1998. (3) J. K. Brown et al. Plant Dis. 84:809, 2000.

scholarly journals Biotic, Molecular, and Phylogenetic Characterization of Bean Calico Mosaic Virus, a Distinct Begomovirus Species with Affiliation in the Squash Leaf Curl Virus Cluster

1999 ◽  
Vol 89 (4) ◽  
pp. 273-280 ◽  
Author(s):  
J. K. Brown ◽  
Kristin M. Ostrow ◽  
Ali M. Idris ◽  
Drake C. Stenger
Keyword(s):  
Nucleotide Sequence ◽  
Mosaic Virus ◽  
Common Ancestor ◽  
Begomovirus Species ◽  
Recent Common Ancestor ◽  
Progeny Virus ◽  
Most Recent Common Ancestor ◽  
Squash Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

Bean calico mosaic virus (BCMoV), a whitefly-transmitted geminivirus from Sonora, Mexico, was purified, and the genome components were cloned and sequenced. Purified viral fractions and cloned genome components were infectious by biolistic inoculation to bean, completing Koch's postulates for both. The B biotype of the whitefly Bemisia tabaci efficiently transmitted both native virus and progeny virus derived from cloned DNA inoculum. Host ranges of native virus and of progeny virus derived from cloned DNA were identical based upon whitefly and biolistic mediated transmission, respectively. BCMoV has a relatively wide experimental host range among begomoviruses known to infect bean, encompassing genera and species within the Fabaceae, Malvaceae, and Solanaceae. BCMoV has a bipartite genome, as do other New World begomoviruses. BCMoV DNA-A shared highest nucleotide sequence identities with squash leaf curl virus-E strain (SLCV-E) and cabbage leaf curl virus (CaLCV) at 80.1 and 80.7%, respectively. BCMoV DNA-B shared highest nucleotide sequence identity with SLCV-E at 70.7%. The common region (CR) sequences of BCMoV and SLCV-E are 73 to 76% identical; however, modular cis-acting elements within the CR involved in replication origin function and recognition are 100% conserved. Phy-logenetic analysis indicated that BCMoV DNA-A shares a most recent common ancestor with the DNA-A of two viruses that also occur in the Sonoran Desert, SLCV-E and Texas pepper virus (TPV-TAM), and CaLCV from Florida. In contrast, a phylogenetic analysis indicated that BCMoV DNA-B shares a most recent common ancestor with SLCV-E; whereas DNA-B of CaLCV clustered in a separate clade with pepper hausteco virus. Collectively, biological and molecular characteristics indicate that BCMoV is a distinct begomovirus species with the northernmost distribution of any begomovirus isolated from bean in the Americas. Furthermore, the phylogenetic relationships of begomovirus cognate components are not necessarily identical, suggesting that DNA-A and DNA-B of some begomoviruses may have different evolutionary histories.

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