Molecular Characterization of Tomato and Chili Leaf Curl Begomoviruses from Pakistan

Abstract

Leaf curl or yellowing symptoms, typical of those caused by begomovirus infection, are commonly observed in chili (Capsicum annuum) and tomato (Lycopersicon esculentum) plantings in Pakistan. One chili sample with leaf curl symptoms was collected in 1998 in Multan (Punjab Province), and two tomato samples with leaf curl and yellowing symptoms were collected from Islamabad and Dargai (North West Frontier Province) in 2000 and 2001, respectively. Virus DNA was first amplified by polymerase chain reaction using the degenerate primer pair PAL1v1978/PAR1c715 (3). The expected 1.4-kb PCR products were obtained from the three samples. Based on the sequences of the 1.4-kb DNA products, specific primers were designed to complete each of the DNA-A sequences. Two primer pairs, DNABLC1/DNABLV2 and DNABLC2/DNABLV2, were used for the detection of DNA-B (2). The genome of the tomato leaf curl isolate from Islamabad contained a DNA-A of 2,739 nucleotides (GenBank Accession No. AF448059), a DNA-B of 2,728 nucleotides (GenBank Accession No. AY150304), and had 94% nucleotide identity in the common region. The genome of the tomato leaf curl isolate from Dargai contained a DNA-A of 2,740 nucleotides (GenBank Accession No. AF448058), a DNA-B of 2,686 nucleotides (GenBank Accession No. AY150305), and had 96% nucleotide identity in the common region. Each of the tomato isolates contained eight predicted open-reading frames (ORFs) (AV1, AV2, AV3, AC1, AC2, AC3, AC4, and AC5) in the DNA-A and two predicted ORFs (BV1 and BC1) in the DNA-B. The DNA-A nucleotide sequence identity of the Islamabad isolate and Dargai tomato isolate is 96% and that of DNA-B is 88%. Sequence comparisons with begomovirus sequences available in the GenBank sequence database showed that these two tomato virus isolates had the highest sequence identity with Tomato leaf curl New Delhi virus-Severe (GenBank Accession No. U15015) from northern India (more than 95% for DNA-A and less than 90% for DNA-B). The DNA-A of the virus associated with chili leaf curl from Pakistan (GenBank Accession No. AF336806) consists of 2,754 nucleotides, containing six predicted ORFs (AV1, AV2, AC1, AC2, AC3, and AC4). The chili virus was unrelated to the two tomato begomovirus isolates from Pakistan, with which it shares less than 75% nucleotide identity. Sequence comparisons show highest sequence identity (87%) with Tomato leaf curl Bangladesh virus (GenBank Accession No. AF188481). DNA-beta of 1.3 kb was detected in the chili begomovirus isolate using Beta01/Beta02 primers (1). There was no evidence for the presence of a DNA-B in the chili begomovirus isolate when tested by the two DNA-B specific primer pairs. Based on DNA sequence comparisons, the chili leaf curl virus from Pakistan, to our knowledge, constitutes a distinct, new monopartite begomovirus. References: (1) R. W. Briddon et al. Mol. Biotechnol. 20:315, 2002. (2) S. K. Green et al. Plant Dis. 85:1286, 2001. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993.

Genetic diversity and distribution of a distinct strain of Chili leaf curl virus and associated betasatellite infecting tomato and pepper in Oman

“…The results presented here show that a fourth species of begomovirus, ChLCV, also causes the disease in Oman. ChLCV has previously only been reported in Pakistan and India (Senanayake et al, 2006(Senanayake et al, , 2012Shih et al, 2003;Mubin et al, 2009).…”

“…Chili leaf curl disease was first recorded in 1960s in India (Mishra et al, 1963;Dhanraj and Seth, 1968). Later a begomovirus, now known as ChLCV, was shown to be associated with leaf curl disease of pepper in Pakistan, India and Oman (Shih et al, 2003;Khan et al, 2006;Senanayake et al, 2006). In common with all monopartite begomoviruses, ChLCV encodes six genes; two in the virion-sense and four in the complementary-sense.…”

IDM Module for the Management of Leaf Curl Disease in Chilli

“…The chilli leaf curl virus (CLCV) disease on chilli was first reported in Pakistan by Shih et al, (2003) and in India by Senanayake et al, (2006). Chilli crop suffers from a large number of viral, fungal, bacterial, nematode and phytoplasma diseases.…”

A Single Species of Betasatellite is Prevalent in Chilli across North Central Pakistan and Shows Phylogeographic Segregation

“…Among the factors responsible for the low yields of chillies across the Indian subcontinent are diseases of viral aetiology. The most important viral disease of chillies, chilli leaf curl disease, is caused by begomoviruses in association with a symptom modulating satellite (Briddon et al., 2003; Shih et al., 2003).…”