Host range and genetic diversity of croton yellow vein mosaic virus, a weed-infecting monopartite begomovirus causing leaf curl disease in tomato.

Croton yellow vein mosaic virus (CYVMV) is a widely occurring begomovirus in Croton bonplandianum, a common weed in the Indian subcontinent. In this study, CYVMV (genus Begomovirus, family Geminiviridae) was transmitted by whiteflies (Bemisia tabaci) to as many as 35 plant species belonging to 11 families, including many vegetables, tobacco varieties, ornamentals and weeds. CYVMV produced bright yellow vein symptoms in croton, whereas in all the other host species, the virus produced leaf curl symptoms. CYVMV produced leaf curl in 13 tobacco species and 22 cultivars of Nicotiana tabacum and resembled tobacco leaf curl virus (TobLCV) in host reactions. However, CYVMV was distinguished from TobLCV in four differential hosts, Ageratum conyzoides, C. bonplandianum, Euphorbia geniculata and Sonchus bracyotis. The complete genome sequences of four isolates originating from northern, eastern and southern India revealed that a single species of DNA-A and a betasatellite, croton yellow vein mosaic betasatellite (CroYVMB) were associated with the yellow vein mosaic disease of croton. The sequence identity among the isolates of CYVMV DNA-A and CroYVMB occurring in diverse plant species was 91.8-97.9 % and 83.3-100 %, respectively. The CYVMV DNA-A and CroYVMB generated through rolling-circle amplification of the cloned DNAs produced typical symptoms of yellow vein mosaic and leaf curling in croton and tomato, respectively. The progeny virus from both the croton and tomato plants was transmitted successfully by B. tabaci. The present study establishes the etiology of yellow vein mosaic disease of C. bonplandianum and provides molecular evidence that a weed-infecting monopartite begomovirus causes leaf curl in tomato.

Comparative epidemiology of three tropical whitefly-transmitted geminiviruses.

Spatial and temporal patterns of spread of African cassava mosaic, okra leaf curl and tobacco leaf curl viruses in West Africa, East Africa and India share some general characteristics. By comparing the results and running new analyses on available data, it is shown that the epidemiology of these viruses is controlled by the same key variables. For instance, spatial spread is characterised by strong border effects due to accumulation of whitefly vectors (Bemisia tabaci) on the wind-exposed field borders under the influence of the prevailing wind. This results in pronounced environmental gradients of disease. Temporal patterns of virus spread are driven by the sinusoidal fluctuation of temperature over the year which correspond with changes of whitefly populations.

Transmission of Pigeon pea sterility mosaic virus by the Eriophyid Mite, Aceria cajani (Acari: Arthropoda).

The transmission characteristics of Pigeon pea sterility mosaic virus (PPSMV) to pigeon pea (Cajanus cajan) by its eriophyid mite vector, Aceria cajani, were studied. Nonviruliferous A. cajani colonies were established on detached healthy leaflets of a PPSMV-immune pigeon pea cultivar floating on water. The transmission efficiency of single A. cajani was up to 53% but was 100% when >5 mites per plant were used. A. cajani acquired PPSMV after a minimum acquisition access period (AAP) of 15 min and inoculated virus after a minimum inoculation access period (IAP) of 90 min. No latent period was observed. Starvation of A. cajani prior to, or following, PPSMV acquisition reduced the minimum AAP and IAP periods to 10 min and 60 min, respectively, and mites retained virus for up to 13 h. None of the mites that developed from eggs taken from PPSMV-infected leaves transmitted the virus, indicating that it is not transmitted transovarially. Taken together, these data suggest a semipersistent mode of transmission of PPSMV by A. cajani.

First Report of the Bemisia tabaci B Biotype in India and an Associated Tomato leaf curl virus Disease Epidemic.

In May 1999, in the Kolar district of Karnataka State, Bemisia tabaci numbers on tomato increased by approximately 1,000-fold that observed previously (3). This was associated with an epidemic of severe tomato leaf curl disease that caused complete crop failure. DNAs extracted from 35 symptomatic tomato leaf samples collected within the epidemic region all gave the expected 500 to 600 bp amplicon with begomovirus-specific primers A/B (1). These primers amplify from the conserved nonanucleotide TAATATTAC in the common region of DNA-A to the conserved amino acid sequence CEGPCKYG within the coat protein gene. AluI and TaqI restriction patterns of all 35 polymerase chain reaction (PCR) products were identical. One PCR product from an epidemic (GenBank no. AF321929) and a non-epidemic (AF321930) site (Bangalore) were cloned and sequenced. The two 531-bp inserts showed 96% nucleotide identity to each other and 94% nucleotide identity to the equivalent region of Tomato leaf curl Bangalore virus (ToLCBV-Ban-4) (AF165098), suggesting that the epidemic was caused by an indigenous ToLCBV strain. Adult B. tabaci were collected from tomato plants at nine sites within the epidemic. DNA was extracted from 9 to 13 individuals per site and analyzed by RAPD-PCR using primers OpB20 and OpB11. Eighty to 100% of individuals per site had identical patterns to those of B biotype individuals from Israel and Florida, which were different to the patterns produced by the indigenous Indian B. tabaci. Adult B. tabaci from the epidemic and nonepidemic (Bangalore) regions were cultured separately on zucchini plants (n = 20) vars. Fordhook and Ambassador. Distinct silverleaf symptoms appeared in all plants fed on by the epidemic B. tabaci, but not on those fed on by the nonepidemic whiteflies. Irregular ripening of tomatoes was also a widespread problem in the epidemic area. Cytochrome oxidase I (COI) (720 bp) gene sequences were obtained for epidemic (AF321927) and nonepidemic (AF321928) B. tabaci, which had only 80% nucleotide identity to each other. A GenBank BLAST search showed that the former were most similar to B biotype whitefly from Israel (AF164667; 97%) and Texas (AF164675; 99%). The B biotype transmits Indian ToLCBV (2) and its introduction into India is of great concern as it is already associated with a devastating plant-disease epidemic. References: (1) D. Deng et al. Ann. App. Biol. 125:327, 1994. (2) P. F. McGrath and B. D. Harrison. Ann. App. Biol. 126:307, 1995. (3) H. K. Ramappa et al. Ann. App. Biol. 133:187, 1998.

Diversity and distribution of begomoviruses infecting tomato in India.

Leaf curl begomoviruses cause serious yield losses to Indian tomato crops. Total DNAs were extracted from leaves of 69 tomato plants and 34 weeds or neighbouring crops collected from all the major tomato producing areas of India. Eighty-one of the 103 samples were positive by PCRs using begomovirus genus-specific primers. Coat protein (CP) genes from 29 samples were PCR amplified, cloned and sequenced. Phylogenetic analyses of the CP sequences revealed five different tomato leaf curl begomovirus (TLCB) clusters each <88% identity to the others. Four clusters represented known Indian TLCBs, whereas one cluster contained sequences originating from Haryana State with most identity (89%) to the provisional Begomovirus species Croton yellow vein mosaic virus.Sixty-five begomovirus positive samples were characterised further by PCR with DNA-beta, DNA-B, four Indian TLCB species, PALIc1960/PARIv722 (universal begomovirus primers), and by sequencing. The majority of samples represented monopartite TLCBs associated with DNA-beta components. All four known TLCBs appeared to be present throughout India. TLCBs were also present in chilli, cowpea, okra and tobacco crops, as well as in some common weeds. Papaya leaf curl virus and Pepper leaf curl Bangladesh virus sequences were detected in tomato. Mixed begomovirus infections, a prerequisite for recombination, were evident in 13 samples.

Tomato leaf curl virus from Bangalore (ToLCV-Ban4): sequence comparison with Indian ToLCV isolates, detection in plants and insects, and vector relationships.

Tomato leaf curl virus (ToLCV) is a whitefly (Bemisia tabaci) transmitted geminivirus (family Geminiviridae, genus Begomovirus) causing a destructive disease of tomato in many regions of India, East Asia and Australia. While ToLCV isolates from Australia and Taiwan have a single genomic component (designated DNA-A), those from Northern India have two components (DNA-A and DNA-B). The ToLCV isolates from Southern India (Bangalore) previously cloned seem to have a DNA-A-like monopartite genome. We have used degenerate DNA-A-specific PCR primers to clone the genome of a ToLCV isolate (named ToLCV-Ban4) from field-infected tomato plants growing in Bangalore, India, in 1997. Degenerate DNA-B-specific PCR primers have not allowed to amplify a putative DNA-B from infected tomato, at the time when DNA-B fragments were amplified from plants infected by known bipartite begomoviruses. The full-length 2759 nucleotide-long DNA-A-like viral genome was sequenced. Similarly to other monopartite ToLCV and TYLCV isolates, ToLCV-Ban4 contains six open reading frames, two on the virion strand and four on the complementary strand. Sequence comparisons indicated that ToLCV-Ban4 is similar to the other three isolates from Bangalore previously sequenced, and is closely related to ToLCV-Ban2 (approximately 91% nucleotide sequence identity). Phylogenetic analysis showed that the ToLCV isolates from Bangalore constitute a group of viruses separated from those of Northern India. ToLCV-Ban4 was detected in tomato and in its whitefly vector Bemisia tabaci by one or by a combination of ELISA, Southern blot hybridization and PCR. Parameters of virus acquisition, retention and transmission by the whitefly vector were investigated in the laboratory. Single whiteflies were able to acquire ToLCV-Ban4 from infected tomato and to transmit the virus to tomato test plants, but five insects were necessary to achieve 100% transmission. Minimum acquisition access and inoculation access periods were 10 min and 20 min, respectively. A latent period of 6 h was required for B. tabaci to efficiently infect tomato test plants. Following a 24 h acquisition access period the insect retained its ability to infect tomato test plants for 12 days, but not for its entire life. In one insect/one plant inoculation tests, female whiteflies were more efficient (approximately 95%) than males (approximately 25%) in transmitting the virus.