No abstract

More than 1200 cultivars, breeding lines, and wild species of Lycopersicon were screened for resistance to the tomato leaf curl virus under field conditions. All accessions of Lycopersicon esculentum were found susceptible to tomato leaf curl virus. Three lines of Lycopersicon hirsutum and one of Lycopersicon peruvianum accessions showed apparent resistance to the tomato leaf curl virus owing to the failure of virus transmission by whiteflies. Morphological analysis by light and scanning electron microscopy revealed distributions of different types of trichomes on leaf surfaces. Lycopersicon hirsutum proved to be the least affected by the leaf curl virus because it did not support whitefly infestation to any extent. Lycopersicon hirsutum differed from other species by the presence of glandular trichomes designated type VIc. Whiteflies became entrapped in the exudate of type VIc trichome glands before they could transmit the virus. Thus, it may be possible to control tomato leaf curl virus transmission by breeding plants with certain types of trichomes, especially trichome type VIc. Key words: leaf curl virus, nonpreference, physical resistance, glandular exudate, trichomes, whitefly.

Tomato yellow leaf curl virus (TYLCV), a heterogeneous complex of whitefly-vectored geminiviruses, is a serious production constraint of tomato (Lycopersicon esculentum Mill.) in Asia, the Middle East, and the Americas. In this study we report on mapping of a DNA fragment introgressed into cultivated tomato presumably from the wild species L. hirsutum Humb. and Bonpl. and found to be associated with TYLCV resistance. To locate introgressions of wild tomato alleles in TYLCV-resistant tomato line H24, its DNA was digested with six restriction enzymes and probed with 90 RFLP markers evenly spaced throughout the genome. This polymorphism survey revealed the presence of one wild tomato introgression each on chromosomes 8 and 11. Plants of a F2 cross between H24 and a susceptible tomato line were probed with randomly amplified polymorphic DNA (RFLP) markers linked to the targeted regions and F3 families were developed by self-pollination of F2 plants that carried none, one, or both introgressions in either homozygous or heterozygous states. Plants of F3 families, parents, and control tomato line Ty52 (homozygous for the Ty-1 allele for TYLCV tolerance) were exposed to viruliferous whiteflies (Bemisia tabaci Gennadius) in greenhouses at the Asian Vegetable Research and Development Center, Taiwan, and the University of Agricultural Sciences, Bangalore, India. Results indicated that F3 families homozygous for the introgression on chromosome 11 were resistant to TYLCV at both locations. Additional probing showed that the chromosome 11 introgression spanned markers TG36 to TG393, covering a distance of at least 14.6 centimorgans. This is the first report of TYLCV resistance in tomato mapped to chromosome 11.

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.

The whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae), is generally considered to have originated from the Indian subcontinent, although little information has so far been collected on the molecular diversity of populations present in this region. The genetic diversity of B. tabaci populations from Karnataka State, south India was analysed using the random amplified polymorphic DNA‐polymerase chain reaction (RAPD‐PCR) technique and partial mitochondrial cytochrome oxidase I (mtCOI) gene sequences (689 bases) of 22 selected samples. A total of 108 whitefly samples analysed by RAPD‐PCR produced 89 polymorphic bands, and cluster analyses grouped them according to their geographic origin into ‘north’ and ‘south’ Karnataka. Phylogenetic analysis of mtCOI gene sequences with reference B. tabaci sequences from other Asian countries divided them into three genotypic clusters. Each cluster was supported with high bootstrap values (82–100%) and the individuals belonging to each cluster shared high nucleotide identities (up to 100%). This indicated at least three distinct genotypes, apparently indigenous to India, which are also present in China, Malaysia, Nepal, Pakistan, and Thailand. These coexist with the B biotype, which was first reported in India in 1999, and has since spread rapidly to other states in south India. The B biotype was more common than the indigenous B. tabaci, in locations where it had been present for more than 2 years. This is reminiscent of the situation in the Americas during the early 1990s, where the B biotype replaced existing biotypes and caused unprecedented losses to agriculture.