The pepper L gene conditions the plant's resistance to Tobamovirus spp. Alleles L(1), L(2), L(3), and L(4) confer a broadening spectra of resistance to different virus pathotypes. In this study, we report the genetic basis for the hierarchical interaction between L genes and Tobamovirus pathotypes. We cloned L(3) using map-based methods, and L(1), L(1a), L(1c), L(2), L(2b), and L(4) using a homology-based method. L gene alleles encode coiled-coil, nucleotide-binding, leucine-rich repeat (LRR)-type resistance proteins with the ability to induce resistance response to the viral coat protein (CP) avirulence effectors by themselves. Their different recognition spectra in original pepper species were reproduced in an Agrobacterium tumefaciens-mediated transient expression system in Nicotiana benthamiana. Chimera analysis with L(1), which showed the narrowest recognition spectrum, indicates that the broader recognition spectra conferred by L(2), L(2b), L(3), and L(4) require different subregions of the LRR domain. We identified a critical amino acid residue for the determination of recognition spectra but other regions also influenced the L genes' resistance spectra. The results suggest that the hierarchical interactions between L genes and Tobamovirus spp. are determined by the interaction of multiple subregions of the LRR domain of L proteins with different viral CP themselves or some protein complexes including them.

Thirty-seven Capsicum accessions containing cultivated and wild species were screened for resistance to Cucumber mosaic virus (CMV), and were also investigated for their response to Tomato aspermy virus (TAV), Tomato mosaic virus (ToMV), Pepper mild mottle virus (PMMoV), and Tomato spotted wilt virus (TSWV). C. baccatum PI 439381-1-3 (PI 439381-1-3), C. frutescens LS 1839-2-4 (LS 1839-2-4), and C. frutescens cv. Tabasco (cv. Tabasco) showed a hypersensitive reaction against CMV-Y, and thus were not systemically infected. Only inoculated leaves of C. annuum cv. Sapporo-oonaga and cv. Nanbu-oonaga were infected with CMV-Y, and viral infection did not spread systemically. These five accessions (PI 439381-1-3, LS 1839-2-4, cv. Tabasco, cv. Sapporo-oonaga, and cv. Nanbu-oonaga) were considered resistant to CMV-Y. These accessions were also resistant to other CMV isolates, but not to the TAV isolate. PI 439381-1-3, LS1839-2-4, cv. Sapporo-oonaga, and cv. Nanbu-oonaga were susceptible to PMMoV, while PI 439381-1-3 and LS1839-2-4 showed systemic necrosis. All CMV-resistant accessions were susceptible to TSWV. Field tests of eight Capsicum accessions, including CMV, PMMoV, and/or TSWV-resistant accessions, demonstrated that most of the PI 439381-1-3 plants were not infected with CMV and PMMoV among the virus-infested fields. As occurred with mechanical inoculation, LS 1839-2-4, cv. Tabasco, cv. Sapporo-oonaga, and cv. Nanbu-oonaga were hard to infect with CMV in the field.

The tobamovirus resistance gene L 3 of Capsicum chinense was mapped using an intra-specific F2 population (2,016 individuals) of Capsicum annuum cultivars, into one of which had been introduced the C. chinense L 3 gene, and an inter-specific F2 population (3,391 individuals) between C. chinense and Capsicum frutescence.Analysis of a BAC library with an AFLP marker closely linked to L 3 -resistance revealed the presence of homologs of the tomato disease resistance gene I2. Partial or fulllength coding sequences were cloned by degenerate PCR from 35 different pepper I2 homologs and 17 genetic markers were generated in the inter-specific combination. The L 3 gene was mapped between I2 homolog marker IH1-04 and BAC-end marker 189D23M, and located within a region encompassing two different BAC contigs consisting of four and one clones, respectively. DNA fiber FISH analysis revealed that these two contigs are separated from each other by about 30 kb. DNA fiber FISH results and Southern blotting of the BAC clones suggested that the L 3 locus-containing region is rich in highly repetitive sequences. Southern blot analysis indicated that the two BAC contigs contain more than ten copies of the I2 homologs. In contrast to the inter-specific F2 population, R. Tomita and J. Murai contributed equally to this work. Communicated by J.S. (Pat) Heslop-Harrison.Electronic supplementary material The online version of this article (doi:10.1007/s00122-008-0848-6) contains supplementary material, which is available to authorized users. 123Theor Appl Genet (2008) 117:1107-1118 DOI 10.1007 no recombinant progeny were identified to have a crossover point within two BAC contigs consisting of seven and two clones in the intra-specific F2 population. Moreover, distribution of the crossover points differed between the two populations, suggesting linkage disequilibrium in the region containing the L locus.