Transmissibility of Citrus leprosis virus by Brevipalpus phoenicis to Solanum violaefolium.

Citrus leprosis virus (CiLV) constitutes one of the most important viruses in citrus in the areas where it occurs. Two morphological types of virus particles have been described from associated leprosis symptoms, nuclear (CiLV-N) and cytoplasmic (CiLV-C) (4). The CiLV-C is more common, representing more than 99% of samples collected from South and Central America (E. W. Kitajima and J. C. V. Rodrigues, unpublished). Both virus types are associated with the mite vector, Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae). So far, CiLV-C has only been naturally transmitted by these mites to citrus (3). Plants of Solanum violaefolium Schott (Solanaceae) and ornamental and sweet orange seedlings were infested with viruliferous adult female mites, colony no. 61 (GenBank Accession No. AY320027) that were previously maintained on citrus seedlings infected with CiLV-C according to Rodrigues et al. (3). Fifteen days after the mites were transferred, spotted yellowish symptoms were observed on leaves of plants of S. violaefolium and similar symptoms were observed after 25 days on citrus leaves. The symptomatic tissues were studied using transmission electron microscopy. Particles typical of CiLV-C were observed in samples from both plant species. To our knowledge, this is the first reported case of mites transmitting CiLV-C to a noncitrus host. Subsequent experiments showed that mites were able to transmit the virus between plants of S. violaefolium. Attempts to transmit the virus by mites from S. violaefolium to citrus were unsuccessful. The dsRNA viral electrophoresis profile showed differences between the two host plants. Reverse transcription-polymerase chain reaction (RT-PCR) and sequencing assays with primers designed to detect CiLV-C (2) amplified DNA fragments of the expected size and base composition. These data suggest the loss or alteration of some viral components from the Solanum sp. host that might be essential for the transmission or infection in citrus. Such a mechanism may explain why, despite the ever-increasing number of Brevipalpus-transmitted viruses in a large number of different host plant species (1), cross transmission is not common. This information shows one of the potential routes for CiLV to invade citrus orchards, and suggests one alternative host plant that allows rapid multiplication of the virus for characterization. References: (1) E. W. Kitajima et al. Exp. Appl. Acarol. 30:135, 2003. (2) E. C. Locali et al. Plant Dis. 87:1317, 2003. (3) J. C. V. Rodrigues et al. Proc. Int. Org. Citrus Virol. 174, 2000 (4) J. C. V. Rodrigues et al. Exp. Appl. Acarol. 30:161, 2003.

First Report of Citrus leprosis virus on Citrus in Santa Cruz, Bolivia.

The citrus crop is rapidly expanding in the Province of Santa Cruz de la Sierra, Bolívia. Citrus, mostly planted by small growers, currently comprises approximately 15,000 ha. Sweet oranges (Citrus sinensis) and mandarins (C. reticulate) are the main citrus-types grown primarily for internal consumption. Recently, there has been an increase in incidence of leprosis-like symptoms (round to elliptic lesions on the leaves, chlorotic to necrotic lesions in young twigs, and depressed small lesions on the fruits). These symptoms were associated with infestations by the tenuipalpid mite Brevipalpus sp. To verify if Citrus leprosis virus was the causal agent of the observed symptoms, leaf and fruit samples (mostly from Valencia sweet orange) were collected from commercial groves in El Torno, 32 km south of Santa Cruz, and Yapacani and Colónia San Juan, 130 km northwest of Santa Cruz. Small fragments of these samples were placed immediately in a mixture of glutaraldehyde and paraformaldehyde in cacodylate buffer and later processed with transmission electron microscopy at ESALQ, Piracicaba, SP, Brazil. Some of the leaf samples were dried at 35°C and used for reverse transcription-polymerase chain reaction (RT-PCR) with primers that specifically amplify portions of the genome of Citrus leprosis virus, cytoplasmic type (CiLV-C) (1) at Centro APTA Citros, Cordeirópolis, SP, Brazil. Brevipalpus sp. mites were also collected and kept in 90% ethanol for further identification at the University of Florida, Gainesville and ESALQ. In the samples from the three surveyed areas, transmission electron microscopy confirmed the presence of short bacilliform particles within endoplasmic reticulum cisternae and electron dense viroplasms in the cytoplasm, typical of infection by CiLV-C (2). CiLV-C specific primers amplified DNA fragments of expected sizes in RT-PCR from dried leaf samples that came from these three localities. Direct sequencing of at least three amplicons of each sample confirmed the identity of the virus. The consensus sequence of the putative movement protein gene in samples from Yapacani and Colónia San Juan (GenBank Accessions Nos. AY960216 and AY960215, respectively) were identical and exhibited 99% nucleotide and 98% amino acid homology with the Brazilian isolate sequence available at GenBank (Accession No. AY289190). The consensus sequence of the putative replicase gene found in the sample from El Torno (GenBank Accession No. AY960214) exhibited 96 and 93% nucleotide and amino acid homology, respectively with the Brazilian isolate (GenBank Accession No. AY289191). Sampled mites were identified as B. phoenicis (Geijskes), the known vector of CiLV-C (2). The symptomatology, particle morphology and cytopathology, detection by molecular methods and the association with infestation by B. phoenicis, together indicate that the foliar, stem, and fruit lesions in sweet orange observed in the Santa Cruz region were caused by CiLV-C. To our knowledge, this is the first report of this virus in Bolivia. References: (1) E. C. Locali et al. Plant Dis. 87:1317, 2003. (2) J. C. V. Rodrigues et al. Exp. Appl. Acarol. 30:161, 2003.

Comparison of the genomes of two Xanthomonas pathogens with differing host specificities.

The genus Xanthomonas is a diverse and economically important group of bacterial phytopathogens, belonging to the gamma-subdivision of the Proteobacteria. Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, which affects most commercial citrus cultivars, resulting in significant losses worldwide. Symptoms include canker lesions, leading to abscission of fruit and leaves and general tree decline. Xanthomonas campestris pv. campestris (Xcc) causes black rot, which affects crucifers such as Brassica and Arabidopsis. Symptoms include marginal leaf chlorosis and darkening of vascular tissue, accompanied by extensive wilting and necrosis. Xanthomonas campestris pv. campestris is grown commercially to produce the exopolysaccharide xanthan gum, which is used as a viscosifying and stabilizing agent in many industries. Here we report and compare the complete genome sequences of Xac and Xcc. Their distinct disease phenotypes and host ranges belie a high degree of similarity at the genomic level. More than 80% of genes are shared, and gene order is conserved along most of their respective chromosomes. We identified several groups of strain-specific genes, and on the basis of these groups we propose mechanisms that may explain the differing host specificities and pathogenic processes.