Swinglea glutinosa (Blanco) Merr., a perennial plant in the family Rutaceae, is originally from southeast Asia but which is now grown worldwide. In Colombia, it is used as an ornamental and principally as a living fence around rural properties and farms in several regions of the country. Citrus leprosis virus cytoplasmic type (CiLV-C) was recently detected in orange groves of the Colombian Piedmont eastern plains, an area known as the Llanos Orientales (2). Because of the potential for country-wide infection of citrus, some measures are being taken to avoid CiLV-C spread to other regions of Colombia. Further surveys made from June to December 2005 to evaluate the extent of the spread of CiLV-C in the Llanos Orientales revealed some plants in S. glutinosa hedges surrounding citrus orchards exhibiting chlorotic spots and ringspots of varied size on the leaves, similar to those caused by CiLV-C on sweet oranges leaves. These plants were found near citrus orchards in the municipalities of Guamal and in some urban areas of Villavicencio City in the Meta Department. The possibility that these symptoms were caused by CiLV-C was investigated soon after sample collection by the same procedures as described previously for sweet orange (2). In the leaf lesions of S. glutinosa, typical bacilliform particles and dense cytoplasmic viroplasm were found with electron microscopy. Total RNA extracted from symptomatic leaves was subjected to reverse transcription-PCR (RT) using primers (Fwd. 5′GATACGGGACGCATAACA-3′/Rev. 5′-TTCTGGCTCAACATCTGG-3′) that specifically amplify a region within the CiLV-C putative methyltransferase gene and this yielded a single fragment of the expected 402 bp (3). Analysis of the consensus sequence derived from 20 RT-PCR products (GenBank Accession No. EU689106) showed 96% nucleotide and 92% amino acid sequence identity to the sequence of a Brazilian CiLV-C isolate (GenBank Accession Nos. DQ352194.1 and YP_654565.1), respectively. Recently, published work described mite transmission of CiLV-C to some nonrutaceous plants (1), but to our knowledge, this is the first report of a nonCitrus rutaceous plant naturally infected by CiLV-C. Mites found in citrus orchards and previously identified as Brevipalpus phoenicis (Geijskes) (2), which are likely the most important vector of CiLV-C in citrus in Colombia, were observed feeding on healthy and symptomatic S. glutinosa, indicating that S. glutinosa is a host for B. phoenicis. Because the use of S. glutinosa as a living fence or hedge is a common practice in Colombia, CiLV-C-infected S. glutinosa plants may play a role in the epidemiology of leprosis in commercial citrus by serving as an inoculum source for this lethal virus. References: (1) M. Bastianel et al. Summa Phytopathol. 32:211, 2006. (2) G. A. León et al. Plant Dis. 90:682, 2006. (3) E. C. Locali et al. Plant Dis. 87:1317, 2003.

turer's instruction. The cDNA was then used as template in the PCR assay using primers CMLV-5F (5'-GGATCCATGTCGGCGCGATTGAATC-3', sequence in italics indicates restriction site BamHT) and CMLV-3R, which amplify the genome fragment including the capsid protein gene of CMLV. The expected PCR product ~590 bp was amplified from all five sympto matic samples, while no such PCR product was amplified from the symp tomless samples. The PCR products were cloned into pMD18-T vector (TaKaRa, Dalian, China). Three positive clones for each of the five amplicons were sequenced in both directions. Sequence alignment and nucleo tide BLAST analysis of the sequences revealed that they were 99% to 100% identical to the corresponding capsid protein gene sequence of a cherry isolate of CMLV (GenBank Accession No. AF170028) and 85% identical with that of the peach wart strain of CMLV (KC207480). Our results confirm the infection of cherry trees by CMLV in Shandong. To our knowledge, this is the first report of CMLV on cherry in China. As the spread of CMLV by mite vector in the field is rare (1), and no bud mite outbreak had occurred in this orchard in the past years, so it is possible that virus-infected propagation materials are largely responsible for the spread of this virus. Considering the importance of cherry cultivation in China, this report prompts the need to survey the occurrence of this virus in Shandong and other provinces, and the need to develop more effective management strategies such as the use of certified virus-free nursery stocks to reduce the impact of CMLV.

CiLV-C is a quarantine pest which causes an economically important disease, reported only on the American continent. During the past 15 years, it has caused economic losses in Brazil, Argentina, Paraguay, Uruguay, Venezuela, Costa Rica, Panamá and Honduras. The disease was recently reported in Guatemala, Bolivia, México, Colombia and Belize. It is a threat to citrus-producing countries where the disease has not been reported. The disease can cause 100% yield loss (Rodrigues et al., 2000).

The newly discovered Citrus leprosis virus cytoplasmic type 2 (CiLV-C2) is one of the causal virus of citrus leprosis disease complex; which leads to substantial loss of citrus production in the states of Meta and Casanare of Colombia. Specific and sensitive detection methods are needed to monitor the dissemination of CiLV-C2 in Colombia, and to prevent introduction of CiLV-C2 to other citrus growing countries. Toward this end, putative coat protein gene (CPG) of CiLV-C2 was amplified from CiLV-C2 infected citrus tissues. The CPG was cloned, expressed and purified a recombinant coat protein of ∼31kDa which used to generate monoclonal antibodies and polyclonal antisera. Four monoclonal antibodies and two polyclonal antisera were selected as being specific following Western blotting. The monoclonal antibody MAb E5 and polyclonal antiserum PAb UF715 were selected testing with an extract of CiLV-C2 infected leaves using triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA). In addition, an immunocapture RT-PCR was standardized using MAb E5 for specific and sensitive detection of CiLV-C2. The standardized TAS-ELISA and IC-RT-PCR were able to detect CiLV-C2 in the extracts of symptomatic citrus leprosis tissues up to the dilutions of 1:160 and 1:2580, respectively. Result demonstrated that CiLV-C2 is present in citrus orchards in Meta and Casanare citrus growing areas of Colombia. TAS-ELISA could be used for routine detection of CiLV-C2, epidemiological studies, and for border inspections for quarantine purposes. IC-RT-PCR could be valuable for CiLV-C2 validation and viral genome analysis.

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