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Infected with Sweet Potato Leaf Curl Virus Revista Mexicana De Fitopatología, Vol Revista Mexicana de Fitopatología ISSN: 0185-3309 [email protected] Sociedad Mexicana de Fitopatología, A.C. México Valverde, Rodrigo A.; Clark, Christopher A.; Fauquet, Claude M. Properties of a Begomovirus Isolated from Sweet Potato [Ipomoea batatas (L.) Lam.] Infected with Sweet potato leaf curl virus Revista Mexicana de Fitopatología, vol. 21, núm. 2, julio-diciembre, 2003, pp. 128-136 Sociedad Mexicana de Fitopatología, A.C. Texcoco, México Available in: http://www.redalyc.org/articulo.oa?id=61221206 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative 128 / Volumen 21, Número 2, 2003 Properties of a Begomovirus Isolated from Sweet Potato [Ipomoea batatas (L.) Lam.] Infected with Sweet potato leaf curl virus Pongtharin Lotrakul, Rodrigo A. Valverde, Christopher A. Clark, Department of Plant Pathology and Crop Physiology, Louisiana Agricultural Experiment Station, Louisiana State University Agricultural Center, Baton Rouge, Louisiana 70803, USA; and Claude M. Fauquet, ILTAB/Donald Danford Plant Science Center, UMSL, CME R308, 8001 Natural Bridge Road, St. Louis, MO 63121, USA. GenBank Accession numbers for nucleotide sequence: AF326775. Correspondence to: [email protected] (Received: November 6, 2002 Accepted: February 12, 2003) Lotrakul, P., Valverde, R.A., Clark, C.A., and Fauquet, C.M. potato leaf curl virus (SPLCV). Por medio de la reacción en 2003. Properties of a Begomovirus isolated from sweet potato cadena de la polimerasa (PCR), utilizando oligonucleótidos [Ipomoea batatas (L.) Lam.] infected with Sweet potato leaf específicos para SPLCV, se confirmó la presencia de SPLCV. curl virus. Revista Mexicana de Fitopatología 21:128-136. El análisis de clones con la secuencia parcial del gene AC1, Abstract. Scions from the sweet potato (Ipomoea batatas) indicó que el genotipo GA90-16 estaba doblemente infectado breeding line GA90-16 graft inoculated to I. setosa caused con SPLCV y un segundo geminivirus, que resultó symptoms similar to those caused by the begomovirus, Sweet emparentado con el primero, al que proponemos designar potato leaf curl virus (SPLCV). Polymerase chain reaction como Ipomoea leaf curl virus (ILCV). Se efecturaron (PCR) using SPLCV-specific primers confirmed the presence transmisiones individuales con moscas blancas (Bemisia of SPLCV. Analyses of partial clones of the AC1 open reading tabaci biotipo B) para separar ambos virus, los cuales fueron frame sequences indicated that GA90-16 was doubly infected diferenciados, analizando los productos de PCR (AV1 OR), with SPLCV and a related geminivirus, which we propose to después de un ensayo de restricción utilizando la enzima designate as Ipomoea leaf curl virus (ILCV). Individual EcoR1. A diferencia de SPLCV, ILCV no causó whitefly (Bemisia tabaci biotype B) transmission was amarillamiento de las venas en I. aquatica e I. cordatotriloba, conducted to separate these two viruses which could be pero causó enrollamiento de las hojas en varias especies de differentiated by restriction enzyme digestion of PCR (AV1 Ipomoea. El componente A de ILCV fue clonado y OR) products utilizing EcoR1. ILCV caused leaf curl secuenciado completamente. La organización del genoma symptoms in several Ipomoea species. Unlike SPLCV, ILCV de este virus es similar a los begomovirus monopartitas del did not cause yellow vein symptoms on I. aquatica and I. Viejo Mundo. La comparación de las secuencias nucleotídicas cordatotriloba. DNA A of ILCV was cloned and sequenced. de los componentes A de ambos virus, demostró que ILCV The genome organization of the virus was typical of other es similar a SPLCV y a Ipomoea yellow vein virus (IYVV), Old World monopartite begomoviruses. Sequence que infecta a una especie de Ipomoea en España, con comparisons showed that ILCV was similar to SPLCV and aproximadamente 76 y 78% de similitud, respectivamente. to another Ipomoea-infecting begomovirus, Ipomoea yellow Por otro lado, la comparación de la secuencia del gene de la vein virus (IYVV) from Spain with about 76 and 78% proteína de la cápside del ILCV, resultó casi idéntica a SPLCV nucleotide sequence identity, respectively. Although the coat e IYVV, mientras las secuencias de la región común y de los protein of ILVC was nearly identical to that of SPLCV and genes AC1, AC2, AC3 y AC4 fueron lo suficientemente IYVV, the sequence of the common region, and the AC1, diferentes, para proponer al ILCV como una nueva especie AC2, AC3, and AC4 ORFs were sufficiently different to perteneciente al género Begomovirus. warrant the nomination of a new begomovirus species. Palabras clave adicionales: Bemisia tabaci, Geminivirus, Additional keywords: Bemisia tabaci, Geminivirus, Ipomoea Ipomoea yellow vein virus, PCR, mosca blanca. yellow vein virus, PCR, whitefly. The family Geminiviridae comprises a group of plant viruses Resumen. Inoculaciones mediante injertos del genotipo de which have small twin- isometric particles containing closed camote (Ipomoea batatas) GA90-16 a I. setosa, causaron circular, single-stranded DNA genomes (Hanley-Bowdoin et síntomas similares a los causados por el begomovirus Sweet al., 1999). This family can be classified into four genera Revista Mexicana de FITOPATOLOGIA/ 129 based on their hosts, insect vector, and genome organization. one SalI (in the case of ILCV) and one Eco RV (in the case of Members of the genus Begomovirus are those containing SPLCV) restriction site. Thus, the two viruses could be mono- or bipartite genomes which infect dicotyledonous differentiated by digesting the amplified DNAs with these plants, and can be transmitted by Bemisia tabaci. Viruses in two enzymes. the genus Mastrevirus infect mainly monocots, have Whitefly transmissions. Colonies of the sweet potato monopartite genomes, and are transmitted by leafhoppers. whitefly (B. tabaci biotype B) were reared in the laboratory Curtoviruses infect a wide range of dicotyledonous plants, as described previously (Lotrakul et al., 2000). Individual have monopartite genomes and are transmitted by whiteflies were used in all transmission experiments except leafhoppers. The genus Topocuvirus has only one member, in host range trials. Transmission experiments were conducted Tomato pseudo-curly top virus (TPCTV) which has a in the laboratory using SO as the acquisition and transmission monopartite genome and is transmitted by tree hoppers to host. For some transmission experiments, SO doubly infected dicotyledonous plants. During the spring of 2000, sweet with SPLCV and ILCV was used as virus acquisition host. potato [Ipomoea batatas (L.) Lam.] cuttings from research Additional transmission experiments were conducted with plots in Athens, GA, were collected for a virus disease survey. the individual virus isolates. Groups of 25 whiteflies were Scions were grafted to I. setosa Ker. (Brazilian morning glory) used in attempts to transmit ILCV to other plant species. In to detect virus infection. A scion from the sweet potato all experiments, whiteflies were caged with infected SO plants breeding line GA90-16 caused leaf curling symptoms on I. for 48 h and then transferred to caged healthy plants for 48 h. setosa similar to those caused by the United States isolate of Test plants were then sprayed with imidacloprid and Sweet potato leaf curl virus (SPLCV) (Lotrakul et al., 1998). transferred to the greenhouse. Plants were evaluated for visual Polymerase chain reaction (PCR) assays confirmed the symptoms 2-3 wk later. PCR amplification using SPLCV- presence of SPLCV in the infected plants. After sequence specific primers was performed to confirm the presence of analysis of the PCR products (partial AC1 ORF), two distinct the virus in tested plants. sequences were consistently obtained. One was nearly Host range. In order to determine the host range of ILCV, identical to that of SPLCV whereas the other was distinct. graft and whitefly inoculations were conducted. In all cases, This suggested a double infection of SPLCV and a related at least three plants of the same species were inoculated. Graft geminivirus which was provisionally designated Ipomoea leaf inoculations were conducted using infected SO as source of curl virus (ILCV). In this investigation, ILCV was separated scions to inoculate I. alba L., I. aquatica, I. batatas (cvs from a double infection with SPLCV by an individual whitefly Beauregard, Excel, Jonathan, L94-96, NC-262, Regal, transmission and the biological and molecular properties were Resisto, Sumor, Wagabolige, Tanzania, W244, W-285, and determined and compared with those of SPLCV. Xushu-18), I. cordatotriloba Ell., I. hederaceae (L.) Jacq., I. incarnata (Vahl) Choisy, I. lacunosa L., I. setosa, and I. MATERIALS AND METHODS tiliaceae Willd. Groups of 25 whiteflies were used in each Plant materials and virus isolates. Cuttings from the attempt to transmit ILCV to Nicotiana benthamiana Domin., breeding line GA90-16 were collected from the field, rooted, Datura stramonium L., Physalis ixocarpa Brot., and and maintained in a greenhouse under natural light. Scions Capsicum frutescens L. cv Tabasco. Preliminary tests from these plants were grafted to I. setosa and PCR was indicated that I. aquatica and I. cordatotriloba could be used performed as described previously (Lotrakul, 2000). Sweet to differentiate SPLCV from ILCV. In order to confirm these potato
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