ANNOTATED SEQUENCE RECORD Molecular characterization of a new begomovirus associated with leaf yellow mosaic disease of Jatropha curcas in India Ashish Srivastava • S. Kumar • Meraj Jaidi • S. K. Raj Received: 8 December 2014 / Accepted: 15 February 2015 Ó Springer-Verlag Wien 2015 Abstract During a survey in June 2011, severe leaf yel- low mosaic disease was observed on about 45 % plants of Jatropha curcas growing in the Katerniaghat wildlife sanctuary in India. An association of a begomovirus with disease was detected in 15 out of 20 samples by PCR using begomovirus genus-specific primers and total DNA iso- lated from symptomatic leaf samples. For identification of the begomovirus, the complete genome was amplified us- ing a Phi-29 DNA-polymerase-based rolling-circle ampli- fication kit and total DNA from five representative samples and then digested with BamHI. The linearized RCA prod- ucts were cloned and sequenced. Their GenBank accession numbers are JN698954 (SKRK1) and JN135236 (SKRK2). The sequences of the two begomovirus isolates were 97 % identical to each other and no more than 86 % to those of jatropha mosaic India virus (JMIV, HM230683) and other begomoviruses reported worldwide. In phylogenetic ana- lysis, SKRK1 and SKRK2 clustered together and showed distant relationships to jatropha mosaic India virus, Jatro- pha curcas mosaic virus, Indian cassava mosaic virus, Sri Lankan cassava mosaic virus and other begomoviruses. Based on 86 % sequence identities and distant phyloge- netic relationships to JMIV and other begomoviruses and the begomovirus species demarcation criteria of the ICTV ( \ 89 % sequence identity of complete DNA-A genome), the begomovirus isolates associated with leaf yellow mosaic disease of J. curcas were identified as members of a new begomovirus species and provisionally designated as jatropha leaf yellow mosaic Katerniaghat virus (JLYMKV). Agroinfectious clones of the DNA molecule of the begomovirus isolate were also generated, and the fulfillment of Koch’s postulates was demonstrated in J. curcas plants. Jatropha curcas (family Euphorbiaceae) is a shrub plant that is grown mainly for biodiesel production in India. It survives in various climatic conditions and has the ability to grow in almost any type of soil, especially on drought soil, and produce 30-40 % of its seed weight in biodiesel [1]. Widespread mosaic disease of J. curcas caused by begomoviruses was first investigated in 2004 in southern India (Karnataka) [2, 3] and then in northern India [4, 5]. Since then, many new begomoviruses have been reported on J. curcas from India and other countries [6–10]. Based on earlier reports of begomoviruses on Jatropha and the widespread cultivation of this plant in India, the Jatropha plots at wild life sanctuary, Katerniyaghat, Uttar Pradesh, India were surveyed during June, 2011 for bego- movirus-disease-like symptoms. Severe leaf yellow mosaic disease was noted on J. curcas plants with an incidence of about 45 %. The severely infected plants exhibited yellow mosaic accompanied by blistering and curling on leaves and stunting of plants as compared to non-symptomatic ones (Fig. 1a, b). Leaf and stems samples from 20 diseased plants were collected for detection of begomoviruses, and stem cuttings were planted in pots and maintained in a glass house for propagation of virus cultures. Based on typical begomovirus-disease-like yellow mo- saic symptoms and the population of whiteflies in the vicinity, begomovirus infection was suspected. Therefore, Electronic supplementary material The online version of this article (doi:10.1007/s00705-015-2375-5) contains supplementary material, which is available to authorized users. A. Srivastava Á S. Kumar Á M. Jaidi Á S. K. Raj (&) Plant Molecular Virology Laboratory, CPMB Division, CSIR-National Botanical Research Institute, Rana Pratap Street, Lucknow 226001, U.P., India e-mail: skraj2@rediffmail.com 123 Arch Virol DOI 10.1007/s00705-015-2375-5