Partial characterization and molecular cloning of sweetpotato feathery mottle virus (SPFMV)

• Published in: Journal of Tropical Plant Pathology (Philippines) Vol. 40; no. 1-2
• Main Authors: Dolores, L.M, Yebron, G.N, Colle, M.G.,Philippines Univ. Los Banos, College, Laguna (Philippines). Inst. of Plant Breeding
• Format: Journal Article
• Language: English
• Published: 01.01.2004
• Subjects: CAESIUM; CESIO; CESIUM; CHLORE; CHLORINE; CLONACION MOLECULAR; CLONAGE MOLECULAIRE; CLORO; FILIPINAS; http://www.fao.org/aos/agrovoc#c_1182; http://www.fao.org/aos/agrovoc#c_1568; http://www.fao.org/aos/agrovoc#c_24357; http://www.fao.org/aos/agrovoc#c_27503; http://www.fao.org/aos/agrovoc#c_34079; http://www.fao.org/aos/agrovoc#c_3937; http://www.fao.org/aos/agrovoc#c_5783; http://www.fao.org/aos/agrovoc#c_5985; IPOMOEA BATATAS; MOLECULAR CLONING; PCR; PHARBITIS NIL; PHILIPPINES; PLANT VIRUSES; VIRUS DE LAS PLANTAS; VIRUS DES VEGETAUX
• ISSN: 0115-0804

Sweetpotato feathery mottle virus (SPFMV) has been identified as one of the most important constraints in sweetpotato production. The virus has been noted in large sweetpotato fields in Central Luzon [Philippines]. Spreading over most of the sweetpotato areas, it has led to substantial yield losses and the loss of an important variety called 'Bureau'. Several SPFVM isolates have been characterized based on differential reactions to diagnostic hosts, Ipomoea setosa and I. nil. The virus was purified from mechanically inoculated I. nil using Cessium chloride (CScl) step gradient centrifugation giving a faint opalescent band near the bottom of the centrifuge tube. This virus yield ranged from 9-30 mg/kg with A260nm/A280nm ratio of around 1.2. The purified virus was infectious and exhibited flexous rod particles typical of a potyvirus under an electron microscope. Further characterization of the SPFMV isolates by reverse transcriptase polymerase chain reaction (RT PCR) of the purified virus resulted to the amplification of the coat protein gene of SPFMV using 2 sets of primers designed to amplify the partial and full length coat protein gene of SPFMV. The expected PCR product sizes of 400 bp and 1.0 kb for partial and full length CP, respectively were obtained and successfully cloned using the TOPO TA cloning kit of INVITROGEN. Such results on the characterization and cloning of SPFMV would be very useful in illuminating its position within the potyvirus group. Moreover, information on the molecular aspects of the virus would help facilitate the development of rapid and sensitive techniques for virus detection and identification that are important in monitoring virus infection in the field.