International Journal of Agriculture, Environment & Biotechnology
Citation: IJAEB: 7(4): 765-767 December 2014
DOI Number: 10.5958/2230-732X.2014.01385.0
©2014 New Delhi Publishers. All rights reserved
Polymerase chain reaction based detection of banana bunchy
top virus using coat protein based primers
R. K. Ponsubrya, Swapna Alex*, K. Rajmohan and K. B. Soni
Department of Plant Biotechnology, College of Agriculture, Vellayani, Thiruvananthapuram, Kerala-695 522, India.
*Corresponding author:
Paper No. 266
Received: 12 February, 2014
Accepted: 16 November, 2014
Published: 23 December, 2014
Banana Bunchy Top virus (BBTV) is a virus seriously affecting banana cultivation. Primers were
designed for the amplification of coat protein gene for the diagnosis of BBTV in the commonly grown
banana cultivars of Kerala, India, viz., Red Banana (AAA), Palayankodan (AAB), Dwarf Cavendish
(AAA), Motta Poovan (AAB) and Ney Poovan (AB). PCR detection using these primers at an early stage
can aid in disease free planting material production.
  • Primers designed for the early detection of Banana bunchy top viral infection using PCR produced
  • an amplicon of size 500 bp in all the infected samples of the five varieties of banana.
    Keywords: PCR primers, BBTV detection, banana
    Banana Bunchy Top Virus (BBTV) is one among
    Chain Reaction (PCR) can detect the virus even at
    the four major viruses seriously affecting banana
    very minute concentrations (Galal, 2007; Joshi and
    plantations in India (Selvarajan et al ., 2010; Banerjee et
    Deshpande, 2011).
    al .,2014).ThevirusbelongstothefamilyNanoviridae
    In the present study, primers were designed for the
    and contains a multi-component genome of circular
    amplification of coat protein gene for the diagnosis
    ssDNA (Vishnoi et al ., 2009). The virus is transmitted
    of BBTV in the commonly grown banana cultivars
    by an aphid vector, Pentalonia nigronervosa Coq. (Hu
    of Kerala, viz., Red Banana (AAA), Palayankodan
    et al ., 1996, Hooks et al ., 2009; Chen and Hu, 2013). The
    (AAB), Dwarf Cavendish (AAA), Motta Poovan
    infection is a great hindrance in the multiplication
    (AAB) and Ney Poovan (AB).
    and supply of quality planting material both by
    conventional and in vitro propagation approaches.
    Materials and Methods
    Since the primary source of infection of BBTV is
    through infected suckers, it is necessary to detect
    Primer Designing and Synthesis
    the presence of the virus in suckers before they are
    planted or used for micropropagation. Diagnosis of
    The sequences of the coat protein gene with the
    this viral disease is usually carried out by Enzyme-
    accession numbers EF584544, AY272038, AY953428,
    linked immunosorbent assay (ELISA). However
    DQ515970 and AY534140 coding for BBTV coat
    this technique is not sufficient to detect the virus in
    protein gene were retrieved from the NCBI Genbank.
    the very early stages. Techniques like Polymerase
    Conserved regions were obtained using multiple
    sequence alignment tool Clustal X (1.8). The primers