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

GENETICS AND PLANT BREEDING

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: alexswapna@hotmail.com

Paper No. 266

Received: 12 February, 2014

Accepted: 16 November, 2014

Published: 23 December, 2014

Abstract

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.

Highlights

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