Banana bunchy top virus (BBTV) affects all varieties of banana plants and causes heavy economic loss in most of the banana cultivating areas. The BBTV genome comprises of six DNA components; in this study, we have cloned the six BBTV-DNA components from one of the BBTV-infected plants (Tri-8) and were submitted to GenBank. Analysis of the BBTV DNA-R component showed that it belonged to south Pacific group. Resistance against BBTV has not been observed so far in banana plants and removal and killing of the infected plants has been routinely practiced. Hence, early detection of BBTV infection would be desirable and various detection methods routinely employed include enzyme linked immunosorbent assay (antigen–antibody based) and molecular-based methods such as polymerase chain reaction (PCR), qPCR, or LAMP PCR. Most of these methods require enzymes or antibodies for detection and hence are expensive. Here, we report a visual detection method (AuNP probe assay) using gold nanoparticles (AuNPs) functionalized with an ssDNA-thiolated probe (CR1). This method is based on the hybridization of the functionalized AuNPs with the target DNA (BBTV). In the AuNP probe assay, the functionalized AuNPs retains red colour when BBTV DNA is present, and in the absence of BBTV DNA, the colour of the functionalized AuNPs changes to purple when salt is added. The AuNP probe assay was compared with PCR for the detection of banana plants and it was found that AuNP probe assay was better than PCR in detecting BBTV infection (86.5% for AuNP probe assay and 65% for PCR). The AuNP probe assay was found to be highly specific to BBTV and was found to detect up to 1 pg/μl of the plasmid (pTZBBTri 4, BBTV DNA) mixed with healthy banana DNA.
This is a preview of subscription content, log in to check access.
We thank Sri Ramachandra University for providing all facilities and support to carry out this research work. Authors thank Kalpana A and Dhanushya S and Vignesh for their immense help during the study. We thank Munmi Borah, PhD scholar (Assam Agricultural University, Jorhat) for the samples as well as Jain irrigation systems for the Healthy, diseased and BSV-infected samples for the study.
Compliance with ethical standards
Conflict of interest
Authors declare no conflict of interest.
Anandhi J, Vijila C, Viswanath GS, Lokeswari TS (2007) Screening banana plants for banana bunchy top virus with primers specific to Indian isolates. J Plant Dis Prot 114:101–107. doi:10.1007/bf03356715CrossRefGoogle Scholar
Andreadou M, Margarita A, Emmanouil L et al (2014) A novel non-amplification assay for the detection of Leishmania spp. in clinical samples using gold nanoparticles. J Microbiol Methods 96:56–61. doi:10.1016/j.mimet.2013.10.011CrossRefGoogle Scholar
Baptista PV, Koziol-Montewka M, Paluch-Oles J et al (2006) Gold-nanoparticle-probe-based assay for rapid and direct detection of Mycobacterium tuberculosis DNA in clinical samples. Clin Chem 52:1433–1434. doi:10.1373/clinchem.2005.065391CrossRefGoogle Scholar
Dale JL (1987) Banana bunchy top: an economically important tropical plant virus disease. Adv Virus Res. 33:301–325CrossRefGoogle Scholar
Dharanivasan G, Mohammed Riyaz SU, Michael Immanuel Jesse D et al (2016) DNA templated self-assembly of gold nanoparticle clusters in the colorimetric detection of plant viral DNA using a gold nanoparticle conjugated bifunctional oligonucleotide probe. RSC Adv 6:11773–11785. doi:10.1039/c5ra25559gCrossRefGoogle Scholar
Dietzgen RG, Thomas JE, Smith GR, Maclean DL (1999) PCR-based detection of viruses in banana and sugarcane. Curr Topics Virol 1:105–118Google Scholar
Furuya N, Somowiyarjo S, Natsuaki KT (2004) Virus detection from local banana cultivars and the first molecular characterization of Banana bunchy top virus in Indonesia. J Agri Sci Tokyo. 49(3):75–81Google Scholar
Geering A, Thomas JE (1996) A comparison of four serological tests for the detection of banana bunchy top virus in banana. Aust J Agric Res 47:403–412CrossRefGoogle Scholar
Magee CJP (1927) Investigation on the bunchy top disease of the banana. Bull Council Sci Ind Res Aust 30:64Google Scholar
Mansoor S, Qazi J, Amin I et al (2005) A PCR-based method, with internal control, for the detection of Banana bunchy top virus in banana. Mol Biotechnol 30:167–170. doi:10.1385/MB:30:2:167CrossRefGoogle Scholar
Mirkin CA, Letsinger RL, Mucic RC, Storhoff JJ (1996) A DNA-based method for rationally assembling nanoparticles into macroscopic materials. Nature 382:607–609. doi:10.1038/382607a0CrossRefGoogle Scholar
Oben TT, Hanna R, Ngeve J et al (2009) Occurrence of banana bunchy top disease caused by the banana bunchy top virus on banana and plantain (Musa sp.) in Cameroon. Plant Dis 93:1076–1076. doi:10.1094/PDIS-93-10-1076CCrossRefGoogle Scholar
Soo P-C, Horng Y-T, Chang K-C et al (2009) A simple gold nanoparticle probes assay for identification of Mycobacterium tuberculosis and Mycobacterium tuberculosis complex from clinical specimens. Mol Cell Probes 23:240–246. doi:10.1016/j.mcp.2009.04.006CrossRefGoogle Scholar
Stainton D, Kraberger S, Walters M et al (2012) Evidence of inter-component recombination, intra-component recombination and reassortment in banana bunchy top virus. J Gen Virol 93:1103–1119. doi:10.1099/vir.0.040337-0CrossRefGoogle Scholar
Stainton D, Martin DP, Muhire BM et al (2015) The global distribution of banana bunchy top virus reveals little evidence for frequent recent, human-mediated long distance dispersal events. Virus Evolut. doi:10.1093/ve/vev009Google Scholar
Su HJ, Tsao LY, Wu ML, Hung TH (2003) Biological and molecular categorization of strains of banana bunchy top virus. J Phytopathol 151:290–296CrossRefGoogle Scholar
Thomas JE (1991) Virus indexing procedures for banana in AustraliaGoogle Scholar
Thomas JE, Dietzgen RG (1991) Purification, characterization and serological detection of virus-like particles associated with banana bunchy top disease in Australia. J Gen Virol 72(Pt 2):217–224. doi:10.1099/0022-1317-72-2-217CrossRefGoogle Scholar
Watanabe S, Bressan A (2013) Tropism, compartmentalization and retention of banana bunchy top virus (Nanoviridae) in the aphid vector Pentalonia nigronervosa. J Gen Virol 94:209–219. doi:10.1099/vir.0.047308-0CrossRefGoogle Scholar
Wei J, Liu H, Liu F et al (2014) Miniaturized paper-based gene sensor for rapid and sensitive identification of contagious plant virus. ACS Appl Mater Interfaces 6:22577–22584. doi:10.1021/am506695gCrossRefGoogle Scholar