Rapid, simple detection of banana bract mosaic virus in abaca using a one-step reverse transcription loop-mediated isoth
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Rapid, simple detection of banana bract mosaic virus in abaca using a one‑step reverse transcription loop‑mediated isothermal amplification (RT‑LAMP) assay Rhosener Bhea L. Koh1 · Cris Francis C. Barbosa2 · Vermando M. Aquino1 · Leny C. Galvez2 Received: 23 November 2019 / Accepted: 8 April 2020 © The Phytopathological Society of Japan and Springer Japan KK, part of Springer Nature 2020
Abstract Banana bract mosaic virus (BBrMV) causes one of the major viral diseases of the fiber crop abaca at a great economic loss in the Philippines. A one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect BBrMV. A set of six primer pairs was designed based on the conserved coat protein gene sequences of BBrMV. The RT-LAMP can be completed in 30 min and was 100,000-fold more sensitive than RT-PCR. The RT-LAMP assay is suitable for field detection of BBrMV using crude RNA extracts and fluorescent DNA intercalating dyes that produce easily visualized results. Keywords Musa textilis Née · Abaca · BBrMV · RT-LAMP · Nucleic acid-based detection · Field detection
Introduction Abaca (Musa textilis Née) is an indigenous fiber crop grown in the Philippines (Spencer 1951). The fiber, with its high mechanical strength and resistance to saltwater damage, is used as raw material in the production of cordage, paper products, and fibercrafts (Armecin et al. 2014). It is also considered a renewable bioresource material and has been used as an alternative raw material for the production of fiber composites in the automotive industry, thus making the abaca plant one of the country’s most economically important crops (Armecin et al. 2014). However, the abaca fiber industry is threatened by four major viral diseases: the abaca bunchy top virus (ABTV), banana bunchy top Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10327-020-00949-9) contains supplementary material, which is available to authorized users. * Leny C. Galvez [email protected] 1
National Institute of Molecular Biology and Biotechnology (NIMBB), University of the Philippines Diliman, 1101 Quezon City, Philippines
Philippine Fiber Industry Development Authority (PhilFIDA), DA‑PCAF Bldg, Department of Agriculture Compound, Diliman, 1101 Quezon City, Philippines
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virus (BBTV), banana bract mosaic virus (BBrMV), and sugarcane mosaic virus (SCMV). All these abaca-infecting viruses have debilitating effects on the fiber yield and quality, thereby causing significant economic losses (Bajet and Magnaye 2002). BBrMV, which causes the banana bract mosaic disease (BBrMD), is the most prevalent of the four viruses having been reported in most abaca-growing regions in the country. BBrMV is a member of the genus Potyvirus, family Potyviridae. In the Philippines, it was first reported in bananas in 1979 (Magnaye and Espino 1990) and in abaca in 2000 (Sharman et al. 2000). The virus can be transmitted through a nonpersistent manner by Pentalonia nigronervosa, A
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