Abstract
Banana mild mosaic virus (BanMMV) (Betaflexiviridae, Quinvirinae, unassigned species) is a filamentous virus belonging to the Betaflexiviridae family. It infects Musa spp. with a very wide geographic distribution. The genome variability of plant viruses, including the members of the Betaflexiviridae family, makes their molecular detection by specific primers particularly challenging. During routine indexing of the Musa germplasm accessions, a discrepancy was observed between electron microscopy and immunocapture (IC) reverse transcription (RT) polymerase chain reaction (PCR) test results for one asymptomatic accession. Filamentous viral particles were observed while molecular tests failed to amplify any fragment. The accession underwent high-throughput sequencing and two complete genomes of BanMMV with 75.3% of identity were assembled. Based on these sequences and on the 54 coat protein sequences available from GenBank, a new forward primer, named BanMMV CP9, compatible with Poty1, an oligodT reverse primer already used in diagnostics, was designed. A retrospective analysis of 110 different germplasm accessions from diverse origins was conducted, comparing BanMMCP2 and BanMMV CP9 primers. Of these 110 accessions, 16 tested positive with both BanMMCP2 and BanMMV CP9, 3 were positive with only BanMMCP2 and 2 tested positive with only BanMMV CP9. Otherwise, 89 were negative with the two primers and free of flexuous virions. Sanger sequencing was performed from purified PCR products in order to confirm the amplification of the BanMMV sequence for the five accessions with contrasting results. It is highly recommended to use the two primers successively to improve the inclusiveness of the protocol.
Highlights
Banana (Musa spp.) is a perennial herbaceous plant and is among the most important staple food crops worldwide
This study aimed to describe the identification and genome sequencing of two isolates of Banana mild mosaic virus (BanMMV), and, based on the virus sequences available in GenBank, to design and test a new diagnostic primer for a routine indexing use
Flexuous rod-shaped virions were detected in a partial purification of banana accession ITC0763 (Figure 1), while negative results were obtained by targeted IC-reverse transcription (RT)-polymerase chain reaction (PCR)
Summary
Banana (Musa spp.) is a perennial herbaceous plant and is among the most important staple food crops worldwide. With worldwide annual production reaching 144 million tons, it represents an important contribution to the economies of many countries in Asia, Africa, Latin-America and the Pacific Islands [2]. Bananas are vegetatively propagated, which involves a high risk of virus spread through the movement of infected planting material [3]. There is a requirement for virus-tested planting material for guaranteeing the exchange of germplasm and for disease control [4]. In this context, the International Plant Protection Convention
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