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Abacá Mosaic Virus: a Distinct Strain of Sugarcane Mosaic Virus

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Abacá Mosaic Virus: a Distinct Strain of Sugarcane Mosaic Virus CSIRO PUBLISHING www.publish.csiro.au/journals/app Australasian Plant Pathology, 2004, 33, 475–484 Abaca´ mosaic virus: a distinct strain of Sugarcane mosaic virus C. F.GambleyA, J. E. ThomasA,C, L. V.MagnayeB and L. HerraduraB ADepartment of Primary Industries and Fisheries, Horticulture and Forest Sciences, 80 Meiers Road, Indooroopilly, Qld 4068, Australia. BDepartment of Agriculture, Bureau of Plant Industry, Bago-Oshiro 8000, Davao City, Philippines. CCorresponding author. Email: [email protected] Abstract. Abaca´ mosaic virus (AbaMV) is related to members of the sugarcane mosaic virus subgroup of the genus Potyvirus. The ∼2kb3 terminal region of the viral genome was sequenced and, in all areas analysed, found to be most similar to Sugarcane mosaic virus (SCMV) and distinct from Johnsongrass mosaic virus (JGMV), Maize dwarf mosaic virus (MDMV) and Sorghum mosaic virus (SrMV). Cladograms of the 3 terminal region of the NIb protein, the coat protein core and the 3 untranslated region showed that AbaMV clustered with SCMV, which was a distinct clade and separate from JGMV,MDMV and SrMV.The N-terminal region of the AbaMV coat protein had a unique amino acid repeat motif different from those previously published for other strains of SCMV. The first experimental transmission of AbaMV from abaca(´ Musa textilis) to banana (Musa sp.), using the aphid vectors Rhopalosiphum maidis and Aphis gossypii, is reported. Polyclonal antisera for the detection of AbaMV in western blot assays and ELISA were prepared from recombinant coat protein expressed in E. coli. A reverse transcriptase PCR diagnostic assay, with microtitre plate colourimetric detection, was developed to discriminate between AbaMV and Banana bract mosaic virus, another Musa-infecting potyvirus. Sequence data, host reactions and serological relationships indicate that AbaMV should be considered a distinct strain of SCMV, and the strain designation SCMV-Ab is suggested. Additional keywords: abaca,´ banana, RT-PCR diagnosis, ELISA, aphid transmission, potyvirus. Introduction vectors include Aphis gossypii (Ocfemia and Celino The Philippines produces about 75% of the world’s abaca´ 1938), Rhopalosiphum nympheae (Celino 1940) and or Manila hemp (Musa textilis), an important fibre crop Rhopalosiphum (Aphis) maidis (Celino and Ocfemia 1941) grown for both domestic and export purposes (FAOSTAT but AbaMV is not transmitted by Pentalonia nigronervosa 2002). A mosaic disease affecting abaca´ was first described (Celino 1940; Ocfemia et al. 1947). AbaMV is not by Calinisan (1934) from the island of Mindanao in the transmitted through true seed of abaca´ (Calinisan 1939), Philippines. It seriously affected abaca´ production (Eloja but is transmitted mechanically and through vegetative and Tinsley 1963) and by 1955, nearly half the area in propagules such as suckers, corm sections and tissue culture the Philippines under abaca´ production was affected, with (Diekmann and Putter 1996). estimated losses of 25–50% in new plantings common Initially, AbaMV was considered to be a strain of (Stover 1972). The disease remains a serious constraint to Sugarcane mosaic virus (SCMV) (Eloja et al. 1962) as abaca´ production today (Raymundo and Bajet 2000). The it was sap transmissible to maize (Celino and Ocfemia disease agent was first identified as a virus (Abaca´ mosaic 1941), had morphologically similar particles and was virus, AbaMV) by aphid transmission experiments from serologically related (Eloja and Tinsley 1963). More abaca´ to abaca´ and from abacato´ Canna indica (Ocfemia recently, the serological relationship with SCMV was and Celino 1938). confirmed, and other potyviruses including Maize dwarf AbaMV naturally infects M. textilis (Musaceae), C. indica mosaic virus (MDMV), Sorghum mosaic virus (SrMV) and (Cannaceae) and Maranta arundinacea (Marantaceae) Dasheen mosaic virus (DsMV) shown to be serologically (Velasco-Magnaye and Eloja 1966) and experimentally, related (Thomas et al. 1997). AbaMV was also shown to the host range also includes species in the Poaceae (Celino be distinct from Banana bract mosaic virus (BBrMV), and Ocfemia 1941; Eloja et al. 1962; Velasco-Magnaye another Musa-infecting potyvirus, both serologically and in and Eloja 1966) and banana (Thomas et al. 1997). Aphid nucleotide sequence (Thomas et al. 1997). The taxonomy © Australasian Plant Pathology Society 2004 10.1071/AP04050 0815-3191/04/040475 476 Australasian Plant Pathology C. F. Gambley et al. Table 1. Potyvirus isolates used in PTA-ELISA and RT–PCR assays Virus Acronym DPI ref no. Host Origin Supplier Johnson grass mosaic virus JGMV 340A Sorghum Queensland, Australia D. M. Persley Maize dwarf mosaic virus MDMV 343A Johnson grass Illinois, USA R. E. Ford Sorghum mosaic virus SrMV 344A Sorghum Illinois, USA R. E. Ford Sugarcane mosaic virus SCMV 366A Sugarcane Queensland, Australia BSESC Banana bract mosaic virus BBrMV 509B Banana cv. Cardaba Mindanao, Philippines Authors Banana bract mosaic virus BBrMV 513B Banana cv. Nendran India K. Jagadish Chandra Banana bract mosaic virus BBrMV Q1107B Abaca´ Luzon, Philippines E. O. Oloteo, R. V.J. Abgona Abaca´ mosaic virus AbaMV 543B Unknown Luzon, Philippines N. Bajet Abaca´ mosaic virus AbaMV 720B Abaca´ Mindanao, Philippines Authors Abaca´ mosaic virus AbaMV 905B Abaca´ Mindanao, Philippines Authors Abaca´ mosaic virus AbaMV 515.3B Abaca´ Mindanao, Philippines Authors Abaca´ mosaic virus AbaMV 730B Abaca´ Mindanao,Philippines Authors Mixed sample AbaMV + 904B Abaca´ Mindanao, Philippines Authors BBrMV AFreeze dried tissue. BFresh or frozen tissue. CBureau of Sugar Experiment Stations, Brisbane. of potyviruses infecting the Poaceae has recently undergone and PCR assays for the detection of AbaMV, and its revision. SCMV was originally thought to be a single virus differentiation from BBrMV. comprising a number of host-adapted strains, especially Methods from sugarcane, sorghum and maize. However, Shukla et al. (1992) considered these strains to form a subgroup of the Virus isolates genus Potyvirus comprising SCMV, MDMV, SrMV and The details of potyviruses used in this work are listed in Table 1. Johnson grass mosaic virus (JGMV). This was based on host Transmission experiments with SCMV (isolate 366) were done in Australia using glasshouse-inoculated plants only. Transmission reactions, cytopathology, amino acid sequencing and peptide experiments with AbaMV were conducted in the Philippines, using field profiling of coat proteins, cell-free translations of RNAs and samples or glasshouse-inoculated source plants. nucleotide sequence analyses. A further novel member of Aphid transmission the subgroup, Zea mosaic virus (ZeMV), has recently been described (Seifers et al. 2000). The known host range of AbaMV. Aphids (R. maidis and A. gossypii) were starved for 1–3 h before an acquisition access period (AAP) of 15–30 min on infected SCMV has expanded with the identification of a new strain tissue and were transferred in groups of approximately 20 aphids per test (SCMV-AOP) that infects African oil palm, in addition to plant, for an inoculation access period (IAP) of 24–48 h. Virus sources hosts in the Poaceae (Morales et al. 2002). The designation for the experiments were either field isolates or infected plants from of AbaMV as a strain of SCMV is inconclusive using only previous aphid transmission experiments. Field isolates of AbaMV were the previously published serological and host range data. indexed for the presence of BBrMV by Southern blot analysis (Thomas et al. 1997). Test plants included abaca,´ and banana cultivars Grand AbaMV cannot be reliably diagnosed by symptoms alone. Nain (Musa AAA group), Lakatan (Musa AAA group), Buhutan (Musa In recent infections in Musa, both AbaMV and BBrMV can balbisiana, BB), Umalag (Musa AAA group) and Latundan (Musa AAB produce similar symptoms of chlorotic areas with rusty-red group). borders (Thomas and Magnaye 2000). A range of symptoms SCMV. Aphids (R. maidis and Myzus persicae) were starved for has been described for abaca´ mosaic disease, including 2–3 h before an AAP of 30–60 s on SCMV-infected sweet corn (Zea mays) and an IAP of 12 h on healthy test plants. Five banana plants mosaics and various coloured midrib and petiole streaks (cv. Williams, Musa AAA group, Cavendish subgroup) and five sweet (Ocfemia and Celino 1938; Eloja et al. 1962; Stover 1972). corn plants were each inoculated with approximately 50 aphids. The However, BBrMV, first described by Magnaye and Espino same numbers of aphids and plants were used as healthy controls, with (1990), is now known to also infect abaca´ (Sharman et al. omission of the AAP following the starvation period. 2000a) and some symptoms described for abaca´ mosaic Virus indexing disease are reminiscent of those caused by BBrMV Positive transmission of AbaMV and SCMV to test plants was and Cucumber mosaic virus in banana. Thus, symptom initially based on symptoms. Transmission of AbaMV was confirmed descriptions reported in the older literature must be viewed by Southern blot analysis or microtitre plate colourimetric detection with caution. (MTPD) of PCR products using an AbaMV specific probe and/or by This paper investigates the relationship between AbaMV plate-trapped antigen ELISA (PTA-ELISA) (Sharman et al. 2000b; and viruses in the SCMV subgroup using serological Thomas et al. 1997). SCMV transmission was confirmed by PTA- ELISA as described below. Absence of BBrMV was monitored by techniques and nucleotide and amino acid sequence MTPD of PCR products, using a BBrMV-specific
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