Maize (Zea mays L.) is a major food and fodder crop cultivated on 1.54 million ha in the Democratic Republic of the Congo (DRC). In December 2013, unusually severe chlorotic mottle symptoms and pale green streaks were observed in local varieties (Mudishi 1 and 2, Bambou, Kasayi, H614, H613, and Mugamba) and exotic varieties (H520, H624, H403, HDK8031, and ZM607) in Beni, Lubero, and Rutshuru territories at 1,015 to 1,748 m elevation in North Kivu Province. Symptoms were prominent on newly emerging leaves that later developed marginal necrosis resembling the symptoms of maize lethal necrosis (MLN), caused by a dual infection of Maize chlorotic mottle virus (MCMV, genus Machlomovirus) and Sugarcane mosaic virus (SCMV, genus Potyvirus). Each of these viruses, but particularly MCMV, is also known to cause severe mosaic and mottling symptoms in maize (4). In January 2014, symptomatic and asymptomatic samples (n = 20) from disease-affected fields in Beni and Lubero provinces were collected for virus testing using Whatman FTA Classic Cards (1) and analyzed for MCMV (2681F: 5′-ATGAGAGCAGTTGGGGAATGCG and 3226R: 5′-CGAATCTACACACACACACTCCAGC) and SCMV (8679F: 5′-GCAATGTCGAAGAAAATGCG and 9595R: 5′-GTCTCTCACCAAGAGACTCGCAGC) by reverse transcription (RT)-PCR (4). Samples were also analyzed for Maize streak virus (MSV, genus Mastrevirus), an endemic virus in DRC, by PCR using MSV specific primers (MSV215-234: CCAAAKDTCAGCTCCTCCG and MSV1770-1792: TTGGVCCGMVGATGTASAG) (3). A DNA product of expected size (~520 bp) resulted only for MCMV in all the symptomatic plant samples. None of the samples tested positive for SCMV or MSV. RT-PCR analyses were performed to ascertain the absence of potyviruses using the degenerate potyvirus primers (CIFor: 5′GGIVVIGTIGGIWSIGGIAARTCIAC and CIRev: 5′ACICCRTTYTCDATDATRTTIGTIGC3′) (2) were also negative. Occurrence of MCMV in symptomatic samples was further confirmed by antigen-coated plate (ACP)-ELISA using anti-MCMV rabbit polyclonal antibodies produced at the Virology Unit, IITA, Ibadan, Nigeria. The RT-PCR product of MCMV was purified and sequenced in both directions (GenBank Accession No. KJ699379). Pairwise comparison of 518 bp nucleotide sequence corresponding to p32 and p37 open reading frames of MCMV by BLASTn search revealed 99.8% nucleotide sequence identity with an MCMV isolate from Kenya (JX286709), 98 to 99% identity with the isolates from China (JQ982468 and KF010583), and 96% identity with the isolates from the United States (X14736 and EU358605). MCMV is a newly emerging virus in Africa, first detected during a severe MLND outbreak in 2011 in Kenya (4). This disease has since become a serious threat to maize production in East Africa. MCMV has been reported in maize from Kenya, Rwanda, Tanzania, and Uganda. To our knowledge, this is the first report of MCMV occurrence in DRC. This finding confirms the further geographic expansion of MCMV and illustrates the need for further studies to identify vectors and also create awareness about the disease and to strengthen surveillance to prevent its further spread in the continent. References: (1) O. J. Alabi et al. J. Virol. Met. 154:111, 2008. (2) C. Ha et al. Arch. Virol. 153:25, 2008. (3) K. E. Palmer and E. P. Rybicki. Arch. Virol. 146:1089, 2001. (4) A. Wangai et al. Plant Dis. 96:1582, 2012.

No abstract

Banana (including plantain; Musa spp.) is an important vegetatively propagated food staple grown as a semi-perennial crop in fields and backyard gardens in Tanzania. Banana bunchy top disease (BBTD), caused by the banana bunchy top virus (BBTV, genus Babuvirus), is the most economically important viral disease of banana, infection of which results in severe stunting and reduction in fruit production by 90-100% within two seasons. The virus is spread by the banana aphid, Pentalonia nigronervosa, and through vegetative propagation of infected sources. BBTV is an introduced virus first reported in sub-Saharan Africa (SSA) in the 1960s in the Democratic Republic of Congo. Since then, BBTV spread was confirmed in 15 countries in Central, Southern, and Western African regions but was not detected in any previous surveys in the East African sub-region. During banana pests and disease surveys conducted in December 2020 – January 2021 in Buhigwe District in the Kigoma Region of Tanzania revealed banana plants with typical BBTV symptoms (severe stunting, leaves with shortened petioles, chlorotic streaks, and yellow leaf margins) in several banana fields in Muhinda (lon. 29.78662, lat. -4.53672) and Mwayaya (lon. 29.8218, lat. -4.49203) villages. Most of the affected plantations were 5 to 15 years old. Leaf samples (N=21) from symptomatic (N=6) and asymptomatic (N=15) banana plants were collected and used for total DNA extraction and BBTV detection by polymerase chain reaction (PCR) using the primer pair BBTV-1 and BBTV-2 to amplify ~240 bp sequence of DNA-R encoding for core master replication initiator protein gene. All samples from symptomatic plants tested positive and asymptomatic plants were negative. To further confirm the virus identity, four samples, each from symptomatic (PCR positive) and asymptomatic (PCR negative) plants from Muhinda and Mwayaya villages, were tested by Triple Antibody Sandwich-Enzyme-linked Immunosorbent Assay (TAS-ELISA) using BBTV ELISA reagent set (Cat. # SRA24700-1000, Agdia, France) following the manufacturer's protocol. Samples from symptomatic plants reacted positively in TAS-ELISA, and asymptomatic plants were negative. The 240-bp PCR product of two isolates was purified, and both strands were sequenced. A BLAST search of the nucleotide sequences (NCBI GenBank Acc.# MW711671 and MW711672) revealed 99% identity with DNA-R sequences of several other BBTV isolates from Africa (Acc. No# JF755994). Further analysis of the 240-bp nucleotide sequences with Maximum-likelihood phylogenetic analysis using MEGA-X software has grouped the two BBTV sequence isolates with the SSA sub-clade of the South-Pacific group. To our knowledge, this is the first report of BBTV infecting bananas in Tanzania, and East Africa endowed with rich banana diversity and popular East African Highland banana clone. BBTV presents a new threat to banana production in this sub-region due to the high risk of further spread through vegetative propagation, traditional planting material exchange practices, and the ubiquitous banana aphid vector. This study warrants delimitation surveys to assess the extent of spread, with simultaneous efforts to raise awareness about BBTD recognition and control measures among banana growers, including eradicating infected mats and replanting with healthy planting material to recover banana production.

No abstract

No abstract

Banana (including plantain; Musa spp.) is a vegetatively propagated semi-perennial crop in fields and backyard gardens in Togo. Banana bunchy top disease (BBTD), caused by banana bunchy top virus (BBTV, genus Babuvirus) is the most economically important viral disease, infection of which causes severe stunting and production losses of 90-100% within two seasons. The virus is spread by banana aphid, Pentalonia nigronervosa, and through vegetative propagation from infected sources. BBTV occurrence was first reported in West Africa in 2011 with confirmation in Republic of Benin and in Nigeria in 2012 . A regional alliance (www.bbtvalliance.org) has been established for BBTV surveillance through frequent surveys in countries neighboring those affected, such as Togo. The surveys conducted in September 2018 in banana growing areas in Togo revealed plants with typical symptoms (severe stunting, bunchy growth with shortened petioles with chlorotic streaks and yellow leaf margins) in three banana fields. Locations were Tsévié, Préfecture de Zio, (6.44°N, 1.21028°E), Lilicope, Préfecture de Zio in Maritime region (6.56583°N, 1.18639°E), and Amoutchou, Préfecture de l’Ogou in Plateaux region (7.3775°N, 1.17472°E). Leaf samples were collected from symptomatic (N=8) and asymptomatic plants (N=30) and used for DNA extraction followed by a polymerase chain reaction (PCR) for BBTV detection to amplify ~240 bp sequence of DNA-R encoding for core replicase gene. All samples from symptomatic plants (N=8) tested positive and asymptomatic plants were negative. To ascertain virus identity the 240-bp PCR product was purified and sequenced in both directions. A BLAST search of the sequence (NCBI GenBank Acc.# MK073116) revealed 99% identity with DNA-R sequences of BBTV isolates from Africa (e.g., JQ437549-Benin, JN290301-Nigeria). Further analysis of the 240-bp nucleotide sequence with Maximum-likelihood phylogenetic analysis using MEGA-X software has grouped the BBTV isolate with sub-Saharan African sub-clade of the South Pacific group. To further confirm the virus identity, two samples from symptomatic (PCR positive) and asymptomatic (PCR negative) plants from Tsévié were tested by TAS-ELISA using BBTV ELISA reagent set (Cat. No. SRA24700-1000, Agdia, France) following the manufacturers’ protocol. Only samples from two symptomatic plants that were positive in PCR reacted positively in TAS-ELISA; asymptomatic plants were negative. BBTV was not observed in any of the 22 locations surveyed as a follow-up in banana producing areas. To our knowledge, this is the first report of BBTV infecting banana in Togo. The plants detected in the three sites were eradicated in the follow-up action implemented by the alliance team together with the Direction de la Protection des Végétaux of Togo. Follow-up surveys were conducted in the same regions in 2019 and 2020 to ensure disease-free status in these sites and other banana producing regions in Togo. Efforts have been made to raise awareness about BBTD recognition, diagnosis, and eradication. To the best of our knowledge this is the first case of rapid detection and eradication of BBTD in sub-Saharan Africa. This study illustrates the importance of regular surveillance for early detection of invasive virus threats and the value of rapid eradication to contain viruses before spread and establishment in a new territory.

Antigen-coated plate enzyme-linked immunosorbent assay (ACP-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the presence and seed transmissibility of bean common mosaic virus-blackeye cowpea mosaic (BCMV-BICM) in farm- retained cowpea seed lots obtained from 46 locations, including markets and farms in major cowpea growing areas in the Ashanti and Brong Ahafo regions of Ghana. In the growout tests, virus symptomatic plants were observed in seedlings of 19 of the 46 seed lots tested under insect-proof screen-house conditions. All the symptomatic plants tested positive to polyclonal antiserum raised against BCMV-BICM in ACP-ELISA. The seed transmission rates based on symptoms ranged from 0 to 37.8 %. RT-PCR with primer pair designed to amplify the potyvirus Cylindrical Inclusion (CI) region resulted in an expected 720 bp DNA segment in 19 seed lots as a further confirmation of virus in the seed lots. The remaining 27 lots were asymptomatic and tested negative to BCMV-BlCM in both ACP-ELISA and RTPCR. The findings of this study revealed seed as the source of primary inoculum in the farmers’ fields and may aid in the implementation of control strategies such as discouraging farmers from retaining their own seeds for subsequent sowing and encouraging them to take appropriate measures in obtaining virus-free cowpea seeds from other sources.