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Sentinel plantations are a promising tool to collect valuable information on plant-pathogen interactions. While it has proven its efficiency to discover new significant threatening organisms (Kenis et al., 2018; Vettraino et al., 2017), this technique has also the potential to identify new host plants for known pathogens. The objective of this study was to investigate the susceptibility of selected Belgian potential host plant species to Xylella fastidiosa by establishing and monitoring ex-patria plantings (EPPO 2020) in a Xylella-contaminated area, the campus of the University of the Balearic Islands (UIB) situated in Palma de Mallorca. First, official approval was obtained from the local government and from the UIB Authorities. Then, selected plant species (Prunus domestica cv. opal, Salix alba, Quercus petraea), were acquired from Belgian producers. Although they were provided with phytosanitary certificates, they were also checked for X. fastidiosa before 27 plants of each species were brought to Spain and planted in March 2018. Later on, they were regularly submitted to regular symptom observations and molecular analyses. Simultaneously, within a 100 meters radius area around the plantation, the abundance and the infectivity of the potential insect vectors, as well as the sanitary state of the local vegetation, were monitored. The survey is still ongoing. So far, no infection by X. fastidiosa has been observed, possibly due to the young age of the trees and the recent establishment of the assay, as well as to the scarcity of the ground herbaceous vegetation and vectors during the first year. This study underlines the hurdles that may be encountered with sentinel plants. Yet it provides a methodology highlighting key features to address X. fastidiosa studies through sentinel plantations. Reference : European and Mediterranean Plant Protection Organization (EPPO) (2020). PM 3/91(1) Sentinel woody plants: concepts and application. Bulletin OEPP/EPPO Bulletin, 50(3), 429–436. Kenis, M., Li, H., Fan, J. T., ...

Viral diseases of banana are one of the major constraints to the production and distribution of germoplasm of banana in the world. To date, six viruses have been listed on banana: Banana bunchy top virus (BBTV), Banana streak virus (BSV), Banana bract mosaic virus (BBrMV), Cucumber mosaic virus (CMV), Banana mild mosaic virus (BMMV), Banana virus x (BVX). In Democratic Republic of Congo (DRC), the first viral disease of banana was found, for the first time, in 1950 in Yangambi, Kisangani region. To date, the distribution of BBTV remains mostly unknown and other banana viruses (BSV, CMV, BBrMV, BMMV, BVX) have not yet been reported. With the aim to describe and establish the frequency of the viruses involved to major viral diseases on banana (Banana bunchy top virus, Banana streak virus, Banana bract mosaic virus, Cucumber mosaic virus) in the Democratic Republic of Congo, epidemiological surveys were conducted in 2011, 2012 and 2013 in the South West of the Democratic Republic of Congo. For BBTV at least 216 samples were collected in 2011 and 2012, for BS, CMV, BBrMV and 450 samples were collected and 306 samples were analyzed using PCR, IC - PCR, IC-One-step-RT-PCR and RT - PCR techniques to detect respectively BBTV, BSV, CMV, and BBrMV. Results indicate that Banana bunchy top virus is the most prevalent virus of banana, in South Western of Democratic Republic of Congo (frequency = 44,6%). The highest frequency was observed in Bas Congo province of Bas Congo with a frequency of 17.8% in contrast, the lowest frequency was observed in Katanga province. Banana streak virus exists in natural condition and express symptoms of Banana streak disease. The most widespread BSV specie is Goldfinger (BSGFV) with a frequency of 64%. This study reveals that there is a transmission of Banana streak virus from banana containing genome B with eBSV integration to Musa acuminata strict, without integrated sequences. Furthermore, the frequency of vector is 9.3%. These results suggest an emergence of Banana streak disease in the study area. Analysis of the frequency of the Cucumber mosaic virus shows that the virus is widespread in all types of cropping systems including vegetables spaces whose N'Djili brasserie, Kimpoko and Kisantu. Very surprisingly CMV is missing from Mbanzangungu one of the largest vegetable crop production site. Performant analyses for the detection of Banana bract mosaic virus have proved negative. These results suggest that the BBrMV remains absent from the study area.

Banana bunchy top virus (BBTV) is one of the most severe and widespread virus limiting production and distribution of planting material of banana (Musa spp.) crops in the world. In Democratic Republic of Congo (DRC), these crops play a major role in daily life of almost 70% of citizen. Many factors influence banana production negatively such as Banana bunchy top disease. Epidemiological survey was conducted in experimental stations and farmers' fields for two consecutive seasons covering 72 sites in five provinces of south western of RDC. The objective of this study was to evaluate the presence and distribution of the Banana bunchy top virus in five provinces of South Western of DRC, with emphasis on the agro-ecological factors. A total of 174 Musa spp. leaves samples were collected and analyzed by PCR. The results revealed the presence of BBTV in all provinces investigated. The frequency of BBTV was 6.3% in Bandundu, 12.1% in Kasaï Oriental, 17.8% Bas Congo, 1.1% in Katanga and 7.5% Kinshasa Urban and Peri-urban. Results also revealed that BBTV occurred in experimental station and farmers' fields, both having all cooking and dessert bananas. The high prevalence of BBTV seemed to be linked to multiple introductions of planting materials in the Bas Congo province during 1990 and 2002. However, the province of Katanga had not experienced the introduction of planting material. This factor would explain the lowest prevalence of Banana bunchy top virus in this province. The results indicated that there was a real need to facilitate access to genetically improved and healthy certified planting material in these provinces.

Banana bunchy top virus (BBTV) is one of the most severe and widespread virus limiting produc- tion and distribution of planting material of banana (Musa spp.) crops in the world. In Democratic Republic of Congo (DRC), these crops play a major role in daily life of almost 70% of citizen. Many factors influence banana production negatively such as Banana bunchy top disease. Epidemiol- ogical survey was conducted in experimental stations and farmers' fields for two consecutive sea- sons covering 72 sites in five provinces of south western of RDC. The objective of this study was to evaluate the presence and distribution of the Banana bunchy top virus in five provinces of South Western of DRC, with emphasis on the agro-ecological factors. A total of 174 Musa spp. leaves samples were collected and analyzed by PCR. The results revealed the presence of BBTV in all provinces investigated. The frequency of BBTV was 6.3% in Bandundu, 12.1% in Kasaï Oriental, 17.8% Bas Congo, 1.1% in Katanga and 7.5% Kinshasa Urban and Peri-urban. Results also re- vealed that BBTV occurred in experimental station and farmers' fields, both having all cooking and dessert bananas. The high prevalence of BBTV seemed to be linked to multiple introductions of planting materials in the Bas Congo province during 1990 and 2002. However, the province of Katanga had not experienced the introduction of planting material. This factor would explain the lowest prevalence of Banana bunchy top virus in this province. The results indicated that there was a real need to facilitate access to genetically improved and healthy certified planting material in these provinces.

Banana bunchy top virus (BBTV) is one of the most severe and widespread virus limiting produc- tion and distribution of planting material of banana (Musa spp.) crops in the world. In Democratic Republic of Congo (DRC), these crops play a major role in daily life of almost 70% of citizen. Many factors influence banana production negatively such as Banana bunchy top disease. Epidemiol- ogical survey was conducted in experimental stations and farmers' fields for two consecutive sea- sons covering 72 sites in five provinces of south western of RDC. The objective of this study was to evaluate the presence and distribution of the Banana bunchy top virus in five provinces of South Western of DRC, with emphasis on the agro-ecological factors. A total of 174 Musa spp. leaves samples were collected and analyzed by PCR. The results revealed the presence of BBTV in all provinces investigated. The frequency of BBTV was 6.3% in Bandundu, 12.1% in Kasaï Oriental, 17.8% Bas Congo, 1.1% in Katanga and 7.5% Kinshasa Urban and Peri-urban. Results also re- vealed that BBTV occurred in experimental station and farmers' fields, both having all cooking and dessert bananas. The high prevalence of BBTV seemed to be linked to multiple introductions of planting materials in the Bas Congo province during 1990 and 2002. However, the province of Katanga had not experienced the introduction of planting material. This factor would explain the lowest prevalence of Banana bunchy top virus in this province. The results indicated that there was a real need to facilitate access to genetically improved and healthy certified planting material in these provinces.

This work is partially supported by the Scottish Government's Rural and Environment Science and Analytical Services (RESAS) Division ; Potato mop-top pomovirus (PMTV) is one of a few viruses that can move systemically in plants in the absence of the capsid protein (CP). Pomoviruses encode the triple gene block genetic module of movement proteins (TGB 1, 2, and 3) and recent research suggests that PMTV RNA is transported either as ribonucleoprotein (RNP) complexes containing TGB1 or encapsidated in virions containing TGB1. Furthermore, there are different requirements for local or systemic (long-distance) movement. Research suggests that nucleolar passage of TGB1 may be important for the long-distance movement of both RNP and virions. Moreover, and uniquely, the long-distance movement of the CP-encoding RNA requires expression of both major and minor CP subunits and is inhibited when only the major CP sub unit is expressed. This paper reviews pomovirus research and presents a current model for RNA movement. ; Publisher PDF ; Peer reviewed

BACKGROUND and OBJECTIVES Viruses are major constraints to the production and exchange of banana and plantain (Musa spp.) germplasm in the world. Several viruses are recognized to affect Musa spp: Banana bunchy top virus (BBTV), Banana streak GF virus (BSGFV), Banana streak OL virus (BSOLV), Banana streak IM virus (BSIMV), Banana streak MY virus (BSMYV), Cucumber mosaic virus (CMV), Banana bract mosaic virus, (BBrMV). To date, in Democratic Republic of Congo (DRC), except BBTV, others banana viruses are not yet reported. The main aim of this work was to elucidate the epidemiological situation, genetic variability and origin of Musa spp. viruses in DRC. MATERIALS and METHODS Epidemiological surveys were conducted from 2011-2013, in DRC, to search main viruses of Musa spp. Up to 666 samples collected from 122 locations; Samples were analysed using PCR based method. Detected viruses were sequenced and phylogenetic studies were performed. RESULTS BBTV is the most widespread virus (frequency=44,8%) in DRC. Molecular analyses support infrequent BBTV dispersal events between continents. This study reveals a low genetic variability of DRC BBTV isolates. They are close, gather in the South Pacific group and are linked with isolates of other sub-Saharan Africa (SSA) countries, but no with Egyptian isolates. Since the earliest BBTV report in SSA was in DRC in 1950, it is likely that BBTV spread from DRC to other SSA countries rather than from Egypt and that at least two BBTV introductions occurred in Africa. Haplotypes analysis based on the coding region m-Rep protein showed 38 haplotypes (haplotype diversity value of 0.944±0.013). In Bas Congo, symptoms of Banana streak disease were associated to the presence of BSV species (frequency=19,9%); the most widespread were BSGFV (frequency=67,2%) and (BSOLV) (frequency=24,2%). The sequenced BSOLV isolate from Musa acuminata, gather in the clade I recognized to gathering the BSV species with endogenous counterpart. CMV was also detected (frequency=9,3%). Genetic diversity analysis reveals the high similarity between DRC CMV isolates with a South African isolate). BBrMV was not detected in collected samples. CONCLUSION DRC is a place of high prevalence of banana viruses, indicating the need for an improved quality of planting material. DRC seems to be the primary center of BBTV spread in Africa. BSV species data revealed the activation of endogenous BSV and the transmission of BSV species from plantain (B genome) to banana (A genome), in natural conditions. While for CMV, data suggest the hypothesis of a unique introduction through infected planting material and the distribution of a single strain of CMV. The BBrMV still absent in DRC. (Texte intégral)

BACKGROUND and OBJECTIVES Banana bunchy top virus (BBTV), is the most devastating and widespread banana virus. Banana and plantain (Musa spp.) are major crops in terms of household income and food security in Democratic republic of Congo (DRC) (1). Despite the large territory under banana and plantain cultivation in the country, the genetic characterization of BBTV isolates has been undertaken for two provinces. The main objectives of this work were to understand the genetic variability of DRC BBTV isolates and to determine their origin. MATERIALS and METHODS In this study, genetic variation in BBTV was assessed from 52 BBTV isolates collected in five out of 11 provinces in DRC (Bandundu, Bas-Congo, Katanga, Kinshasa and Kasai Oriental). 52 full sequences of DNAR and 30 full sequences of all BBTV components were analysed. RESULTS Full genome sequencing of DNA-R components was performed revealing a low genetic variation (98-100% nucleotide identity) amongst the BBTV isolates detected in seven of 11 DRC provinces. The phylogenetic tree showed that all DRC isolates gather a unique clade in the South Pacific group of BBTV. Based on the coding region for replication initiator protein, haplotype diversity was estimated to be 0.944±0.013, with 30 different haplotypes from 68 isolates in DRC. In addition, fives isolates were selected from each province for total genome sequencing, confirming low genetic variation among isolates from seven provinces (97-100% nucleotide identity). CONCLUSION This study strengthens the hypothesis of a single BBTV introduction some time ago, followed by the spread of the virus in the country. Analysis of the geographical dispersion reveals specific haplotypes groups according to the different agro-environmental conditions.

International audience ; Cassava is one of the most important staple crops in Africa and its production is seriously damaged by viral diseases. In this study, we identify for the first time and characterize the genome organization of novel ampeloviruses infecting cassava plants in diverse geographical locations using three high-throughput sequencing protocols [Virion-Associated Nucleotide Acid (VANA), dsRNA and total RNA], and we provide a first analysis of the diversity of these agents and of the evolutionary forces acting on them. Thirteen new Closteroviridae isolates were characterized in field-grown cassava plants from the Democratic Republic of Congo (DR Congo), Madagascar, Mayotte, and Reunion islands. The analysis of the sequences of the corresponding contigs (ranging between 10,417 and 13,752 nucleotides in length) revealed seven open reading frames. The replication-associated polyproteins have three expected functional domains: methyltransferase , helicase, and RNA-dependent RNA polymerase (RdRp). Additional open reading frames code for a small transmembrane protein, a heat-shock protein 70 homolog (HSP70h), a heat shock protein 90 homolog (HSP90h), and a major and a minor coat protein (CP and CPd respectively). Defective genomic variants were also identified in some cassava accessions originating from Madagascar and Reunion. The isolates were found to belong to two species tentatively named Manihot esculenta-associated virus 1 and 2 (MEaV-1 and MEaV-2). Phylogenetic analyses showed that MEaV-1 and MEaV-2 belong to the genus Ampelovirus, in particular to its subgroup II. MEaV-1 was found in all of the countries of study, while MEaV-2 was only detected in Madagascar and Mayotte. Recombination analysis provided evidence of intraspecies recombination occurring between the isolates from Madagascar and Mayotte. No clear association with visual symptoms in the cassava host could be identified.

International audience ; Cassava is one of the most important staple crops in Africa and its production is seriously damaged by viral diseases. In this study, we identify for the first time and characterize the genome organization of novel ampeloviruses infecting cassava plants in diverse geographical locations using three high-throughput sequencing protocols [Virion-Associated Nucleotide Acid (VANA), dsRNA and total RNA], and we provide a first analysis of the diversity of these agents and of the evolutionary forces acting on them. Thirteen new Closteroviridae isolates were characterized in field-grown cassava plants from the Democratic Republic of Congo (DR Congo), Madagascar, Mayotte, and Reunion islands. The analysis of the sequences of the corresponding contigs (ranging between 10,417 and 13,752 nucleotides in length) revealed seven open reading frames. The replication-associated polyproteins have three expected functional domains: methyltransferase , helicase, and RNA-dependent RNA polymerase (RdRp). Additional open reading frames code for a small transmembrane protein, a heat-shock protein 70 homolog (HSP70h), a heat shock protein 90 homolog (HSP90h), and a major and a minor coat protein (CP and CPd respectively). Defective genomic variants were also identified in some cassava accessions originating from Madagascar and Reunion. The isolates were found to belong to two species tentatively named Manihot esculenta-associated virus 1 and 2 (MEaV-1 and MEaV-2). Phylogenetic analyses showed that MEaV-1 and MEaV-2 belong to the genus Ampelovirus, in particular to its subgroup II. MEaV-1 was found in all of the countries of study, while MEaV-2 was only detected in Madagascar and Mayotte. Recombination analysis provided evidence of intraspecies recombination occurring between the isolates from Madagascar and Mayotte. No clear association with visual symptoms in the cassava host could be identified.

International audience ; Cassava is one of the most important staple crops in Africa and its production is seriously damaged by viral diseases. In this study, we identify for the first time and characterize the genome organization of novel ampeloviruses infecting cassava plants in diverse geographical locations using three high-throughput sequencing protocols [Virion-Associated Nucleotide Acid (VANA), dsRNA and total RNA], and we provide a first analysis of the diversity of these agents and of the evolutionary forces acting on them. Thirteen new Closteroviridae isolates were characterized in field-grown cassava plants from the Democratic Republic of Congo (DR Congo), Madagascar, Mayotte, and Reunion islands. The analysis of the sequences of the corresponding contigs (ranging between 10,417 and 13,752 nucleotides in length) revealed seven open reading frames. The replication-associated polyproteins have three expected functional domains: methyltransferase , helicase, and RNA-dependent RNA polymerase (RdRp). Additional open reading frames code for a small transmembrane protein, a heat-shock protein 70 homolog (HSP70h), a heat shock protein 90 homolog (HSP90h), and a major and a minor coat protein (CP and CPd respectively). Defective genomic variants were also identified in some cassava accessions originating from Madagascar and Reunion. The isolates were found to belong to two species tentatively named Manihot esculenta-associated virus 1 and 2 (MEaV-1 and MEaV-2). Phylogenetic analyses showed that MEaV-1 and MEaV-2 belong to the genus Ampelovirus, in particular to its subgroup II. MEaV-1 was found in all of the countries of study, while MEaV-2 was only detected in Madagascar and Mayotte. Recombination analysis provided evidence of intraspecies recombination occurring between the isolates from Madagascar and Mayotte. No clear association with visual symptoms in the cassava host could be identified.

International audience ; Cassava is one of the most important staple crops in Africa and its production is seriously damaged by viral diseases. In this study, we identify for the first time and characterize the genome organization of novel ampeloviruses infecting cassava plants in diverse geographical locations using three high-throughput sequencing protocols [Virion-Associated Nucleotide Acid (VANA), dsRNA and total RNA], and we provide a first analysis of the diversity of these agents and of the evolutionary forces acting on them. Thirteen new Closteroviridae isolates were characterized in field-grown cassava plants from the Democratic Republic of Congo (DR Congo), Madagascar, Mayotte, and Reunion islands. The analysis of the sequences of the corresponding contigs (ranging between 10,417 and 13,752 nucleotides in length) revealed seven open reading frames. The replication-associated polyproteins have three expected functional domains: methyltransferase , helicase, and RNA-dependent RNA polymerase (RdRp). Additional open reading frames code for a small transmembrane protein, a heat-shock protein 70 homolog (HSP70h), a heat shock protein 90 homolog (HSP90h), and a major and a minor coat protein (CP and CPd respectively). Defective genomic variants were also identified in some cassava accessions originating from Madagascar and Reunion. The isolates were found to belong to two species tentatively named Manihot esculenta-associated virus 1 and 2 (MEaV-1 and MEaV-2). Phylogenetic analyses showed that MEaV-1 and MEaV-2 belong to the genus Ampelovirus, in particular to its subgroup II. MEaV-1 was found in all of the countries of study, while MEaV-2 was only detected in Madagascar and Mayotte. Recombination analysis provided evidence of intraspecies recombination occurring between the isolates from Madagascar and Mayotte. No clear association with visual symptoms in the cassava host could be identified.

Cassava is one of the most important staple crops in Africa and its production is seriously damaged by viral diseases. In this study, we identify for the first time and characterize the genome organization of novel ampeloviruses infecting cassava plants in diverse geographical locations using three high-throughput sequencing protocols [Virion-Associated Nucleotide Acid (VANA), dsRNA and total RNA], and we provide a first analysis of the diversity of these agents and of the evolutionary forces acting on them. Thirteen new Closteroviridae isolates were characterized in field-grown cassava plants from the Democratic Republic of Congo (DR Congo), Madagascar, Mayotte, and Reunion islands. The analysis of the sequences of the corresponding contigs (ranging between 10,417 and 13,752 nucleotides in length) revealed seven open reading frames. The replication-associated polyproteins have three expected functional domains: methyltransferase , helicase, and RNA-dependent RNA polymerase (RdRp). Additional open reading frames code for a small transmembrane protein, a heat-shock protein 70 homolog (HSP70h), a heat shock protein 90 homolog (HSP90h), and a major and a minor coat protein (CP and CPd respectively). Defective genomic variants were also identified in some cassava accessions originating from Madagascar and Reunion. The isolates were found to belong to two species tentatively named Manihot esculenta-associated virus 1 and 2 (MEaV-1 and MEaV-2). Phylogenetic analyses showed that MEaV-1 and MEaV-2 belong to the genus Ampelovirus, in particular to its subgroup II. MEaV-1 was found in all of the countries of study, while MEaV-2 was only detected in Madagascar and Mayotte. Recombination analysis provided evidence of intraspecies recombination occurring between the isolates from Madagascar and Mayotte. No clear association with visual symptoms in the cassava host could be identified.

Cassava is one of the most important staple crops in Africa and its production is seriously damaged by viral diseases. In this study, we identify for the first time and characterize the genome organization of novel ampeloviruses infecting cassava plants in diverse geographical locations using three high-throughput sequencing protocols [Virion-Associated Nucleotide Acid (VANA), dsRNA and total RNA], and we provide a first analysis of the diversity of these agents and of the evolutionary forces acting on them. Thirteen new Closteroviridae isolates were characterized in field-grown cassava plants from the Democratic Republic of Congo (DR Congo), Madagascar, Mayotte, and Reunion islands. The analysis of the sequences of the corresponding contigs (ranging between 10,417 and 13,752 nucleotides in length) revealed seven open reading frames. The replication-associated polyproteins have three expected functional domains: methyltransferase, helicase, and RNA-dependent RNA polymerase (RdRp). Additional open reading frames code for a small transmembrane protein, a heat-shock protein 70 homolog (HSP70h), a heat shock protein 90 homolog (HSP90h), and a major and a minor coat protein (CP and CPd respectively). Defective genomic variants were also identified in some cassava accessions originating from Madagascar and Reunion. The isolates were found to belong to two species tentatively named Manihot esculenta-associated virus 1 and 2 (MEaV-1 and MEaV-2). Phylogenetic analyses showed that MEaV-1 and MEaV-2 belong to the genus Ampelovirus, in particular to its subgroup II. MEaV-1 was found in all of the countries of study, while MEaV-2 was only detected in Madagascar and Mayotte. Recombination analysis provided evidence of intraspecies recombination occurring between the isolates from Madagascar and Mayotte. No clear association with visual symptoms in the cassava host could be identified. ; Peer reviewed