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Pseudomonas isolates from tropical environments have been underexplored and may form an untapped reservoir of interesting secondary metabolites. In this study, we compared Pseudomonas and cyclic lipopeptide (CLP) diversity in the rhizosphere of a cocoyam root rot disease (CRRD) suppressive soil in Boteva, Cameroon with those from four conducive soils in Cameroon and Nigeria. Compared with other soils, Boteva andosols were characterized by high silt, organic matter, nitrogen and calcium. Besides, the cocoyam rhizosphere at Boteva was characterized by strains belonging mainly to the P. koreensis and P. putida (sub)groups, with representations in the P. fluorescens, P. chlororaphis, P. jessenii and P. asplenii (sub)groups. In contrast, P. putida isolates were prominent in conducive soils. Regarding CLP diversity, Boteva was characterized by strains producing 11 different CLP types with cocoyamide A producers, belonging to the P. koreensis group, being the most abundant. However, putisolvin III-V producers were the most dominant in the rhizosphere of conducive soils in both Cameroon and Nigeria. Furthermore, we elucidated the chemical structure of putisolvin derivatives-putisolvin III-V, and described its biosynthetic gene cluster. We show that high Pseudomonas and metabolic diversity may be driven by microbial competition, which likely contributes to soil suppressiveness to CRRD.

Pseudomonas isolates from tropical environments have been underexplored and may form an untapped reservoir of interesting secondary metabolites. In this study, we compared Pseudomonas and cyclic lipopeptide (CLP) diversity in the rhizosphere of a cocoyam root rot disease (CRRD) suppressive soil in Boteva, Cameroon with those from four conducive soils in Cameroon and Nigeria. Compared with other soils, Boteva andosols were characterized by high silt, organic matter, nitrogen and calcium. Besides, the cocoyam rhizosphere at Boteva was characterized by strains belonging mainly to the P. koreensis and P. putida (sub)groups, with representations in the P. fluorescens, P. chlororaphis, P. jessenii and P. asplenii (sub)groups. In contrast, P. putida isolates were prominent in conducive soils. Regarding CLP diversity, Boteva was characterized by strains producing 11 different CLP types with cocoyamide A producers, belonging to the P. koreensis group, being the most abundant. However, putisolvin III-V producers were the most dominant in the rhizosphere of conducive soils in both Cameroon and Nigeria. Furthermore, we elucidated the chemical structure of putisolvin derivatives-putisolvin III-V, and described its biosynthetic gene cluster. We show that high Pseudomonas and metabolic diversity may be driven by microbial competition, which likely contributes to soil suppressiveness to CRRD.

International audience; Cocoyam (Xanthosoma sagittifolium (L.)), an important tuber crop in the tropics, is severely affected by the cocoyam root rot disease (CRRD) caused by Pythium myriotylum. The white cocoyam genotype is very susceptible while the red cocoyam has some field tolerance to CRRD. Fluorescent Pseudomonas isolates obtained from the rhizosphere of healthy red and white cocoyams from three different fields in Cameroon were taxonomically characterized. The cocoyam rhizosphere was enriched with P. fluorescens complex and P. putida isolates independent of the plant genotype. LC–MS and NMR analyses revealed that 50% of the Pseudomonas isolates produced cyclic lipopeptides (CLPs) including entolysin, lokisin, WLIP, putisolvin and xantholysin together with eight novel CLPs. In general, CLP types were linked to specific taxonomic groups within the fluorescent pseudomonads. Representative CLP‐producing bacteria showed effective control against CRRD while purified CLPs caused hyphal branching or hyphal leakage in P. myriotylum. The structure of cocoyamide A, a CLP which is predominantly produced by P. koreensis group isolates within the P. fluorescens complex is described. Compared with the white cocoyam, the red cocoyam rhizosphere appeared to support a more diverse CLP spectrum. It remains to be investigated whether this contributes to the field tolerance displayed by the red cocoyam.

Retrotransposon-based molecular markers were applied for the first time within the genera Xanthosoma and Colocasia to assess intraspecific variability among 27 accessions of cocoyam (Xanthosoma sagittifolium) and taro (Colocasia esulenta). Over 16 distinct retrotransposon fragments were isolated, sequenced, and LTR primers were designed to obtain Inter-Retrotransposon Amplified Polymorphism (IRAP) fingerprints. The set of six polymorphic LTR primers yielded 433 reproducible bands across a set of 20 X. sagittifolium samples. Out of the 433 bands, 400 fragments (92%) were polymorphic. In seven C. esculenta accessions, the six primers amplified a total of 354 reproducible, informative data points, of which 285 (80.5%) were polymorphic. Data concerning the number of polymorphic bands and Shannon’s index in X. sagittifolium accessions suggest that retrotransposon activity continued after Xanthosoma speciation. Cluster analysis placed all the accessions in two groups according to their species delimitation. The accessions of X. sagittifolium were further divided into two subgroups corresponding to their ploidy level. Moreover, our results showed that the genetic variability accessed by IRAP markers allows separation of X. sagittifolium and C. esculenta accessions according to their type and botanical variety respectively. These data provide a basis for better germplasm management, future systematic studies and genetic improvement, as well as for exploration of the role of retrotransposons in cocoyam and taro polyploid formation and genome dynamics.

In Cameroon, andosols are suspected to be suppressive to cocoyam ( Xanthosoma sagittifolium ) root rot disease (CRRD) caused by the Oomycete pathogen Pythium myriotylum . To determine factors involved in disease suppressiveness, andosols were studied in comparison to ferralsols known to be disease-conducive. Soil samples were collected from six sites of which three were in andosols around Mount Cameroon (Boteva, Njonji, and Ekona) and the three others in ferralsols (Bakoa, Lapkwang, and Nko’o canane). Greenhouse plant experiments were used to assess soil suppressiveness. Soils were artificially infested with two levels of P. myriotylum inoculum (100 and 300 mycelia strands g −1 soil) prior to planting cocoyam. Disease severity was significantly higher in ferralsols than in andosols. Andosols partly lost their suppressiveness as a result of autoclaving and could recover suppressiveness following recolonisation by their original microflora. Soil microbial groups implicated in the disease suppression were investigated by assessing the effect of fungicide, bactericide, and pasteurisation on andosol suppressiveness. Andosols suppressiveness was significantly reduced following pasteurisation and treatment with fungicide and bactericide. The possible influence of microbial biomass on andosol suppressiveness was investigated by comparing microbial populations of suppressive andosols to those in andosols that had lost suppressiveness. A comparative analysis of suppressive and conducive soil properties was performed to identify soil variables, which may contribute to soil suppressiveness. Soil chemical analysis results showed that organic matter content was higher in andosols than in ferralsols. In addition, the content of mineral nutrients such as Ca, K, Mg and N, was higher in andosols than in ferralsols. These soil variables negatively correlated with disease severity. By contrast, sand and clay, which were higher in ferralsols than in andosols, were positively related to disease severity. This study has confirmed the suppressive nature of andosols from Mount Cameroon to CRRD. The results suggest that high organic matter content is likely mediating P. myriotylum suppression in andosols by improving soil structure, increasing soil nutrient content and microbial biomass, and sustaining microbial activity.

Intraspecific variability within 51 isolates of Pythium myriotylum from cocoyam ( Xanthosoma sagittifolium ) and other host crops was analysed using optimum growth temperature, esterase banding patterns, AFLPs, rDNA–ITS sequencing, and virulence to cocoyam. P. myriotylum isolates virulent to cocoyam could easily be differentiated from other isolates of P. myriotylum by their optimum growth temperature. Isolates from cocoyam grew best at 28 °C with no growth at 37 °C, while P. myriotylum isolates from other host crops had their optimum growth temperature at 37 °C. Esterases produced consistent zymograms with 18 discrete esterase markers, but no monomorphic markers were produced for isolates virulent to cocoyam. Isozyme profiles based on esterase analysis showed that isolates that infect cocoyam plantlets formed a related group, irrespective of their geographic origin. P. myriotylum isolates from other host plants also grouped together, but could clearly be distinguished from the cocoyam cluster. AFLPs produced 189 scorable bands for the cocoyam isolates, of which 77 % are monomorphic. Phenetic analysis of AFLP data grouped all isolates originating from cocoyam together except for the isolates C103-04, CMR17, CMR22, and CMR25. These isolates regrouped with isolates of Pythium myriotylum from other host crops or the outgroup and were found not to be pathogenic for cocoyam. ITS sequences of isolates of P. myriotylum from cocoyam were 99.1–99.7 % identical to sequences deposited in GenBank. However, alignments of ITS sequences revealed a base transition at position 824 from adenine in typical isolates of P. myriotylum to guanine in isolates that could infect cocoyam plantlets. In a limited pathogenicity test, all isolates from cocoyam having guanine at position 824 were able to infect tissue culture derived cocoyam but not those exhibiting adenine. This study demonstrates for the first time, molecular evidence that isolates of P. myriotylum that infect cocoyam are distinct from P. myriotylum isolates from other crops and have developed a certain degree of host adaptation.

In this study, the putative role of phenazines and rhamnolipid-biosurfactants, antagonistic metabolites produced by Pseudomonas aeruginosa PNA1, was tested in the biological control of Pythium splendens on bean (Phaseolus vulgaris L) and Pythium myriotylum on cocoyam (Xanthosoma sagittifolium L Schott). A rhamnolipid-deficient and a phenazine-deficient mutant of PNA1 were used either separately or jointly in plant experiments. When the mutants were applied separately, no disease-suppressive effect was observed, although both mutants still produced one of the antagonistic compounds (phenazines or rhamnolipids). When the mutants were concurrently introduced in the soil, the biocontrol activity was restored to wild-type levels. Bean seeds developed significantly less pre-emergence damping-off caused by P. splendens when treated with a mixture of purified phenazine-1-carboxamide and rhamnolipids than with any of the chemicals alone. When phenazines and rhamnolipids were combined at concentrations that had no observable effects when the metabolites were applied separately, mycelial growth of P. myriotylum was significantly reduced. In addition, microscopic analysis revealed substantial vacuolization and disintegration of Pythium hyphae after incubation in liquid medium amended with both metabolites. Results of this study indicate that phenazines and biosurfactants are acting synergistically in the control of Pythium spp.

Aim: To screen for novel antagonistic Pseudomonas strains producing both phenazines and biosurfactants that are as effective as Pseudomonas aeruginosa PNA1 in the biocontrol of cocoyam root rot caused by Pythium myriotylum. Material and Results: Forty pseudomonads were isolated from the rhizosphere of healthy white and red cocoyam plants appearing in natural, heavily infested fields in Cameroon. In vitro tests demonstrated that Py. myriotylum antagonists could be retrieved from the red cocoyam rhizosphere. Except for one isolate, all antagonistic isolates produced phenazines. Results from whole-cell protein profiling showed that the antagonistic isolates are different from other isolated pseudomonads, while BOX-PCR revealed high genomic similarity among them. 16S rDNA sequencing of two representative strains within this group of antagonists confirmed their relatively low similarity with validly described Pseudomonas species. These antagonists are thus provisionally labelled as unidentified Pseudomonas strains. Among the antagonists, Pseudomonas CMR5c and CMR12a were selected because of their combined production of phenazines and biosurfactants. For strain CMR5c also, production of pyrrolnitrin and pyoluteorin was demonstrated. Both CMR5c and CMR12a showed excellent in vivo biocontrol activity against Py. myriotylum to a similar level as Ps. aeruginosa PNA1. Conclusion: Pseudomonas CMR5c and CMR12a were identified as novel and promising biocontrol agents of Py. myriotylum on cocoyam, producing an arsenal of antagonistic metabolites. Significance and Impact of the Study: Present study reports the identification of two newly isolated fluorescent Pseudomonas strains that can replace the opportunistic human pathogen Ps. aeruginosa PNA1 in the biocontrol of cocoyam root rot and could be taken into account for the suppression of many plant pathogens.

After the closure of Cameroon’s National Produce Marketing Board in 1990, rigorous measures no longer existed to control production of quality cocoa. This study assesses farmers’ knowledge of postharvest practices that could influence cocoa bean quality (CBQ), their perception of quality cocoa and the factors that hinder good farming and postharvest practices (GFPHP). We also make suggestions on how quality control can be incorporated in the production chain to improve on CBQ produced. In a cross-sectional, purposive study in Fako Division, South West Region of Cameroon, 100 farmers were enrolled from 10 villages (10 from each village) which make up the major production area in Agro-ecological Zone IV of Cameroon. Farmers were administered a semi-structured questionnaire in the language they best understood. Cocoa farming in this area is a tradition and our results show that 97 % of farmers are males and acquired farming practices mainly through their family member (43 %) and/or friends (34 %). Farms are self-owned, 76 and 85 %, respectively, are less than 5Ha and have been cultivated for more than 20 years. Only 10 % of farmers could perceive quality cocoa using physical characterization. Although more than 95 % of farmers knew GFPHP, only 48 % undertake these practices. Farmers’ focus was on bean weight and not on CBQ and 64 % of farmers think quality has little impact on the price. Most farmers (71 %) sell to “buyam-sellams” who care little about quality. The proportion of respondents who acquired knowledge through formal learning was significantly higher than those who did so from informal learning with regards to both good cocoa production practice (p = 0.036) and appreciation of quality attributes (p = 0.043). Subsidizing inputs, access to credits, creating cooperatives, regulating price of cocoa and training on postharvest management in decreasing order could enhance practices that will ensure quality cocoa production. GFPHP is important in the production of quality cocoa. Left alone, farmers will not pay attention to these quality attributes especially as premium quality is not rewarded. Organizing farmers into cooperatives will reduce the link between farmers and companies exporting cocoa and alleviate most bottlenecks in the sector.