Diversity of entomopathogenic fungi associated with Mediterranean fruit fly (Ceratitis capitata (Diptera: Tephritidae)) in Moroccan Argan forests and nearby area: impact of soil factors on their distribution.

BACKGROUND: Studying the ecology of biocontrol-agents is a prerequisite to effectively control medfly (Ceratitis capitata (Diptera: Tephritidae)) with entomopathogenic fungi. In this context, factors affecting the occurrence and distribution of medfly-associated entomopathogenic-fungi were studied. Soil samples (22) were collected from natural and cultivated areas of Souss-region Morocco. RESULTS: A total of 260 fungal isolates belonging to 22 species and 10 genera were obtained by using medfly pupae as bait. Medfly-associated fungi were detected in all studied soils and pupae infection percentages ranged from 3.33% to 48%. Two genera, Fusarium and Beauveria were the most frequent with 83 isolates (32%) and 50 isolates (19.23%) respectively. Pathogenicity test of isolated species against medfly pupae showed high mortality rates up to 91% for some strains. Principal component analysis (PCA) demonstrated a strong influence of origin, physical, and chemical properties of soil on the abundance of these fungi. In general, medfly-associated fungi were more abundant in soils with moderate pH (7.5 to 8) having high sand and organic content. High relative humidity negatively influenced the abundance of these fungi. Both factors directly affected the fungal infection percentages in pupae. The response of fungi to these parameters varied among species. According to principal component analysis (PCA), the soils of argan fields and forests were more suitable for the development of medfly-associated fungi than citrus orchards. CONCLUSIONS: These results guide identifying suitable soils for the effective application of entomopathogenic fungi as biological control agents. In summary, isolated indigenous strains seem to be a promising option to control C. capitata.

Diversity of root-knot nematodes in Moroccan olive nurseries and orchards: does Meloidogyne javanica disperse according to invasion processes?

BACKGROUND: Root-knot nematodes (RKN) are major pest of olive tree (Olea europaea ssp. europaea), especially in nurseries and high-density orchards. Soil samples were collected from main olive growing areas of Morocco, to characterize Meloidogyne species and to discuss the contribution of biotic and abiotic factors in their spatial distribution. RESULTS: RKN were found in 159 soil samples out of 305 from nurseries (52.1% occurrence) and in 11 out of 49 soil samples from orchards (23.2% occurrence). Biochemical and molecular characterisation (PAGE esterase and SCAR) revealed the dominance of M. javanica both in nurseries and orchards with minor presence of M. incognita only in nurseries, and M. arenaria in only one nursery. RKN were distributed on aggregated basis. Frequent presence of M. javanica in orchards might have come from nurseries. In contrast, the detection of M. incognita in nurseries alone suggests that this species could not reproduce in orchards because of either the competition with other plant-parasitic nematodes or unfit local habitats. The impact of environmental variables (climate, habitat origin and physicochemical characteristics of the substrates) on the distribution of Meloidogyne species is also discussed. CONCLUSION: Olive nurseries in Morocco are not able to guarantee the safety of rooted plants. As a result, olive production systems are exposed to strong RKN invasion risks. Consequently, the use of healthy substrates in nurseries may prevent plant-parasitic nematode induction in orchards.

How anthropogenic changes may affect soil-borne parasite diversity? Plant-parasitic nematode communities associated with olive trees in Morocco as a case study.

BACKGROUND: Plant-parasitic nematodes (PPN) are major crop pests. On olive (Olea europaea), they significantly contribute to economic losses in the top-ten olive producing countries in the world especially in nurseries and under cropping intensification. The diversity and the structure of PPN communities respond to environmental and anthropogenic forces. The olive tree is a good host plant model to understand the impact of such forces on PPN diversity since it grows according to different modalities (wild, feral and cultivated olives). A wide soil survey was conducted in several olive-growing regions in Morocco. The taxonomical and the functional diversity as well as the structures of PPN communities were described and then compared between non-cultivated (wild and feral forms) and cultivated (traditional and high-density olive cultivation) olives. RESULTS: A high diversity of PPN with the detection of 117 species and 47 genera was revealed. Some taxa were recorded for the first time on olive trees worldwide and new species were also identified. Anthropogenic factors (wild vs cultivated conditions) strongly impacted the PPN diversity and the functional composition of communities because the species richness, the local diversity and the evenness of communities significantly decreased and the abundance of nematodes significantly increased in high-density conditions. Furthermore, these conditions exhibited many more obligate and colonizer PPN and less persister PPN compared to non-cultivated conditions. Taxonomical structures of communities were also impacted: genera such as Xiphinema spp. and Heterodera spp. were dominant in wild olive, whereas harmful taxa such as Meloidogyne spp. were especially enhanced in high-density orchards. CONCLUSIONS: Olive anthropogenic practices reduce the PPN diversity in communities and lead to changes of the community structures with the development of some damaging nematodes. The study underlined the PPN diversity as a relevant indicator to assess community pathogenicity. That could be taken into account in order to design control strategies based on community rearrangements and interactions between species instead of reducing the most pathogenic species.

Taxonomic identification of Morocco scorpions using MALDI-MS fingerprints of venom proteomes and computational modeling.

The venom of scorpions has been the subject of numerous studies. However, their taxonomic identification is not a simple task, leading to misidentifications. This study aims to provide a practical approach for identifying scorpions based on the venom molecular mass fingerprint (MFP). Specimens (251) belonging to fifteen species were collected from different regions in Morocco. Their MFPs were acquired using MALDI-MS. These were used as a training dataset to generate predictive models and a library of mean spectral profiles using software programs based on machine learning. The computational model achieved an overall recognition capability of 99 % comprising 32 molecular signatures. The models and the library were tested using a new dataset for external validation and to evaluate their capability of identification. We recorded an accuracy classification with an average of 97 % and 98 % for the computational models and the library, respectively. To our knowledge, this is the first attempt to demonstrate the potential of MALDI-MS and MFPs to generate predictive models capable of discriminating scorpions from family to species levels, and to build a library of species-specific spectra. These promising results may represent a proof of concept towards developing a reliable approach for rapid molecular identification of scorpions in Morocco. SIGNIFICANCE OF THE STUDY: With their clinical importance, scorpions may constitute a desirable study model for many researchers. The first step in studying scorpion is systematically identifying the species of interest. However, it can be a difficult task, especially for the non-experts. The taxonomy of scorpions is primarily based on morphometric characters. In Morocco, the high number of species and subspecies mainly endemic, and the morphological similarities between different species may result in false identifications. This was observed in many reports according to the scorpion experts. In this study, we describe a reliable practical approach for identifying scorpions based on the venom molecular mass fingerprints (MFPs). By using two software programs based on machine learning, we have demonstrated that these MFPs contains sufficient inter-specific variation to differentiate between the scorpion species mentioned in this study with a good accuracy. Using a drop of venom, this new approach could be a rapid, accurate and cost saving method for taxonomic identification of scorpions in Morocco.

Molecular diversity assessed by MALDI mass spectrometry of two scorpion species venom from two different locations in Morocco.

Scorpion venom is a cocktail of molecules whose composition is remarkably plastic, controlled by several factors. The Moroccan scorpion fauna is characterized by its richness and high rate of endemism and the venom molecular variability of many species is not yet well characterized. The aim of the present study was to highlight the molecular variability of the venom composition of Androctonus amoreuxi and Buthacus stockmanni (endemic species), both belonging to the Buthidae family, collected from two Moroccan regions, Zagora and Tan-tan. Characterization of the molecular mass fingerprints (MFPs) of each specimen was performed by Matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) using a sandwich (Sand) and a dried-droplet (DD) sample preparation and dilutions. Considering these two methods, a total of 828 ion signals were detected, and Sand method produced more adducts (56%) than DD (44%). We observed interspecific variations in the venom composition between these two species showing they share 235 ion signals, while 226 and 367 are specific for these two species, respectively. Moreover, B. stockmanni specimens showed a clear difference in their MFPs between the two geographical areas studied, suggesting intraspecific variations. Moreover, specimens from each population also show an intraspecific variability. In addition, for the same individual, a variation in the venom composition was also recorded depending on the milking frequency. Our results confirmed the presence of characteristic components in each extracted venom sample. In conclusion, MFPs assessed by MALDI-MS represent a fast, non-supervised, sensitive, reliable and cost-efficient approach for taxonomic identification and molecular variability characterization. This study undoubtedly represents a step forward for understanding the scorpion venom plasticity, intra/inter variations, and their temporal and geographical variability.

Phytoparasitic nematodes of organic vegetables in the Argan Biosphere of Souss-Massa (Southern Morocco).

Agroecological productivity of the Arganeraie Biosphere Reserve of Morocco is limited by the wide spread and dynamics of plant parasitic nematodes (PPN). Ecological studies of nematode communities are required to develop effective biological management of these bioagressors as conventional control methods of PPN are inadequate and have persistent harmful effects. Fifty-nine organic vegetable soils in Souss-Massa were nematologically sampled, and assessment of taxonomic proliferation was made in relation to host species, geographical origin, and climatic and microclimatic factors. Twenty-four nematode genera were identified as obligate and facultative plant feeders. Taxonomic diversity increased from Chtouka to Taroudant and Tiznit provinces. Soil texture, organic matter, pH, nitrogen, zinc, magnesium, copper, altitude, and humidity and temperature were seen to effect driving roles in the abundance, distribution, and community structures of nematodes. The most prevalent taxa posing a high risk to organic agriculture of Souss Massa were needle nematodes (Longidorus spp.) and root-knot nematodes (Meloidogyne spp.). Edaphic and climatic variables effected nematode populations greatly. A combination of biological treatments and appropriate agroecological practices restricting important economic PPN growth and enhancing soil quality are required to achieve sustainable management in the area.