Beneficial Microorganisms as Bioprotectants against Foliar Diseases of Cereals: A Review.

Cereal production plays a major role in both animal and human diets throughout the world. However, cereal crops are vulnerable to attacks by fungal pathogens on the foliage, disrupting their biological cycle and photosynthesis, which can reduce yields by 15-20% or even 60%. Consumers are concerned about the excessive use of synthetic pesticides given their harmful effects on human health and the environment. As a result, the search for alternative solutions to protect crops has attracted the interest of scientists around the world. Among these solutions, biological control using beneficial microorganisms has taken on considerable importance, and several biological control agents (BCAs) have been studied, including species belonging to the genera Bacillus, Pseudomonas, Streptomyces, Trichoderma, Cladosporium, and Epicoccum, most of which include plants of growth-promoting rhizobacteria (PGPRs). Bacillus has proved to be a broad-spectrum agent against these leaf cereal diseases. Interaction between plant and beneficial agents occurs as direct mycoparasitism or hyperparasitism by a mixed pathway via the secretion of lytic enzymes, growth enzymes, and antibiotics, or by an indirect interaction involving competition for nutrients or space and the induction of host resistance (systemic acquired resistance (SAR) or induced systemic resistance (ISR) pathway). We mainly demonstrate the role of BCAs in the defense against fungal diseases of cereal leaves. To enhance a solution-based crop protection approach, it is also important to understand the mechanism of action of BCAs/molecules/plants. Research in the field of preventing cereal diseases is still ongoing.

Counter-attack of biocontrol agents: Environmentally benign Approaches against Root-knot nematodes (Meloidogyne spp.) on Agricultural crops.

Root-knot nematodes (Meloidogyne spp.) are obligate sedentary endoparasites, considered severe crop-damaging taxa among all plant-parasitic nematodes globally. Their attacks through parasitic proteins alter the physiology and machinery of the host cells to favour parasitism and reduction in crop yield. Currently, the use of excessive pesticides as a fast remedy to manage this pest is hazardous for both the environment and humans. Keeping this view in mind, there is an urgent need for developing efficient eco-friendly strategies. Bio-control as an eco-friendly is considered the best approach to manage nematodes without disturbing non-target microbes. In bio-control, living agents such as fungi and bacteria are the natural enemies of nematodes and the best substitute for pesticides. Fungi, including nematode-trapping fungi, can sense host signals and produce special trapping devices viz., constricting rings and adhesive knobs/loops, to capture nematodes and kill them. Whereas, endo-parasitic fungi kill nematodes by enzymatic secretions and spore adhesion through their hyphae. Bacteria can also control nematodes by producing antibiotic compounds, competing for nutrients and rhizosphere, production of hydrolytic enzymes viz., chitinases, proteases, lipases, and induction of systemic resistance (ISR) in host plants. Scientists throughout the world are trying to evolve environmentally benign methods that sustain agricultural production and keep nematodes below a threshold level. Whatever methods evolve, in the future the focus should be on important aspects like green approaches for managing nematodes without disturbing human health and the environment.

Occurrence and Geographic Distribution of Plant-Parasitic Nematodes Associated with Citrus in Morocco and Their Interaction with Soil Patterns.

Plant-parasitic nematodes (PPNs) are found in citrus plantations throughout the world, but they are considered to be the most problematic pest in Morocco. Citrus fruit quality and yield have been adversely affected by PPNs. Due to data unavailability of nematodes associated with citrus, a detailed survey was conducted in the main citrus-growing regions of Morocco during 2020-2021 to assess the occurrence, distribution, and diversity of PPNs associated with rhizospheres of citrus trees. In addition, some soil properties have also been assessed for their impact on soil properties. Plant-parasitic nematode diversity was calculated using two ecological indexes, the Shannon diversity index (H') and the Evenness index (E). The collected soil and root samples were analyzed, and eleven genera and ten species of plant-parasitic nematodes were identified. The results show that the most predominant PPN species were Tylenchulus semipenetrans (88%), Helicotylenchus spp. (75%), Pratylenchus spp. (47%), Tylenchus spp. (51%), and Xiphinema spp. (31%). The results showed that PPN distributions were correlated with soil physicochemical properties such as soil texture, pH levels, and mineral content. Based on the obtained result, it was concluded that besides the direct effects of the host plant, physicochemical factors of the soil could greatly affect PPN communities in citrus growing orchards.

Moroccan entomopathogenic nematodes as potential biocontrol agents against Dactylopius opuntiae (Hemiptera: Dactylopiidae).

Dactylopius opuntiae (Cockerell) (Hemiptera: Dactylopiidae) or prickly pear cochineal, is the most damaging pest on cactus species with heavy economic losses worldwide. The efficacy of two Moroccan EPN isolates; Steinernema feltiae (Filipjev) (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora (Poinar) (Rhabditida: Heterorhabditidae) (applied at 25, 50, and 75 IJs cm-2) against D. opuntiae nymphs and young females were evaluated under both laboratory bioassays and field conditions. Results showed that S. feltiae was more effective, causing higher mortality of nymphs and adult females (98.8% and 97.5%, respectively) after 8 days of exposure, resulting in an LT50 value of 5.9 days (nymph) and 6.0 days (young female). While, H. bacteriophora had lower mortalities (83.8% for nymph and 81.3% for adult females). For the cochineal nymphs and adult females, no significant difference was observed among S. feltiae at 25, 50, and 75 IJs cm-2, and the positive control, D-limonene applied at 0.5 g/L which was used due to its high effectiveness against nymphs and females of D. opuntiae. In the field experiment, D-limonene at 0.5 g/L and S. feltiae applied at 75 IJs cm-2 were effective in reducing nymph and adult female populations by 85.3-93.9% at 12 days of post exposure period. To our knowledge, this work is the first report on the use of EPNs to control D. opuntiae. Thus, in addition to D-limonene, both Moroccan EPN isolates S. feltiae, and H. bacteriophora could be used as part of the integrated pest management strategy against D. opuntiae. Many factors such as temperature can affect the establishment and effectiveness of EPNs under field conditions. Therefore, additional studies under field conditions are needed.

Antagonistic potential of Moroccan entomopathogenic nematodes against root-knot nematodes, Meloidogyne javanica on tomato under greenhouse conditions.

The root-knot nematode, Meloidogyne javanica is a devastating pest affecting tomato production worldwide. Entomopathogenic nematodes (EPNs) are considered very promising biocontrol agents that could be used to effectively manage plant-parasitic nematode. The antagonistic activity of five EPN strains isolated from different fields in Morocco was evaluated against juvenile (J2s) antagonism in soil, the number of egg masses, and the galling index of M. javanica and J2s reproduction in the root. In greenhouse experiments, Steinernema feltiae strains (EL45 and SF-MOR9), Steinernema sp. (EL30), and those of Heterorhabditis bacteriophora (HB-MOR7 and EL27) were applied to the soil alongside RKN J2s. There was a significant reduction in M. javanica densities in the soil and roots by EPNs treatments when compared to the positive control. The EPNs decreased both egg masses formation and galling index by 80% compared to the positive control. The application of EPNs at a rate of 50 and 75 infective juveniles (IJs) cm-2 gave significant control of all studied nematological parameters compared to the positive control, which confirmed the importance of the doses applied. The applied dose was significantly correlated with M. javanica parameters according to polynomial regression models. The results also showed that S. feltiae strain (EL45) significantly increased plant height and root length, while H. bacteriophora strain (HB-MOR7) only enhanced root fresh weight. Therefore, both indigenous EPN strains; EL45 and SF-MOR9 have eco-friendly biological potential against M. javanica in vegetable crops.

Monitoring of Host Suitability and Defense-Related Genes in Wheat to Bipolaris sorokiniana.

Spot blotch caused by Bipolaris sorokiniana is a destructive disease of wheat worldwide. This study investigated the aggressiveness of B. sorokiniana isolates from different wheat-growing areas of Bolu province in Turkey on the cultivar Seri-82. Host susceptibility of 55 wheat cultivars was evaluated against the most aggressive isolate. Our results indicated that the cultivars Anafarta and Koç-2015 were the most resistant. A specific and sensitive qPCR assay was developed for detecting the pathogen in plant tissues and evaluating wheat plants with different resistance levels. Three primer sets, BsGAPDHF/BsGAPDHR, BsITSF/BsITSR, and BsSSUF/BsSSUR, were designed based on glyceraldehyde-3-phosphate dehydrogenase, internal transcribed spacers, and 18S rRNA loci of B. sorokiniana with detection limits of 1, 0.1, and 0.1 pg of pathogen DNA, respectively. The qPCR assay was highly sensitive and did not amplify DNA from the other closely related fungal species and host plants. The protocol differentiated wheat plants with varying degrees of resistance. The assay developed a useful tool for the quantification of the pathogen in the early stages of infection and may provide a significant contribution to a more efficient selection of wheat genotypes in breeding studies. In the present study, expression levels of PR proteins, phenylalanine ammonia-lyase, catalase, ascorbate peroxidase, and superoxide dismutase enzymes were upregulated in Anafarta (resistant) and Nenehatun (susceptible) cultivars at different post-infection time points, but more induced in the susceptible cultivar. The results showed considerable variation in the expression levels and timing of defense genes in both cultivars.

High-throughput molecular technologies for unraveling the mystery of soil microbial community: challenges and future prospects.

Soil microbial communities play a crucial role in soil fertility, sustainability, and plant health. However, intensive agriculture with increasing chemical inputs and changing environments have influenced native soil microbial communities. Approaches have been developed to study the structure, diversity, and activity of soil microbes to better understand the biology and plant-microbe interactions in soils. Unfortunately, a good understanding of soil microbial community remains a challenge due to the complexity of community composition, interactions of the soil environment, and limitations of technologies, especially related to the functionality of some taxa rarely detected using conventional techniques. Culture-based methods have been shown unable and sometimes are biased for assessing soil microbial communities. To gain further knowledge, culture-independent methods relying on direct analysis of nucleic acids, proteins, and lipids are worth exploring. In recent years, metagenomics, metaproteomics, metatranscriptomics, and proteogenomics have been increasingly used in studying microbial ecology. In this review, we examined the importance of microbial community to soil quality, the mystery of rhizosphere and plant-microbe interactions, and the biodiversity and multi-trophic interactions that influence the soil structure and functionality. The impact of the cropping system and climate change on the soil microbial community was also explored. Importantly, progresses in molecular biology, especially in the development of high-throughput biotechnological tools, were extensively assessed for potential uses to decipher the diversity and dynamics of soil microbial communities, with the highlighted advantages/limitations.

Potential of Moroccan entomopathogenic nematodes for the control of the Mediterranean fruit fly Ceratitis capitata Wiedemann (Diptera: Tephritidae).

The Mediterranean fruit fly, Ceratitis capitata Wiedemann, is a deleterious pest worldwide affecting fruit production. The entomopathogenic nematodes (EPNs) are a potential biocontrol agent that could be effectively used to control this Mediterranean fruit fly. In this study, five EPN strains reported from different fields in Morocco were evaluated for their efficacy against C. capitata. In laboratory assays, Steinernema feltiae-SF-MOR9, S. feltiae-SF-MOR10 and Heterorhabditis bacteriophora-HB-MOR7 strains showed significantly higher infectivity and penetration rates when compared to the other strains. S. feltiae-SF-MOR9 caused the highest larval mortality rate (80%) at 50 infective juveniles (IJs) cm-2. However, additional results showed that both S. feltiae strains were significantly effective in controlling C. capitata larvae in apricot (Prunus armeniaca) fruits on soil surface with high mortality rate at 50 and 100 IJs cm-2. Different soil textures and moisture levels resulted in a significant variation in EPN strain virulence against C. capitata. Sandy clay loam soil in combination with 50 IJs cm-2 of S. feltiae (SF-MOR9 or SF-MOR10) caused a higher mortality rate of C. capitata larvae. Furthermore, applying these EPN strains at 50-100 IJs cm-2 in combination with 10-15% moisture level showed optimal results against C. capitata larvae. Therefore, those two Moroccan EPN strains could be used as promising eco-friendly biological agents against C. capitata.

First report of the dagger nematode Xiphinema pachtaicum on onion in Morocco.

In 2018, during a survey in Souss region of Morocco, the dagger nematode Xiphinema pachtaicum was found parasitizing onion, (Allium cepa L.). The populations of dagger nematode were identified on the basis of their morphological and morphometric characters and by sequencing and analysing the D2D2 region of 28S rRNA. Microscopic observation of females demonstrated the occurrence of Xiphinema pachtaicum on onion. The 28S D2D3 sequences shared 99 to 100% similarity with sequence corresponding to X. pachtaicum in GenBank. To our knowledge this is the first report of Xiphinema pachtaichum infecting onion in Morocco.In 2018, during a survey in Souss region of Morocco, the dagger nematode Xiphinema pachtaicum was found parasitizing onion, (Allium cepa L.). The populations of dagger nematode were identified on the basis of their morphological and morphometric characters and by sequencing and analysing the D2D2 region of 28S rRNA. Microscopic observation of females demonstrated the occurrence of Xiphinema pachtaicum on onion. The 28S D2D3 sequences shared 99 to 100% similarity with sequence corresponding to X. pachtaicum in GenBank. To our knowledge this is the first report of Xiphinema pachtaichum infecting onion in Morocco.

Occurrence of the Root-Knot Nematode species in Vegetable Crops in Souss Region of Morocco.

Root-knot nematodes 'Meloidogyne spp' are the most destructive group of plant parasitic nematodes causeing serious losses in vegetables crops and this damages worsened when crops grown under greenhouses conditions. In this sutdy, the distribution and characterization of root-knot nematode species collected from the Souss region of Morocco where vegetables crops intensively cultivated were determined by using both morphological and molecular tools. Out of the 110 samples collected from different greenhouses 91 (81.7%) were found to be infested with root-knot nematodes. Thirty-seven populations of root-knot nematodes were morphologically identified based on perineal patterns as well as molecularlly using species-specific primers. The obtained results indicated that Meloidogyne javanica and M. incognita were identified in 86.4% and 13.5% of the total populations, respectively. The lowest incidence of root-knot nematodes (64%) was found in Toussous, whereas the highest frequencies of 100% and 90% were detected in Taddart and Biogra, respectively. As the majority of the samples have been infested with Meloidogyne species; this indicates that there is an urgent need to provide farmers with a proper control strategy.