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Safflower is known as a tolerant plant to abiotic stress factors. This study was conducted to introduce some physiological indices to improve drought-tolerant safflower genotypes for cultivation in marginal and arid areas. Six safflower genotypes were studied for two years (2017–2019) in the East Azarbaijan Agricultural and Natural Resources Research and Education Centre of Iran under non-stressed and low-water conditions from flowering to seed maturity. The occurrence of water deficits led to a significant decrease in relative water content (RWC), stomatal conductance (gs), osmotic adjustment (Oadj), water potential (WP) and agronomic water use efficiency (WUEa) and an increase in the water stress index (CWSI). In addition, the values of these traits differed significantly between the safflower genotypes. The correlations between the physiological traits and seed yield were significant. The regression relationships between seed yield and the above traits showed that CWSI, WP and WUEa had a strong relationship with seed yield under normal (R2 = 0.854, 0.801 and 0.856, respectively) and water deficit conditions (R2 = 0.931, 0.877 and 0.900, respectively). It can be concluded that the CWSI, WP and WUEa indices are able to select high-yielding and drought-tolerant safflower genotypes for the late season. Among the components of seed yield, the number of capitula per plant (r = 0.86) and seeds per capitula (r = 0.92), which were positively and significantly correlated with seed yield, played the main roles in the formation of seed yield. The Golemehr and Mec.295 genotypes achieved higher seed yields under normal (4676 and 4961 kg h−1, respectively) and water deficit conditions (3211 and 3385 kg h−1, respectively) and can be recommended for cultivation in marginal and arid areas. [ABSTRACT FROM AUTHOR]

Simple Summary: The study investigated how different types of fertilizers affected wheat aphids, a harmful pest for wheat plants. The nano-Cu fertilizer had negative effects on the aphids, leading to shorter lifespans and fewer babies. In contrast, the nitrogen fertilizer was not effective in controlling the aphids. The nano-Fe and nano-Zn fertilizers had fewer negative impacts compared to nano-Cu. While nitrogen treatment had positive effects on wheat aphid lifespan. Overall, the study suggests that incorporating nano-Cu into pest control strategies for wheat aphids could be a promising approach. In conclusion, choosing the right fertilizer can make a big difference in managing pests like aphids on wheat plants, and using nano-Cu could be a valuable tool in protecting crops. The use of nanofertilizers has both advantages and concerns. One benefit is that nano-fertilizers can enhance plant resistance against insect pests, making them a valuable strategy in integrated pest management (IPM). This study focused on the effect of wheat leaves treated with nano-chelated fertilizers and nitrogen (N) fertilizer on the wheat aphid (Schizaphis graminum Rondani), a harmful pest of wheat plants that transmits dangerous viruses. The nano-Cu treatment showed the longest pre-adult longevity. Additionally, the nano-Cu treatment resulted in the lowest adult longevity, fecundity, nymphoposition day number, intrinsic rate of population growth (r), finite rate of population increase (λ), and net reproductive rate (R0) and gross reproductive rate (GRR). Also, nano-Cu treatment led to the highest amount of (T). The N treatment led to the highest levels of fecundity, nymphoposition days, r, λ, and R0. Nano-Fe and nano-Zn demonstrated fewer negative effects on S. graminum life table parameters than nano-Cu. Our results indicate that N treatment yielded numerous advantageous effects on the wheat aphid while simultaneously impeding the efficacy of the aphid control program. Conversely, nano-Cu treatment exhibited a detrimental influence on various parameters of the aphid's life table, resulting in a reduction in the pest's fitness. Consequently, the integration of nano-Cu should be seriously considered as a viable option in the IPM of the wheat aphid. [ABSTRACT FROM AUTHOR]

Plant disease outbreaks are increasingly exacerbated by climate change and the conditions of stress combinations. They are negatively affecting crop yield and driving threats to food security in many areas of the world. Although synthetic pesticides offer relative success in the control of pests and plant diseases, they are often overused, and this method faces numerous drawbacks, including environmental toxicity, soil degradation, and adverse effects on human health. Therefore, alternatives are being developed and examined, including the biocontrol of pests and pathogens and biomass pyrolysis leading to wood vinegar that has shown great promise in agriculture and organic farming. However, while wood vinegar use is expanding and allows the control of numerous pests and bacterial and fungal diseases, its application to control oomycete diseases is limited. This study aimed to test wood vinegar for the control of oomycete plant pathogens from which six wood vinegars of pistachio, pomegranate, almond, pine, cypress, and walnut were produced. The inhibitory effects of volatile metabolites (semi-VOCs) of different wood vinegars concentrations (100%, 50%, 25%, 12.5%, and 6.25%) were examined against the hyphal growth of Globisporangium ultimum and Pythium aphanidermatum isolates. An in vitro analysis unambiguously demonstrated that for Globisporangium ultimum, the wood vinegar semi-VOCs of almond, pistachio (C 100% and 50%), and walnut (C 100%) totally inhibited mycelial growth. On the other hand, Pythium aphanidermatum, pistachio (C 100%, 50%, and 25%), and cypress (C 100%) expressed their abilities to completely inhibit the mycelial growth. Other treatments, including relevant concentrations of pine and pomegranate significantly inhibited the growth of mycelia of both species compared to the control (p ≤ 0.05). Therefore, wood vinegar could be considered a natural and organic product to use in agriculture to cope not only against pests, bacterial and fungal pests but also against emerging oomycete plant diseases. [ABSTRACT FROM AUTHOR]

The vascular wilt of date palm (Phoenix dactylifera L.), also known as Bayoud and caused by Fusarium oxysporum f.sp. albedinis (FOA), is the most destructive disease in North Africa. The disease has resulted in huge economic losses due to declining crop yield and quality. The use of potential biocontrol agents is a sustainable and environmentally friendly strategy compared to synthetic fungicides. The use of date palm-associated microflora for the screening of native antagonistic bacteria with potential applications is the most promising way to control this disease. Thus, the epidemic palm groves (in the valley of M'zab-Ghardaia-Algeria) were chosen for the isolation of rhizospheric bacteria and endophytes from the soil and roots of healthy or infected plants. A total of 8 bacterial isolates (83, 84, 300, 333, 322, 260, 249, and 227) selected from 75 FOA-active strains, showed strong activity against several strains of FOA and other major plant pathogens. Their phylogenetic identification proved they belong to the genus Bacillus (Bacillus sp., B. subtilis, B. atrophaeus, B. halotolerans, B. amyloliquefasiens, and B. paralicheformis). Bacterial cultures and a cell-free culture supernatant tested alone or in co-culture showed FOA inhibitory activity. However, the effect of the co-culture did not show any synergy in both cases. Unlike diffusible compounds, volatile organic compounds did not show a significant antifungal ability. The concept of selecting biocontrol agents in vitro does not always guarantee in vivo performance. In addition to antibiosis, other strategies such as competition and resistance induction are required for biocontrol agent efficacy. To evaluate the biocontrol effect in vivo, germinated seeds of date palm were treated with bacteria, infected with the pathogen, and then incubated for 45 days at room temperature in the dark. The majority of the strains (84, 300, and 333) tested showed moderate fungistatic effects and the protection rate reached an average of 60%. In particular, promising results were obtained with B. paralicheniformis strain 260 which significantly protected palm seeds up to 85%, compared to its in vitro test where a low inhibition rate (27.77%) was recorded. Screening methods should be equipped with multifactorial or plant-mediated control mechanisms. Furthermore, these bacteria have shown other potentialities to improve plant growth and resist stressful conditions. Thus, an effective biocontrol agent must combine several beneficial characteristics to be used successfully in situ. In this respect, Bacillus remains the best candidate for biological control. [ABSTRACT FROM AUTHOR]

In addition to changing global demography and global warming, agricultural production systems around the world are threatened by intensive agricultural practices (overuse of land and excessive use of chemical fertilizers and pesticides) that deplete soils by affecting their dynamics and their fertility, pollute the environment, lower production, and alter biodiversity on a large scale. The use of bioformulations based on PGPMs (plant growth-promoting microorganisms) seems to be a promising and sustainable strategy to overcome these threats, thanks to their tolerance to various biotic and abiotic stresses and via their beneficial effects in promising plant growth, pest protection, bioremediation, and restoration of degraded lands. In recent years, particular attention has been paid to encapsulated formulations because they offer several advantages over conventional bioformulation (liquid and solid) related to shelf life, problems of survival and viability in the environment, and the efficiency of rhizospheric colonization. This review focuses on the types of encapsulations and the different technologies used in this process as well as the most commonly used substrates and additives. It also provides an overview on the application of encapsulated bioformulations as biofertilizers, biopesticides, or other biostimulators and summarizes the knowledge of the scientific literature on the development of nanoencapsulation in this sector. [ABSTRACT FROM AUTHOR]

Halotolerant plant-growth-promoting rhizobacteria (PGPR) could not only promote plant growth, but also help in counteracting the detrimental effects of salt stress. In the present study, a total of 76 bacteria were isolated from the rhizosphere, non-rhizospheric soil and endophytes of the halophyte Salsola tetrandra, collected from natural saline soils in Algeria. Phylogenetic analysis based on the 16S rDNA sequence of Gram-negative bacteria (n = 51) identified, showed seventeen representative isolates grouped into four genera (Pseudomonas, Acinetobacter, Enterobacter, and Providencia). These bacterial isolates that exhibited different PGPR traits were selected and tested for their ability to tolerate different abiotic stress (NaCl, PEG8000, and pH). The majority of isolates were drought tolerant (60% of PEG8000) and had an optimal growth at high pH values (pH 9 and 11) and some strains tolerated 2 M of NaCl. Strains identified as Enterobacter xiangfangensis BE1, Providencia rettgeri BR5 and Pseudomonas stutzeri MLR6 showed high capacity of adaptation on their PGP traits. The salt-tolerant isolates were finally chosen to promote growth and enhance salt tolerance, separately or combined, of Arabidopsis thaliana (Col-0) exposed or not to 0.1 M NaCl, by following fresh and root weight, primary root elongation and lateral root number. The best bacterial effect was recorded for the MLR6 strain in increasing shoot fresh weight and for BE1 in terms of root fresh weight in the absence of salt stress. At stressed conditions, all growth parameters declined. However, inoculation of Arabidopsis thaliana with the three bacterial strains (MLR6, BE1 and BR5), single or in co-culture, conferred an increase in the shoot weight, primary root length and lateral root number. The use of these strains separately or combined as biofertilizers seems to be a powerful tool in the development of sustainable agriculture in saline soils. [ABSTRACT FROM AUTHOR]

Natural dyes have been used from ancient times for multiple purposes, most importantly in the field of textile dying. The increasing demand and excessive costs of natural dye extraction engendered the discovery of synthetic dyes from petrochemical compounds. Nowadays, they are dominating the textile market, with nearly 8 × 105 tons produced per year due to their wide range of color pigments and consistent coloration. Textile industries consume huge amounts of water in the dyeing processes, making it hard to treat the enormous quantities of this hazardous wastewater. Thus, they have harmful impacts when discharged in non-treated or partially treated forms in the environment (air, soil, plants and water), causing several human diseases. In the present work we focused on synthetic dyes. We started by studying their classification which depended on the nature of the manufactured fiber (cellulose, protein and synthetic fiber dyes). Then, we mentioned the characteristics of synthetic dyes, however, we focused more on their negative impacts on the ecosystem (soil, plants, water and air) and on humans. Lastly, we discussed the applied physical, chemical and biological strategies solely or in combination for textile dye wastewater treatments. Additionally, we described the newly established nanotechnology which achieves complete discharge decontamination. [ABSTRACT FROM AUTHOR]

Diverse vegetable oils are extracted from oleagenic trees and plants all over the world. In particular, olive oil represents a strategic socio-economic branch in the Mediterranean countries. These countries use either two or three-phase olive oil extraction systems. In this work, we focus on the by-products from three-phase olive oil extraction, which are the liquid olive mill wastewater (OMW) and the solid olive mill pomace (OMP) rejected in evaporative ponds. The disposal of this recalcitrant waste poses environmental problems such as the death of different species of insects and animals. In-depth ICP-OES analysis of the heavy metal composition of OMW and OMP revealed the presence of many metals ranging from non-toxic to highly toxic. The LC-HRMS characterization of these by-products indicated the presence of several secondary metabolites harmful to humans or to the environment. Thus, we aimed to identify OMW and OMP indigenous microbiota through metagenomics. The bacterial population was dominated by the Acetobacter (49.7%), Gluconobacter (17.3%), Gortzia (13.7%) and Nardonalla (5.3%) genera. The most abundant fungal genera were Nakazawaea, Saccharomyces, Lachancea and Candida. These microbial genera are responsible for OMW, OMP and soil toxicity alleviation. [ABSTRACT FROM AUTHOR]

There is a great interest in mitigating soil salinity that limits plant growth and productivity. In this study, eighty-nine strains were isolated from the rhizosphere and endosphere of two halophyte species (Suaeda mollis and Salsola tetrandra) collected from three chotts in Algeria. They were screened for diverse plant growth-promoting traits, antifungal activity and tolerance to different physico-chemical conditions (pH, PEG, and NaCl) to evaluate their efficiency in mitigating salt stress and enhancing the growth of Arabidopsis thaliana and durum wheat under NaCl–stress conditions. Three bacterial strains BR5, OR15, and RB13 were finally selected and identified as Bacillus atropheus. The Bacterial strains (separately and combined) were then used for inoculating Arabidopsis thaliana and durum wheat during the seed germination stage under NaCl stress conditions. Results indicated that inoculation of both plant spp. with the bacterial strains separately or combined considerably improved the growth parameters. Three soils with different salinity levels (S1 = 0.48, S2 = 3.81, and S3 = 2.80 mS/cm) were used to investigate the effects of selected strains (BR5, OR15, and RB13; separately and combined) on several growth parameters of wheat plants. The inoculation (notably the multi-strain consortium) proved a better approach to increase the chlorophyll and carotenoid contents as compared to control plants. However, proline content, lipid peroxidation, and activities of antioxidant enzymes decreased after inoculation with the plant growth-promoting rhizobacteria (PGPR) that can attenuate the adverse effects of salt stress by reducing the reactive oxygen species (ROS) production. These results indicated that under saline soil conditions, halotolerant PGPR strains are promising candidates as biofertilizers under salt stress conditions. [ABSTRACT FROM AUTHOR]

The genusPythium, with slightly over 140 described species, has been classified traditionally with other filamentous, coenocytic, sporangia-producing fungi as “Phycomycetes”. However, with recent advances in chemical, ultrastructural and molecular studies,Pythiumspp. are now considered as “fungus-like organisms” or “pseudo-fungi” and are placed in the kingdom Chromista or kingdom Stramenopila, distinct from the true fungi or the kingdom Fungi. There is no comprehensive web-based identification key available for the identification of all describedPythiumspecies. This article describes a web-based identification tool (which has been described by the authors under the name of Fungid) that uses morphological features. For the isolate in question, the software compares the unknown isolate with near to 90 known species submitted in the database. One or more than one species is suggested to the user by the software. [ABSTRACT FROM PUBLISHER]

World population growth and modernization have engendered multiple environmental problems: the propagation of humans and crop diseases and the development of multi-drug-resistant fungi, bacteria and viruses. Thus, a considerable shift towards eco-friendly products has been seen in medicine, pharmacy, agriculture and several other vital sectors. Nowadays, studies on endophytic fungi and their biotechnological potentials are in high demand due to their substantial, cost-effective and eco-friendly contributions in the discovery of an array of secondary metabolites. For this review, we provide a brief overview of plant–endophytic fungi interactions and we also state the history of the discovery of the untapped potentialities of fungal secondary metabolites. Then, we highlight the huge importance of the discovered metabolites and their versatile applications in several vital fields including medicine, pharmacy, agriculture, industry and bioremediation. We then focus on the challenges and on the possible methods and techniques that can be used to help in the discovery of novel secondary metabolites. The latter range from endophytic selection and culture media optimization to more in-depth strategies such as omics, ribosome engineering and epigenetic remodeling. [ABSTRACT FROM AUTHOR]

Forests are an essential component of the natural environment, as they support biodiversity, sequester carbon, and play a crucial role in biogeochemical cycles—in addition to producing organic matter that is necessary for the function of terrestrial organisms. Forests today are subject to threats ranging from natural occurrences, such as lightning-ignited fires, storms, and some forms of pollution, to those caused by human beings, such as land-use conversion (deforestation or intensive agriculture). In recent years, threats from pests and pathogens, particularly non-native species, have intensified in forests. The damage, decline, and mortality caused by insects, fungi, pathogens, and combinations of pests can lead to sizable ecological, economic, and social losses. To combat forest pests and pathogens, biocontrol may be an effective alternative to chemical pesticides and fertilizers. This review of forest pests and potential adversaries in the natural world highlights microbial inoculants, as well as research efforts to further develop biological control agents against forest pests and pathogens. Recent studies have shown promising results for the application of microbial inoculants as preventive measures. Other studies suggest that these species have potential as fertilizers. [ABSTRACT FROM AUTHOR]

Plant growth promoting bacteria (PGPB) have been the target of intensive research studies toward their efficient use in the field as biofertilizers, biocontrol, and bioremediation agents among numerous other applications. Recent trends in the field of PGPB research led to the development of versatile multifaceted PGPB that can be used in different field conditions such as biocontrol of plant pathogens in metal contaminated soils. Unfortunately, all these research efforts lead to the development of PGPB that failed to perform in salty environments. Therefore, it is urgently needed to address this drawback of these PGPB toward their efficient performance in salinity context. In this paper we provide a review of state-of-the-art research in the field of PGPB and propose a road map for the development of next generation versatile and multifaceted PGPB that can perform in salinity. Beyond soil desalinization, our study paves the way towards the development of PGPB able to provide services in diverse salty environments such as heavy metal contaminated, or pathogen threatened. Smart development of salinity adapted next generation biofertilizers will inevitably allow for mitigation and alleviation of biotic and abiotic threats to plant productivity in salty environments. [ABSTRACT FROM AUTHOR]

The phosphogypsum (PG) endogenous bacterial community and endophytic bacterial communities of four plants growing in phosphogypsum-contaminated sites, Suaeda fruticosa (SF), Suaeda mollis (SM), Mesembryanthmum nodiflorum (MN) and Arthrocnemum indicum (AI) were investigated by amplicon sequencing. Results highlight a more diverse community of phosphogypsum than plants associated endophytic communities. Additionally, the bacterial culturable communities of phosphogypsum and associated plant endophytes were isolated and their plant-growth promotion capabilities, bioremediation potential and stress tolerance studied. Most of plant endophytes were endowed with plant growth-promoting (PGP) activities and phosphogypsum communities and associated plants endophytes proved highly resistant to salt, metal and antibiotic stress. They also proved very active in bioremediation of phosphogypsum and other organic and inorganic environmental pollutants. Genome sequencing of five members of the phosphogypsum endogenous community showed that they belong to the recently described species Bacillus albus (BA). Genome mining of BA allowed the description of pollutant degradation and stress tolerance mechanisms. Prevalence of this tool box in the core, accessory and unique genome allowed to conclude that accessory and unique genomes are critical for the dynamics of strain acquisition of bioremediation abilities. Additionally, secondary metabolites (SM) active in bioremediation such as petrobactin have been characterized. Taken together, our results reveal hidden untapped valuable bacterial actors for waste remediation. [ABSTRACT FROM AUTHOR]

Oomycete and fungal pathogens, mainly Phytophthora and Fusarium species, are notorious causal agents of huge economic losses and environmental damages. For instance, Phytophthora ramorum, Phytophthora cryptogea, Phytophthora plurivora and Fusarium solani cause significant losses in nurseries and in forest ecosystems. Chemical treatments, while harmful to the environment and human health, have been proved to have little or no impact on these species. Recently, biocontrol bacterial species were used to cope with these pathogens and have shown promising prospects towards sustainable and eco-friendly agricultural practices. Olive trees prone to Phytophthora and Fusarium disease outbreaks are suitable for habitat-adapted symbiotic strategies, to recover oomycetes and fungal pathogen biocontrol agents. Using this strategy, we showed that olive trees-associated microbiome represents a valuable source for microorganisms, promoting plant growth and healthy benefits in addition to being biocontrol agents against oomycete and fungal diseases. Isolation, characterization and screening of root microbiome of olive trees against numerous Phytophthora and other fungal pathogens have led to the identification of the Bacillus velezensis OEE1, with plant growth promotion (PGP) abilities and strong activity against major oomycete and fungal pathogens. Phylogenomic analysis of the strain OEE1 showed that B. velezensis suffers taxonomic imprecision that blurs species delimitation, impacting their biofertilizers' practical use. Genome mining of several B. velezensis strains available in the GenBank have highlighted a wide array of plant growth promoting rhizobacteria (PGPR) features, metals and antibiotics resistance and the degradation ability of phytotoxic aromatic compounds. Strain OEE1 harbours a large repertoire of secreted and volatile secondary metabolites. Rarefaction analysis of secondary metabolites richness in the B. velezenis genomes, unambiguously documented new secondary metabolites from ongoing genome sequencing efforts that warrants more efforts in order to assess the huge diversity in the species. Comparative genomics indicated that B. velezensis harbours a core genome endowed with PGP features and accessory genome encoding diverse secondary metabolites. Gas Chromatography-Mass Spectrometry (GC-MS) analysis of OEE1 Volatile Organic Compounds (VOCs) and Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS) analysis of secondary metabolites identified numerous molecules with PGP abilities that are known to interfere with pathogen development. Moreover, B. velezensis OEE1 proved effective in protecting olive trees against F. solani in greenhouse experiments and are able to inhabit olive tree roots. Our strategy provides an effective means for isolation of biocontrol agents against recalcitrant pathogens. Their genomic analysis provides necessary clues towards their efficient implementation as biofertilizers. [ABSTRACT FROM AUTHOR]

Halophyte Limoniastrum monopetalum, an evergreen shrub inhabiting the Mediterranean region, has well-documented phytoremediation potential for metal removal from polluted sites. It is also considered to be a medicinal halophyte with potent activity against plant pathogens. Therefore, L. monopetalum may be a suitable candidate for isolating endophytic microbiota members that provide plant growth promotion (PGP) and resistance to abiotic stresses. Selected for biocontrol abilities, these endophytes may represent multifaceted and versatile biocontrol agents, combining pathogen biocontrol in addition to PGP and plant protection against abiotic stresses. In this study 117 root culturable bacterial endophytes, including Gram-positive (Bacillus and Brevibacillus), Gram-negative (Proteus, Providencia, Serratia, Pantoea, Klebsiella, Enterobacter and Pectobacterium) and actinomycete Nocardiopsis genera have been recovered from L. monopetalum. The collection exhibited high levels of biocontrol abilities against bacterial (Agrobacterium tumefaciens MAT2 and Pectobacterium carotovorum MAT3) and fungal (Alternaria alternata XSZJY-1, Rhizoctonia bataticola MAT1 and Fusarium oxysporum f. sp. radicis lycopersici FORL) pathogens. Several bacteria also showed PGP capacity and resistance to antibiotics and metals. A highly promising candidate Bacillus licheniformis LMRE 36 with high PGP, biocontrol, metal and antibiotic, resistance was subsequently tested in planta (potato and olive trees) for biocontrol of a collection of 14 highly damaging Fusarium species. LMRE 36 proved very effective against the collection in both species and against an emerging Fusarium sp. threatening olive trees culture in nurseries. These findings provide a demonstration of our pyramiding strategy. Our strategy was effective in combining desirable traits in biocontrol agents towards broad-spectrum resistance against pathogens and protection of crops from abiotic stresses. Stacking multiple desirable traits into a single biocontrol agent is achieved by first, careful selection of a host for endophytic microbiota recovery; second, stringent in vitro selection of candidates from the collection; and third, application of the selected biocontrol agents in planta experiments. That pyramiding strategy could be successfully used to mitigate effects of diverse biotic and abiotic stresses on plant growth and productivity. It is anticipated that the strategy will provide a new generation of biocontrol agents by targeting the microbiota of plants in hostile environments. [ABSTRACT FROM AUTHOR]

To explore proteolytic activity of endophytic fungi inhabiting date palm roots, a Penicillium bilaiae isolate, displaying the highest level of protease production, has been recovered. Response surface methodology (RSM) was applied to optimize culture conditions for protease production by the fungus. Plackett-Burman design allowed for screening of variables effective in protease production. Results indicated that temperature, initial pH and glucose concentration dramatically affect protease yield. These factors were further optimized using a Box-Behnken design and RSM. A combination of initial pH (6.26), temperature (24.5 °C), glucose (13.75 g/L), NaNO3 (1.5 g/L), MgSO4 (0.2 g/L), KH2PO4 (0.5 g/L) and KCl (0.5 g/L) were optimum for maximum production of protease. A 1086-fold enhancement of protease production was gained after optimization. Biochemical properties of fungal protease including the effect of pH and temperature on the activity and the stability of proteolytic enzyme were determined. Moreover, the influence of carbon and nitrogen sources, metal ions, detergents as well as enzyme inhibitors was investigated. Our results highlighted that protease of Penicillium bilaiae isolate TDPEF30 could be considered as a promising candidate for industrial applications. [ABSTRACT FROM AUTHOR]

Endophytic fungi of healthy and brittle leaf diseased (BLD) date palm trees (Phoenix dactylifera L.) represent a promising source of bioactive compounds with biomedical, industrial, and pharmaceutical applications. The fungal endophytes Penicillium citrinum isolate TDPEF34, and Geotrichum candidum isolate TDPEF20 from healthy and BLD date palm trees, respectively, proved very effective in confrontation assays against three pathogenic bacteria, including two Gram-positive bacteria Bacillus thuringiensis (Bt) and Enterococcus faecalis (Ef), and one Gram-negative bacterium Salmonella enterica (St). They also inhibited the growth of three fungi Trichoderma sp. (Ti), Fusarium sporotrichioides (Fs), Trichoderma sp. (Ts). Additionally, their volatile organic compounds (VOCs) were shown to be in part responsible for the inhibition of Ti and Ts and could account for the full inhibition of Fs. Therefore, we have explored their potential as fungal cell factories for bioactive metabolites production. Four extracts of each endophyte were prepared using different solvent polarities, ethanol (EtOH), ethyl acetate (EtOAc), hexane (Hex), and methanol (MetOH). Both endophyte species showed varying degrees of inhibition of the bacterial and fungal pathogens according to the solvent used. These results suggest a good relationship between fungal bioactivities and their produced secondary metabolites. Targeting the discovery of potential anti-diabetic, anti-hemolysis, anti-inflammatory, anti-obesity, and cytotoxic activities, endophytic extracts showed promising results. The EtOAc extract of G. candidum displayed IC50 value comparable to the positive control diclofenac sodium in the anti-inflammatory assays. Antioxidant activity was evaluated using α,α-diphenyl-β-picrylhydrazyl (DPPH), β-carotene bleaching, reducing power (RP), and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonique) (ABTS) radical scavenging assays. The findings revealed strong anti-oxidant power with an IC50 of 177.55 µg/mL for G. candidum EtOAc extract using DPPH assay, probably due to high polyphenol and flavonoid content in both fungal extracts. Finally, LC-HRMS (Liquid Chromatography–High Resolution Mass Spectrometry) and GC-MS (Gas Chromatography–Mass Spectrometry) analysis of G. candidum and P. citrinum extracts revealed an impressive arsenal of compounds with previously reported biological activities, partly explaining the obtained results. Finally, LC-HRMS analysis indicated the presence of new fungal metabolites that have never been reported, which represent good candidates to follow for the discovery of new bioactive molecules. [ABSTRACT FROM AUTHOR]

Rotavirus, one of the main pathogens causing morbidity and mortality in neonatal dairy calves worldwide, is responsible for 30-44% of cattle deaths. It is considered to be the most common etiologic agent of diarrhea in neonatal dairy calves and children, the dominant type being group A. Two hundred seventy animals from 27 farms from 2 regions of Kuwait were tested for the presence of Rotavirus serogroup A (RVA) using latex agglutination test (LAT) and reverse transcription-polymerase chain (RT-PCR) testing. RVA non-structural proteins NSP1-2, NSP4-5 and capsid protein genes VP1-7 were characterized by next generation sequencing. LAT was positive in 15.56% of the animals, and RT-PCR in 28.89%. Using RT-PCR as a reference method, LAT was 100% specific but only 83.33% sensitive. ANOVA analysis showed correlation only with the location of the farms but no significant correlation with the age and sex of the animals. Although there was a tendency of clustering of RVA positive animals, it did not reach statistical significance ( p = 0.035 for LAT). The phylogenetic analysis showed that Kuwaiti isolates of group A rotavirus clustered with human rotaviruses. Taken together, it seems that rotavirus was present in most of the dairy farms in Kuwait. The high occurrence of the virus in calves in Kuwaiti dairy farms and the close phylogenetic affinity with human isolates warrants urgent action to minimize and control its spread between calves in farms., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Alotaibi, Al-Amad, Chenari Bouket, Al-Aqeel, Haider, Hijji, Belbahri and Alenezi.)

Date palm ( Phoenix dactylifera L.) plantations in North Africa are nowadays threatened with the spread of the Bayoud disease caused by Fusarium oxysporum f. sp. albedinis , already responsible for destroying date production in other infected areas, mainly in Morocco. Biological control holds great promise for sustainable and environmental-friendly management of the disease. In this study, the additional benefits to agricultural ecosystems of using plant growth promoting rhizobacteria (PGPR) or endophytes are addressed. First, PGPR or endophytes can offer an interesting bio-fertilization, meaning that it can add another layer to the sustainability of the approach. Additionally, screening of contrasting niches can yield bacterial actors that could represent wardens against whole genera or groups of plant pathogenic agents thriving in semi-arid to arid ecosystems. Using this strategy, we recovered four bacterial isolates, designated BFOA1, BFOA2, BFOA3 and BFOA4, that proved very active against F. oxysporum f. sp. albedinis . BFOA1-BFOA4 proved also active against 16 Fusarium isolates belonging to four species: F. oxysporum (with strains phytopathogenic of Olea europaea and tomato), F. solani (with different strains attacking O. europaea and potato), F. acuminatum (pathogenic on O. europaea ) and F. chlamydosporum (phytopathogenic of O. europaea ). BFOA1-BFOA4 bacterial isolates exhibited strong activities against another four major phytopathogens: Botrytis cinerea , Alternaria alternata , Phytophthora infestans , and Rhizoctonia bataticola. Isolates BFOA1-BFOA4 had the ability to grow at temperatures up to 35°C, pH range of 5-10, and tolerate high concentrations of NaCl and up to 30% PEG. The isolates also showed relevant direct and indirect PGP features, including growth on nitrogen-free medium, phosphate solubilization and auxin biosynthesis, as well as resistance to metal and xenobiotic stress. Phylogenomic analysis of BFOA1-BFOA4 isolates indicated that they all belong to Bacillus halotolerans , which could therefore considered as a warden against Fusarium infection in plants. Comparative genomics allowed us to functionally describe the open pan genome of B. halotolerans and LC-HRMS and GCMS analyses, enabling the description of diverse secondary metabolites including pulegone, 2-undecanone, and germacrene D, with important antimicrobial and insecticidal properties. In conclusion, B. halotolerans could be used as an efficient bio-fertilizer and bio-control agent in semi-arid and arid ecosystems.

The Gram positive, non-pathogenic endospore-forming soil inhabiting prokaryote Bacillus amyloliquefaciens is a plant growth-promoting rhizobacterium. Bacillus amyloliquefaciens processes wide biocontrol abilities and numerous strains have been reported to suppress diverse bacterial, fungal and fungal-like pathogens. Knowledge about strain level biocontrol abilities is warranted to translate this knowledge into developing more efficient biocontrol agents and bio-fertilizers. Ever-expanding genome studies of B. amyloliquefaciens are showing tremendous increase in strain-specific new secondary metabolite clusters which play key roles in the suppression of pathogens and plant growth promotion. In this report, we have used genome mining of all sequenced B. amyloliquefaciens genomes to highlight species boundaries, the diverse strategies used by different strains to promote plant growth and the diversity of their secondary metabolites. Genome composition of the targeted strains suggest regions of genomic plasticity that shape the structure and function of these genomes and govern strain adaptation to different niches. Our results indicated that B. amyloliquefaciens : (i) suffer taxonomic imprecision that blurs the debate over inter-strain genome diversity and dynamics, (ii) have diverse strategies to promote plant growth and development, (iii) have an unlocked, yet to be delimited impressive arsenal of secondary metabolites and products, (iv) have large number of so-called orphan gene clusters, i.e., biosynthetic clusters for which the corresponding metabolites are yet unknown, and (v) have a dynamic pan genome with a secondary metabolite rich accessory genome.

The soil-borne gram-positive bacteria Aneurinibacillus migulanus strain Nagano shows considerable potential as a biocontrol agent against plant diseases. In contrast, A. migulanus NCTC 7096 proved less effective for inhibition of plant pathogens. Nagano strain exerts biocontrol activity against some gram-positive and gram-negative bacteria, fungi and oomycetes through the production of gramicidin S (GS). Apart from the antibiotic effects, GS increases the rate of evaporation from the plant surface, reducing periods of surface wetness and thereby indirectly inhibiting spore germination. To elucidate the molecular basis of differential biocontrol abilities of Nagano and NCTC 7096 , we compared GS production and biosurfactant secretion in addition to genome mining of the genomes. Our results proved that: (i) Using oil spreading, blood agar lysis, surface tension and tomato leaves wetness assays, Nagano showed increased biosurfactant secretion in comparison with NCTC 7096 , (ii) Genome mining indicated the presence of GS genes in both Nagano and NCTC 7096 with two amino acid units difference between the strains: T342I and P419S. Using 3D models and the DUET server, T342I and P419S were predicted to decrease the stability of the NCTC 7096 GS synthase, (iii) Nagano produced two additional GS-like molecules GS-1155 (molecular weight 1155) and GS-1169 (molecular weight 1169), where one or two ornithine residues replace lysine in the peptide. There was also a negative correlation between surface tension and the quantity of GS-1169 present in Nagano , and (iv) the Nagano genome had a full protein network of exopolysaccharide biosynthesis in contrast to NCTC 7096 which lacked the first enzyme of the network. NCTC 7096 is unable to form biofilms as observed for Nagano . Different molecular layers, mainly gramicidin secondary metabolite production, account for differential biocontrol abilities of Nagano and NCTC 7096 . This work highlighted the basis of differential biological control abilities between strains belonging to the same species and demonstrates techniques useful to the screening of effective biocontrol strains for environmentally friendly secondary metabolites that can be used to manage plant pathogens in the field.

In this study, we aimed to explore and compare the composition, metabolic diversity and antimicrobial potential of endophytic fungi colonizing internal tissues of healthy and brittle leaf diseased (BLD) date palm trees ( Phoenix dactylifera L.) widely cultivated in arid zones of Tunisia. A total of 52 endophytic fungi were isolated from healthy and BLD roots of date palm trees, identified based on internal transcribed spacer-rDNA sequence analysis and shown to represent 13 species belonging to five genera. About 36.8% of isolates were shared between healthy and diseased root fungal microbiomes, whereas 18.4 and 44.7% of isolates were specific to healthy and BLD root fungal microbiomes, respectively. All isolates were able to produce at least two of the screened enzymes including amylase, cellulase, chitinase, pectinase, protease, laccase and lipase. A preliminary screening of the isolates using disk diffusion method for antibacterial activity against four Gram-positive and three Gram-negative bacteria and antifungal activities against three phytopathogenic fungi indicated that healthy and BLD root fungal microbiomes displayed interesting bioactivities against examined bacteria and broad spectrum bioactivity against fungal pathogens. Some of these endophytic fungi (17 isolates) were fermented and their extracts were evaluated for antimicrobial potential against bacterial and fungal isolates. Results revealed that fungal extracts exhibited antibacterial activities and were responsible for approximately half of antifungal activities against living fungi. These results suggest a strong link between fungal bioactivities and their secondary metabolite arsenal. EtOAc extracts of Geotrichum candidum and Thielaviopsis punctulata originating from BLD microbiome gave best results against Micrococcus luteus and Bacillus subtilis with minimum inhibitory concentration (MIC, 0.78 mg/mL) and minimum bactericidal concentration (6.25 mg/mL). G. candidum gave the best result against Rhizoctonia solani with MIC 0.78 mg/mL and minimum fungicidal concentration (MFC, 6.25 mg/mL). In conclusion, using plant microbiomes subjected to biotic stresses offers new endophytes with different bioactivities than those of healthy plants. Therefore, date palm endophytic fungi represent a hidden untapped arsenal of antibacterial and broad spectrum antifungal secondary metabolites and could be considered promising source of bioactive compounds with industrial and pharmaceutical applications.

Pythiumkandovanense sp. nov. (ex-type culture CCTU 1813T = OPU 1626T = CBS 139567T) is a novel oomycete species isolated from Lolium perenne with snow rot symptoms in a natural grassland in East-Azarbaijan province, Iran. Phylogenetic analyses based on sequence data from internal transcribed spacer (ITS)-rDNA, coxI and coxII mitochondrial genes clustered our isolates in Pythium group E as a unique, well supported clade. Pythium kandovanense sp. nov. is phylogenetically and morphologically distinct from the other closely related species in this clade, namely Pythium rostratifingens and Pythium rostratum. Pythium kandovanense sp. nov. can be distinguished from these two species by its cylindrical sporangia and lower temperatures for optimum and maximum growth rate. The development of zoospores released through a shorter discharge tube is an additional morphological feature which can be used to differentiate Pythium kandovanense sp. nov. from Pythium rostratifingens. Laboratory inoculation tests demonstrated the pathogenicity of Pythium kandovanense sp. nov. to L. perenne under wet cold (0-3 °C) conditions.