Rhizospheric Actinomycetes Revealed Antifungal and Plant-Growth-Promoting Activities under Controlled Environment

Actinomycetes has large habitats and can be isolated from terrestrial soil, rhizospheres of plant roots, and marine sediments. Actinomycetes produce several bioactive secondary metabolites with antibacterial, antifungal, and antiviral properties. In this study, some Actinomycetes strains were isolated from the rhizosphere zone of four different plant species: rosemary, acacia, strawberry, and olive. The antagonistic activity of all isolates was screened in vitro against Escherichia coli and Bacillus megaterium. Isolates with the strongest bioactivity potential were selected and molecularly identified as Streptomyces sp., Streptomyces atratus, and Arthrobacter humicola. The growth-promoting activity of the selected Actinomycetes isolates was in vivo evaluated on tomato plants and for disease control against Sclerotinia sclerotiorum. The results demonstrated that all bacterized plants with the studied Actinomycetes isolates were able to promote the tomato seedlings' growth, showing high values of ecophysiological parameters. In particular, the bacterized seedlings with Streptomyces sp. and A. humicola showed low disease incidence of S. sclerotiorum infection (0.3% and 0.2%, respectively), whereas those bacterized with S. atratus showed a moderate disease incidence (7.6%) compared with the positive control (36.8%). In addition, the ability of the studied Actinomycetes to produce extracellular hydrolytic enzymes was verified. The results showed that A. humicola was able to produce chitinase, glucanase, and protease, whereas Streptomyces sp. and S. atratus produced amylase and pectinase at high and moderate levels, respectively. This study highlights the value of the studied isolates in providing bioactive metabolites and extracellular hydrolytic enzymes, indicating their potential application as fungal-biocontrol agents

Applications of Absorbent Polymers for Sustainable Plant Protection and Crop Yield

Natural strategies for protecting the environment as well as plant, animal and human health is considered one of the main goals of developed countries. Recently, the use of absorbent polymers and hydrogel in agriculture has demonstrated several benefits for soil amendments, saving water content, reducing the consumption of soil nutrients, minimizing the negative impacts of dehydration and moisture stress in crops and controlling several phytopathogens. The seed-coating technology for establishing the crops is a recent common practice used for improving seed protection and enhancing plant growth. Coating materials include absorbent polymers and hydrogels based on growth regulators, pesticides, fertilizers and antagonist microorganisms. The current review has highlighted the importance of different types of superabsorbent polymers and hydrogels in an integrated strategy to protect seeds, plants and soil in a balanced manner to preserve the ecosystem

An Overview of Metabolic Activity, Beneficial and Pathogenic Aspects of Burkholderia Spp

Burkholderia is an important bacterial species which has different beneficial effects, such as promoting the plant growth, including rhizosphere competence for the secretion of allelochemicals, production of antibiotics, and siderophores. In addition, most of Burkholderia species have demonstrated promising biocontrol action against different phytopathogens for diverse crops. In particular, Burkholderia demonstrates significant biotechnological potential as a source of novel antibiotics and bioactive secondary metabolites. The current review is concerned with Burkholderia spp. covering the following aspects: discovering, classification, distribution, plant growth promoting effect, and antimicrobial activity of different species of Burkholderia, shedding light on the most important secondary metabolites, their pathogenic effects, and biochemical characterization of some important species of Burkholderia, such as B. cepacia, B. andropogonis, B. plantarii, B. rhizoxinica, B. glumae, B. caryophylli and B. gladioli

Investigating the Effects of Plant Essential Oils on Post-Harvest Fruit Decay

Essential oils are one of the most important natural products derived from plants, due to their various biological properties and their medicinal and nutritional uses. This chapter provides an overview of several different aspects relating to essential oils including a historical perspective, the uses of essential oils, their main sources and antifungal activity, their bioactive single constituents and their modes of action. The chapter will also give an insight into the chemical measures necessary for controlling plant pathogens and their negative impact on human health and/or the environment. It will also review the different sources of essential oils such as sage, oregano, thyme and marjoram from the Lamiaceae family, vervain from the Verbanacae family, and magnolia from the Magnoliaceae family. The antimicrobial activity of essential oils is reviewed, with particular emphasis on the antifungal properties exhibited against some serious pathogenic fungi and post-harvest disease. Moreover, various antimicrobial tests and techniques, such as various kill-time studies, killing time determination, LD-50 and growth curve recording, poisoned food techniques, spore germination and measurement of metabolic CO2 are included. Finally, five case studies relating to the antifungal activity of some plant essential oils, either in vitro or in vivo, against post-harvest pathogenic fungi are reviewed at the end of this chapter

Detection and identification of Phytophthora species in southern Italy by RFLP and sequence analysis of PCR-amplified nuclear ribosomal DNA.

In four neighbouring regions of southern Italy, Basilicata, Campania, Apulia and Calabria, pepper and zucchini plants showing Phytophthora blight symptoms, tomato plants with either late blight or buckeye rot symptoms, plants of strawberry showing crown rot symptoms and declining clementine trees with root and fruit rot were examined for Phytophthora infections by means of polymerase chain reaction (PCR) assays, using primers directed to nuclear ribosomal DNA (rDNA) repeat sequences. All diseased plants and trees examined tested positive. The detected fungal-like organisms were differentiated and characterized on the basis of primer specificity as well as through extensive restriction fragment length polymorphism (RFLP) and sequence analysis of PCR-amplified rDNA. Phytophthora capsici was identified in diseased pepper and zucchini plants, P. infestans was identified in tomato with late blight symptoms whereas buckeye rot-affected tomatoes and diseased strawberry plants proved to be infected by P. nicotianae and P. cactorum, respectively. Declining clementine trees were infected with P. citrophthora and P. nicotianae in about the same proportion. Also, thirty-one pure culture-maintained isolates of Phytophthora which had previously been identified in southern Italy by traditional methods but were never examined molecularly, were examined by RFLP and sequence analysis of PCR-amplified nuclear rDNA. Among these, an isolate from gerbera which had previously been identified by traditional methods only at genus level, was assigned to P. tentaculata. For the remaining pure culture-maintained isolates examined, the molecular identification data obtained corresponded with those delineated by traditional methods. Most of the diseases examined were already known to occur in southern Italy but the pathogens were molecularly detected and fully characterized at nuclear rDNA repeat level only from other geographic areas, very often outside Italy. A new disease to southern Italy was the Phytophthora blight of zucchini. This is also the first report on the presence and molecular identification of P. tentaculata from Italy

A Comprehensive Review on the Biological, Agricultural and Pharmaceutical Properties of Secondary Metabolites Based-Plant Origin

: Natural products are compounds produced by living organisms and can be divided into two main categories: primary (PMs) and secondary metabolites (SMs). Plant PMs are crucial for plant growth and reproduction since they are directly involved in living cell processes, whereas plant SMs are organic substances directly involved in plant defense and resistance. SMs are divided into three main groups: terpenoids, phenolics and nitrogen-containing compounds. The SMs contain a variety of biological capabilities that can be used as flavoring agents, food additives, plant-disease control, strengthen plant defenses against herbivores and, additionally, it can help plant cells to be better adapted to the physiological stress response. The current review is mainly focusing on certain key elements related to the significance, biosynthesis, classification, biochemical characterization and medical/pharmaceutical uses of the major categories of plant SMs. In addition, the usefulness of SMs in controlling plant diseases, boosting plant resistance and as potential natural, safe, eco-friendly substitutes for chemosynthetic pesticides were also reported in this review

Chemical Composition and Antimicrobial Properties of Mentha x piperita cv. ‘Kristinka’ Essential Oil

Several economically important crops, fruits and vegetables are susceptible to infection by pathogenic fungi and/or bacteria postharvest or in field. Recently, plant essential oils (EOs) extracted from different medicinal and officinal plants have had promising antimicrobial effects against phytopathogens. In the present study, the potential microbicide activity of Mentha x piperita cv. ‘Kristinka’ (peppermint) EO and its main constituents have been evaluated against some common phytopathogens. In addition, the cell membrane permeability of the tested fungi and the minimum fungicidal concentrations were measured. The antifungal activity was tested against the following postharvest fungi: Botrytis cinerea, Monilinia fructicola, Penicillium expansum and Aspergillus niger, whereas antibacterial activity was evaluated against Clavibacter michiganensis, Xanthomonas campestris, Pseudomonas savastanoi and P. syringae pv. phaseolicola. The chemical analysis has been carried out using GC-MS and the main components were identified as menthol (70.08%) and menthone (14.49%) followed by limonene (4.32%), menthyl acetate (3.76%) and -caryophyllene (2.96%). The results show that the tested EO has promising antifungal activity against all tested fungi, whereas they demonstrated only a moderate antibacterial effect against some of the tested bacteria

Investigations on Fungi Isolated from Apple Trees with Die-Back Symptoms from Basilicata Region (Southern Italy)

Val d'Agri is an important orchard area located in the Basilicata Region (Southern Italy). A phenomenon affecting cv. "Golden Delicious" apples which lead to tree death has been observed in the past several years in this area. This phenomenon has already been detected in about 20 hectares and is rapidly expanding. The symptoms observed were "scaly bark" and extensive cankers, mainly located in the lower part of the trunk, associated with wood decay. Dead plants ranged from 20% to 80% and, in many cases, trees were removed by farmers. In order to identify the causes of this phenomenon, investigations were started in autumn/winter 2019. In order to determine the possible causal agents, fungal and bacterial isolations, from symptomatic tissues, were performed in laboratory. Bacterial isolations gave negative results, whereas pure fungal cultures (PFCs) were obtained after 3-4 passages on potato dextrose agar (PDA) media. Genetic material was extracted from each PFC and amplified by PCR using three pairs of primers: ITS5/4, Bt2a/Bt2b and ACT-512F/ACT-783R. The amplicons were directly sequenced, and nucleotide sequences were compared with those already present in the NCBI GenBank nucleotide database. All isolated fungi were identified based on morphological features and multilocus molecular analyses. Neofusicoccum parvum, Diaporthe eres and Trametes versicolor were most frequently isolated, while Pestalotiopsis funerea, Phomopsis spp. and Diaporthe foeniculina were less frequently isolated. All nucleotide sequences obtained in this study have been deposited into the EMBL database. Pathogenicity tests showed that N. parvum was the most pathogenic and aggressive fungus, while Phomopsis sp. was demonstrated to be the less virulent one. All the investigated fungi were repeatedly reisolated from artificially inoculated twigs of 2-year-old apple trees, cv. "Golden Delicious", and subsequently morphologically and molecularly identified. The role played by the above-mentioned fungi in the alterations observed in field is also discussed

Metagenomic Analysis of Bacterial Community Structure and Dynamics of a Digestate and a More Stabilized Digestate-Derived Compost from Agricultural Waste

Recycling of different products and waste materials plays a crucial role in circular economy, where the anaerobic digestion (AD) constitutes an important pillar since it reuses nutrients in the form of organic fertilizers. Knowledge about the digestate and compost microbial community structure and its variations over time is important. The aim of the current study was to investigate the microbiome of a slurry cow digestate produced on a farm (ADG) and of a more stabilized digestate-derived compost (DdC) in order to ascertain their potential uses as organic amendments in agriculture. The results from this study, based on a partial fragment of 16S bacterial rRNA NGS sequencing, showed that there is a greater microbial diversity in the DdC originated from agricultural waste compared to the ADG. Overall, the existence of a higher microbial diversity in the DdC was confirmed by an elevated number (1115) of OTUs identified, compared with the ADG (494 OTUs identified). In the DdC, 74 bacterial orders and 125 families were identified, whereas 27 bacterial orders and 54 families were identified in the ADG. Shannon diversity and Chao1 richness indexes were higher in DdC samples compared to ADG ones (Shannon: 3.014 and 1.573, Chao1: 68 and 24.75; p< 0.001 in both cases). A possible association between the microbiome composition at different stages of composting process and the role that these microorganisms may have on the quality of the compost-like substrate and its future uses is also discussed

An attempt of biocontrol the tomato-wilt disease caused by Verticillium dahliae using Burkholderia gladioli pv. agaricicola and its bioactive secondary metabolites

There is a great interest in discovering new microbial natural biocides such as microbial secondary metabolites to reduce the environmental pollution due to the excessive use of synthetic pesticides. Verticillium wilt, caused by the soil-borne Verticillium dahliae, is a widespread disease in tomato growing in many parts of the world. Burkholderia gladioli pv. agaricicola produces some antimicrobial substances and extracellular hydrolytic enzymes which exhibited promising antimicrobial activity towards several phytopathogens. The aims of the current research are to assess in vitro fungicidal effect of 4 strains of B. gladioli pv. agaricicola (ICMP11096, 11097, 12220 and 12322) against V. dahliae using culture or cell-free culture filtrate. In situ assay was performed to evaluate the biocontrol effect of the most efficient bacterial strain on wilt disease caused by V. dahliae in tomato plants. Results demonstrated that the studied bacterial strain ICMP12322 exerted the highest in vitro antifungal activity against V. dahliae which correlated with its ability to produce extracellular hydrolytic enzymes. Furthermore, in situ results showed that the selected bacterial strain significantly minimized the disease incidence