Green synthesis of Pleurotus eryngii-Derived Nanomaterials for Phytopathogen Control.

Growing concerns over the human health and environmental impacts of conventional fungicides, coupled with the escalating challenge of microbial resistance, have fueled the search for sustainable biocontrol strategies against plant pathogens. This study reports, for the first time, the green synthesis and characterization of a novel, eco-friendly nanomaterial, designated Pleurotus eryngii-Lecithin-Chitosan Nanomaterial (PEELCN), derived from P. eryngii extract (PEE), lecithin (L), and chitosan (C). The structural attributes of PEELCN were elucidated using Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), Thermogravimetric Analysis (TGA), X-ray Diffraction (XRD), and zeta potential measurements, confirming the successful formation of a stable and uniform nanostructure. The antifungal activity of PEELCN, and PEE, was assessed against five economically important phytopathogenic fungi: Neoscytalidium dimidiatum, Alternaria alternata, Verticillium dahliae, Bipolaris sorokiniana, and Fusarium oxysporum. Both PEE and PEELCN exhibited significant inhibitory effects on the mycelial growth of V. dahliae, B. sorokiniana, and N. dimidiatum, with varying degrees of efficacy. The differential antifungal activity suggests a species-specific mode of action. The findings highlight the promising potential of PEELCN as a sustainable, biocompatible, and cost-effective nanofungicide for the management of plant diseases, with the potential for development into a commercially viable biofungicide for sustainable agriculture.

New Detection Methods for Cryphonectria Hypovirus 1 (CHV1) through SYBR Green-Based Real-Time PCR and Loop-Mediated Isothermal Amplification (LAMP).

Some mycoviruses can be considered as effective biocontrol agents, mitigating the impact of phytopathogenic fungi and consequently reducing disease outbreaks while promoting plant health. Cryphonectria parasitica, the causal agent of chestnut blight and a highly destructive pathogen, experienced a notable decrease in its virulence with the identification of cryphonectria hypovirus 1 (CHV1), a naturally occurring biocontrol agent. In this study, two innovative diagnostic protocols designed for the accurate and efficient detection of CHV1 are introduced. The ORF A and ORF B regions of CHV1 are targeted by these techniques, which employ colorimetric loop-mediated isothermal amplification (LAMP) with 2 Colorimetric LAMP Master Mix and real-time quantitative PCR (qPCR) with SYBR Green chemistry, respectively. The LAMP assay presents a discernible color transition, changing from pink to yellow after a 35 min incubation period. Comparative analysis, when assessed against two established reverse transcription-PCR (RT-PCR) techniques, reveals a significant enhancement in sensitivity for both the LAMP approach, which offers a tenfold increase, and the qPCR method, which showcases a remarkable 100-fold sensitivity improvement. Throughout the comparison phase, it was evident that the RT-PCR, LAMP, and qPCR procedures displayed superior performance compared to the Bavendamm test, relying on phenol oxidase activity, effectively distinguishing hypovirulent strains. Consequently, this study introduces two pioneer diagnostic assays for highly sensitive CHV1 detection, representing a substantial advancement in the realm of CHV1 surveillance techniques. These methodologies hold significant promise for enhancing research endeavors in the domain of the biological control of C. parasitica.

Plant-Associated Neoscytalidium dimidiatum-Taxonomy, Host Range, Epidemiology, Virulence, and Management Strategies: A Comprehensive Review.

Neoscytalidium dimidiatum, a plant- and human-associated fungus, has emerged as a substantial global ecological and agricultural threat aggravated by global warming. It inflicts various diseases, including canker, blight, dieback, leaf spot, root rot, and fruit rot, across a wide spectrum of fruit trees, field crops, shrubs, and arboreal species, with a host range spanning 46 plant families, 84 genera, and 126 species, primarily affecting eudicot angiosperms. Six genera are asymptomatic hosts. Neoscytalidium dimidiatum exhibits worldwide distribution, with the highest prevalence observed in Asia and North America, notably in Iran, Turkey, and California. Rising disease prevalence and severity, aggravated by climate change, particularly impact tropical arid places across 37 countries spanning all 7 continents. This comprehensive review encapsulates recent advancements in the understanding of N. dimidiatum, encompassing alterations in its taxonomic classification, host range, symptoms, geographic distribution, epidemiology, virulence, and strategies for effective management. This study also concentrates on comprehending the taxonomic relationships and intraspecific variations within N. dimidiatum, with a particular emphasis on N. oculus and N. hylocereum, proposing to consider these two species as synonymous with N. dimidiatum. Furthermore, this review identifies prospective research directions aimed at augmenting our fundamental understanding of host-N. dimidiatum interaction.

Molecular phylogeny of plant pathogenic fungi based on start codon targeted (SCoT) polymorphism.

BACKGROUND: A number of molecular marker systems have been developed to assess genetic diversity, carry out phylogenetic analysis, and diagnose and discriminate plant pathogenic fungi. The start codon targeted (SCoT) markers system is a novel approach used here to investigate intra and interspecific polymorphisms of phytopathogenic fungi. MATERIALS AND METHODS: This study assessed genetic variability between and within 96 isolates of ten fungal species associated with a variety of plant species using 36 SCoT primers. RESULTS: The six primers generated 331 distinct and reproducible banding patterns, of which 322 were polymorphic (97.28%), resulting in 53.67 polymorphic bands per primer. All primers produced informative amplification profiles that distinguished all fungal species. With a resolving power of 10.65, SCoT primer 12 showed the highest polymorphism among species, followed by primer 33 and primer 29. Polymorphic loci (PPL), Nei's diversity index (h), and Shannon index (I) percentages were 6.25, 0.018, and 0.028, respectively. UPGMA analysis separated all isolates based on morphological classification and revealed significant genetic variation among fungal isolates at the intraspecific level. PCoA analysis strongly supported fungal species discrimination and genetic variation. The other parameters of evaluation proved that SCoT markers are at least as effective as other DNA markers. CONCLUSIONS: SCoT markers were effective in identifying plant pathogenic fungi and were a powerful tool for estimating genetic variation and population structure of different fungi species.

Fungal Pathogens Associated with Crown and Root Rot in Wheat-Growing Areas of Northern Kyrgyzstan.

Fungal species associated with crown and root rot diseases in wheat have been extensively studied in many parts of the world. However, no reports on the relative importance and distribution of pathogens associated with wheat crown and root rot in Kyrgyzstan have been published. Hence, fungal species associated with wheat crown/root rot were surveyed in three main wheat production regions in northern Kyrgyzstan. Fungal species were isolated on 1/5 strength potato-dextrose agar amended with streptomycin (0.1 g/L) and chloramphenicol (0.05 g/L). A total of 598 fungal isolates from symptomatic tissues were identified using morphological features of the cultures and conidia, as well as sequence analysis of the nuclear ribosomal internal transcribed spacer (ITS) region, the translation elongation factor 1α (TEF1), and the RNA polymerase II beta subunit (RPB2) genes. The percentage of fields from which each fungus was isolated and their relative percentage isolation levels were determined. Bipolaris sorokiniana, the causal agent of common root rot, was the most prevalent pathogenic species isolated, being isolated from 86.67% of the fields surveyed at a frequency of isolation of 40.64%. Fusarium spp. accounted for 53.01% of all isolates and consisted of 12 different species. The most common Fusarium species identified was Fusarium acuminatum, which was isolated from 70% of the sites surveyed with an isolation frequency of 21.57%, followed by Fusarium culmorum, Fusarium nygamai, Fusarium oxysporum, and Fusarium equiseti, all of which had a field incidence of more than 23%. Inoculation tests with 44 isolates representing 17 species on the susceptible Triticum aestivum cv. Seri 82 revealed that Fusarium pseudograminearum and F. culmorum isolates were equally the most virulent pathogens. The widespread distribution of moderately virulent B. sorokiniana appears to be a serious threat to wheat culture, limiting yield and quality. With the exception of F. culmorum, the remaining Fusarium species did not pose a significant threat to wheat production in the surveyed areas because common species, such as F. acuminatum, F. nygamai, F. oxysporum, and F. equiseti, were non-pathogenic but infrequent species, such as Fusarium redolens, Fusarium algeriense, and F. pseudograminearum, were highly or moderately virulent. Curvularia inaequalis, which was found in three different fields, was mildly virulent. The remaining Fusarium species, Fusarium solani, Fusarium proliferatum, Fusarium burgessii, and Fusarium tricinctum, as well as Microdochium bolleyi, Microdochium nivale, and Macrophomina phaseolina, were non-pathogenic and considered to be secondary colonizers. The implications of these findings are discussed.

Clonostachys rosea Strain ST1140: An Endophytic Plant-Growth-Promoting Fungus, and Its Potential Use in Seedbeds with Wheat-Grain Substrate.

In this study, Clonostachys rosea strain ST1140, a naturally occurring endophyte in healthy roots of a pistachio tree, was identified morphologically and molecularly through DNA sequencing, and its endophytic ability and growth effect in/on three solanaceous plant species were tested. Three different organic substrates (bread wheat-grain, sawdust, and leonardite) were also investigated for their utility in cultivating and multiplying the strain. In-tray and pot experiments, the rates of seed germination and vegetative development of pepper, tomato, and eggplant inoculated with C. rosea at planting were compared to those of non-inoculated controls. In pot experiments, inoculating seedbeds with increasing doses of C. rosea strain ST1140 with wheat-grain substrate resulted in higher plant height values for all plant species, and the strain endophytically colonized the roots of all plant species. In-tray experiments revealed that ST1140 inoculation resulted in 2-4 days earlier and 10% higher germination rates for all plant species, as well as more vigorous and accelerated seedling growth (10-13 days earlier for seedlings at the 4-5 true leaf stage) on all inoculated plant species. Among three different organic substrates, wheat-grain was found to be the most effective for long-term cultivation and multiplication of the fungus, which could be of interest for its development as a commercial product. These results promised the strain ST1140's use as a biofertilizer in seedbeds with a wheat-grain substrate.

Fungal Pathogens Associated with Crown and Root Rot of Wheat in Central, Eastern, and Southeastern Kazakhstan.

Kazakhstan is the fourteenth largest wheat producer in the world. Despite this fact, there has not been a comprehensive survey of wheat root and crown rot. A quantitative survey was conducted for the purpose of establishing the distribution of fungi associated with root and crown rot on wheat (Triticum spp.). During the 2019 growing season, samples were taken from the affected plants' roots and stem bases. A total of 1221 fungal isolates were acquired from 65 sites across the central (Karagandy region), eastern (East Kazakhstan region), and southeastern (Almaty region) parts of the country and identified using morphological and molecular tools. The internal transcribed spacer (ITS), translation elongation factor 1-alpha (EF1-α), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sequences were successfully used to identify the species of fungal isolates. It was found that Bipolaris sorokiniana (44.80%) and Fusarium acuminatum (20.39%) were the most predominant fungal species isolated, which were present in 86.15 and 66.15% of the fields surveyed, respectively, followed by F. equiseti (10.16%), Curvularia spicifera (7.62%), F. culmorum (4.75%), F. oxysporum (4.10%), F. redolens (2.38%), Rhizoctonia solani AG2-1 (1.06%), Nigrospora oryzae (0.98%), C. inaequalis (0.90%), F. pseudograminearum (0.74%), F. flocciferum (0.74%), Macrophomina phaseolina (0.66%), F. cf. incarnatum (0.33%), Fusarium sp. (0.25%), and F. torulosum (0.16%). A total of 74 isolates representing 16 species were tested via inoculation tests on the susceptible Triticum aestivum cv. Seri 82 and the results revealed that F. culmorum and F. pseudograminearum, B. sorokiniana, Fusarium sp., R. solani, F. redolens, C. spicifera, C. inaequalis, and N. oryzae were virulent, whereas others were non-pathogenic. The findings of this investigation demonstrate the presence of a diverse spectrum of pathogenic fungal species relevant to wheat crown and root rot in Kazakhstan. To the best of our knowledge, this is the first report of F. pseudograminearum, Fusarium sp., C. spicifera, and C. inaequalis as pathogens on wheat in Kazakhstan.

Characterization and Pathogenicity of Pythium-Like Species Associated with Root and Collar Rot of Kiwifruit in Turkey.

During the period of June to October in 2018, a widespread decline was observed on kiwifruit vines in the vineyards located in the Altinordu, Fatsa, and Persembe districts of Ordu province in Turkey. The symptoms were associated with reddish-brown rots expanding from the root to the collar with sparse off-color foliage. Based on the percentage of the total infected samples across 18 vineyards, the most common oomycete species were Globisporangium intermedium (37.1%), Phytopythium vexans (34.3%), Globisporangium sylvaticum (14.3%), Globisporangium heterothallicum (11.4%), and Pythium dissotocum (2.9%). The morphological identification of isolates was confirmed based on partial DNA sequences containing the nuclear rDNA internal transcribed spacer region gene and the mitochondrial cytochrome c oxidase subunit II gene. The optimum growth temperature and the optimum pH values of the five species ranged from 22.98 to 28.25°C and 5.67 to 8.51, respectively. Pathogenicity tests on the seedlings of kiwifruit cv. Hayward revealed significant differences in virulence among isolates. Phytopythium vexans and Globisporangium sylvaticum isolates caused severe root and collar rot resulting in seedling death, while Globisporangium heterothallicum and Globisporangium intermedium isolates had relatively lower virulence. All Globisporangium spp. and Phytopythium vexans isolates significantly decreased plant growth parameters (plant height, shoot and root dry weights, and root length); however, Pythium dissotocum caused very mild symptoms and did not affect these parameters of growth. To our knowledge, this is the first study reporting Globisporangium sylvaticum, Globisporangium heterothallicum, and Globisporangium intermedium causing root and collar rot on kiwifruit not only in Turkey but also in the world.

Phytopythium litorale: A Novel Killer Pathogen of Plane (Platanus orientalis) Causing Canker Stain and Root and Collar Rot.

Decline symptoms associated with lethal stem and branch canker stain along with root and collar rots were observed on 5- to 7-year-old roadside oriental plane trees (Platanus orientalis) in Diyarbakir, Turkey. Above-ground symptoms included leaf necrosis, leaf curling, extensive bluish or blackish staining of shoots, branches, stem bark, and wood surfaces, as well as stem cankers and exfoliation of branch bark scales. A general decline of the trees was distinctly visible from a distance. A Phytophthora/Pythium-like oomycete species with globose to ovoid, often papillate and internally proliferating sporangia was consistently isolated from the fine and coarse roots and stained branch parts and shoots. The pathogen was identified as Phytopythium litorale based on several morphological features. Partial DNA sequences of three loci, including nuclear rDNA internal transcribed spacer (ITS) and the large ribosomal subunit (LSU), and mitochondrial cytochrome c oxidase subunit II (coxII) confirmed the morphological identification. All P. litorale isolates were homothallic, developing gametangia, ornamented oogonia with elongate to lobate antheridia. Pathogenicity of P. litorale was tested by inoculation on excised shoots and by root inoculation on seedlings. P. litorale produced large lesions and blights on shoots in just 5 days and killed 100% of the seedlings in a month. This paper presents the first confirmed report of P. litorale as an important pathogen on a plant species causing branch and stem cankers, and root and collar rot, in and on P. orientalis, resulting in a rapid decline of trees and suggesting a threat to plane.