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Severe leaf spot disease was observed on Aloe vera plants in the winters of 2011 and 2012 during a survey of various nurseries of Gwalior, India. Irregular, sunken, dark creamish brown spots having reddish brown margin were noticed on both surfaces of the leaves. The causal organism was consistently isolated from symptomatic leaves on potato dextrose agar media (PDA). A total 59 isolates of fungi were recovered from diseased A. vera leaves, and 37 isolates were identified as belonging to the genus Fusarium. On the basis of morphological characteristics and internal transcribed spacer (ITS) region of rDNA amplified using the primers ITS4/ITS5 the pathogen was identified as Fusarium proliferatum (Matsushima) Nirenberg and pathogenicity of the isolate was confirmed by using Koch’s postulates. To the best of our knowledge, this is the first report of leaf spot disease caused by Fusarium proliferatum on A. vera plants in India.

Potato stands first as a non-cereal food crop for global consumption purposes. During the year 2019–20, dry rot disease symptoms were observed on potato tubers of the cultivar ‘Kufri Pukhraj’ kept in a cold store in the Moga district of Punjab State of India. Initial symptoms were observed as wrinkled and shriveled tubers with necrotic lesions on the tuber skin. The cottony white mycelial growth was also evident on some cracked tubers. During the incubation of these tubers at 25 °C for 7 days, the symptoms were more prominent with intensive whitish mycelial growth throughout the tuber. The pure culture of isolated fungus showed sparsely white colonies, which turned violet after 2 weeks. The oval or club-shaped microconidia were abundantly present on monophialides and multiphialides. The slender or straight macroconidia having 3–5 septa were also observed. The long and short chains of microconidia were observed in the fresh colonies. The molecular identification based on translation elongation factor 1 α (TEF1 α) and internal transcribed spacer region primers revealed the fungus identity as Fusarium proliferatum. The pathogenicity test was also performed to prove the Koch postulates on the initially identified host cultivar. To the best of our knowledge, this is the first report of dry rot disease caused by Fusarium proliferatum in India.

Cucurbits include the major vegetables group cultivated widely in India during summer and rainy seasons. Out of different cucurbitaceous crops grown, bottle gourd (Lagenaria siceraria) occupies a highest place in human diet but prevalence of various mycoflora acts as a major limiting factor in deteriorating the crop qualitatively as well as quantitatively. In the present study, mycoflora associated with seeds and fruits of bottle gourd was isolated using different methods and characterized on the basis of colony colour, type of spore, spore size and spore shape. The identity was further confirmed through microbial identification services of CABI (Centre for Agriculture and Biosciences International). The isolated fungal flora was enumerated on the basis of per cent isolation frequency of individual mycoflora. A total of fourteen fungal species belonging to eight genera were isolated with prevalence of Fusarium proliferatum as external as well as internal seed borne and Talaromyces pinophilus as external seed borne mycoflora. Amongst three methods used for seed mycoflora isolation, moist blotter paper method was found to be the best followed by deep freezing and agar plate methods, respectively.

The endophytic fungi are an important source of bioactive natural products. This study includes evaluation of antimicrobial activity and phytochemical analysis of endophytic fungus ACQR8, recovered from the root tissues of a folk medicinal plant Cissus quadrangularis L., which was identified as Fusarium proliferatum by microscopic and molecular methods. The fungus showed broad spectrum antibacterial and antifungal activities against variety of important plant and human pathogens. To the best of our knowledge, this is the first study to record antifungal activity of any endophytic Fusarium sp. against phytopathogens Rhizoctonia solani, Fusarium oxysporum and the dermatophytes Trichophyton mentagrophyte and Microsporum gypseum. The minimum inhibitory concentration of the crude extract ranged from 40–120 µg/ml against pathogenic bacteria and 0.2–2.5 mg/ml against fungal pathogens. Besides, this study probably is also the first to document the phytochemical constituents of an endophytic Fusarium sp. Phytochemical testing and GC-MS (gas chromatography- mass spectrometry) analysis revealed the presence of fatty acids and its derivatives, phenolics, terpenoids and unsaturated alkenes in the crude extract which scientifically validates the highly promising antibacterial and antifungal activity exhibited by endophytic isolate ACQR8.

Relevance and methods. Losses of agricultural crops are associated not only with the development of diseases during the growing season, but also during post-harvest storage. Garlic is a popular vegetable and aromatic crop in world. Significant losses in garlic yield during cultivation and storage are associated with fungal pathogens, the most harmful of which are representatives of the genus Fusarium. In the Moscow region, the defeat of garlic by Fusarium occurs annually, but with varying intensity. At the Federal Scientific Vegetable Center (FSVC), it was shown that rot and wilting of garlic plants is caused by a complex of pathogenic fungi, including mainly different species of Fusarium. At the same time, the ratio of Fusarium species in the pathogenic complex changes from year to year, new Fusarium species and their races are registered. The aim of this study was to identify fungal phytopathogens causing dry rot of garlic cloves during post-harvest storage. To carry out the work, garlic bulbs of cultivars Dubkovsky and Strelets were taken from the FSVC storage.Results. As a result of visual examination, cloves with symptoms of dry rot were identified. The diseased cloves tissues were plated on potato dextrose agar to obtain fungal colonies. Analysis of the morphological and cultural characteristics of fungal isolates, as well as the nucleotide sequences of four DNA regions (ITS spacers, genes EF1α, RPB1, and RPB2) showed that the causative agent of dry rot of garlic cloves is the pathogenic fungus Fusarium proliferatum. In addition, in the field, identification was carried out based on the analysis of the sequences of spacers ITS and the EF1αgene of phytopathogenic fungi inhabiting the root zone of garlic plants. As a result, two species of fungi of the genus Fusarium (F. proliferatum and F. oxysporum f. sp. cepae), as well as the species Rhizoctonia solani, Volutella rosea, and Ceratobasidium sp. were found in the root zone of garlic cultivars.

Sugars and organic acids are important factors associated with the postharvest quality of fruits and vegetables, but respiratory metabolism effects the metabolite contents. The objectives of this study were to explore the affects of Fusarium proliferatum infection on the postharvest quality and respiratory metabolism of green asparagus. It was found that F. proliferatum caused the soluble sugar concentration of asparagus to increase over the first 2 d of storage and then decrease again to become significantly lower than the control at 4 d. The titratable acidity (TA) of infected asparagus also increased from day 2 to day 6 and then decreased to become significantly lower than the control. Similarly, the key enzyme activities of the Embden-Meyerhof-Parnas (EMP) pathway, tricarboxylic acid (TCA) cycle, and pentose phosphate pathway (PPP) increased in pathogen-infected asparagus at the beginning of storage and then decreased to become lower than the control at the end. These findings demonstrate that F. proliferatum caused quality deterioration and an impairment of respiratory metabolism in postharvest asparagus. This research helps to clarify the relationships between pathogen infection, physiological metabolism, and asparagus quality and will provide valuable information for the control of postharvest diseases of asparagus.

Symptoms of dry clove rot of Allium sativum produced in Slovakia were noticed during storage in 2019. In 2020, garlic bulbs of eight cultivars were analysed and fungal isolates were obtained. Morphological and molecular identification confirmed the isolation of Fusarium proliferatum from all cultivars. The mean proportion of symptomatic cloves in evaluated samples reached 78.68%, and the mean area of clove surface damaged by the disease ranged from 5.97% to 14.83% 5 weeks after the harvest. The pathogenicity of the selected F. proliferatum isolate was verified by inoculation of garlic cloves in laboratory bioassays. This is the first report of F. proliferatum causing dry rot of garlic bulbs in Slovakia.

The diversity and genetic differentiation of populations of Fusarium species associated with sorghum fields, both endophytes obtained from sorghum performing and non performing plants and isolates obtained from two sampling periods were investigated. Fusarium specific Internal Transcribed Spacer 2 (FITS2) primers set were used to assess genetic variability of 32 isolates from susceptible Fusarium spp. endophytes from Sorghum tissues. Fusarium thapsinum (Gibberella thapsina) with 68.75% of the isolates constituted the majority of Fusarium spp. isolated in performing plants. Gibberella thapsina species identified are described as non-pathogenic and associated to performing plant of sorghum. Previously, some species of Fusarium thapsinum have been recognized as pathogenic and responsible for yield losses in several cereal crops including Sorghum bicolor produced in Burkina Faso. The other Fusarium spp. identified in this study including Fusarium subglutinans, Fusarium chlamydosporum, Gibberella intermedia, Fusarium dlaminii, Fusarium oxysporum, Fusarium proliferatum, and Fusarium spp. An additional unknown fungi species were also identified. A diverse population of 10 sequence types was found, although 8 sequence types represented nearly two-thirds of the isolates studied. The sequence types were placed in different phylogenetic clades within Fusarium spp., and endophytic isolates were not monophyletic. Phylogenetic analysis from Neighbor-Joining/UnWeighted Neighbor-Joining showed a high genetic relationship among these 32 isolates of Fusarium spp. and high variation in FITS sequence of them. The use of specific phylomarker of the genus Fusarium allowed to identify the endophytic species of this genus and to establish the phylogenetic relationships between the endophytic species of Fusarium. The phylogenetic analysis revealed three groups of the fungi. However, no relationship between these groups and the geographical origins of these fungi has been established. Key words: Fusarium thapsinum, endophyte, FITS2 marker, sorghum.

The study was conducted in the laboratories of Faculty of Agriculture/University of Kufa for the period from 24/12/2017 to 24/2/2019 to isolate and diagnose fungi associated with imported bananas fruits (Musa spp.) in local markets. The study included five varieties of bananas for different origins to identify the ability of isolated fungi and their production for mycotoxins, including F. proliferatum. and evaluation of the effectiveness ultraviolet radiation uv-c with a wavelength of 254 nm, for 15 minutes and different concentrations of peppermint and aloevera extracts were evaluated their effectiveness in reducing of Fumonisin B1. The results showed the effect of UV-C and different concentrations of hot water extracts of peppermint and aloe vera to inhibit growth fungus F. proliferatum on PSA medium, a significant effect of peppermint extract,as it showed an inhibition of the growth fungus F. proliferatum. Concentration 1.5%, with amounted to 75.52% compared to control treatment, in which the percentage of inhibition was 0.00%. Ultraviolet light (UV-c) had a significant role in inhibiting the growth of the fungus. Inhibition which increasing the duration of the fungus exposure to ultraviolet (UV) radiation, as the duration of 15 minutes gave the highest rate of inhibition amounting to 81.35 % compared to control treatment of 0.00 %. The results also showed the high efficiency of ultraviolet radiation and peppermint extract at concentration of 1.5% in reducing toxicity of Fumonisin B1 and the protecting banana fruits from fungal infection, in addition to the possibility of prolonging the storage period.

International audience; The incidence of garlic rot has constantly increased in France since the early 2000s. To set up an efficient method of garlic protection against this disease, we have clarified the etiology of this disease. This was achieved by surveying garlic from the two main French basins of garlic production during 3 years. Fungi were isolated from 5493 garlic cloves belonging to pink, purple and white garlic types. Sequencing of the translation elongation factor 1 alpha gene of 1171 strains revealed that 94 % of the strains belong to the species Fusarium proliferatum and 6 % belong to F. oxysporum. The pathogenicity of both species on garlic was confirmed by artificial inoculations and re‐isolations. There was significantly more F. oxysporum in symptomatic garlic cloves coming from the southeast basin (9.44 %) than from the southwest basin (2.76 %). This study confirms that garlic rot is present in pink, purple and white types. However, pink type garlic harbors F. oxysporum significantly less frequently (1.59 %) than do white (9.39 %) and purple (7.34 %) types. Sequencing of rpb1, rpb2, ITS and IGS regions of a subsample of strains revealed that there is little genetic diversity of the French population of F. proliferatum.

The aim of the study was to test in vitro and in vivo the efficacy of triazoles and biocontrol agents (BCAs) against Fusarium proliferatum and F. oxysporum, the former signaled as the main causal agent of garlic dry rot and the latter also involved. In vitro trials were organized using potato dextrose agar with added chemicals or BCAs inoculated with selected F. proliferatum and F. oxysporum. Garlic cloves were dipped before sowing in suspensions prepared with the fungicides showing the best performances in vitro; then they were dipped in Fusaria suspension before sowing. In in vitro trials, the maximum Fusaria growth inhibition was performed by Propiconazole + Prochloraz (100%), followed by Tebuconazole (88.9%). BCAs showed great capacity to control Fusaria, with a maximum growth inhibition of 80% (Trichoderma harzianum + T. gamsii). In vivo bacterial BCAs showed a similar capacity to control F. proliferatum and F. oxysporum compared to chemical products (mean of severity index 18.6% and 11.7%, respectively). In vivo results confirmed the in vitro performances, except for Trichoderma, which had the worst performances in vivo. Therefore, the results are preliminary but promising for future field application.

Ambrosia beetles (Coleoptera: Scolytinae) cultivate their fungal symbiont within host substrates as the sole source of nutrition on which the larvae and adults must feed. To investigate a possible role for semiochemicals in this interaction, we characterized electrophysiological and behavioral responses ofXylosandrus germanusto volatiles associated with its fungal symbiontAmbrosiella grosmanniae. During still-air walking bioassays,X. germanusexhibited an arrestment response to volatiles ofA. grosmanniae, but not antagonistic fungiBeauveria bassiana,Metarhizium brunneum,Trichoderma harzianum, the plant pathogenFusarium proliferatum, or malt extract agar. Solid phase microextraction-gas chromatography-mass spectrometry identified 2-ethyl-1-hexanol, 2-phenylethanol, methyl benzoate and 3-methyl-1-butanol in emissions fromA. grosmanniae; the latter two compounds were also detected in emissions fromB. bassiana. Concentration-responses using electroantennography documented weak depolarizations toA. grosmanniaefungal volatiles, unlike the comparatively strong response to ethanol. When tested singly in walking bioassays, volatiles identified fromA. grosmanniaeelicited relatively weak arrestment responses, unlike the responses to ethanol.Xylosandrus germanusalso exhibited weak or no long-range attraction to the fungal volatiles when tested singly during field trials in 2016–2018. None of the fungal volatiles enhanced attraction ofX. germanusto ethanol when tested singly; in contrast, 2-phenylethanol and 3-methyl-1-butanol consistently reduced attraction to ethanol. Volatiles emitted byA. grosmanniaemay represent short-range olfactory cues that could aid in distinguishing their nutritional fungal symbiont from other fungi, but these compounds are not likely to be useful as long-range attractants for improving detection or mass trapping tactics.

Avocado (Persea americana Mill.) is grown for its nutritious fruit. However, the quantity and quality of these fruits are threatened by some fungal organisms which can cause health complications when it is consumed by man. DNA extraction provides a unique tool for identification of organisms. This study was conducted to isolate and identify fungal species associated with avocado fruit using both morphological and molecular techniques. Fungal species were isolated from Persea americana purchased from Choba market, Port Harcourt, Rivers State, Nigeria using Potato Dextrose Agar (PDA) as a growth medium. The morphology of isolated fungi on PDA were cotton-like blackish grey spots, white villous colonies, greyish powdery spores and black spores for isolates 1 to 4 respectively. Extraction of DNA from fungal isolates was carried out using Zymo Fungal/Bacteria DNA Miniprep Kit. PCR amplification of the ITS1-2 regions of isolates was carried out using fungal universal primer pair; ITS4 and ITS5.PCR amplification of the ITS1-2 gene sequences yielded amplicons between 537-580 base pairs. PCR products were sequenced and the sequencing result after BLAST search revealed the identity of the four fungal species as follows: Lasiodiplodia theobromae, Fusarium proliferatum, Penicillium sp. and Aspergillus niger. This study will promote the knowledge of specific fungal species associated with Persea americanna and help plant pathologists to proffer preventive and control measures to enhance fruit protection and yield quality.

Fusarium basal rot (FBR) of onion is a serious disease problem worldwide. The Fusarium species causing FBR can also produce mycotoxins that are potentially harmful to humans and animals. In this study, a multiple reaction monitoring technique with ultra-high-performance liquid chromatography–tandem mass spectrometry (MRM UHPLC-MS/MS) was developed and validated for onion matrix to study Fusarium mycotoxins in the harvested onions. This study was focused on fumonisins B1, B2, and B3 (FB1, FB2, and FB3), beauvericin (BEA), and moniliformin (MON), which are the main mycotoxins produced by Fusarium oxysporum and Fusarium proliferatum. In the in-house validated protocol, the onion samples were extracted with methanol:water (3:1) using magnetic stirring for 15 min. FBs and BEA were determined directly from the filtered extracts, whereas MON required sample concentration prior to analysis. No cleanup of extracts was needed prior to analysis. The target mycotoxins were separated on an Acquity UPLC system BEH C18 column with gradient elution. Mycotoxins were identified and quantified using 13C-FB1 as internal standard. Minor matrix effect was compensated using multi-point matrix-matched calibration curves with uninfected onion sample. For the mycotoxins studied, a good linearity was obtained (R2 ≥ 0.99) and the recoveries were in the range of 67–122%, with the highest standard deviation for MON, 22%. The limits of quantification were from 2.5 to 10 ng g−1 in onion matrix. The method was successfully employed for the analysis of mycotoxins in harvested onions showing FBR symptoms and found to be infected with F. oxysporum and F. proliferatum.

The aim of the current study was to isolate endophytes from the roots of Rhazya stricta and assess their potential to improve the growth of drought-stressed sunflower seedlings. The potential role of the isolated endophytic fungus was initially screened by using two rice varieties (mutant rice cultivar Waito-C: Gibberellins deficient; and Dongji byeo: Gibberellins-producing normal cultivar). A significant (p < 0.05) increase in various growth attributes of both rice varieties associated with one of the isolates i.e., N4 was noticed. Furthermore, the N4 isolate was tested for its role in improving the agronomic attributes of sunflowers under drought stress. The symbiotic association significantly (p < 0.05) improved the host growth and protection from PEG-induced drought stress. The drought mitigation in N4-associated sunflower seedlings can be linked with the regulation of phytohormones, stress-related metabolites, low-molecular-weight proteins and sugars, and scavenging of reactive oxygen species (ROS). Microscopic analysis revealed that the potent endophytic fungal strain consisted of thin and hyaline hyphae, forming dense olive-green mycelia (4–5 cm in dm) with black flask-shaped fruiting bodies. Based on the ITS sequence homology and phylogeny, the strain was identified as Fusarium proliferatum (MG251448). The results of this study concluded that this phytohormone-secreting endophyte can improve crop productivity in dry areas where drought stress is the main challenge faced by crops.

The purpose of these studies was to study the interaction of pathogens of corn root rot in the agricultural sector of the Northern region of Krasnodar region. Corn root rot in the agricultural farms of the region annually cause significant damage to the harvest of silage and corn grain. In the course of research, the biological feature of the relationship between two species of the genus Fusarium: Fusarium concentricum Nirenberg & O'donnell and Fusarium proliferatum Matsush., optimizing the five-fold increase in the first species of mushroom compared to the growth of the second. PCR analysis revealed Fusarium oxysporum strain IMI 58289 with increased ability to exhibit elements of aggressive synergism. These fungi belong to the Department Ascomycota, order Hypocreales. In the soil of the studied rhizosphere, a natural hyperparasite – Trichoderma was detected, which can be used to minimize Fusarium infection.

BACKGROUND Rice spikelet rot disease (RSRD) is an emerging disease that significantly reduces rice yield and quality. In this study, we evaluated the potential use of the broad-spectrum endophytic fungus Phomopsis liquidambaris B3 as a biocontrol agent against RSRD. We also compared the control effects of different treatments, including chemical fungicides and treatment with multiple strains and single strains in combination or individually, against RSRD. The objective of this study was to find an effective and environmentally friendly control strategy to reduce the occurrence of RSRD and improve the rice yield. RESULTS In pot experiments, the effect of B3 alone was better than that of fungicide or combined measures. The results showed that root colonization by B3 significantly reduced the incidence and disease index of RSRD by 41.0% and 53.8%, respectively. This was related to enhanced superoxide dismutase (SOD), peroxidase (POD), and polyphenol oxidase (PPO) activity, and to significantly upregulated expression levels of OsAOX, OsLOX, OsPAL, and OsPR10 in rice. Moreover, B3 improved the diversity of the bacterial community rather than the fungal community in the rice rhizosphere. It also led to a decrease in Fusarium proliferatum colonization and fumonisin content in the grain. Finally, root development was markedly promoted after B3 inoculation, and the yield improved by 48.60%. The result of field experiments showed that the incidence of RSRD and the fumonisin content were observably reduced in rice receiving B3, by 24.41% and 37.87%, respectively. CONCLUSION The endophytic fungus Phomopsis liquidambaris B3 may become an effective tool to relieve rice spikelet rot disease. © 2020 Society of Chemical Industry.

Grain storage plays a crucial role in ensuring food security to Thai farmers so sustainable protection methods against deleterious microorganisms, such as fungi, are necessary. Essential oils (EOs) have demonstrated broad-spectrum fumigant antifungal activity against most storage fungi that are problematic in Thailand. Four storage fungi (Aspergillus flavus, A. niger, Curvularia lunata and Fusarium proliferatum) were isolated from dried rice and corn grains (stored for at least six months). EOs were extracted by hydrodistillation from clove buds (Syzygium aromaticum), fruit peel and leaves of makrut lime (Citrus hystrix), eucalyptus leaves (Eucalyptus sp.) and lemongrass stems (Cymbopogon citratus). The fungi inoculated on PDA in plastic cups were exposed to each EO vapour originating from paper disc attached in the lids in an inverted position. The minimum inhibitory concentration (MIC) for each EO was determined. Selected MICs were combined in a binary manner and similarly tested against the fungi. Fractional inhibitory concentration indices (FICI) were determined for each combination. Lemongrass and makrut lime leaf EOs were the most effective with MICs of 0.09 µL/mL against Curvularia lunata and 0.19-0.28 µL/mL against A. flavus, A. niger and F. proliferatum. Eucalyptus oil produced the least effective vapour (MIC 0.56-0.74 µL/mL) against all tested pathogens. A combination of lemongrass and makrut lime leaf EOs was partially synergistic against A. niger (FICI=0.75) but was fully synergistic against the other three fungi tested (FICI=0.5). The EOs from lemongrass and makrut lime leaf have potential to suppress the growth of the four grain-storage fungi tested.

Fumonisin B1 (FB1) is a prevelant mycotoxin in our alimentary chain. It was produced by the fungi of the genus Fusarium verticillioides and Fusarium proliferatum. FB1 was playing as a competitive inhibitor of ceramide synthase; a key enzyme in the biosynthesis of sphingolipids. Indeed, it was associated with several affects in humans and livestock animals. The aim of our report was elucidated to evaluate the protective effects of Lactobacillus paracasei BEJ01 (LP) isolated from the traditional butter of Tunisia against the FB1 genotoxicity, hematoxicity, oxidative stress and histological damage in liver and kidney of BALB/c mice. Forty old week mice were randomly divided into four treatment groups (10 mice/group): Group 1: control; Group 2: LP (2 × 109 CFU/ml ~ 2 mg/kg p.c); Group 3: FB1 (100 μg/kg p.c.); Group 4: LP (2 × 109 CFU/ml ~ 2 mg/kg p.c) + FB1 (100 μg/kg p.c.). 48 h after the end of the treatment (10 days), the mice were sacrificed and the blood, liver and kidney were collected. The blood was used for hematological and biochemical studies. Liver and kidney samples were the target for evaluation of oxidative stress enzymes status and histopathological damage. The results indicated that FB1 was induced toxicities in the blood parameters and in liver and kidney tissues as well as in the profile of oxidative stress enzymes levels. The co-treatment with LP was found able to mitigate the FB1 toxicities by reduction of its bioavailability in the gastro intestinal tract. However, treatment with LP alone was safe and no sign of toxicity was showed. In Summary, the LP strain was able to prevent FB1 toxicities and indeed it could be exploited as one of the biological strategies for foodstuffs decontamination.

Cannabis (Cannabis sativa L., marijuana) plants grown under greenhouse or controlled environments with symptoms of leaf yellowing, leaf necrosis and defoliation were observed during 2018–2019. Addi...

Fusarium proliferatum and F. verticillioides are mycotoxin-producing, seedborne pathogens of maize. They are often asymptomatic in seed, eluding symptom-based detection. Experiments were conducted in nonsterile soil to determine whether interspecific competition influenced establishment in maize plants of an introduced isolate of F. proliferatum or F. verticillioides. Hygromycin-resistant, green fluorescent protein (GFP) transformed (GFP-tagged) F. proliferatum (F. proliferatum-green) and hygromycin-resistant, monomeric red fluorescent protein (mRFP) transformed (mRFP-tagged) F. verticillioides (F. verticillioides-red) strains were developed to provide molecular markers to track fungal establishment. Heat-killed Fusarium-free maize seed, colonized with F. proliferatum-green or F. verticillioides-red by immersion in a spore suspension for 16 h, served as the source of inoculum. The ability of F. proliferatum-green and F. verticillioides-red to colonize viable maize plants already colonized by the other species was determined. Maize plants were retrieved from soil after 14 days and DNA was extracted from three consecutive root segments and three consecutive stem segments. A TaqMan multiplex real-time quantitative PCR protocol was developed to identify and quantify F. proliferatum-green and F. verticillioides-red from each plant segment from each treatment; the experiment was repeated three times. This experiment confirmed that F. proliferatum-green and F. verticillioides-red effectively colonized roots and stems of the maize plant already colonized with the other species. Prior colonization of maize tissues by F. verticillioides-red (P = 0.6749) and other seedborne microorganisms (P = 0.1910) reduced but did not prevent subsequent colonization by F. proliferatum-green. Similarly, prior colonization of maize tissues by F. proliferatum-green (P = 0.7032) and other seedborne microorganisms (P = 0.1447) reduced but did not prevent subsequent colonization by F. verticillioides-red.

Sulfatases catalyze the hydrolysis of sulfuric acid esters and play a key role in a number of biological processes of both higher eukaryotes and prokaryotes. According to literature data, for the implementation of catalysis, some representatives of this group of enzymes require posttranslational modification of serine or cysteine residues in the active center into the unique amino acid Cα-formylglycine. Nevertheless, it is confirmed that even in the absence of this modification, some sulfatases are capable of catalyzing the hydrolysis of sulfoesters. In this work, we studied the structural and functional features of active recombinant sulfatase from the mycelial fungus Fusarium proliferatum LE1, which contains a cysteine residue in the active center. A theoretical atomic model of the enzyme was first constructed, the structural organization of its active center was determined, and key amino acid residues involved in the binding of p-nitrophenyl sulfate and in the reaction of its hydrolysis were identified. Point amino acid substitutions of the identified residues led to inactivation of the enzyme. In particular, mutant forms of the enzyme with the replacement of catalytic cysteine by serine and threonine containing a hydroxyl group in the side chain completely lost activity, which indicates the direct participation of the mercapto group of cysteine in the hydrolysis reaction.

Fumonisin B1 is a mycotoxin produced by Fusarium verticillioides and Fusarium proliferatum found in various crops, particularly maize. Besides carcinogenicity, other manifestations have been registered in different animals and in humans. In the case of humans, epidemiological studies have reported high prevalence of esophageal cancer in populations exposed to fumonisins. This study aimed to evaluate the minimum concentration of FB1 capable of inducing cytotoxicity (cell viability test), genotoxicity (comet assay) and mutagenicity (micronucleus) in cultured human leukocytes and to evaluate the effectiveness of in silico tests to predict FB1 toxicity. All concentrations analyzed (200; 100; 50; 5; 0.5; 0.05; 0.005 μg/mL and 300; 30; 3; 1; 0.1; 0.01 fg/mL) except the lowest demonstrated dose-dependent toxicity in all parameters analyzed (p

Chlorosis on rachis followed by plant wilting of Syagrus romanzoffiana (Queen Palm) was observed during the survey of commercial ornamental plant growing nurseries and landscapes of sixteen locations from five districts of Punjab province. About 25% plants were found affected by the disease. Symptomatic root and leaf tissues were sliced, surface sterilized, transferred on PDA medium and incubated at 26 °C. Frequently whitish to purple fungal colony pigmentation was observed with club shaped microconidia and slender shaped macroconidia. Based on morpho-cultural characters the fungal isolate was identified as Fusarium proliferatum. The internal transcribed spacer (ITS) region and translation elongation factor 1-α (TEF1-α) region were sequenced for molecular characterization. Phylogenetic trees were constructed based on the sequences of both investigated loci to explore the place of investigated fungal isolate FMB-F-P-QP in evolutionary hierarchy of Fusarium species. In phylograms, this isolate was clearly nested with the strains of Fusarium proliferatum. The morphogenomic studies proved this isolate as F. proliferatum. The characterized culture deposited in FMB-Culture Collection-UAF (FMB 0133) and generated sequences of both loci were submitted to GenBank (accession No. MH557215 for ITS; accession No. MH568689 for TEF1-α). Koch’s postulates were accomplished by applying F. proliferatum and yellowing of leaves started three weeks post inoculation while chlorosis on one side of rachis or complete wilting appeared after six months. This is the first reported study of Syagrus romanzoffiana wilt caused by F. proliferatum in Pakistan. Four strains of three Trichoderma species (one strain of T. harzianum, two strains of T. atroviridae and one strain of T. viridae) were evaluated for their antagonistic potential against F. proliferatum. T. harzianum showed higher antifungal potential 85%, as compared to the other tested strains.

Fungal wilt and root rot diseases affecting tomato have become prevalent in China in recent years and have caused considerable damage. In 2016 to 2018, symptoms of putative wilt and root rot diseases were observed in several locations in tomato cultivars with resistance to Fusarium oxysporum f. sp. lycopersici races 1 and 2. The objective of this study was to identify the causative agents of wilt and root rot of tomato in China and provide a basis for disease prevention and resistance breeding programs. Based on DNA sequence analyses of the internal transcribed spacer (ITS) region, 91 isolates from the roots of tomato plants showing symptoms of wilt and root rot were identified, including F. oxysporum (64 isolates), Fusarium solani (11 isolates), Fusarium proliferatum (2 isolates), Fusarium graminearum (2 isolates), Fusarium equiseti (1 isolate), Pythium aphanidermatum (6 isolates), Ascomycota sp. (2 isolates), and Plectosphaerella cucumerina (3 isolates). F. oxysporum accounted for 70.33% of the isolates obtained. In this case, using PCR-based methods for differentiation of F. oxysporum, we identified several formae speciales and races of F. oxysporum: 7 isolates were identified as F. oxysporum f. sp. lycopersici race 1, 2 isolates as F. oxysporum f. sp. lycopersici race 2, 35 isolates as F. oxysporum f. sp. lycopersici race 3, and 13 isolates as F. oxysporum f. sp. radicis-lycopersici. Pathogenicity tests revealed 55 isolates of tomato wilt and root rot pathogens to be virulent. This study demonstrated that F. oxysporum f. sp. lycopersici race 3 was the most widespread and highly virulent race among these tomato pathogens in China, followed by F. oxysporum f. sp. radicis-lycopersici. Therefore, the development of resistant varieties of tomato against F. oxysporum f. sp. lycopersici race 3 and F. oxysporum f. sp. radicis-lycopersici would aid efforts to develop effective disease management strategies.

Postharvest fruit rots are of the most important diseases of fruit after picking. One of the main aims of this study was to investigate the pathogens causing apple fruit rots in Naoussa Imathia and Ptolemaida Kozanis. The results showed that the fungus Penicillium was responsible for about 78% and 84% in 2016 and 2017 respectively of the postharvest fruit rots of apples, while the fungus B. cinerea caused about 17% and 12% respectively. The fungi Monilinia laxa, Fusarium avenaceum, Fusarium proliferatum, Alternaria tenuissima, Truncatella angustata and Boeremia exigua were responsible for less than 5% of the total rotted fruit in both years. The sensitivity of 100 P. expansum isolates to the commercially available fludioxonil and the mixture fludioxonil + cyprodinil was also tested. None of the isolates was resistant to the fungicides tested. Finally, the efficacies of the above fungicides against P. expansum were examined. The results showed that both fludioxonil and the mixture fludioxonil + cyprodinil were appropriate to control P. expansum in apples. Both protected fruit for at least 30 days.

Fusarium proliferatum is a globally distributed fungal pathogen that affects a range of crop hosts and is one of the main producers of mycotoxins, such as fumonisins, in foods. Specific PCR primers are commonly used for detection and identification of this pathogen. The aim of this study was to validate previously published F. proliferatum‐specific primers targeting the intergenic spacer (IGS) region and characterize intraspecific variation and homologous recombination events for isolates obtained from diseased garlic bulbs in Spain. Sixty‐nine isolates were morphologically identified as F. proliferatum, and their identity was confirmed by sequencing of the translation elongation factor; however, specific IGS primers did not result in an amplification product for nine of these isolates. Further analysis showed that this was due to polymorphism in the IGS region and six isolates were classified as IGS type I, while the remaining isolates were type II. Sequencing of the complete IGS region revealed numerous sequence polymorphisms amongst F. proliferatum isolates, and regions of recombination. Duplication and deletion events may have occurred via unequal crossing over during mitotic or meiotic recombination. These results suggest that the IGS region may be too variable as a reliable target for F. proliferatum‐specific identification.\ud \ud

Crop fungal infections such as those caused by Fusarium species often result in yield loss pre- and post-harvest and during storage, contaminate food and exacerbate food insecurity. Synthetic fungicides are becoming less attractive as they pose health risks to consumers and are not environmentally friendly. As an alternative to synthetic fungicides, medicinal plant species may present sustainable and affordable sources of environmentally friendly fungicides for controlling plant diseases. This study was aimed at evaluating the in vitro antifungal activity of different medicinal plant extracts individually or in combination against Fusarium proliferatum, F. solani, F. verticillioides and F. graminearum. Different solvent (water, acetone and ethyl acetate) extracts obtained from the leaves of thirteen medicinal plant species were evaluated for antifungal activity using the micro-plate dilution assay. Their combined effect was evaluated by determining their fractional inhibitory concentration index (FICI). When evaluated individually, acetone extracts obtained from Combretum erythrophyllum, Harpephyllum caffrum and Quercus acutissima were the most active extracts inhibiting the growth of all the four pathogens with minimum inhibitory concentration (MIC) values less than 0.1 mg/ml. Out of 204 extract combinations evaluated, 150 extract combinations demonstrated either synergistic or additive antifungal activity. among the best combinations, combined acetone extracts of Harpephyllum caffrum and Combretum erythrophyllum showed strong synergistic antifungal activity against F. graminearum, F. proliferatum and F. verticillioides (MIC values of 0.02 mg/ml, 0.002 mg/ml and 0.001 mg/ml, respectively), and strong additive antifungal activity against F. solani (MIC = 0.02 mg/ml). The approach of using medicinal plant extracts from renewable plant parts either individually or in combination is sustainable, affordable, environmentally friendly and may be more beneficial in the fight against crop pathogenic diseases, particularly in organic farming.

Papain-like cysteine proteases (PLCPs) in plants are essential to prevent phytopathogen invasion. In order to search for cysteine protease inhibitors and to investigate compounds that could be associated to pineapple Fusarium disease, a chemistry investigation was performed on Fusarium proliferatum isolated from Ananas comosus (pineapple) and cultivated in Czapek medium. From F. proliferatum extracts, nine secondary metabolites were isolated and characterized by nuclear magnetic resonance spectroscopy and mass spectrometry experiments: beauvericin (1), fusaric acid (2), N-ethyl-3-phenylacetamide (3), N-acetyltryptamine (4), cyclo(L-Val-L-Pro) cyclodipeptide (5), cyclo(L-Leu-L-Pro) cyclodipeptide (6), cyclo(L-Leu-L-Pro) diketopiperazine (7), 2,4-dihydroxypyrimidine (8), and 1H-indole-3-carbaldehyde (9). Compounds 1, 3, and 6 showed significant inhibition of papain, with IC(50) values of 25.3 ± 1.9, 39.4 ± 2.5, and 7.4 ± 0.5 μM, respectively. Compound 1 also showed significant inhibition against human cathepsins V and B with IC(50) of 46.0 ± 3.0 and 6.8 ± 0.7 μM, respectively. The inhibition of papain by mycotoxins (fusaric acid and beauvericin) may indicate a mechanism of Fusarium in the roles of infection process. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s42770-020-00256-7) contains supplementary material, which is available to authorized users.

Maize is currently the most important crop in China. A major concern in maize production is maize ear rot caused by Fusarium spp., which results in yield losses, reduction of seed quality and the accumulation of mycotoxins in the harvested grains. To identify the importance of the different Fusarium species in maize infection, we performed a comprehensive survey on 9,000 asymptomatic and randomly collected maize kernels. Seeds were collected from 12 different provinces covering all major maize growing areas in China and included five maize varieties. In total 1,022 Fusarium isolates were retrieved that were identified based on morphological characteristics, by species specific diagnostic PCRs and by EF1-α gene sequencing. Eight different species were identified: Fusarium verticillioides (75.34%), Fusarium graminearum (8.32%), Fusarium proliferatum (7.14%), Fusarium subglutinans (4.11%), Fusarium meridionale (1.57%), Fusarium oxysporum (1.37%), Fusarium semitectum (1.17%), and Fusarium asiaticum (0.98%). The distribution of Fusarium species was found to be different in different regions with the largest diversity observed in Hubei province, where all eight Fusarium species were isolated. Genetic chemotyping within the F. graminearum species complex indicated that all of the 85 F. graminearum isolates showed the 15-acetyldeoxynivalenol chemotype, whereas all F. asiaticum (n=10) and F. meridionale (n=16) isolates had the nivalenol chemotype even when isolated from the same maize field. To our knowledge this is the largest collection of Fusarium isolates from maize and further exploitations of this collection are discussed.

Seafood is part of a healthful diet, but seafood consumption is not risk-free because of sea contaminations by anthropogenic activities and environmental factors. Their microbiological quality is of major concern to the food processors, consumers and public health authorities hence the need for proper assessment of the microbial quality. This study therefore characterizes pathogenic microorganisms isolated from fresh sea snail and shrimp in Ugbo nla, Igbokoda, Ondo State, Nigeria. Isolation and molecular identification of microorganisms from snail and shrimp using 16sRNA and 18sRNA gene sequence for bacteria and fungi respectively, effects of salt concentration and temperature on isolated microorganisms were carried out using standard microbiological methods. Molecular identification revealed the presence different pathogens namely; Proteus mirabilis LZPM-03, Pantoea agglomerans BJCP2, Pseudomonas aeruginosa R8-589, Klebsiella pneumoniae 2N3, Serratia marcescens, Alcaligenes faecalis ACBC1, Yersinia. pseudotubercules YPIII, Escherichia coli 2A, Lysinibacillus sphaericus C3-41, Listeria monocytogen J1776, Staphylococcus aureus and Proteus mirabilis LCX20, Fusarium proliferatum FproStRINI, Aspergillus flavus NRRL3357, Aspergillus oryzae RIB40 and Aspergillus peniciliodes. Also, 4% salt concentration at 45°C effectively inhibited the pathogens isolated from seafood after 18 h. The presence of pathogenic microorganisms in the sea foods could make them a potential health-risk when consumed without proper cooking. Salting, a traditional technique could be used to eliminate these pathogens before consumption. Key words: Seafood, molecular identification and salt concentration.

Many fungi limit onion production in Burkina Faso. This study aims to identify the main Fusarium species associated with onion plant in field in order to determine those involved in seedling damping-off and bulb rot, and develop adequate management strategies of these diseases. For this purpose, 36 isolates of Fusarium were isolated from onion plants in 17 sites and subjected to molecular analysis and biometric characterization. The results revealed that the isolates belong to five Fusarium species: Fusarium oxysporum (44.44% of the isolates), Fusarium proliferatum (41.66%), Fusarium solani (5.55%), Fusarium fujikuroi (5.55%) and Fusarium thapsinum (2.77%). Fusarium oxysporum, F. proliferatum, F. solani and F. fujikuroi had faster mycelial development, with a growth rate of 7.72 - 8.27 mm/d, than F. thapsinum (6.52 mm/d). Conidia of F. oxysporum, F. proliferatum and F. solani were longer (4.74 - 5.96 μm) than those of F. fujikuroi and F. thapsinum (3.20 - 4.04 μm). Fusarium solani and F. oxysporum, respectively, had the largest and most partitioned conidia.

A novel visual detection of Fusarium proliferatum species through recombinase polymerase amplification and rolling circle amplification was established. Single-stranded circle DNA was produced based on one strand of RPA product, which used as a template for rolling circle amplification.

Background: Bottle gourd is a cucurbitaceous vegetable of culinary and medicinal importance cultivated in various tropical and sub-tropical regions of world. This crop is exposed to a wide variety of seed and soil mycoflora, out of which Fusarium proliferatum is utmost important as far as seed germination, viability and seedling vigour are concerned. Methods: Study was taken up to evaluate different fungicides and bioagents for their efficacy against the fungus Fusarium proliferatum under in vitro through spore germination inhibition technique. Result: Spore germination inhibition of 86.00%, 85.00% and 81.33% was recorded with hexaconazole (5% SC) @ 0.2% (C3), mancozeb (75% WP) @ 0.3% (C3) and Pseudomonas fluorescens (1% WP) @ 2% (C3), respectively. The inhibition in spore germination by mancozeb (75% WP) and Pseudomonas fluorescens (1% WP) was upto 77.33% and it was 61.78% and 67.33% in treatments involving carbendazim (50% WP) and Trichoderma harzianum (1% WP) that could be exploited to devise integrated approach for disease management.

THE HETEROCYTOUS cyanobacterium Westiellopsis prolifica has been recently documented in Egypt; however, our understanding of its diverse biotechnological and biomedical applications remains a limited and challenging topic. In this study, a W. prolifica strain isolated from hyper-arid desert habitats was investigated to test its antimicrobial potential against six foodborne pathogenic bacteria (Bacillus cereus, Staphylococcus aureus, Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, and Klebsiella pneumoniae) and eight mycotoxigenic fungi (Aspergillus carbonarius, A. flavus, A. niger, A. ochraceus, A. parasiticus, A. westerdijikia, Fusarium proliferatum, and Penicillium verrucosum), as well as to evaluate its probable toxicity and biosynthesis of the most common microcystin variant (MC–LR) using high-performance liquid chromatography (HPLC). The antimicrobial activity of various W. prolifica extracts was assessed using the disk–diffusion method, and the minimum inhibitory concentrations (MICs) were also determined. Our findings revealed that the extracts of dryland-dwelling W. prolifica had significant antimicrobial activities against all tested bacterial and fungal strains, with average MIC values of 0.4–6.7 and 0.3–9.2mg.mL−1, respectively. This antimicrobial potential can be ascribed to the combined synergistic effects of its biologically active and structurally diverse metabolites. Moreover, this species was a non-producer of MC–LR and was non-toxic based on the brine shrimp bioassay. Thus, the cyanoprokaryote W. prolifica can be considered a sustainable multifunctional candidate in the development of the pharmaceutical industry in Egypt.

Apple replant disease (ARD) is a common problem in major apple planting areas, and biological factors play a leading role in its etiology. Here, we isolated the bacterial strain QSB-6 from the rhizosphere soil of healthy apple trees in a replanted orchard using the serial dilution method. Strain QSB-6 was provisionally identified as Bacillus amyloliquefaciens based on its morphology, physiological and biochemical characteristics, carbon source utilization, and chemical sensitivity. Maximum likelihood analysis based on four gene sequences [16S ribosomal RNA gene (16S rDNA), DNA gyrase subunit A (gyrA), DNA gyrase subunit B (gyrB), and RNA polymerase subunit B (rpoB)] from QSB-6 and other strains indicated that it had 100% homology with B. amyloliquefaciens, thereby confirming its identification. Flat standoff tests showed that strain QSB-6 had a strong inhibitory effect on Fusarium proliferatum, Fusarium solani, Fusarium verticillioides, Fusarium oxysporum, Alternaria alternata, Aspergillus flavus, Phoma sp., Valsa mali, Rhizoctonia solani, Penicillium brasilianum, and Albifimbria verrucaria, and it had broad-spectrum antibacterial characteristics. Extracellular metabolites from strain QSB-6 showed a strong inhibitory effect on Fusarium hyphal growth and spore germination, causing irregular swelling, atrophy, rupture, and cytoplasmic leakage of fungal hyphae. Analysis of its metabolites showed that 1,2-benzenedicarboxylic acid and benzeneacetic acid, 3- hydroxy-, methyl ester had good inhibitory effects on Fusarium, and increased the length of primary roots and the number of lateral roots of Arabidopsis thaliana plantlet. Pot experiments demonstrated that a QSB-6 bacterial fertilizer treatment (T2) significantly improved the growth of Malus hupehensis Rehd. seedlings. It increased root length, surface area, tips, and forks, respiration rate, protective enzyme activities, and the number of soil bacteria while reducing the number of soil fungi. Fermentation broth from strain QSB-6 effectively prevented root damage from Fusarium. terminal restriction fragment length polymorphism (T-RFLP) and quantitative PCR (qPCR) assays showed that the T2 treatment significantly reduced the abundance of Fusarium in the soil and altered the soil fungal community structure. In summary, B. amyloliquefaciens QSB-6 has a good inhibitory effect on Fusarium in the soil and can significantly promote plant root growth. It has great potential as a biological control agent against ARD.

Aims: This article explored the effects and possible mechanism of different inoculation patterns of endophytic fungi Phomopsis liquidambaris B3 on the plant immune response and defense results to pathogen Fusarium. proliferatum.Methods: Rice seedlings pre-inoculated, instant-inoculated and non-inoculated with B3 were infected with F. proliferatum and grown in controllable conditions. The disease severity, F. proliferatum infection and plant growth were evaluated, and the activity of defensive enzymes, defense-related substances, reactive oxygen content, and defense hormone SA content were determined. Chemical treatment experiments and plant cell staining to verify plant cell death.Results: Pre-inoculation of endophytic fungus Ph. liquidambaris B3 may triggered the SA-dependent defense pathways of plants, increased defense-related enzyme activities and up-regulated the expression of defense genes, and thus decreased F. proliferatum colonization. However, instant-inoculation Ph. liquidambaris B3 inhibited its own root colonization, caused excessive burst of ROS and leaves cell death. Conclusion: For a specific plant pathogen, the instant-inoculation Ph. liquidambaris B3 when F. proliferatum infected leads to excessive burst of reactive oxygen species (ROS) and plant cell death. We recommend pre-inoculation of biological control agent and other prevention and control measures before pathogen infection to achieve the best control effect rather than instant control method applicated in the agricultural crop management.

In Bangladesh, Fusarium proliferatum is a prevalent pathogen of maize plants that poses a significant threat to human health and livestock by producing carcinogenic fumonisin. However, the use of the Bacillus species, which can colonize the infected plant parts, has been practiced globally to mitigate fumonisin contamination in maize. In this study, B. subtilis MMM1, a native isolate from the maize rhizosphere, was found to be the most effective antagonist against F. proliferatum, which reduced their mycelial growth and total fumonisin accumulation by 85.31% and 85.12%, respectively, over control through a fivefold reduction in fungal population (1.1 × 104 CFU/g maize grain) in vitro. Furthermore, out of the five bacilli used in this study, B. subtilis MMM1 was able to increase the root and, shoot length, and the vigor index of maize seedlings, by colonizing the fresh roots at 82 × 106 CFU/g root tissue. This suggests that B. subtilis MMM1 might be put forth both as a biocontrol agent and a plant growth promoter in Bangladesh to combat F. proliferatum. Nevertheless, evaluation of its efficacy in reducing fumonisin contamination in maize should be investigated under field conditions.

Maize stalk rot caused by Fusarium species is one of the most important fungal diseases of maize throughout the world. The disease is responsible for considerable yield losses and has also been associated with mycotoxin contamination of the crop. In this study, a survey of maize stalk rot was performed in seven locations of Yunnan Province in China during the cropping season of 2015 and 2016. Based on morphological and molecular characteristics, 204 isolates belonging to 12 Fusarium spp. from symptomatic stalks of maize were identified. Among the isolated strains, 83 were identified as Fusarium meridionale (40.5%), 46 as Fusarium boothii (22.5%), 34 as Fusarium temperatum (16.5%), 12 as Fusarium equiseti (5.9%), 10 as Fusarium asiaticum (4.9%), six as Fusarium proliferatum (3.0%), four as Fusarium verticillioides (2.0%), four as Fusarium incarnatum (2.0%), two as Fusarium avenaceum (1.0%), one as Fusarium cerealis (0.5%), one as Fusarium graminearum (0.5%), and one as Fusarium cortaderiae (0.5%). Fusarium cortaderiae was the first report on the causal agent of maize stalk rot disease in China. These isolates were divided into five chemotypes: nivalenol (NIV), deoxynivalenol (DON), beauvericin (BEA), zearalenone (ZEN), and fumonisin (FUM). Phylogenetic analysis based on partial sequences of the translation elongation factor 1α (TEF1-α) showed a high degree of interspecific polymorphisms among the isolates. Pathogenicity analysis on maize stalks indicated that all the 12 species of Fusarium were able to cause the disease symptoms with different aggressiveness. This study on population, pathogenicity, and toxigenic chemotypes of Fusarium species associated with maize stalk rot in Yunnan Province of southwest China, will help design an effective integrated control strategy for this disease.

Date palm (Phoenix dactylifera L.) is one of the world's oldest cultivated fruit crops. In Jordan, date palm farming started in the 1990s. The major date palm planting areas are Jordan valley, Aqaba, and Azraq (Al Antary et al., 2015). 'Medjool' and 'Barhi' are the two major cultivars in Jordan. In early 2018, some 18- to 24- month old date palm trees (cv 'Medjool') showed light brownish discoloration and dryness symptoms on the leaves and branches of infected date palm trees at the Jordan University Agricultural Research Station (JUARS) at the Jordan Valley (GPS coordinates 32.086871, 35.597219) (Figure 1). All the leaf parts including leaf base, spines, and leaflets were wrinkled and malformed. The infection led to a loss of 1-2% out of 1100 total Medjool trees at the station. Similar symptoms were observed in many date palm farms in the Jordan Valley. Diseased samples from rachis tissue from the JUARS were collected, surface sterilized with 5% sodium hypochlorite for five minutes, rinsed with distilled water for three times, dried, and plated on potato dextrose agar (PDA) medium (HIMEDIA). The plates were incubated at 25°C for seven days. After that, different fungal colonies were purified using the hyphal tip method. Mycelium of a representative isolate (FpDP2018JO-01) was harvested, DNA extracted using the CTAB protocol (Doyle and Doyle, 1990), amplified with three primers: ITS1/4 (White et al., 1990), β-tubulin and the elongation factor 1-alpha (EF1) gene regions. Amplicons were sequenced at Macrogen Inc, South Korea. Sequences were edited via MEGA 7 software (Kumar et al., 2016) and Blastn at the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov) which was used to search for similar accessions. The sequences were submitted to the GenBank and accession numbers were received for ITS1/2 (MK522076), β-tubulin (MK720958) and elongation factor 1 alpha (MW533146). The sequences were further used at the Fusarium MLST (https://fusarium.mycobank.org/) for identity confirmation. ITS1/4 and β-tubulin could not discriminate the species Fusarium proliferatum but EF1 - alpha could (Figure 2a-c; Supplement 1). For morphological identification, four representative F. proliferatum isolates (FpDP2018JO-01- FpDP2018JO-04) were used. Mycelium were white to dark purple in color, macroconidia (20.5 - 44.5 × 3.3 - 7.5 μm) were thin, slender, with 3-5 septa, and microconidia (4.3 - 12.1 × 2.5 - 4.3 μm) were thin and aseptate (Figure 3). Koch's postulate was performed on one-year-old seedlings according to Abdalla et al., 2000 method using the same sequenced isolate (FpDP2018JO-01). Five plants were inoculated by injecting 2 mlof inoculum into the crown area using a hypodermic needle and syringe. The inoculum was prepared according to Abdalla et al. (2000). The control set of seedlings (n=5) were injected with sterile distilled water. The experiment was arranged in a CRD design. Symptoms were evaluated three months after inoculation. On seedlings, yellowing of leaflets, discoloration of spines and rachis, and dryness of leaves were observed. Control seedlings showed no symptoms. Re-isolation form the detached leaves and infected seedlings was conducted to satisfy Koch's postulates. Fusarium sp. was confirmed to be F. proliferatum based on their microscopic characteristics. To our knowledge, this is the first record of F. proliferatum on date palm in Jordan. Date palm in Jordan especially 'Medjool' is an important cash crop. Fusarium spp. is an important pathogen that could cause huge losses on date palm and other crops. In Jordan, the pathogen has been isolated from samples from six farms so far, but detailed studies have not been conducted. It would be of importance to survey date palm farms for fungal diseases, test their pathogenicity using several isolates, and characterize them for proper management strategies. F. proliferatum was isolated from roots and leaves of declining date palm trees from many regions of Saudi Arabia and caused symptoms similar to those of F. oxysporum f. sp. albedinis, the causal agent of Bayoud (Abdalla et al. 2000; Saleh et al. 2016). Notonly that, but F. proliferatum was found to have the highest colonization abilities on date palm leaflets and is becoming serious pathogen on date palm (Saleh et al. 2016.

This study evaluates the influence of crop seasons, cultivation regions, and traditional agricultural practices on the occurrence of F. proliferatum and A. flavus in the rice chain in the Mekong Delta, Vietnam. A survey on pre- and post-harvest practices was performed from 2017 to 2019 in parallel with sampling. Results showed that F. proliferatum (36.3%) and A. flavus (10%) were predominantly present throughout the rice chain. These fungi frequently occurred in winter–spring and autumn–winter crops in Can Tho paddy. Especially, F. proliferatum appeared both on the field and during transportation (50–100%), while A. flavus presented at all stages (10–33%). The occurrence of F. proliferatum reduced 70–27% after drying, depended on the seasons, compared to field and transportation stages and could not be detected anymore at further stages. Applying poor pre-harvest agricultural practices such as the use of certain varieties (Jasmine, DT8 varieties), combination of fertilizers (organic–inorganic), fields with crop debris, unhygienic boats, and delayed drying time of 8–12 h or 12–28 h resulted in an increase in fungal contamination on paddy. This study provides a detailed description of fungi contamination in crop seasons, cultivation regions, and agricultural practices, which may help in understanding the fungal dynamic and allow identification of good agricultural practices to mitigate the fungal contamination and potential mycotoxin production.

In January 2018, gerbera plants showed root rot symptoms were found in a greenhouse at Guizhou Horticultural Institute in Guiyang city, Guizhou Province in southwest China. Morphological and molecular identifications confirmed the isolation of Fusarium proliferatum. The pathogenicity was verified with Koch’s postulates. To our knowledge, this is the first report confirming root rot of gerbera caused by Fusarium proliferatum in China.

A total of 26 fungal strains were isolated from aphids and tomato leaf miner as indigenous species and from red palm weevil and peach fruit fly as an invasive insect. Fungal strains tested for their abilities to attack both bean and wheat aphids. The mortality of aphids due to thesefungi was estimated. Results showed that eight entomopathogenic fungi Scopulariopsis brevicaulis, Verticillium sp., Fusarium chlamydosporum, Fusarium proliferatum, Fusarium semitectum, Pochonia chlamydosporia var. catenulata, Fusarium solaniand Fusarium verticillioides exhibited high virulence 60-100%. However, Scopulariopsis brevicaulis, Verticillium sp., Fusarium chlamydosporum and Fusarium solani proved to be the most virulent species against the target aphids inducing 90%-100% mortality.

Symptoms of vegetative malformation were observed on coconut palms (Cocos nucifera L.) in the Qeshm Island, Bandar Abbas and Minab, in Hormozgan province, southern Iran. The symptoms included misshapen and dwarfed leaves with shortened, thickened and tightened leaflets in wavy and zigzag form. The aim of this study was to identify the causal pathogen of coconut palm malformation and complete Koch's postulates for putative pathogen. Small pieces of surface‐disinfested malformed vegetative tissues of coconut palms were cultured on potato dextrose agar (PDA) medium. Fusarium isolates were permanently obtained from the symptomatic tissues. Sequence data from the internal transcribed spacer region (ITS1–5.8S‐ITS2) and translation elongation factor 1 alpha (TEF‐1α) gene were used for molecular identification of the isolates. BLAST search of the sequences showed 99%–100% identity to several Fusarium proliferatum strains in the GenBank, FUSARIUM‐ID and Fusarium MLST databases. A phylogeny inferred using individual sequence data from ITS region and TEF‐1α gene placed our isolates together with the other F. proliferatum sequences retrieved from the GenBank. Pathogenicity tests were carried out using one‐year‐old healthy coconut palm seedlings and conidial suspensions (10⁶ conidia/ml) of the F. proliferatum isolates. The first visible symptoms appeared on newly produced leaves of the inoculated seedlings during the 16th week after inoculation, wherease no disease symptoms were observed on the control plants until the end of the experiment. Reisolation from symptomatic tissues of the inoculated seedlings yielded isolates of F. proliferatum with morphological and molecular characteristics identical to those of the isolates used for inoculations. This is the first report of coconut palm malformation caused by F. proliferatum worldwide.

Fusarium is a saprophytic fungus which is broadly disseminated in soil, plant debris and other organic substrates. These fungi can affect indigenous tissue in immunocompromised patients and result in some fungal infections such as Onychomycosis, bone and joint disease, or sinusitis. In this paper, we report a 56-year-old female has Diabetes milletus who established to disseminate Fusarium infection in her left first toenail from the Dermatology Department / Khabat Skin Center, Sulaimani Province, Iraq. To study of these fungi as the true causative agents of nail infection, we describe a PCR/sequencing assay to confirm routinely diagnosis of the infecting fungi in this case with toenail Onychomycosis. Designation of the ordinary laboratory of Fusarium is very challenging because of the septate hyphae of Fusarium are hard to discriminate from those of Aspergillus, which has a more favourable consequence and this time-consuming. Therefore, molecular tools were used for identification of F. proliferatum from this case by amplifying ITS region of rDNA using a pair of universal primers ITS1 and ITS4. Infectious fungal molecular detection advances the select of right antifungal therapy, in that way improving the medication rate of nails infections. DNA sequence analysis and Phylogenetic tree of the amplified ITS-rDNA was used for species identification of this strain of F. proliferatum. For final identification sequencing of DNA has been carried out by [Macrogen Korea] using forward primer ITS1, The obtained consensus of query sequence was compared with the ITS DNA database on the BLAST homepage, aligned and diagnosed strain deposited to the GenBank and we received an accession number (MK112619). The sequence of the F. proliferatum amplicon was aligned using ClustalW and the alignment was used to make phylogenetic analysis using MEGA software version X. To the best of our knowledge, the isolated Fusarium in this paper is the first case of its kind to be sequenced and reported by the molecular method in Kurdistan region of Iraq

Fusarium proliferatum is one of the most pathogenic Fusarium species on date palm worldwide. We evaluated the colonization ability of 42 F. proliferatum strains collected from date palm trees with disease symptoms on detached leaflets of cvs Sheeshee and Sukkary. Cell wall degrading enzyme (CWDE) production by these strains was assessed with multiple relative enzyme activity (RA) indices. The ability of F. proliferatum strains to colonize leaflet cuttings varied, and there were significant differences (P

Onychomycosis is a fungal nail infection caused mainly by dermatophytes. Non-dermatophytes such as yeasts, Acremonium, Aspergillus, and Fusarium species account for only 10% of onychomycosis, and Fusarium species are relatively common amongst these pathogens. However, the main Fusarium species pathogens are Fusarium solani and Fusarium oxysporum. Fusarium proliferatum is a rare pathogen in onychomycosis, and only a few cases have been reported worldwide. We report a case of a 68-year-old woman presenting with a white- to yellow-colored patch on her left third fingernail plate, which was diagnosed by fungus culture and sequencing as onychomycosis caused by Fusarium proliferatum.