Fusarium Lateritium Research Articles

New insights into decoding the lifestyle of endophytic Fusarium lateritium Fl617 via comparing genomes

Fungal-plant interactions have persisted for 460 million years, and almost all terrestrial plants on Earth have endophytic fungi. However, the mechanism of symbiosis between endophytic fungi and host plants has been inconclusive. In this dissertation, we used a strain of endophytic Fusarium lateritium (Fl617), which was found in the previous stage to promote disease resistance in tomato, and selected the pathogenic Fusarium oxysporum Fo4287 and endophytic Fusarium oxysporum Fo47, which are in the same host and the closest relatives of Fl617, to carry out a comparative genomics analysis of the three systems and to provide a new perspective for the elucidation of the special lifestyle of the fungal endophytes. We found that endophytic F. lateritium has a smaller genome, fewer clusters and genes associated with pathogenicity, and fewer plant cell wall degrading enzymes (PCWDEs). There were also relatively fewer secondary metabolisms and typical Fusarium spp. toxins, and a lack of the key Fusarium spp. pathogenicity factor, secreted in xylem (SIX), but the endophytic fungi may be more sophisticated in their regulation of the colonization process. It is hypothesized that the endophytic fungi may have maintained their symbiosis with plants due to the relatively homogeneous microenvironment in plants for a long period of time, considering only plant interactions and discarding the relevant pathogenicity factors, and that their endophytic evolutionary tendency may tend to be genome streamlining and to enhance the fineness of the regulation of plant interactions, thus maintaining their symbiotic status with plants.

Interaction of a bioherbicidal fungus with a phenoxy herbicide for controlling velvetleaf (Abutilon theophrasti)

ABSTRACT The fungus, Fusarium lateritium Nees ex Fr. (FL), has shown potential as a bioherbicide for velvetleaf (Abutilon theophrasti Medik.) control. However, to achieve optimal infection and weed mortality, bioherbicide applications must be applied to weeds that are in the cotyledonary growth stage, followed by a dew treatment of at least 16 h. Greenhouse and field experiments were established to discover possible synergistic disease interactions with a phenoxy herbicide to improve the bioherbicidal potential of this fungus, and to mitigate some of the physical and environmental restrictions that limit the utility of this fungus as a bioherbicide. In greenhouse experiments, mortalities of 94% and 94% were recorded from sequential applications of 2,4-DB [4-(2,4-dichlorophenoxy)butyric acid] (0.02 kg ae ha−1) followed by FL (1.5 × 106 spores ml−1) at 5 min or 7 days after herbicide treatment. Velvetleaf plants in the fifth-to-seventh leaf growth stage were infected and killed with < 8 hrs. of dew. Similar results occurred under field conditions. Infection and weed control were inhibited by tank mixtures of 2,4-DB and FL, and by sequential applications of FL followed by 2,4-DB. These results suggest that timely applications of 2,4-DB followed by F. lateritium may provide effective control of velvetleaf.

Distribution of the invasive ambrosia beetle Anisandrus maiche (Coleoptera, Scolytinae) in Switzerland and first record in Europe of its ambrosia fungus Ambrosiella cleistominuta

Ambrosia beetles are highly successful as invaders because they are often transported internationally with wood packaging and other wood products and because their inbreeding mating systems facilitates establishment of invading populations. In 2022, two independent insect surveys in canton Ticino (southern Switzerland) revealed the widespread occurrence of the invasive ambrosia beetle Anisandrus maiche (Kurentzov, 1941) from southern to central-upper Ticino. This species is native to east Asia and has previously been found as a non-native invasive species in the United States, Canada, western Russia, Ukraine and, in 2021, in northern Italy. Here, we present the results of several trapping studies using different trap types (bottle traps, funnel traps and Polytrap intercept traps) and attractants and a map of the distribution of the species. In total, 715 specimens of A. maiche, all female, were trapped, and the identity of selected individuals was confirmed by morphological and molecular identification based on three mitochondrial and nuclear markers (COI, 28S and CAD). Trap samples from early April to early September 2022 in intervals of two to four weeks showed that flights of A. maiche occurred mainly from June to mid-August. Isolation of fungal associates of A. maiche from beetles trapped alive revealed the presence of four fungal species, including the ambrosia fungus Ambrosiella cleistominuta, the known mutualist of A. maiche. The identity of A. cleistominuta was confirmed by comparing DNA sequences of its nuclear, internal transcribed spacer (ITS) gene with reference sequences in NCBI and BOLDSYSTEMS. This represents the first record of A. cleistominuta in Europe. Of the other fungal associates isolated from A. maiche in Ticino, Fusarium lateritium is of note as there is a possibility that A. maiche could act as a vector of this plant pathogen. We highlight several research needs that should be addressed to gain insight into the potential impact of these non-native species and to overcome problems with heteroplasmy in COI sequences in studies of invasion and population genetics of ambrosia beetles.

Known from trees and the tropics: new insights into the Fusarium lateritium species complex

Known from trees and the tropics: new insights into the Fusarium lateritium species complex

Antifungal activities of leaf extracts from the Capsicum genus

The emergence of microorganisms resistant to antimicrobials has driven the search for new substances aimed at the control and prevention of pathogens. Plant antimicrobial peptides have biotechnological applications in the development of drugs and agricultural inputs for this purpose. The present work aims to evaluate the antifungal activity of extracts containing peptides obtained from the leaves of Capsicum plants. The accessions UENF 1626, 1632 and 1381 and the cultivar Carioquinha were used. The leaves were macerated, and then extraction was performed using a 60% methanol and dichloromethane solution in a 1:1 ratio. The hydromethanolic phase was separated and used for testing after evaporation of methanol. All extracts have a similar electrophoretic profile on tricine gel stained with Coomassie Blue, showing 2 bands of approximately 6.5 and 3.5 kDa. Inhibition of fungal growth with 24 h of culture was performed in liquid culture medium in 96-well plates with reading at 620 nm using extracts with a protein concentration of 200 mg mL-1. The filamentous fungi Fusarium solani, Fusarium lateritium, and Colletotrichum lindemuthianum and the yeasts Candida albicans, Candida buinensis, Candida tropicalis, Saccharomyces cerevisiae, Pichia membranifaciens and Kluyveromyces marxianus were used. The extract with the greatest antifungal potential was the leaves of Capsicum annuum cv. Carioquinha, inhibiting the growth of F. solani by 29%, C. lindemuthianum by 15%, Candida buinensis by 22% and K. marxianus by 24%. Therefore, the extract has peptides with biotechnological potential.

First report of rot disease in Prunus salicina var. taoxingli fruit caused by Fusarium lateritium in China.

Shengzhou nane (Prunus salicina var. taoxingli) as main cultivated plum in Zhejiang province, China, ripened in summer and was susceptible to microbial infection during the maturation stages because of high ambient temperature. In July 2022, Shengzhou nane fruits were found with symptoms of dark skin and grey-white lesions on surface, and an average incidence of approximate 16% (data obtained from investigation of ten trees in each orchard, and fifty fruits in each tree) in four orchards in Shengzhou city, Zhejiang province, China. To isolate the pathogen, four symptomatic partially rotted fruits (about 25%) from four orchards were collected. The plum tissue from the margin of lesions was excised into small blocks (4×4×3 mm). Eight blocks were disinfested with 70% ethanol for 30 seconds and 2% NaClO for 3 minutes, rinsed at least twice with distilled water, and incubated on PDA medium supplemented with 30.0 μg/mL Chloramphenicol at 25°C for 5 days in darkness until sporulation. Using single-spore isolation, four uniform isolates were obtained. The purified mycelium of isolates on PDA were milky white, the reverse on agar was dark pink. Colonies grew with a mean mycelium growth rate of 4.62 ± 0.38 mm per day on PDA. Macroconidia was shaped like a sickle and the size was 27.27 ± 3.43×4.54 ± 0.39 μm (n=50) with 2 to 3 septate. No microconidia and chlamydospores were observed. These morphological characteristics were consistent with previous descriptions of Fusarium lateritium (F. lateritium) species (Yun et al. 2013). Molecular identification was performed by sequencing nuclear ribosomal internal transcribed spacer region (ITS) gene with ITS1/ITS4 (White et al. 1990) and translation elongation factor-1α (TEF) with EF1-728F/EF1-986R (Carbone and Kohn. 1999). The representative sequences were submitted to Genbank (Accession Nos. OP315081 for the ITS and OP394153 for the TEF-1α). Blastn result indicated that ITS and TEF-1a sequences shared 99.28% and 100% similarity with two strains of F. lateritium (GenBank Accession Nos. MK311296.1 and JF740854.1). To fulfill Koch's postulates, the isolated F. lateritium (TXL-IT5) with a conidial suspension (1×106 spores per mL) was inoculated on ten fresh harvested healthy mature Shengzhou nane fruits (20 μL per fruits) after surface sterilization. The control fruits were inoculated with 20 μL distilled water. The experiment was repeated three times.Fruits were kept in a sealed plastic boxes at 25 °C with 90% relative humidity in dark. Four days after inoculation, all inoculated fruits were infected and appeared the same symptoms which were found in naturally infected fruits while those in the control group remained symptomless. The pathogen was re-isolated from symptomatic rotten fruits using above-mentioned method and was identified asF. lateritiumbased on morphological characteristics as well asITS and TEFgene sequences. In addition, this pathogen was reported to infect Dalbergia tonkinensisin north Vietnam (Nhung et al. 2018), causing shoot dieback of Acer negundo in northeast Poland (Patejuk et al. 2022), and rot of Chinese cherry during postharvest storage (Wang et al. 2020). However, to the best of our knowledge, this is the first report of F. lateritium causing rot on Shenshou nane fruits. Our findings serve as a warning for scientists and growers who should pay attention to this disease and take effective control strategies to reduce epidemics of this disease in China and other regions.

Fusarium endophyticum sp. nov. (Nectriaceae, Hypocreales), a new endophytic fungus from northern Thailand

A new endophytic ascomycete, described herein as Fusarium endophyticum, was isolated from leaves of Camellia sinensis var. assamica collected from Nan Province, northern Thailand. This species is described using morphological characteristics and molecular multi-gene analyses. Multi-gene phylogenetic analyses of a combination of internal transcribed spacer region of the rDNA (ITS), translation-elongation factor 1 alpha (tef-1), RNA polymerase II second largest subunit (rpb2), and RNA polymerase II largest subunit (rpb1) genes revealed that F. endophyticum belonged to the Fusarium lateritium species complex (FLSC) and was distinct from all other known species. A full description, illustrations and a phylogenetic tree showing the position of F. endophyticum are provided.

A Novel Effector FlSp1 Inhibits the Colonization of Endophytic Fusarium lateritium and Increases the Resistance to Ralstonia solanacearum in Tobacco.

Effectors are crucial for the interaction between endophytes and their host plants. However, limited attention has been paid to endophyte effectors, with only a few reports published. This work focuses on an effector of Fusarium lateritium, namely FlSp1 (Fusarium-lateritium-Secreted-Protein), a typical unknown secreted protein. The transcription of FlSp1 was up-regulated after 48 h following fungal inoculation in the host plant, i.e., tobacco. The inactivation of FlSp1 with the inhibition rate decreasing by 18% (p < 0.01) resulted in a remarkable increase in the tolerance of F. lateritium to oxidative stress. The transient expression of FlSp1 stimulated the accumulation of reactive oxygen species (ROS) without causing plant necrosis. In comparison with the wild type of F. lateritium (WT), the FlSp1 mutant of the F. lateritium plant (ΔFlSp1) reduced the ROS accumulation and weakened the plant immune response, which resulted in significantly higher colonization in the host plants. Meanwhile, the resistance of the ΔFlSp1 plant to the pathogenic Ralstonia solanacearum, which causes bacterial wilt, was increased. These results suggest that the novel secreted protein FlSp1 might act as an immune-triggering effector to limit fungal proliferation by stimulating the plant immune system through ROS accumulation and thus balance the interaction between the endophytic fungi and their host plants.

Diversity and Pathogenicity of Fungi Associated with Fruit Rot of Winter Jujube in Shandong Province, China.

Winter jujube originated from China and had an extremely high nutritional value. In 2021, symptomatic winter jujube fruits were collected from eight locations in Zhanhua District of Binzhou City, Shandong Province. In total, 108 fungal isolates were obtained and grouped into 11 species based on morphological characteristics and multilocus phylogenetic analysis, including Nothophoma quercina (43.52%), Fusarium lateritium (20.37%), Alternaria alternata (12.03%), F. proliferatum (7.41%), F. graminearum (4.63%), Botryosphaeria dothidea (3.70%), Fusarium sp. (2.78%), A. tenuissima (2.78%), Diaporthe eres (1.85%), Nigrospora oryzae (0.93%), and Cercospora nicotianae (0.93%). All fungal isolates obtained in this study showed aggressiveness on detached winter jujube fruits except N. oryzae and C. nicotianae isolates, of which F. proliferatum was the most virulent, while A. alternata isolates, which have been considered the major pathogen of winter jujube fruit rot, showed a relatively low-level virulence in this study. Furthermore, D. eres, F. graminearum, F. lateritium, and an unclassified Fusarium species were first reported as causal agents of winter jujube fruit rot. The typical symptoms of winter jujube fruit rot observed in this study could be distinguished into two types. N. quercina, A. alternata, A. tenuissima, Fusarium sp., D. nobilis, and F. lateritium isolates caused reddish brown to dark gray lesions on the peel, while B. dothidea, F. graminearum, and F. proliferatum isolates caused peel and pulp decay, resulting in red to reddish brown and water-soaked lesions. In addition, haplotype analysis of N. quercina isolates obtained in this study and validly published articles showed that there were 11 haplotypes worldwide; the isolates obtained in the current study were grouped into three haplotypes (Hap 1, Hap 2, and Hap 11), and two of them (Hap 2 and Hap 11) were confirmed as new haplotypes.

Root and rhizosphere fungi associated with the yield of diverse Brassica napus genotypes

The plant microbiome is a key driver of plant health and agroecosystem functioning. This study used internal transcribed spacer (ITS) amplicon sequencing to examine the dynamics of root-associated fungal microbiomes in genetically-diverse Brassica napus genotypes following highly resolved sampling across growth stages at multiple sites in Saskatchewan, Canada. Across all samples including individual B. napus genotypes, sampling times and sites (n = 936 for both roots and rhizosphere), genotype was associated with only 2% of variance in fungal community structure, which was mostly driven by site-year (67% of variance for root and 79% for rhizosphere). Within a single site, this influence increased to between 4 and 16% of root and 5–13% of rhizosphere community variance, indicating genotype-specific adaptation to site conditions across the growing season. A single amplicon sequence variant (ASV) of Olpidium brassicae accounted for over 88% of the root fungal ASVs at two of the three sites. Despite O. brassicae dominance, a diverse fungal core microbiome consisting of 38 ASVs was detected in all genotypes of each site-year. We also identified 13 ASVs that were highly associated with B. napus yield; common between the rhizosphere and root, Gibberella baccata was negatively correlated with yield while Nectria ramulariae and a Tetracladium sp. were positively correlated with yield. The presence of a diverse core microbiome, dominated by O. brassicae, and identification of taxa associated with B. napus yield performance across a variety of environments reveal strategic targets for better understanding the role of the root microbiome in B. napus production.

Apple Endophytic fungi and their antagonism against apple scab disease.

Endophytic fungi are microorganisms with the ability to colonize plants for the entire or at least a significant part of their life cycle asymptomatically, establishing a plant-fungus association. They play an important role in balancing ecosystems, as well as benefiting host through increasing plant growth, and protecting the host plants from abiotic and biotic stresses using various strategies. In the present study, endophytic fungi were isolated from wild and endemic apple cultivars, followed by characterizing their antifungal effect against Venturia inaequalis. To characterize the endophytic fungi, 417 fungal strains were separated from 210 healthy fruit, leaf, and branch samples collected from the north of Iran. Among the purified fungal isolates, 33 fungal genera were identified based on the morphological characteristics, of which 38 species were detected according to the morphological features and molecular data of ITS, tef-1α, and gapdh genomic regions (related to the genus). The results represented that most of the endophytic fungi belonged to Ascomycota (67.8%), 31.4% of isolates were mycelia sterilia, while the others were Basidiomycota (0.48%) and Mucoromycota (0.24%). Additionally, Alternaria, Cladosporium, and Nigrospora were determined as the dominant genera. The antifungal properties of the identified isolates were evaluated against V. inaequalis in vitro to determine the release of media-permeable metabolites, Volatile Organic Compounds (VOCs), chitinase, and cellulase as antifungal mechanisms, as well as producing phosphate solubilisation as growth-promoting effect. Based on the results of metabolite and VOC tests, the six isolates of Acremonium sclerotigenum GO13S1, Coniochaeta endophytica 55S2, Fusarium lateritium 61S2, Aureobasidium microstictum 7F2, Chaetomium globosum 2S1 and Ch. globosum 3 L2 were selected for greenhouse tests. Further, Co. endophytica 55S2 and F. lateritium 61S2 could solubilize inorganic phosphate. All isolates except Ch. globosum 3 L2 exhibited cellulase activity, while chitinase activity was observed in Ch. globosum 2S1, Ch. globosum 3 L2, and F. lateritium 61S2. Finally, Co. endophytica 55S2 and Ch. globosum 2S1 completely controlled the disease on the apple seedling leaves under greenhouse conditions.

Molecular characterization of endophytic fungi from pine (Pinus oocarpa) in Honduras

Pine is one of the important coniferous genera in America. In Honduras, Pinus oocarpa is the most critical conifer species since it plays an essential role for the country's economy and represents a symbol of national identity. Endophytic fungi are defined as microorganisms that reside in the internal tissues of plants without causing any obvious immediate negative effects and are an integral part of their associated microbial communities. This study aimed to isolate and characterize the endophytic fungi present in the pines of Honduras through the amplification and sequencing of the ribosomal RNA gene's internal transcribed spacer (ITS). A total of 7 pines from the department of El Paraíso in eastern Honduras were analyzed. A total of 14 fungi grouped into 6 genera and 7 species were isolated: Fusarium lateritium, F. pseudocircinatum, Pestalotiopsis pini, P. microspora, Xylaria grammica, Trichoderma atroviride, y Nigrospora oryzae. To our knowledge, this is the first report of endophytic fungal species in pines in Honduras. Although some endophytic fungi may be mutualistic or saprophytic, the present study shows the presence of several genera of endophytic fungi that have been reported as pine pathogens. The presence of these fungi in the pines of Honduras represents a potential threat to the health of the forest. Further research is needed to increase knowledge about the importance of these fungi and the potential impact they could have on pine forests in Honduras. Keywords: Endophytic fungi, Pinus oocarpa, Honduras

Fusarium citri-sinensis sp. nov. (Ascomycota: Nectriaceae) isolated from fruit of Citrus sinensis in China

During an investigation of fungal pathogens associated with Citrus sinensis in Zigui county of Yichang city, China, a new species, Fusarium citri-sinensis sp. nov. was isolated from fruit of C. sinensis. The species is described using morphology and molecular multi-locus analyses based on a combined data of internal transcribed spacer region of the rDNA (ITS), translation-elongation factor 1 alpha (EF-1α), and RNA polymerase second largest subunit (RPB2) genes. It is one of the species in Fusarium lateritium species complex (FLSC).

Culturable Endophytic Fungi in Fraxinus excelsior and Their Interactions with Hymenoscyphus fraxineus

The species diversity of culturable endophytic fungi was studied in the leaves and twigs of symptomatic and asymptomatic Fraxinus excelsior trees. Endophytic mycobiota was dominated by Ascomycota species, with Pleosporales (44.17%) and Diaporthales (23.79%) endophytes being the most frequently observed in the tree samples. The number of endophytic isolates and species richness varied depending on the sampling date (May and October) and tissue location. Of the 54 species identified based on ITS sequences, 14 were classified as dominant. The most frequently isolated species were Diaporthe eres, followed by Alternaria alternata, Dothiorella gregaria, and Fraxinicola fraxini. The inhibitory effect of 41 species (75 isolates) of endophytes on the radial growth of a Hymenoscyphus fraxineus isolate was studied under in vitro conditions (dual cultures). The radial growth of H. fraxineus was the most inhibited by four endophytic fungi from twigs (Fusarium lateritium, Didymella aliena, Didymella macrostoma, and Dothiorella gregaria). The inhibitory effect of the four isolates was also studied under in planta conditions. The isolates artificially inoculated into the trunks of ash trees reduced the length of necroses formed by H. fraxineus co-inoculated in the same trunks. This effect depended on the isolate, and the inhibition was most prominent only on trunks inoculated with F. lateritium and D. aliena. Although the total length of necrotic lesions formed by the H. fraxineus infection was shorter in the ash trunks co-inoculated with the endophytes, the difference was not significant.

Acuminatums E and F, two new cyclic lipopeptides from Fusarium lateritium HU0053 and their antifungal activity

Two new cyclic lipopeptides, acuminatums E (1) and F (2), together with four known cyclic lipopeptides, acuminatums A–D (3–6) were isolated from the corn culture of endophytic Fusarium lateritium HU0053. Their structures were elucidated by spectroscopic and advanced Marfey's amino acid analysis. All compounds were found to exhibit antifungal activities against Penicillium digitatum. Acuminatum F (2), a new cyclic lipopeptide containing an unusual 3, 4-dihydroxy-phenylalanine unit exhibited the strongest antifungal activities with inhibition zone of 6.5 mm at the dose of 6.25 μg. Therefore, acuminatum F might be a potential environmental-friendly preservative for citrus fruits.

Fungal endophytes in Fraxinus excelsior petioles and their in vitro antagonistic potential against the ash dieback pathogen Hymenoscyphus fraxineus.

Fungal endophytes were isolated from 250 asymptomatic leaf petioles of Fraxinus excelsior collected from trees showing symptoms of ash dieback in five forest sites in southern Poland. Fungal isolations yielded 1646 colonies representing 97 taxa, including 92 Ascomycota and 5 Basidiomycota species. The most common Ascomycota comprised Nemania serpens (38.0 % of colonized petioles), Diaporthe eres (33.6 %), Venturia fraxini (26.4 %), Diaporthe sp. 1 (20.4 %), Alternaria sp. 1 (14.8 %), Colletotrichum acutatum (14.8 %), Nemania diffusa (14.0 %), Colletotrichum gloeosporioides (12.4 %) and Colletotrichum sp. (12.4 %). The occurrence of all these taxa except Alternaria sp. 1 was significantly different between the studied forest sites. Two yeast species, Vishniacozyma foliicola (4.8 %) and Cystobasidium pinicola (2.8 %), dominated among the Basidiomycota endophytes detected. All the fungal endophytes were tested in dual culture antagonistic assays against two strains of Hymenoscyphus fraxineus, resulting in the development of four interaction types. The interactions included the physical contact of co-partners' mycelia (41.8 %), development of an inhibition zone (47.4 %), growth of endophyte mycelia over H. fraxineus colonies (9.3 %) and growth of H. fraxineus mycelia over endophyte colonies (1.5 %). The strongest antibiotic activity against H. fraxineus, measured by the width of the inhibition zone, was observed for Cytospora pruinosa, Fusarium lateritium, Phoma sp. 2, Pleosporales sp. 2 and Thielavia basicola. A variety of morphophysiological deformations of H. fraxineus hyphae were observed under endophyte pressure: spiral twist of the hyphae, formation of cytoplasmic extrusions, development of torulose hyphae and excessive lateral branching of the hyphae. The strongest antagonistic effects, coupled with the potential to overgrow H. fraxineus colonies, was shown by Clonostachys rosea, Nemania diffusa, N. serpens, Peniophora cinerea, Rosellinia corticium and Xylaria polymorpha. Some of these species were able to attack H. fraxineus hyphae in a mycoparasitic manner. The antagonistic activities included the physical penetration of H. fraxineus hyphae, dissolution of hyphal cell walls, disappearance of pigmentation, disintegration of hyphae and degradation of other fungal structures. In contrast, one of the most commonly detected endophytes in ash leaves, Venturia fraxini, did not show in vitro antagonistic potential against H. fraxineus. Finally, we discuss the potential of the detected fungal endophytes to combat H. fraxineus invasion, the cause of ash decline in Europe.

Exploring Fungal Species Diversity in the Premature Yeast Flocculation (PYF) of Barley Malt Using Deep Sequencing

Premature yeast flocculation (PYF) is the early flocculation of yeast during fermentation, which causes logistical and financial hardship to the malting and brewing industries. The occurring of PYF is suspected to be caused by the fungal infection of the barley or at least in part. The aim of this study was to perform a non-culture approach to uncover the fungal diversity in PYF malt. The genomic DNA was extracted and analyzed by amplicon sequencing. A total pairs read number of 209,523 sequences was obtained and used for taxonomic classification. The dominant fungal communities on the PYF malt were obtained based on the microbial diversity analysis by species annotation. The internal transcribed spacers sequencing analysis showed a decreased number of fungal species in PYF malt. Results showed that compared with control malt, PYF malt displayed a higher abundance of Ascomycota at the phyla level, and the abundance of Candida and Gibberella was exceptionally high in two PYF malts at the genus level. The absolute abundance of Candida and Gibberella baccata at the species level was also higher in PYF malts. The fungal species diversity within the malt indicated that the fungi community diversity was greater in the control malt than in the PYF malt. Taken together, exploring the diversity of the fungal community could be useful for malt quality control and for eliminating contamination risks.

Draft Genome Sequence of a New Fusarium Isolate Belonging to Fusarium tricinctum Species Complex Collected From Hazelnut in Central Italy.

In summer 2019, during a survey on the health status of a hazelnut orchard located in the Tuscia area (the province of Viterbo, Latium, Italy), nuts showing symptoms, such as brown-grayish spots at the bottom of the nuts progressing upward to the apex, and necrotic patches on the bracts and, sometimes, on the petioles, were found and collected for further studies. This syndrome is associated with the nut gray necrosis (NGN), whose main causal agent is Fusarium lateritium. Aiming to increase knowledge about this fungal pathogen, the whole-genome sequencing of a strain isolated from symptomatic hazelnut was performed using long Nanopore reads technology in combination with the higher precision of the Illumina reads, generating a high-quality genome assembly. The following phylogenetic and comparative genomics analysis suggested that this isolate is caused by the F. tricinctum species complex rather than F. lateritium one, as initially hypothesized. Thus, this study demonstrates that different Fusarium species can infect Corylus avellana producing the same symptomatology. In addition, it sheds light onto the genetic features of the pathogen in subject, clarifying facets about its biology, epidemiology, infection mechanisms, and host spectrum, with the future objective to develop specific and efficient control strategies.

Characterization of Fungi Associated with Olive Fruit Rot and Olive Oil Degradation in Crete, Southern Greece.

In November 2019, a severe outbreak of fruit rot was observed in olive orchards in Crete, southern Greece. Symptoms appeared primarily on fruits and stalks, resembling those caused by anthracnose. Typical symptoms were fruit rot, shrinkage, and mummification, associated commonly with stalk discoloration and fruit drop. Disease incidence was estimated at up to 100% in some cases, and an unprecedented increase in olive oil acidity reaching up to 8% (percentage of oleic acid) in severely affected olive groves was recorded. Thirty-two olive groves were then surveyed, and samples of fruit, stalk, leaf, and shoot were collected. Visual, stereoscopic, and microscopic observations revealed several fungi belonging to the genera Alternaria, Botryosphaeria, Capnodium, Colletotrichum, Fusarium, and Pseudocercospora. Fungal infection in fruits was commonly associated with concomitant infestation by the olive fruit fly Bactrocera oleae along with increased air temperature and relative humidity conditions that prevailed in October and November 2019. Twenty representative fungal strains isolated from symptomatic fruits and stalks were characterized by morphological, physiological, and molecular analyses. By internal transcribed spacer regions of ribosomal DNA region and translation elongation factor 1-α gene sequencing analysis, these isolates were identified as Alternaria spp., A. infectoria, Botryosphaeria dothidea, Colletotrichum boninense sensu lato, Fusarium lateritium, F. solani species complex and Stemphylium amaranthi. Pathogenicity tests on punctured fruits revealed that all isolates were pathogenic; however, F. solani isolates along with B. dothidea were the most virulent, and wounds were necessary for efficient fungal infection. Moreover, as few as 10 spores of F. solani were sufficient to cause significant infection in punctured fruits. F. solani was also capable of infecting olive fruits in the presence of B. oleae, with no additional wounding, in artificial inoculation experiments. Moreover, it was capable of colonizing and affecting olive blossoms. Further analyses of olive oil extracted from fruits artificially inoculated with F. solani indicated a significant increase in oil acidity, K232, K270, and peroxide value, whereas total phenol content was significantly decreased. To the best of our knowledge, this is the first report of F. solani associated with olive fruit rot and olive oil degradation worldwide.

Inhibition of Serine Protease, α-Amylase and Growth of Phytopathogenic Fungi by Antimicrobial Peptides from Capsicum chinense Fruits.

Plant fungal diseases cause major problems for the global economy. Antimicrobial peptides have aroused great interest in the control of phytopathogens, as they are natural molecules and have a broad spectrum of inhibitory activity. Herein, we have tried to identify and characterize antimicrobial peptides present in fruits of Capsicum chinense and to evaluate their enzymatic and antifungal activities. The retained fraction obtained in the anion exchange chromatography with strong antifungal activity was subjected to molecular exclusion chromatography and obtained four fractions named G1, G2, G3, and G4. The 6.0-kDa protein band of G2 showed similarity with protease inhibitors type II, and it was able to inhibit 100% of trypsin and α-amylase activities. The protein band with approximately 6.5kDa of G3 showed similarity with sequences of protease inhibitors from genus Capsicum and showed growth inhibition of 48% for Colletotrichum lindemuthianum, 49% for Fusarium lateritium, and 51% for F. solani and F. oxysporum. Additionally, G3 causes morphological changes, membrane permeabilization, and ROS increase in F. oxysporum cells. The 9-kDa protein band of G4 fraction was similar to a nsLTP type 1, and a protein band of 6.5kDa was similar to a nsLTP type 2. The G4 fraction was able to inhibit 100% of the activities of glycosidases tested and showed growth inhibition of 35 and 50% of F. oxysporum and C. lindemuthianum, respectively. C. chinense fruits have peptides with antifungal activity and enzyme inhibition with biotechnological potential.