Endophytic Species Research Articles

Fatty acid metabolites of Dendrobium nobile were positively correlated with representative endophytic fungi at altitude.

Altitude, as a comprehensive ecological factor, regulates the growth and development of plants and microbial distribution. Dendrobium nobile (D. nobile) planted in habitats at different elevations in Chishui city, also shows metabolic differences and endophytes diversity. What is the triangular relationship between altitude, endophytes, and metabolites? In this study, the diversity and species of endophytic fungi were tested by ITS sequencing and metabolic differences in plants were tested by UPLC-ESI-MS/MS. Elevation regulated the colonization of plant endophytic fungal species and fatty acid metabolites in D. nobile. The results indicate that and high altitude was better for the accumulation of fatty acid metabolites. Therefore, the high-altitude characteristic endophytic floras were screened, and the correlation with fatty acid metabolites of plants was built. The colonization of T. rubrigenum, P. Incertae sedis unclassified, Phoma. cf. nebulosa JZG 2008 and Basidiomycota unclassified showed a significantly positive correlation with fatty acid metabolites, especially 18-carbon-chain fatty acids, such as (6Z,9Z,12Z)-octadeca-6,9,12-trienoic acid, 3,7,11,15-tetramethyl-12-oxohexadeca-2,4-dienoic acid and Octadec-9-en-12-ynoic acid. What is more fascinating is these fatty acids are the essential substrates of plant hormones. Consequently, it was speculated that the D. nobile- colonizing endophytic fungi stimulated or upregulated the synthesis of fatty acid metabolites and even some plant hormones, thus affecting the metabolism and development of D. nobile.

Cryptostroma corticale and fungal endophytes associated with Fraxinus excelsior affected by ash dieback

AbstractWhile assessing the filamentous fungi associated with woody tissues of stem collar rots, necroses, and lesions of European ash trees (Fraxinus excelsior) presenting symptoms of ash dieback in Germany, Cryptostroma corticale was recovered from three different ash trees. These isolated strains were the first report of C. corticale on ash and the first proof of an association of this plant pathogen with woody tissues of other tree species than Acer spp. in Germany. To test the pathogenicity of C. corticale against F. excelsior and to fulfil Koch’s postulates, inoculation tests in planta with strains isolated from Acer pseudoplatanus and F. excelsior were conducted according to Henle–Koch’s postulates in a greenhouse located in Göttingen. The pathogenicity tests were performed with apparently healthy ash saplings from June 2021 until January 2022. After three and seven months, neither necroses or lesions due to C. corticale nor disease symptoms were observed. Mostly, the inoculation wounds healed over, and C. corticale could not be re-isolated from the ash woody tissue. In an attempt to re-isolate the inoculated strains, the filamentous fungal endophytes of the ash woody tissues were isolated and identified. A total of 32 taxa of the Ascomycota were found, where the most common species was Boeremia exigua. Most frequently observed orders were Pleosporales (58.4%), followed by Sordariales (13.5%), Hypocreales (9.4%), and Diaporthales (8.7%). On average, 3.7 endophytic species were recorded on each sapling.

Endophytic Trichoderma species from rubber trees native to the Brazilian Amazon, including four new species.

Fungi belonging to the genus Trichoderma have been widely recognized as efficient controllers of plant diseases. Although the majority of isolates currently deployed, thus far, have been isolated from soil, endophytic Trichoderma spp. is considered to be a promising option for application in biocontrol. In this study, 30 endophytic Trichoderma isolates-obtained from the leaves, stems, and roots of wild Hevea spp. in the Brazilian Amazon-were analyzed using specific DNA barcodes: sequences of internal transcribed spacers 1 and 2 of rDNA (ITS region), genes encoding translation elongation factor 1-α (TEF1-α), and the second largest subunit of RNA polymerase II (RPB2). The genealogical concordance phylogenetic species recognition (GCPSR) concept was used for species delimitation. A phylogenetic analysis showed the occurrence of Trichoderma species, such as T. erinaceum, T. ovalisporum, T. koningiopsis, T. sparsum, T. lentiforme, T. virens, and T. spirale. Molecular and morphological features resulted in the discovery of four new species, such as T. acreanum sp. nov., T. ararianum sp. nov., T. heveae sp. nov., and T. brasiliensis sp. nov. The BI and ML analyses shared a similar topology, providing high support to the final trees. The phylograms show three distinct subclades, namely, T. acreanum and T. ararianum being paraphyletic with T. koningiopsis; T. heveae with T. subviride; and T. brasiliensis with T. brevicompactum. This study adds to our knowledge of the diversity of endophytic Trichoderma species in Neotropical forests and reveals new potential biocontrol agents for the management of plant diseases.

Antibacterial and Antioxidant Activity of Endophytic Fungi Extract Isolated from Leaves of Sungkai (Peronema canescens)

Sungkai is a plant that is widely found in Indonesia. This plant is often used in traditional medicine so the biotechnology of endophytic fungi is highly needed due to natural resources from plants that have been reduced. This study reported on endophytic fungi found in sungkai leaves and the compound produced. Morphological and molecular identification through phylogenetic tree analysis was carried out to determine the endophytic fungal species found. Potato Dextrose Broth media was used for the cultivation process. Ethyl acetate was used as a solvent for the extraction and the evaporation process used a rotary evaporator. Antioxidant and antibacterial tests were carried out using the DPPH method and paper disc diffusion. Chromatographic techniques were used to isolate the compound and spectroscopic analysis was performed to identify its chemical structure. The results of the morphological and molecular analysis showed Trichoderma asperellum as an endophytic fungus identified. The pure compound obtained from this endophytic fungus was 4-hydroxybenzoic acid. The antioxidant and antibacterial activity showed a strong category (IC50 = 43.88 ????g/mL; MIC 64 ????g/mL). This compound was very likely to be a raw material for new antibiotics and antioxidants through further research with various modifications.

Ginger rhizome priming with lipopeptide-producing endophytic Bacillus species to control ginger soft-rot disease caused by Pythium myriotylum

Ginger rhizome priming with lipopeptide-producing endophytic Bacillus species to control ginger soft-rot disease caused by Pythium myriotylum

In vitro efficacy of fungal endophytes and silver pyrazolate against Raffaelea lauricola, causal agent of laurel wilt of avocado

The South Florida avocado industry is being severely impacted by laurel wilt disease. Laurel wilt disease of avocado is caused by the fungal pathogen, Raffaelea lauricola (RL) and is vectored by ambrosia beetle, Xyleborus glabratus. Treatments options are limited, economically not sustainable, and require reapplication fungicides every couple of years. There is a crucial need for developing multiple modes of control using novel biological and chemical agents. The ambrosia beetle associated pathogenic fungi are known to outcompete other microorganisms by taking advantage of ethanol produced by the pathogen and the stressed tree. Endophytes, which reside inside the host plant tissue are part of the plant microbiome represent source of new potential biological control agents. In this study, three ethanol tolerant endophytic fungal species, isolated from avocado bark, were evaluated using in vitro dual culture assay and colonization tube (packed with bark/sapwood shaving) against RL. The endophytic isolates Tricoderma crissum, Tricoderma simmonsii, Lasiodiplodia theobromae were found to be highly capable of suppressing the mycelial colony growth of RL. The results suggest that combined abilities of ethanol tolerance and competitive colonization can provide useful criteria for identifying potential biocontrol agents. In vitro anti-RL activity of silver pyrazolate compound was assessed in both agar and liquid medium. Silver pyrazolate at levels of 30 and 45 ppm were found to be highly effective against RL. Further in planta research is needed to study the effects of endophytic fungal isolates and silver pyrazolate to assess their potential as additional tools for management of laurel wilt.

Endophytic Colletotrichum (Sordariomycetes, Glomerellaceae) species associated with Citrusgrandis cv. "Tomentosa" in China.

Colletotrichum species are well-known plant pathogens, saprobes, endophytes, human pathogens and entomopathogens. However, little is known about Colletotrichum as endophytes of plants and cultivars including Citrusgrandis cv. "Tomentosa". In the present study, 12 endophytic Colletotrichum isolates were obtained from this host in Huazhou, Guangdong Province (China) in 2019. Based on morphology and combined multigene phylogeny [nuclear ribosomal internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (gapdh), chitin synthase 1 (chs-1), histone H3 (his3) actin (act), beta-tubulin (β-tubulin) and glutamine synthetase (gs)], six Colletotrichum species were identified, including two new species, namely Colletotrichumguangdongense and C.tomentosae. Colletotrichumasianum, C.plurivorum, C.siamense and C.tainanense are identified as being the first reports on C.grandis cv. "Tomentosa" worldwide. This study is the first comprehensive study on endophytic Colletotrichum species on C.grandis cv. "Tomentosa" in China.

Production and Characterization of New Biosurfactants/Bioemulsifiers from Pantoea alhagi and Their Antioxidant, Antimicrobial and Anti-Biofilm Potentiality Evaluations.

The present work aimed to develop rapid approach monitoring using a simple selective method based on a positive hemolysis test, oil spreading activity and emulsification index determinations. It is the first to describe production of biosurfactants (BS) by the endophytic Pantoea alhagi species. Results indicated that the new BS evidenced an E24 emulsification index of 82%. Fourier-transform infrared (FTIR) results mentioned that the described BS belong to the glycolipid family. Fatty acid profiles showed the predominance of methyl 2-hyroxydodecanoate in the cell membrane (67.00%) and methyl 14-methylhexadecanoate (12.05%). The major fatty acid in the BS was oleic acid (76.26%), followed by methyl 12-methyltetradecanoate (10.93%). Markedly, the BS produced by the Pantoea alhagi species exhibited antimicrobial and anti-biofilm activities against tested human pathogens. With superior antibacterial activity against Escherchia coli and Staphylococcus aureus, a high antifungal effect was given against Fusarium sp. with a diameter of zone of inhibition of 29.5 mm, 36 mm and 31 mm, obtained by BS dissolved in methanol extract. The DPPH assay indicated that the BS (2 mg/mL) showed a higher antioxidant activity (78.07 inhibition percentage). The new BS exhibited specific characteristics, encouraging their use in various industrial applications.

Biodegradation of Aflatoxin B1 in Maize Grains and Suppression of Its Biosynthesis-Related Genes Using Endophytic Trichoderma harzianum AYM3

Aflatoxin B1 is one of the most deleterious types of mycotoxins. The application of an endophytic fungus for biodegradation or biosuppression of AFB1 production by Aspergillus flavus was investigated. About 10 endophytic fungal species, isolated from healthy maize plants, were screened for their in vitro AFs-degrading activity using coumarin medium. The highest degradation potential was recorded for Trichoderma sp. (76.8%). This endophyte was identified using the rDNA-ITS sequence as Trichoderma harzianum AYM3 and assigned an accession no. of ON203053. It caused a 65% inhibition in the growth of A. flavus AYM2 in vitro. HPLC analysis revealed that T. harzianum AYM3 had a biodegradation potential against AFB1. Co-culturing of T. harazianum AYM3 and A. flavus AYM2 on maize grains led to a significant suppression (67%) in AFB1 production. GC-MS analysis identified two AFB1-suppressing compounds, acetic acid and n-propyl acetate. Investigating effect on the transcriptional expression of five AFB1 biosynthesis-related genes in A. flavus AYM2 revealed the downregulating effects of T. harzianum AYM3 metabolites on expression of aflP and aflS genes. Using HepaRG cell line, the cytotoxicity assay indicated that T. harazianum AYM3 metabolites were safe. Based on these results, it can be concluded that T. harzianum AYM3 may be used to suppress AFB1 production in maize grains.

Molecular characterization of fungal endophyte diversity isolated from Aconitum heterophyllum: a critically endangered medicinal plant of Kashmir Himalaya.

Aconitum heterophyllum is a rare perennial herb from Kashmir Himalayas. Due to its threatened status and dependence on its environment, the plant was examined for any potential endophytes, which is of utmost importance for bioprospection. In the current study, endophytic fungal diversity associated with A. heterophyllum was examined, and 328 fungal isolates were found in the plant's leaf, stem, and root tissues. Twelve (12) endophytic fungal species were identified utilizing, molecular analysis of the nuclear ribosomal DNA Internal Transcribes Spacer (ITS), rLSU, and rSSU sequences. Maximum likelihood analysis was used to determine the phylogenetic connection between each isolate. The genera Arthrinium, Chaetomium, Purpureocillium, Alternaria, Penicillium, Aspergillus, Cladosporium, and Bjerkandera species dominated the ascomycete and basidiomycete fungal endophytes.

Growth-Promoting Characteristics of Fungal and Bacterial Endophytes Isolated from a Drought-Tolerant Mint Species Endostemon obtusifolius (E. Mey. ex Benth.) N. E. Br.

Endophytes are primarily endosymbiotic bacteria and fungi that colonize the interior tissues of their host plant. They enhance the host plant's growth and attenuate adverse effects of biological stress. Endophytic species of many indigenous plants are an untapped resource of plant growth-promoting microorganisms that can mitigate abiotic stress effects. Thus, this study aimed to isolate endophytes from the roots and leaves of the medicinal plant Endostemon obtusifolius to evaluate their in vitro growth-promoting capacities and drought tolerance and to characterize the most promising species. Twenty-six endophytes (fourteen bacteria and twelve fungi) were isolated and cultured from the roots and leaves of E. obtusifolius. All 26 endophytes produced flavonoids, and 14 strains produced phenolic compounds. Of the 11 strains that displayed good free radical scavenging capability (low IC50) in the 1-1-diphenyl-1-picryhydrazyl radical scavenging assay, only three strains could not survive the highest drought stress treatment (40% polyethylene glycol). These 11 strains were all positive for ammonia and siderophore production and only one strain failed to produce hydrogen cyanide and solubilize phosphate. Seven isolates showed aminocyclopropane-1-carboxylate deaminase activity and differentially synthesized indole-3-acetic acid. Using molecular tools, two promising symbiotic, drought stress tolerant, and plant growth-enhancing endophytic species (EORB-2 and EOLF-5) were identified as Paenibacillus polymyxa and Fusarium oxysporum. The results of this study demonstrate that P. polymyxa and F. oxysporum should be further investigated for their drought stress mitigation and plant growth enhancement effects as they have the potential to be developed for use in sustainable agricultural practices.

Hydrolytic Enzyme Production and Susceptibility to Antifungal Compounds of Opportunistic Candida parapsilosis Strains Isolated from Cucurbitaceae and Rosaceae Fruits

Endophytic yeast species were studied in the internal tissues of ripe cultivated vegetables and fruits. A total of 19 yeast species, 11 ascomycete species, and 8 basidiomycete species were observed in the internal tissues of all fruits examined. The opportunistic yeast Candida parapsilosis was present in all plants studied. Several virulence factors (production of hydrolytic enzymes and sensitivity to antifungal agents) were examined in all 107 isolates of C. parapsilosis from the internal tissues of fruits. The most virulent isolates were found in vegetables. C. parapsilosis is widespread in nature and is often isolated from a variety of non-human sources. It is frequently involved in invasive infections that seriously affect human health. This species poses a high risk to immunocompromised individuals, such as HIV patients and surgical patients or children whose immune systems are not sufficiently mature. Since virulent isolates of Candida parapsilosis have been found in vegetables and fruits; their raw consumption may not be safe. Finally, we emphasize the importance of ongoing phenotypic and genetic studies of endophytic isolates of Candida parapsilosis and their comparison with clinical isolates.

Endophytic Penicillium species secretes mycophenolic acid that inhibits the growth of phytopathogenic fungi.

The worldwide demand for reduced and restricted use of pesticides in agriculture due to serious environmental effects, health risks and the development of pathogen resistance calls for the discovery of new bioactive compounds. In the medical field, antibiotic-resistant microorganisms have become a major threat to man, increasing mortality. Endophytes are endosymbiotic microorganisms that inhabit plant tissues without causing any visible damage to their host. Many endophytes secrete secondary metabolites with biological activity against a broad range of pathogens, making them potential candidates for novel drugs and alternative pesticides of natural origin. We isolated endophytes from wild plants in Israel, focusing on endophytes that secrete secondary metabolites with biological activity. We isolated 302 different endophytes from 30 different wild plants; 70 of them exhibited biological activity against phytopathogens. One biologically active fungal endophyte from the genus Penicillium, isolated from a squill (Urginea maritima) leaf, was further examined. Chloroform-based extraction of its growth medium was similarly active against phytopathogens. High-performance liquid chromatography separation followed by gas chromatography/mass spectrometry analysis revealed a single compound-mycophenolic acid-as the main contributor to the biological activity of the organic extract.

Effects of the dual inoculation of dark septate endophytes and Trichoderma koningiopsis on blueberry growth and rhizosphere soil microorganisms.

Blueberry is a shallow root plant in which the absorption of nutrients is inefficient, resulting in slow growth under artificial cultivation conditions. Endophytes play an important role in promoting plant growth; however, the effects of Trichoderma spp. and dark septate endophytes (DSEs) on host plant growth and soil microorganisms are still debatable. We isolated two endophytic fungal species, Trichoderma koningiopsis (TK) and a DSE (Amesia nigricolor; AN), from blueberry roots, which can solubilize insoluble phosphorus and produce amylase and cellulase to promote plant growth. We found that under dual inoculation, the colonization rate and colonization intensity of TK were higher than they were under single inoculation with TK, while the colonization rate and colonization intensity of AN were lower under dual inoculation than under single inoculation with AN. The plant nutrients, root activity, available potassium, and parts of soil phosphatase activities were highest under dual inoculation. TK inoculation resulted in the highest diversity and richness in the soil fungi and bacteria, followed by dual inoculation. The abundance of Ascomycota, Acidobacteriae, Firmicutes, and Actinobacteriota increased significantly, resulting in Trichoderma and Vicinamibacteria inoculated with TK, Chaetomium and Alicyclobacillales inoculated with AN, and Hypocreales and Burkholderiaceae with dual inoculation enriched in the soil.

Insights into the core bacterial consortia of root endophytes in two cultivated varieties of rice in West Bengal

Root endophytes are considered to be one of the potent environment-friendly substitutes for chemical fertilizers, as they possess an ability to induce crosstalk inside the hosts for growth promotion, nitrogen fixation, phosphate solubilization and iron sequestration. This study aimed to explore and evaluate the key root endophytic bacterial consortia of two widely cultivated varieties of rice (Oryza sativa L.), cv. ‘Saraswati’ (OS01) and cv. ‘Kunti’ (OS04). Detailed comparative metagenome data were generated for endophytes of OS01 and OS04 and the species richness was calculated. OS01 showed higher endophyte species richness than OS04, with alpha diversity values of 3.10 and 2.40, respectively. Bacillus, Magnetospirillum, Methanocystis, Desulfomicrobium and Pantoea were identified as common endophyte members for both cultivars. Solibacillus, Paenibacillus, Candidatus, and Melospira were unique members of OS01, and Herbaspirillum, Pandoraea, Anabaenopsis for OS04. Considerable occurrence of nitrogen fixing bacteria and methanogenic bacteria in the cultivars confirmed biological nitrogen fixation, which can contribute to plant development. Core homeotic pathways of amino acid biosynthesis and carbon metabolism were also reflected in endophytes from both cultivars, indicating a supportive environment for microorganisms. Sulfur metabolism pathways were likewise predicted to be active in the niche under study, which may be attributed as a response to arsenic stress. Furthermore, the most abundant genera identified may potentially serve as crucial consortium candidates for host plant development and contribute to better yield in a sustainable manner.

Secondary metabolites of Alternaria: A comprehensive review of chemical diversity and pharmacological properties.

Fungi are considered to be one of the wealthiest sources of bio-metabolites that can be employed for yielding novel biomedical agents. Alternaria, including parasitic, saprophytic, and endophytic species, is a kind of dark fungi that can produce a broad array of secondary metabolites (SMs) widely distributed in many ecosystems. These are categorized into polyketides, nitrogen-containing compounds, quinones, terpenes, and others based on the unique structural features of the metabolites. New natural products derived from Alternaria exhibit excellent bioactivities characterized by antibacterial, antitumor, antioxidative, phytotoxic, and enzyme inhibitory properties. Thus, the bio-metabolites of Alternaria species are significantly meaningful for pharmaceutical, industrial, biotechnological, and medicinal applications. To update the catalog of secondary metabolites synthesized by Alternaria fungi, 216 newly described metabolites isolated from Alternaria fungi were summarized with their diverse chemical structures, pharmacological activity, and possible biosynthetic pathway. In addition, possible insights, avenues, and challenges for future research and development of Alternaria are discussed.

Antifungal potential of endophytic Bacillus species isolated from tomato (Solanum lycopersicum) against Fusarium oxysporum collected from selected farms in Nigeria

ABSTRACT Fusarium wilt, caused by F. oxysporum, is a prevalent destructive tomato (Solanum lycopersicum L.) disease in Nigeria. Development of safe, and efficient biocontrol measures is important in the management of the disease. In this study, incidence of fusarium wilt of tomato in four agricultural zones of Oyo State was established through purposive sampling, and the abundance, and in vitro-biocontrol potential of Bacillus endophytes were determined. Strains of F. oxysporum and Bacillus endophytes were isolated from tomato plants collected from 21 farms. In vitro-inhibition potential of endophytes was determined through a modified, dual-culture assay, involving six endophytes, and carbendazim (fungicide), set up in a completely randomized design. Using a rating-scale of 0–9, the highest wilt severity (6.67) and incidence (39.30%) were observed on Akinware farm in Ibadan/Ibarapa agricultural zone of Oyo State. Fusarium oxysporum IB3q caused severe wilt symptoms, including leaf chlorosis, necrosis, vascular discoloration, and wilt, on infected Alausa and Ibadan-local tomato varieties. Thirty-nine strains of Bacillus endophytes were isolated from the tissues of healthy tomato plants. Six of the isolated Bacillus endophytes inhibited F. oxysporum. Bacillus amyloliquefaciens Sa08 and B. subtilis Og04 had inhibitory zones of 15.67 and 19.43 mm, respectively, against F. oxysporum, and caused hyphal disintegration of the pathogen. They produced cell wall-degrading enzymes, with relative activities ranging from 0.87 to 5.04. Based on their in vitro-inhibition potential, B. amyloliquefaciens Sa08 and B. subtilis Og04 should be deployed, as prospective biocontrol measures against seed-borne fusarium pathogens and fusarium wilt on tomato plants.

Endophytic species of Induratia from coffee and carqueja plants from Brazil and its potential for the biological control of toxicogenic fungi on coffee beans by means of antimicrobial volatiles.

Several endophytic fungi have been reported to have produced bioactive metabolites. Some of them, including the Induratia species, have the capacity to emit volatile compounds with antimicrobial properties with broad spectrum against human and plant pathogens. The present study aimed to prospect the Induratia species producing volatile organic compounds (VOCs), in carqueja plants used in alternative medicine and coffee plants in Brazil. A total of 11 fungal isolates producing volatile metabolites were obtained by a parallel growth technique, using I. alba 620 as a reference strain. Phylogenetic relationships revealed the presence of at least three distinct species, I. coffeana, I. yucatanensis, and Induratia sp. SPME/GC/MS analyses of the VOCs in the headspace above the mycelium from Induratia species cultured for 10days on PDA revealed the volatile profile emitted by I. coffeana CCF 572, I. coffeana COAD 2055, I. yucatanensis COAD 2062, and Induratia sp. COAD 2059. Volatile organic compounds produced by I. coffeana isolates presented antimicrobial activity against Aspergillus ochraceus, A. sclerotiorum, A. elegans, A. foetidus, A. flavus, A. tamari, A. tubingensis, A. sydowii, A. niger, A. caespitosus, A. versicolor, and A. expansum, sometimes by decreasing the growth rate or, mainly, by fully inhibiting colony growth. Fifty-eight percent of the target species died after 6days of exposure to VOCs emitted by I. coffeana CCF 572. In addition, VOCs emitted by the same fungus inhibited the growth in A. ochraceus inoculated into coffee beans, which indicates that plants which have I. coffeana as an endophyte may be protected from attacks by this plant pathogen.

The endophytic Fusarium strains: a treasure trove of natural products.

The complexity and structural diversity of the secondary metabolites produced by endophytes make them an attractive source of natural products with novel structures that can help in treating life-changing diseases. The genus Fusarium is one of the most abundant endophytic fungal genera, comprising about 70 species characterized by extraordinary discrepancy in terms of genetics and ability to grow on a wide range of substrates, affecting not only their biology and interaction with their surrounding organisms, but also their secondary metabolism. Members of the genus Fusarium are a source of secondary metabolites with structural and chemical diversity and reported to exhibit diverse pharmacological activities. This comprehensive review focuses on the secondary metabolites isolated from different endophytic Fusarium species along with their various biological activities, reported in the period from April 1999 to April 2022.

Two novel endophytic Tolypocladium species identified from native pines in south Florida

Supplementary information:<br />Fig. S1. Alignment used to construct the concatenated phylogenetic tree.<br />Fig. S2. Alignment used to construct the ITS phylogenetic tree.<br />Fig. S3. Phylogram of Tolypocladium species collected in this study and previously described generated from Maximum likelihood analysis of ITS sequence alignment. Bootstrap support (BS) values > 75 % are indicated.<br />Table S1. Strains used to construct the multilocus phylogeny.Information on isolation source and GenBank accession numbers.