Edodes Strains Research Articles

Transcriptional variation and RNA polymorphism among different Lentinula edodes (Berk.) Pegler strains.

Lentinula edodes is a commercially important mushroom known for its nutritional and therapeutic values. However, the molecular mechanisms underlying the distinct nutritional and physiological attributes of various L. edodes strains are not well understood. This study focused on three Lentinula strains (DMRO-356, DMRO-623, and DMRO-388s) with different nutritional and productivity profiles. Illumina sequencing was used to perform a whole-transcriptome analysis, conducting 100-base pair paired-end sequencing of total messenger RNA (mRNA) in duplicate, resulting in 28-48 million sequencing reads per strain. After rigorous data filtering, over 99% of high-quality reads were retained, and more than 95% were aligned to the Lentinula genome. Differential gene expression analyses identified 2210 differentially expressed genes between DMRO-356 and DMRO-623, 862 between DMRO-356 and DMRO-388s, and 2212 between DMRO-623 and DMRO-388s. Significant genetic variations were found among the strains, including 7753 single nucleotide polymorphisms (SNPs) in DMRO-356 versus DMRO-623 and 4080 SNPs in DMRO-356 versus DMRO-388s. Additionally, 349 insertions/deletions (InDels) were found in DMRO-356/DMRO-623 and 218 in DMRO-356/DMRO-388 s. Non-synonymous SNPs, which alter amino acid compositions, were analyzed, showing a preference for polar over charged amino acids. These differentially expressed genes were associated with various nutritional and developmental processes, highlighting the importance of genetic variations in shaping amino acid composition and potentially affecting protein function. This study is the first comprehensive exploration of transcriptional differences among Lentinula strains available for its cultivation, providing valuable insights to enhance mushroom quality and productivity. © 2024 Society of Chemical Industry.

Genome Sequencing of Lentinula edodes Revealed a Genomic Variant Block Associated with a Thermo-Tolerant Trait in Fruit Body Formation.

The formation of multicellular fruiting bodies in basidiomycete mushrooms is a crucial developmental process for sexual reproduction and subsequent spore development. Temperature is one of the most critical factors influencing the phase transition for mushroom reproduction. During the domestication of mushrooms, traits related to fruiting bodies have significantly impacted agricultural adaptation and human preferences. Recent research has demonstrated that chromosomal variations, such as structural variants (SVs) and variant blocks (VBs), play crucial roles in agronomic traits and evolutionary processes. However, the lack of high-quality genomic information and important trait data have hindered comprehensive identification and characterization in Lentinula edodes breeding processes. In this study, the genomes of two monokaryotic L. edodes strains, characterized by thermo-tolerance and thermo-sensitivity during fruiting body formation, were reassembled at the chromosomal level. Comparative genomic studies of four thermo-tolerant and thermo-sensitive monokaryotic L. edodes strains identified a 0.56 Mbp variant block on chromosome 9. Genes associated with DNA repair or cellular response to DNA damage stimulus were enriched in this variant block. Finally, we developed eight CAPS markers from the variant block to discriminate the thermo-tolerant traits in L. edodes cultivars. Our findings show that the identified variant block is highly correlated with the thermo-tolerant trait for fruiting body formation and that alleles present in this block may have been artificially selected during L. edodes domestication.

Divergent effects of entomopathogenic fungi and medicinal mushroom species on Drosophila melanogaster lifespan

We tested the effect of entomogenous fungi isolated from the Coleoptera species Ophrida xanthospilota and of the medicinal mushroom Lentinula edodes on the lifespan of Drosophila melanogaster. All the four analyzed strains of entomogenous fungi showed pathogenicity to D. melanogaster by causing 100% mortality of fruit flies within 20 culture days. The shortest lifespan was recorded in flies treated with Fusarium verticillioides (15 days). On the contrary, L. edodes showed a remarkable lifespan-extending effect on the model organism, with fruit fly individuals surviving for 55 culture days. Fungal growth was microscopically observed on the bodies of dead fruit flies treated with the four entomogenous fungal species, confirming that the tested fungi were responsible for the mortality of the model insects. Our findings showed that the four fungal species previously found on the body of Coleoptera individuals are entomopathogenic and could be exploited in agriculture applications for the biocontrol of noxious pests. The significant lifespan extending effect on Drosophila produced by the analyzed L. edodes strain could represent very important experimental evidence of the anti-aging and lifespan prolonging effects of this medicinal mushroom species.

Use of Eucalyptus Charcoal Waste in the Formulation of Substrate for the Cultivation of Two Strains (LED 20/11 and LED 20/12) of Lentinula edodes

The shiitake mushroom (Lentinula edodes) is globally valued for its nutrition and medicinal properties. New technologies aim to increase production with less environmental impact, considering materials such as charcoal for substrate enrichment. This manuscript evaluated the effect of fine charcoal (FC) on the substrate formulation of two L. edodes strains (LED 20/11 and LED 20/12). The substrate consisted of 72% eucalyptus sawdust, 12.5% rice bran, 12.5% wheat bran, and 3% calcium carbonate (control treatment without charcoal). Treatments with FC proportionally reduced the use of sawdust, with doses of 1%, 2%, 4%, 8%, and 16% (relative to the substrate material). Yield, mushroom number, and mushroom weight were evaluated. The concentration of FC significantly affected the parameters analyzed, especially at the 4% dose. A negative correlation between mushroom number and weight was observed. For yield, the control treatment and the lowest dose of FC (1%) had the highest yields for the first harvest. Strain LED 20/12 showed lower yield variability due to the percentage of FC applied to the substrate. The incorporation of FC into the substrate for shiitake cultivation demonstrates efficacy; however, both the concentration and strain used are limiting factors for its applicability.

Near-gapless genome and transcriptome analyses provide insights into fruiting body development in Lentinula edodes

Fruiting body development in macrofungi is an intensive research subject. In this study, high-quality genomes were assembled for two sexually compatible monokaryons from a heterokaryotic Lentinula edodes strain WX1, and variations in L. edodes genomes were analyzed. Specifically, differential gene expression and allele-specific expression (ASE) were analyzed using the two monokaryotic genomes and transcriptome data from four different stages of fruiting body development in WX1. Results revealed that after aeration, mycelia sensed cell wall stress, pheromones, and a decrease in CO2 concentration, leading to up-regulated expression in genes related to cell adhesion, cell wall remodeling, proteolysis, and lipid metabolism, which may promote primordium differentiation. Aquaporin genes and those related to proteolysis, mitosis, lipid, and carbohydrate metabolism may play important roles in primordium development, while genes related to tissue differentiation and sexual reproduction were active in fruiting body. Several essential genes for fruiting body development were allele-specifically expressed and the two nuclear types could synergistically regulate fruiting body development by dominantly expressing genes with different functions. ASE was probably induced by long terminal repeat-retrotransposons. Findings here contribute to the further understanding of the mechanism of fruiting body development in macrofungi.

Production of Extracellular Enzymes by Lentinula edodes Strains in Solid-State Fermentation on Lignocellulosic Biomass Sterilized by Physical and Chemical Methods.

Two methods of sterilization of lignocellulosic biomass were performed in this study. Eucalypt waste (EW) supplemented with rice bran (RB) was added in the proportions 80:20 and 90:10 in dry weight. The compositions were sterilized by physical method (autoclaving) and by chemical method (H2O2). The production of extracellular enzymes by Lentinula edodes strains was compared within the two methods. Inactivation of catalase present in RB was achieved with 250mM H2O2. The use of H2O2, when compared by physical method, favored high production of hydrolytic enzymes such as endoglucanase (1,600IU/kg), twofold higher, β-glucosidase (1,000IU/kg), fivefold higher, xylanase (55,000IU/kg), threefold higher and β-xylosidase (225IU/kg), similar results. Oxidative enzymes, MnP and laccase, were produced within a different profile between strains, with shorter times for laccase (2,200IU/kg) by SJC in 45days and MnP (2,000IU/kg) by CCB-514 in 30days. High production of extracellular enzymes is achieved by the use of the chemical method of sterilization of lignocellulosic biomass; in addition to no energy consumption, this process is carried out in a shorter execution time when compared to the physical process. The use of H2O2 in sterilization does not produce toxic compounds from the degradation of hemicellulose and cellulose such as furfural and hydroxy-methyl-furfural that cause inhibition of microorganisms and enzymes.

Prevalence and diversity of mycoviruses occurring in Chinese Lentinula edodes germplasm resource

Incidence and banding patterns of virus-like dsRNA elements in 215 Chinese genetically diverse Lentinula edodes strains collected from wide geographic distribution (or producing areas) were first investigated, and 17 viruses were identified including eight novel viruses. The results revealed a 63.3% incidence of dsRNA elements in the cultivated strains and a 67.2% incidence in the wild strains. A total of 10 distinguishable dsRNAs ranging from 0.6 to 12 kbp and 12 different dsRNA patterns were detected in the positive strains. The molecular information of these dsRNA elements was characterized, and the molecular information of the other 12 different viral sequences with (+) ssRNA genome was revealed in four L. edodes strains with complex dsRNA banding patterns. RT-PCR was also done to verify the five dsRNA viruses and 12 (+) ssRNA ones. The results presented may enrich our understanding of L. edodes virus diversity, and will promote further research on virus-host interactions. ImportanceViral infections involve complicated interactions including benign, harmful or possibly beneficial to hosts. Sometimes environment could lead to a transition in lifestyles from persistent to acute, resulting in a disease phenotype. The quality of spawn, such as the vulnerability to infection of viruses, is therefore important for mushroom production. Lentinula edodes, a wood rot basidiomycete fungus, was widely cultivated in the world for its edible and medicinal properties. In this study, the profile of dsRNA elements from Chinese genetically diverse L. edodes strains collected from wide geographic distribution or producing areas was first investigated. The molecular information of the dsRNA elements was characterized. Additionally, 12 different viral sequences with (+) ssRNA genome from four L. edodes strains with complex dsRNA banding patterns were identified. The results presented here will broaden our knowledge about mushroom viruses, and promote further studies of L. edodes production and the interaction between viruses and L. edodes.

Using food residues (potato peels) as an alternative to potato dextrose agar in the growth of edible food fungi

Due to the great importance of edible fungi, their rapid life cycle, and the possibility of their production throughout the year, their cultivation has become a vital resource that many countries depend on in their pursuit of economic growth and food security. The present study aimed to devise a local nutrient media for primary mycelium growth of nine species of fungi belonging to Pleurotus eryngii , Pleurotus floridanus , Pleurotus ostreatus (white strain) , Agaricus heterocystis , Agaricus bispous (brown strain ), Agaricus bispous (white strain), Lentinula edodesstrain No. 1, Lentinula edodes strain No. 2, Flammulina velutipe (yellow strain) using culture mycelium inoculation for reproduction and production of edible fungi. Different natural materials used for this purpose involved potato peels in a concentration ranging from 5 to 20 g/ L and compared with the commercial medium Potato Dextrose Agar at a rate of 39 g/Land according to the manufacturer's instructions. The study also focused on the effect of physical factors such as temperature and pH on the growth rate of fungal mycelium. The results showed that the media prepared from potato peels at 5 g/L and 10 g/L concentrations achieved the best growth rate for the studied fungi compared to Peptone Dextrose Agar (PDA). The increase in the growth rate of fungi on media prepared from potato peels was attributed to the effect of nutrients for the prepared medium because it contained effective and necessary compounds for the growth in addition to their ease of absorption and consumption to be ready for food for agricultural fungi. The effect on the growth and production of edible fungi or any other type of fungi has not previously been studied and compared to the commercial medium PDA.

Expression, Characterization and Purification of Latcripin-5 from Lentinula edodes Strain C91-3 and its In Vitro Anticancer Activities

Expression, Characterization and Purification of Latcripin-5 from Lentinula edodes Strain C91-3 and its In Vitro Anticancer Activities

Morphological and transcriptomic analysis revealing morphological variations and genetic clues in one Lentinula edodes abnormal browning strain.

Strain abnormal browning is a common problem during cultivation of Lentinula edodes. In this study, the L. edodes strain mycelia isolated from Le-WB and cultured on MYG (Le-WP) isolated from an abnormal browning bag was compared with its normal control mycelia isolated from Le-BB and cultured on MYG (Le-BP). The aerial hyphae of Le-WP were white, and the hyphal growth was significantly reduced. Morphological observation of Le-WP under scanning electron microscope (SEM) and transmission electron microscopy (TEM) revealed abnormal organelle structures. Through transcriptomic analysis, moredifferentially expressed genes (DEGs) were expressed in the metabolic process and catalytic activity in Le-WP than Le-BP. Two Kyoto encyclopedia of genes and genomes (KEGG) pathways named pentose and glucorunate interconversions, and starch and sucrose metabolism were found to be enriched in Le-WP. The gene expression profiles involved in these two pathways were further analyzed and 12 key genes were selected to be verified by quantitative real-time PCR (qRT-PCR), and the results showed that most of these genes were upregulated in Le-WP. Additionally, the content of 1,3-beta-glucan in Le-WP was also significantly higher than in other samples. This research suggests that abnormal strains may be related to the abnormal synthesis of 1,3-beta-glucan, and it needs further research. This research exhibits possible morphological and genetic clues of Le-WP and lays the foundation for understanding the degeneration of L. edodes strains.

Comparative Transcriptome Analysis Provides Insights Into the Mechanism by Which 2,4-Dichlorophenoxyacetic Acid Improves Thermotolerance in Lentinula edodes.

As the widest cultivated edible mushroom worldwide, Lentinula edodes suffers serious yield and quality losses from heat stress during growth and development, and in our previous study, exogenous 2,4-Dichlorophenoxyacetic acid (2,4-D) was found to improve the thermotolerance of L. edodes strain YS3357, but the molecular mechanism remains unclear. Here, we explored the potential protective mechanism of exogenous 2,4-D against heat stress by transcriptome analysis. 2,4-D possible improve the thermotolerance of L. edodes through regulating antioxidant genes, transcription factors, energy-provision system, membrane fluidity, and cell wall remodeling. Furthermore, 2,4-D was also found to regulate the saturation levels of fatty acids and ATP content in L. edodes mycelium under heat stress. This study proposed a regulatory network of 2,4-D in regulating L. edodes response to heat stress, providing a theoretical basis for improving L. edodes thermotolerance, and facilitating the understanding of the molecular mechanism of exogenous hormones in alleviating abiotic stress damage to macrofungi.

Chromosomal genome and population genetic analyses to reveal genetic architecture, breeding history and genes related to cadmium accumulation in Lentinula edodes

BackgroundLentinula edodes (Berk.) is the second most productive mushroom in the world. It contains compounds effective for antiviral, antitumor, antioxidant and immune regulation. Although genomes have previously been reported for this species, a high-quality chromosome-level reference for L. edodes is unavailable. This hinders detailed investigation of population genetics, breeding history of strains and genes related to environmental stress responses.ResultsA high-quality chromosome-level genome was constructed. We separated a monokaryon from protoplasts of the commercial L. edodes strain L808 and assembled the genome of L. edodes using PacBio long-read and Illumina short-read sequencing, along with the high-throughput chromatin conformation capture (Hi-C) technique. We assembled a 45.87 Mb genome, and 99% of the sequences were anchored onto 10 chromosomes. The contig and scaffold N50 length were 2.17 and 4.94 Mb, respectively. Over 96% of the complete Benchmarking Universal Single-Copy Orthologs (BUSCO) were identified, and 9853 protein-coding genes were predicted. We performed population genome resequencing using 34 wild strains and 65 commercial cultivars of L. edodes originating from China, Japan, the United States and Australia. Based on whole-genome variants, we showed substantial differences in the Chinese wild population, which divided into different branches according to the main areas of their geographical distribution. We also determined the breeding history of L. edodes at the molecular level, and demonstrated that the cultivated strains in China mainly originated from wild strains from China and Northeast Asia. Phenotypic analysis showed that 99 strains exhibited differences on the Cd accumulation. Three significant loci in the of L. edodes genome were identified using the genome-wide association study (GWAS) of Cd accumulation traits. Functional genes associated with Cd accumulation traits were related to DNA ligase and aminoacyl tRNA synthetase, indicating that DNA damage repair and in vivo protein translation may be responses to Cd stress.ConclusionsA high-quality chromosome-level genome and population genetic data of L. edodes provide genetic resources for functional genomic, evolutionary and artificial breeding studies for L. edodes.

Decontamination of Eucalyptus Waste with H2O2: Effect on Activities of Extracellular Enzymes Produced by Lentinula Edodes Strains

Decontamination of Eucalyptus Waste with H2O2: Effect on Activities of Extracellular Enzymes Produced by Lentinula Edodes Strains

Comparison of Cultivation, Mushroom Yield, and Fruiting Body Characteristics of Lentinula edodes Strains according to the Inoculation Method

Comparison of Cultivation, Mushroom Yield, and Fruiting Body Characteristics of Lentinula edodes Strains according to the Inoculation Method

Curing two predominant viruses occurring in Lentinula edodes by chemotherapy and mycelial fragmentation methods

Previous research has established that Lentinula edodes mycovirus HKB (LeV-HKB) and L. edodes partitivirus 1(LePV1) are major mycoviruses identified in L.edodes germplasm. In this paper, two different methods for curing these two dsRNA mycoviruses, ribavirin treatment and mycelial fragmentation, were evaluated for the first time. Mycelial fragmentation was found to resulted in LeV-HKB- and LePV1-cured fungal strains, whereas ribavirin treatment could eliminate LeV-HKB only. Although no LePV1-cured strain was obtained via ribavirin treatment by the end of the experiment, the relative LePV1 concentration in the eighth successive subcultures was lower than that of the untreated control. The culture features of several virus-cured strains had faster mycelial growth rate and higher colony density than the infected ones. It was also suggested that LeV-HKB infection may affect the pigmentation in plate- and bag-cultivated mycelia of L. edodes strain L135.

Population genomics provides insights into the genetic basis of adaptive evolution in the mushroom-forming fungus Lentinula edodes

IntroductionMushroom-forming fungi comprise diverse species that develop complex multicellular structures. In cultivated species, both ecological adaptation and artificial selection have driven genome evolution. However, little is known about the connections among genotype, phenotype and adaptation in mushroom-forming fungi. ObjectivesThis study aimed to (1) uncover the population structure and demographic history of Lentinula edodes, (2) dissect the genetic basis of adaptive evolution in L. edodes, and (3) determine if genes related to fruiting body development are involved in adaptive evolution. MethodsWe analyzed genomes and fruiting body-related traits (FBRTs) in 133 L. edodes strains and conducted RNA-seq analysis of fruiting body development in the YS69 strain. Combined methods of genomic scan for divergence, genome-wide association studies (GWAS), and RNA-seq were used to dissect the genetic basis of adaptive evolution. ResultsWe detected three distinct subgroups of L. edodes via single nucleotide polymorphisms, which showed robust phenotypic and temperature response differentiation and correlation with geographical distribution. Demographic history inference suggests that the subgroups diverged 36,871 generations ago. Moreover, L. edodes cultivars in China may have originated from the vicinity of Northeast China. A total of 942 genes were found to be related to genetic divergence by genomic scan, and 719 genes were identified to be candidates underlying FBRTs by GWAS. Integrating results of genomic scan and GWAS, 80 genes were detected to be related to phenotypic differentiation. A total of 364 genes related to fruiting body development were involved in genetic divergence and phenotypic differentiation. ConclusionAdaptation to the local environment, especially temperature, triggered genetic divergence and phenotypic differentiation of L. edodes. A general model for genetic divergence and phenotypic differentiation during adaptive evolution in L. edodes, which involves in signal perception and transduction, transcriptional regulation, and fruiting body morphogenesis, was also integrated here.

Enhanced Expression of Thaumatin-like Protein Gene (LeTLP1) Endows Resistance to Trichoderma atroviride in Lentinula edodes.

Lentinula edodes (shiitake mushrooms) is heavily affected by the infection of Trichoderma atroviride, causing yield loss and decreases quality in shiitake mushrooms. The selection and breeding of fungal-resistant L. edodes species are an important approach to protecting L. edodes from T. atroviride infection. Herein, a highly resistant L. edodes strain (Y3334) and a susceptible strain (Y55) were obtained by using a resistance evaluation test. Transcriptome analyses and qRT-PCR detection showed that the expression level of LeTLP1 (LE01Gene05009) was strongly induced in response to T. atroviride infection in the resistant Y3334. Then, LeTLP1-silenced and LeTLP1-overexpression transformants were obtained. Overexpression of LeTLP1 resulted in resistance to T. atroviride. Compared with the parent strain Y3334, LeTLP1-silenced transformants had reduced resistance relative to T. atroviride. Additionally, the LeTLP1 protein (Y3334) exhibited significant antifungal activity against T. atroviride. These findings suggest that overexpression of LeTLP1 is a major mechanism for the resistance of L. edodes to T. atroviride. The molecular basis provides a theoretical basis for the breeding of resistant L. edodes strains and can eventually contribute to the mushroom cultivation industry and human health.

Transcriptome Analysis Identified Candidate Genes Involved in Fruit Body Development under Blue Light in Lentinula edodes

Lentinula edodes is an edible mushroom that is rich in polysaccharides, glucan, and lentinan. It is famous for its earthy, sweet, umami flavor, and is used in various foods all over the world. Although Lentinula edodes does not carry out photosynthesis with light, its fruit body development is regulated by light. In this study, we analyzed the morphological changes of L. edodes strain Sanjo701ho and identified the global gene expression patterns using EdgeR of fruit body development under blue light. The phenotype analysis under different light sources revealed that the pileus diameter grew, while the stipe length was suppressed under blue light. To understand the changes in the transcriptome under different light sources in L. edodes, gene set enrichment analysis (GSEA), KOG functional categories, and KEGG pathways were used and compared to the no-light condition. Lignocellulose, CAZyme, and transcription factor classified DEGs (differentially expressed genes) were identified to better understand the significant DEGs affected by light sources in the synthesis, metabolism, and recognition of complex carbohydrates. Six glycoside hydrolases (GHs), four auxiliary activities (AAs), three carbohydrate esterases (CEs), and glycosyltransferases (GTs) were identified as upregulated in the CAZyme DEGs. Furthermore, four β-glucosidase, one glucose oxidase, and one multicopper oxidase-related gene for lignocellulolytic genes were upregulated in the blue light condition, and AT_hook transcription factor, CBFD_NFYB_HMF transcription factor, HMG_box transcription factor, and fungal specific transcription factor were upregulated in the blue light condition. This study helps us understand fruit body development in mushroom-breeding programs.

Chemical and productivity characterization of parental and hybrid strains of Lentinula edodes cultivated in different agricultural residues


 Productivity of parental and hybrid strains of Lentinula edodes cultivated with two mixtures of agricultural wastes (mixture 1: peat moss, banana leaves, oak sawdust, millet seed, cotton seed hull and CaCO3, and mixture 2: wheat straw, oak sawdust, cotton seed hull and CaCO3), the chemical composition of the fruit bodies and also the antioxidant activities of hexanic extracts of the mushrooms were evaluated. The strains of Lentinula edodes (LEN97 and LEN954XLEC241) cultivated on both mixtures produced fruit bodies with highest protein contents since 16.71 to 17.10%. The hybrid strain of Lentinula edodes strain (LEN954XLEC241) cultivated on both mixtures showed the highest biological efficiencies since 74.61 to 82.80% and production rates between 1.21 and 1.30%, and also their hexanic extracts presented the highest antioxidant effects ranged from 8.01 to 9.84 mg mL- 1. The use of hybrid strain of Lentinula edodes can be used to improve the production of this genera.

Pathological Properties of Cryptococcus pseudolongus on the Mycelia and Fruit Body of Lentinula edodes

Recently, Cryptococcus pseudolongus has been reported as a new pathogen of shiitake (Lentinula edodes). However, its pathological properties are not much known. To further understand its impact on the mushroom, we investigated the pathogen’s interactions with the mycelium of shiitake, histopathological properties, host range, and sensitivity to diverse antifungal agents. The strain C. pseudolongus DUCC 4014 inhibited the mycelial growth of L. edodes strain (cultivar Sanjo 701ho) and caused browning in the mycelia confronted with the yeast on PDA. Spray inoculation of the yeast caused an abnormal browning symptom on the cap and/or gills of three shiitake cultivars grown on sawdust media in vinyl bags. Scanning electron microscopic images of the abnormally browned parts of shiitake fruit body illustrated that mushroom tissues were loosed and dispersed in the middle and edge of the cap and the arrangement of basidiospores borne on basidia in the gills was disturbed compared to those of normal shiitake fruit body. Spray inoculation also led to developing abnormal browning on the harvested fruit body, indicating C. pseudolongus could be a problem during mushroom storage. But the yeast was not able to induce abnormal browning on mushrooms of Pleurotus ferulae, Pleurotus fostreatus, and Agaricus bisporus. But it induced browning only on button mushroom (A. bisporus) when they were inoculated after wounding. Tests with 16 kinds of fungicides revealed that the cell growth of C. pseudolongus could be inhibited by benzalkonium chloride at MIC 7 μg/ml and benomyl at MIC 3 μg/ml.