Hyphal Growth Rate Research Articles

Pyriofenone Interacts with the Major Facilitator Superfamily Transporter of Phytopathogenic Fungi to Potentially Control Tea Leaf Spot Caused by Lasiodiplodia theobromae.

Tea leaf spot caused by Lasiodiplodia theobromae is a newly discovered fungal disease in southwest China. Due to a lack of knowledge of its epidemiology and control strategies, the disease has a marked impact on tea yield and quality. Pyriofenone is a new fungicide belonging to the aryl phenyl ketone fungicide group, which has shown marked efficacy in controlling various fungal diseases. However, its mechanism of action is not yet understood. This study found that pyriofenone exhibits strong in vitro inhibitory activity against various phytopathogenic fungi. Specifically, it showed strong inhibitory activity against L. theobromae, with a half-maximal effective concentration (EC50) value of 0.428 μg/ml determined by measuring mycelial growth rate. Morphological observations, using optical, scanning electron, and transmission electron microscopy, revealed that pyriofenone induces morphological abnormalities in L. theobromae hyphae. At lower doses, the hyphae became swollen, the distance between septa decreased, and the hyphal growth rate slowed. At higher doses and longer exposures, the hyphae collapsed. Transcriptomic and bioinformatic analyses indicated that pyriofenone can affect the expression of genes related to membrane transporters. Homology modeling suggested that pyriofenone may bind to a candidate target protein of the major facilitator superfamily (MFS) transporter, with a free binding energy of -7.1 kcal/mol. This study suggests that pyriofenone may potentially regulate the transport of metabolites in L. theobromae, thus affecting hyphal metabolism and interfering with hyphal growth. Pyriofenone exhibits in vitro inhibitory activity against various tea foliar pathogens and holds promise for future applications to the control of tea foliar diseases.

Biological characteristics of Cordyceps militaris single mating-type strains.

Cordyceps militaris has been extensively cultivated as a model cordyceps species for commercial purposes. Nevertheless, the problems related to strain degeneration and breeding technologies remain unresolved. This study assessed the physiology and fertility traits of six C. militaris strains with distinct origins and characteristics, focusing on single mating-type strains. The results demonstrated that the three identified strains (CMDB01, CMSY01, and CMJB02) were single mating-type possessing only one mating-type gene (MAT1-1). In contrast, the other three strains (CMXF07, CMXF09, and CMMS05) were the dual mating type. The MAT1-1 strains sourced from CMDB01, CMSY01, and CMJB02 consistently produced sporocarps but failed to generate ascospores. However, when paired with MAT1-2 strains, the MAT1-1 strains with slender fruiting bodies and normal morphology were fertile. The hyphal growth rate of single mating-type strains (CMDB01, CMSY01, and CMJB02) typically surpassed that of dual mating-type strains (CMXF07, CMXF09, and CMMS05). The growth rates of MAT1-2 and MAT1-1 strains were proportional to their ratios, such that a single mating-type strain with a higher ratio exhibited an increased growth rate. As C. militaris matured, the adenosine content decreased. In summary, the C. militaris strains that consistently produce sporocarps and have a single mating type are highly promising for production and breeding.

Protein kinase A regulatory subunit is required for normal growth, zoosporogenesis, and pathogenicity in Phytophthora sojae

Phytophthora sojae, one of the most devastating Oomycete pathogens, causes severe diseases that lead to economic loss in the soybean industry. The production of zoospores play a crucial role during the development of Phytophthora disease. In this work, CRISPR/Cas9 genome editing technology were used to obtain protein kinase A regulatory subunit (PsPkaR) knockout mutants. The role of PsPkaR in the production of zoospores and pathogenicity of P. sojae was analyzed. The overall findings indicate that PsPkaR is involved in regulating the growth process of P. sojae, primarily affecting the hyphal morphology and growth rate. Additionally, PsPkaR participates in the regulation of the release process of zoospores. Specifically, knocking-out PsPkaR resulted in incomplete cytoplasmic differentiation and uneven protoplast division, leading to abnormal release of zoospores. Furthermore, when the PsPkaR knockout mutants were inoculated on soybean leaves, the pathogenicity was significantly reduced compared to that of the wild-type and control strains. These findings of this study provide important clues and evidence regarding the role of the cAMP-PKA signaling pathway in the interaction between P. sojae and its host. This work contributes to a better understanding of the pathogenic mechanism of P. sojae and the development of corresponding prevention and control strategies.

Oleic acid improves the conidial production and quality of Metarhizium rileyi as a biocontrol agent

ABSTRACT Metarhizium rileyi is a biological agent used in the control of various pests, particularly noctuid insects. However, this fungal species is slow-growing, resulting in a prolonged culture period and limiting its commercial production. In this study, 0.3% (w/w) oleic acid significantly enhanced the rates of hyphal growth, conidial production, and conidial germination by 23%, 9%, and 10%, respectively. The expression of genes related to microbial growth and development (MrPbs2, MrMsn2, MrSwi6, MrNsdD, Mrap1 and MrSte12) was significantly increased in the presence of oleic acid. The M. rileyi conidial production and its effects on the mortality of Spodoptera frugiperda were investigated using rice-vegetable oil solid medium (RVOSM) and its substitute Sabouraud maltose agar plus yeast extract (SMAY) plates containing 0.03% oleic acid. The fungi retained their toxicity to S. frugiperda. Therefore, oleic acid can improve the growth potential of fungi, and RVOSM can be used as a substrate for conidia production for the application of M. rileyi as a biocontrol agent.

Using fimbrin to quantify the endocytic subapical collar during polarized growth in three filamentous fungi

ABSTRACT Filamentous fungi produce specialized cells called hyphae. These cells grow by polarized extension at their apex, which is maintained by the balance of endocytosis and exocytosis at the apex. Although endocytosis has been well characterized in other organisms, the details of endocytosis and its role in maintaining polarity during hyphal growth in filamentous fungi is comparatively sparsely studied. In recent years, a concentrated region of protein activity that trails the growing apex of hyphal cells has been discovered. This region, dubbed the “endocytic collar” (EC), is a dynamic 3-dimensional region of concentrated endocytic activity, the disruption of which results in the loss of hyphal polarity. Here, fluorescent protein–tagged fimbrin was used as a marker to map the collar during growth of hyphae in three fungi: Aspergillus nidulans, Colletotrichum graminicola, and Neurospora crassa. Advanced microscopy techniques and novel quantification strategies were then utilized to quantify the spatiotemporal localization and recovery rates of fimbrin in the EC during hyphal growth. Correlating these variables with hyphal growth rate revealed that the strongest observed relationship with hyphal growth is the distance by which the EC trails the apex, and that measured endocytic rate does not correlate strongly with hyphal growth rate. This supports the hypothesis that endocytic influence on hyphal growth rate is better explained by spatiotemporal regulation of the EC than by the raw rate of endocytosis.

The Small Ras Superfamily GTPase Rho4 of the Maize Anthracnose Fungus Colletotrichum graminicola Is Required for β-1,3-glucan Synthesis, Cell Wall Integrity, and Full Virulence.

Small Ras superfamily GTPases are highly conserved regulatory factors of fungal cell wall biosynthesis and morphogenesis. Previous experiments have shown that the Rho4-like protein of the maize anthracnose fungus Colletotrichum graminicola, formerly erroneously annotated as a Rho1 protein, physically interacts with the β-1,3-glucan synthase Gls1 (Lange et al., 2014; Curr. Genet. 60:343–350). Here, we show that Rho4 is required for β-1,3-glucan synthesis. Accordingly, Δrho4 strains formed distorted vegetative hyphae with swellings, and exhibited strongly reduced rates of hyphal growth and defects in asexual sporulation. Moreover, on host cuticles, conidia of Δrho4 strains formed long hyphae with hyphopodia, rather than short germ tubes with appressoria. Hyphopodia of Δrho4 strains exhibited penetration defects and often germinated laterally, indicative of cell wall weaknesses. In planta differentiated infection hyphae of Δrho4 strains were fringy, and anthracnose disease symptoms caused by these strains on intact and wounded maize leaf segments were significantly weaker than those caused by the WT strain. A retarded disease symptom development was confirmed by qPCR analyses. Collectively, we identified the Ras GTPase Rho4 as a new virulence factor of C. graminicola.

Isolation and identification of two Beauveria bassiana strains from silkworm, Bombyx mori.

Silkworm diseases caused by fungi infection occur frequently in sericulture and brought huge economic loss to sericulture. However, on the other hand, some fungi such as Beauveria bassiana, as an important entomological fungus, play an important role in biological control of insect pests. Here, two fungal pathogens causing yellow muscardine were isolated from the silkworm and named as SZY1 and SZY2. These two strains showed almost the same conidial morphology which were smooth, near-spherical, spherical, or ovoid and 2.7 ± 0.6µm × 2.5 ± 0.9µm in size, and the hyphal growth rate was also similar. However, the conidia production of SZY2 was almost twice as many as that of SZY1. The complete ribosomal RNA gene was sequenced and analyzed. As a result, the gene sequences of internal transcript space 1 (ITS1)-5.8S rRNA-internal transcript space 2 (ITS2) of SZY1 and SZY2 were identical in sequence and size, and for 18S rRNA, 28S rRNA, and intergenic spacer (IGS), the gene identity of SZY1 to SZY2 was 99%, 99%, and 98%, respectively. Results of phylogenetic analysis based on either ITS1-5.8S rRNA-ITS2 or 18S rRNA showed that both SZY1 and SZY2 were closely related to Beauveria bassiana. These results revealed that the pathogens of yellow muscardine SZY1 and SZY2 were identified as two different strains of Beauveria bassiana, which could provide diagnostic evidence for silkworm muscardine and was helpful for the research and development of novel Bombyx batryticatus and fungal biological insecticide.

The C2H2 Transcription Factor SsZFH1 Regulates the Size, Number, and Development of Apothecia in Sclerotinia sclerotiorum.

Sclerotinia sclerotiorum is a notorious phytopathogenic Ascomycota fungus with a host range of >600 plant species worldwide. This homothallic Leotiomycetes species reproduces sexually through a multicellular apothecium that produces and releases ascospores. These ascospores serve as the primary inoculum source for disease initiation in the majority of S. sclerotiorum disease cycles. The regulation of apothecium development for this pathogen and other apothecium-producing fungi remains largely unknown. Here, we report that a C2H2 transcription factor, SsZFH1 (zinc finger homologous protein), is necessary for the proper development and maturation of sclerotia and apothecia in S. sclerotiorum and is required for the normal growth rate of hyphae. Furthermore, ΔSszfh1 strains exhibit decreased H2O2 accumulation in hyphae, increased melanin deposition, and enhanced tolerance to H2O2 in the process of vegetative growth and sclerotia formation. Infection assays on common bean leaves, with thin cuticles, and soybean and tomato leaves, with thick cuticles, suggest that the deletion of Sszfh1 slows the mycelial growth rate, which in turn affects the expansion of leaf lesions. Collectively, our results provide novel insights into a major fungal factor mediating maturation of apothecia with additional effects on hyphae and sclerotia development.

First Report of Pythium Cluster B2a Species Causing Root Rot in Welsh Onion.

Hiroshima Prefecture has the highest production area of hydroponically grown Welsh onions (Allium fistulosum L.) in Japan. Since the cultivation began in 1988, root rot (Fig. 1A) followed by leaf browning (Fig. 1B) has caused significant economic losses. Approximately 80% (loss of 45 million JPY) of plant loss occurred from May to Sep 2009 (Shimizu, unpublished), and the disease was observed again in 2020. Diseased Welsh onions (five to six leaf stage) were collected in 2009. Abundant nonseptate hyphae of Pythium-like organisms were observed in the rotted roots (Fig. 1C). Disinfected symptomatic tissue samples were placed on NARF medium (Morita and Tojo 2007) and incubated at 25°C for 3 to 7 days. Six Pythium-like organisms were isolated, and their morphological features on a grass blade culture, potato carrot agar (PCA) (van der Plaats-Niterink 1981), cornmeal agar (CMA) and V8 juice agar (Miller 1955) were examined. Hyphal growth rates from 1-46°C were measured by culturing on PCA. The ribosomal internal transcribed spacer (ITS) regions and mitochondrial COI of the isolates were amplified and sequenced according to Ueta and Tojo (2016). All six isolates obtained showed similar morphology, hyphal growth rates, and sequences of ITS and COI. Detailed descriptions are provided here for the representative isolate 72 (MAFF246451). The isolate produced asexual structures but did not form sexual structures, including oogonia, antheridia, and oospores on all the media used. Hyphae were up to 6.8 μm wide. Appressoria were knob-like terminations (Fig. 1D). Sporangia were filamentous and indistinguishable from the hyphae. Zoospores (Fig. 1E) were formed at 5-25°C. The diameter of encysted zoospores ranged 7.4-10.1 (av. 8.9) μm (Fig. 1F). Cardinal temperatures for hyphal growth on PCA were 5°C min, 28-31°C opt, and 35°C max. The daily growth rate at 25°C was 15.0 mm per day. The sequence analysis of all isolates, including isolate 72 (GenBank ac nos AB700596 for ITS, LC630955 for COI) showed the present isolates belonged to Pythium Cluster B2a (Robideau et al. 2011) (Fig. 2). Based on these features, the six isolates were identified as Pythium Cluster B2a sp. In the inoculation test, isolate 72 was cultured on CMA at 25°C for 5 days. Mycelium disks (5 mm diam) obtained from the culture were placed on the primary roots of 8-day-old Welsh onion seedlings (cv Koutou), which were grown at a density of six plants on rock wool cubes moistened with tap water in a 50 mL plastic pot. The inoculated and non-inoculated plants were grown at 28°C for 7 days in a growth chamber. The experiment was repeated twice using three pots per replication. The plants inoculated with isolate 72 wilted, and their roots rotted 7 days after inoculation. No disease was found observed on the non-inoculated plants. The isolate of Pythium Cluster B2a sp. was consistently re-isolated from the diseased plants, thus, fulfilling Koch's postulates. Pythium Cluster B2a sp. causing stem rot on lettuce has been recorded in Italy (Garibaldi et al. 2017). To our knowledge, this is the first report of Pythium Cluster B2a sp. on Welsh onions. Since significant losses to root rot of Welsh onion have occurred in Japan, identification of the causal organism will enable the development of management practices to reduces losses.

Genomic regions associated with virulence in Setosphaeria turcica identified by linkage mapping in a biparental population

Northern corn leaf blight (NCLB) and sorghum leaf blight (SLB) are significant diseases of maize and sorghum, respectively, caused by the filamentous fungus Setosphaeria turcica. Strains of S. turcica are typically host-specific and infect either maize or sorghum. Host specificity in this pathogen is attributed to a single locus for maize and a second distinct locus for sorghum. To identify the genetic basis of host specificity in S. turcica, we generated a biparental population of S. turcica by crossing strains specific to maize and sorghum, phenotyped the population for leaf blight on sorghum and maize, genotyped the population to create a linkage map of S. turcica, and located candidate virulence regions. A total of 190 ascospores from 35 pseudothecia were isolated from the cross of maize and sorghum-specific strains. Greenhouse phenotyping of the biparental population (n = 144) showed independent inheritance of virulence, as indicated by a 1:1:1:1 segregation for virulence to maize, sorghum, both maize and sorghum, and avirulence to both crops. The population and host-specific parent strains were genotyped using genome skim sequencing on an Illumina NovaSeq 6000 platform resulting in over 780 million reads. A total of 32,635 variants including single nucleotide polymorphisms and indels were scored. There was evidence for a large deletion in the sorghum-specific strain of S. turcica. A genetic map consisting of 17 linkage groups spanning 3,069 centimorgans was constructed. Virulence to sorghum and maize mapped on distinct linkage groups with a significant QTL detected for virulence to maize. Furthermore, a single locus each for the in vitro traits hyphal growth rate and conidiation were identified and mapped onto two other linkage groups. In vitro traits did not correlate with in planta virulence complexity, suggesting that virulence on both hosts does not incur a fitness cost. Hyphal growth rate and conidiation were negatively correlated, indicating differences in hyphal growth versus dispersal ability for this pathogen. Identification of genetic regions underlying virulence specificity and saprotrophic growth traits in S. turcica provides a better understanding of the S. turcica- Andropogoneae pathosystem.

Culturable Endophytic Fungi from Glycyrrhiza inflata Distributed in Xinjiang, China with Antifungal Activity

A total of 99 endophytic fungal isolates were obtained from the roots of Glycyrrhiza inflata, which was a traditional medicinal plant mainly distributed in Xinjiang, China. Twenty-two distinct isolates were selected for further taxonomical identification by morphological traits and internal transcribed spacer (ITS) rRNA gene sequence analysis. Eleven genera were identified, among which Aspergillus, Alternaria and Fusarium were dominant. The crude extracts of 22 distinct identified fungi were successively evaluated for their antifungal activities on three rice fungal pathogens using the method of hyphal radial growth rate. Among them, the crude extract of Alternaria an-gustiovoidea Glinf007 showed the significantly mycelial growth inhibitory activity. The results demonstrated that G. inflata contained a diversity of culturable endophytic fungi, which could produce natural antimicrobial compounds that might be of great value to the agriculture and pharmaceutical industries.

Occurrence, hyphal growth rate, and carbon source utilization of fungi from continental Antarctica

Reported data on the diversity and physiological capabilities of fungi inhabiting ice-free outcrops of continental Antarctica are limited. The present study aimed at investigating the occurrence of fungi on fragments of microbial mats on the lakeshore and lake water and compared it with fungal occurrence on terrestrial moss, using a culture-dependent method. Another purpose was to evaluate the responses of hyphal growth to temperature and the profiles of substrate utilization of fungal isolates under pure culture conditions. A total of 46 fungal isolates were obtained and grouped into eight taxa, with Phoma herbarum being the most frequent taxon, the occurrence of which was closely related to the nitrogen level of the substrata. The colony diameter growth rates of most fungal isolates under pure culture conditions increased linearly with temperature to reach the maximum at 20–25 °C, indicating that these fungi were mesophilic. Moreover, fungal isolates exhibited diversity and redundancy in their utilization of 71 carbon sources, having potentials to utilize an array of substrates including sugars, sugar alcohols, hexose-phosphates, amino acids, hexose acids, and carboxylic acids.

Sclerotinia sclerotiorum Thioredoxin1 (SsTrx1) is required for pathogenicity and oxidative stress tolerance.

Sclerotinia sclerotiorum infects host plant tissues by inducing necrosis to source nutrients needed for its establishment. Tissue necrosis results from an enhanced generation of reactive oxygen species (ROS) at the site of infection and apoptosis. Pathogens have evolved ROS scavenging mechanisms to withstand host‐induced oxidative damage. However, the genes associated with ROS scavenging pathways are yet to be fully investigated in S. sclerotiorum. We selected the S. sclerotiorum Thioredoxin1 gene (SsTrx1) for our investigations as its expression is significantly induced during S. sclerotiorum infection. RNA interference‐induced silencing of SsTrx1 in S. sclerotiorum affected the hyphal growth rate, mycelial morphology, and sclerotial development under in vitro conditions. These outcomes confirmed the involvement of SsTrx1 in promoting pathogenicity and oxidative stress tolerance of S. sclerotiorum. We next constructed an SsTrx1‐based host‐induced gene silencing (HIGS) vector and mobilized it into Arabidopsis thaliana (HIGS‐A) and Nicotiana benthamiana (HIGS‐N). The disease resistance analysis revealed significantly reduced pathogenicity and disease progression in the transformed genotypes as compared to the nontransformed and empty vector controls. The relative gene expression of SsTrx1 increased under oxidative stress. Taken together, our results show that normal expression of SsTrx1 is crucial for pathogenicity and oxidative stress tolerance of S. sclerotiorum.

Bjerkandera adusta M1 inhibits the growth of Fusarium oxysporum f. sp. conglutinans and fusarium wilt incidence in Brassica napus L.

Bjerkandera adusta degrades polycyclic aromatic compound. However, its effect on preventing plant disease is unknown. We carried out both the confrontation culture and greenhouse pot experiments to address effects of B. adusta M1 strain on the growth of pathogenic Fusarium oxysporum f. sp. conglutinans (FOC) and incidence of Fusarium wilt in Brassica napus L. The results showed the hyphal growth rate of B. adusta M1 was significantly greater than that of FOC. The B. adusta M1 fermentation broth also significantly inhibited the growth of FOC hyphae by 62.79 ± 1.80%, which was greater than an inhibition rate of 40.63 ± 1.68% by the fungicide, carbendazim. The electron microscopic image showed the FOC hyphae was directly penetrated by the B. adusta M1 hyphae, indicating a strong mycoparasitism by B. adusta M1. Both the B. adusta M1 and its fermentation broth reduced the incidence and disease index of the Fusarium wilt in Br. napus, and the control effects of B. adusta M1 and its fermentation broth on Fusarium wilt were comparable or better than carbendazim. Furthermore, both the B. adusta M1 and its fermentation broth increased the activity of the defense enzymes including superoxide dismutase (25.92% and 22.38%), catalase (78.80% and 82.28%), peroxidase (46.64% and 6.83%) and phenylalanine ammonia-lyase (44.23% and 5.77%). Similarly, both the B. adusta M1 and its fermentation broth decreased the cell membrane permeability (48.68% and 33.71%) and malondialdehyde concentration (71.04% and 17.51%) of plant leaf. Therefore, B. adusta M1 can be applied as a potential biocontrol fungus to against Fusarium wilt.

Identification of Fusarium oxysporum as causative agent of rhizome rot disease in seed ginger rhizomes in Wayanad, Kerala

Rhizome rot was reported by ginger farmers of Wayanad, Kerala as a major productivity problem being faced in current growing year (2019–2020). The etiological agent was a fungus with a hyphal growth rate of 83.75 ± 3 mm after 5 doi. Subsequent morphological and molecular characterization by elucidating the fungal colony, conidial morphology, ITS sequencing and phylogenetic analysis identified the isolates as Fusarium oxysporum. Koch’s postulates were fulfilled after inoculating healthy rhizomes with the isolates (1 × 105 conidia/ ml), obtained from symptomatic rhizomes collected from field. Typical rot symptoms were visible after 4 days of infection (doi). Present study identifying the etiological agent provides information to farmers in selecting appropriate disease control measures to prevent further financial loss and opens research avenues for developing sustainable solutions.

Cadmium Stress Reprograms ROS/RNS Homeostasis in Phytophthora infestans (Mont.) de Bary.

Heavy metal pollution causes many soils to become a toxic environment not only for plants, but also microorganisms; however, little is known how heavy metal contaminated environment affects metabolism of phytopathogens and their capability of infecting host plants. In this study the oomycete Phytophthora infestans (Mont.) de Bary, the most harmful pathogen of potato, growing under moderate cadmium stress (Cd, 5 mg/L) showed nitro-oxidative imbalance associated with an enhanced antioxidant response. Cadmium notably elevated the level of nitric oxide, superoxide and peroxynitrite that stimulated nitrative modifications within the RNA and DNA pools in the phytopathogen structures. In contrast, the protein pool undergoing nitration was diminished confirming that protein tyrosine nitration is a flexible element of the oomycete adaptive strategy to heavy metal stress. Finally, to verify whether Cd is able to modify P. infestans pathogenicity, a disease index and molecular assessment of disease progress were analysed indicating that Cd stress enhanced aggressiveness of vr P. infestans towards various potato cultivars. Taken together, Cd not only affected hyphal growth rate and caused biochemical changes in P. infestans structures, but accelerated the pathogenicity as well. The nitro-oxidative homeostasis imbalance underlies the phytopathogen adaptive strategy and survival in the heavy metal contaminated environment.

Rhizoctonia solani AG 11 isolated for the first time from sugar beet in Poland.

Two isolates of Rhizoctonia solani AG11 were isolated from sugar beet seedlings from South-west Poland. Both isolates gave C2 reactions in anastomose pairings with the tester isolates of AG11. The membership of both isolates to AG11 was confirmed by analysis of pectic isozyme profiles, and by verification that the internal transcribed spacer sequences of both isolates matched the references in the GenBank database. Both AG11 isolates formed white-beige to creamy-colored mycelium with wide concentric zonation. One of them formed light-colored sclerotia. The average daily rate of hyphal growth at 21 °C was 22.8 mm and 22.6 mm on PDA. They were mildly pathogenic to sugar beet seedlings due to the mycelial and secondary metabolites’ activity. The sensitivity to fungicides typically used in sugar beet protection was different for each isolate; one of them (isolate ID11) was less sensitive to thiram than the other (isolate ID3). This article discusses the worldwide occurrence of R. solani AG11, expands the currently known host range, shows its broad world distribution in regions of moderate climate, and confirms the isolates’ low frequency.

In Vitro and In Vivo Effect of Poplar Bud Extracts on Phytophthora infestans: A New Effective Biological Method in Potato Late Blight Control.

The effect of populin extract from black poplar (Populus nigra) on seven different late blight strains was tested under laboratory and field conditions. The growth rate of hyphae was found to be significantly lower in vitro after 3 and 4 v/v% populin applications. Stain M16 was resistant to populin treatment under lab conditions, however. Both 5% and 10% concentration populin reduced the M16 strain’s severity on potato leaves under field conditions and proved to be even more effective than conventionally used fungicides Infinito 687 Sc and Valis M. Higher infection intensity at the 1% level was observed after 24 h using Valis M, and the same trend toward 10% infection remained after 48 and 72 h as well. Low, almost-no-infection intensity was detected after populin 5% and 10% treatment under an open field condition. Altogether, it can be concluded that populin extract can be a low-cost option for growers and an environmentally friendly approach in late blight control.

Probing Real-Time Hyphal Growth of Candida albicans using Atomic Force Microscopy: The Effect of Temperature

Candida albicans is an opportunistic human pathogen that colonizes many areas of the human body, including the gastrointestinal (GI) and genitourinary (GU) tracts of healthy individuals. Alterations in host immunity, the resident microbiota, or environmental factors such as increased pH, nutrient limitation, hypoxia and increased CO2 concentration can lead to C. albicans overgrowth, causing either mucosal diseases (e.g. oral thrush) or life-threatening infections. The ability of C. albicans to grow long filamentous hypha is critical for its pathogenic potential as it allows the fungus to invade the underlying substratum, enhances biofilm formation and enables macrophage evasion. In the present work, we introduce an atomic force microscopy (AFM) based method in order to investigate hyphal growth of C. albicans in real-time under physiologically relevant conditions on silicone elastomer substrates, the common material used for intravascular catheters. In particular, we found hyphal growth rates differ significantly for measurements performed at three different temperatures (35°C, 36°C, 37°C).

Multifunction of the ER P-Type Calcium Pump Spf1 During Hyphal Development in Candida albicans.

Candida albicans is one of the most important fungal pathogens. Hyphal development is required for the virulence of this pathogen. Our previous study has revealed that Spf1, an ER P-type calcium pump, plays an important role in hyphal development. However, the detailed mechanisms by which this protein functions in this process remain to be investigated. In this study, we found that loss of Spf1 led to decreased growth biomass under the hypha-inducing condition, suggesting a role of this protein in maintaining hyphal growth rate. Actin staining further revealed that the spf1Δ/Δ mutant showed attenuated tip-localization of actin patches and the defect in transport of both the chitin synthase Chs3 and the hypha-related factor Hwp1, implying that Spf1 functions in polarized growth of the hyphae by regulating actin organization and consequent polarized transport of morphogenesis-associated factors. Moreover, deletion of SPF1 led to abnormal vacuolar morphology under the hypha-inducing condition, which may also contribute to the defect of hyphal development in the spf1Δ/Δ mutant. This study revealed the pleiotropic role of Spf1-regulated calcium homeostasis in controlling hyphal development in C. albicans.