Basal stem rot (BSR) of oil palm roots is due to the invasion of fungal mycelia of Ganoderma species which spreads to the bole of the stem. In addition to root contact, BSR can also spread by airborne basidiospores. These fungi are able to break down cell wall components including lignin. BSR not only decreases oil yield, it also causes the stands to collapse thus causing severe economic loss to the oil palm industry. The transmission and mode of action of Ganoderma, its interactions with oil palm as a hemibiotroph, and the molecular defence responses of oil palm to the infection of Ganoderma boninense in BSR are reviewed, based on the transcript profiles of infected oil palms. The knowledge gaps that need to be filled in oil palm-Ganoderma molecular interactions i.e. the associations of hypersensitive reaction (HR)-induced cell death and reactive oxygen species (ROS) kinetics to the susceptibility of oil palm to Ganoderma spp., the interactions of phytohormones (salicylate, jasmonate and ethylene) at early and late stages of BSR, and cell wall strengthening through increased production of guaiacyl (G)-type lignin, are also discussed.
Basidiospores were isolated from the fruiting bodies of Ganoderma infecting oil palms from an estate in Johor and from ornamental palms (including oil palms) from Singapore. The spores were then germinated to obtain homokaryotic mycelia. Based on clamp connection formation in paired hyphal fusions, tester strains were identified from the homokaryons isolated. Compatibility tests were then carried out using these testers to determine the relatedness of the homokaryotic Ganoderma isolates, both from Johor and from Singapore. Results from the compatibility tests showed that Ganoderma from both locations belong to the same species, while the Ganoderma isolates from Singapore share some common alleles. The pathogenicity tests carried out on Chrysalidocarpus lutescens seedlings using inoculum growing on rubber wood blocks showed that dikaryotic mycelia can cause basal stem rot infection.
In 1994 Corner published five new species within the genus Psathyrella, all having been collected on the Malay Peninsula between 1929 and 1930. Three of these species belong to the genus Hebeloma and with their vinaceous colored lamellae and spore print, when fresh, they belong to H. sect. Porphyrospora. Of these three species, only one, P. flavidifolia, was validly published and thus we herewith recombine it as H. flavidifolium. The other two species, P. splendens and P. verrucispora, are synonyms of H. parvisporum and H. lactariolens, respectively. We also describe a new Malayan species, H. radicans, which also belongs to H. sect. Porphyrospora. These findings confirm the western Pacific Rim as a diversity hotspot for H. sect. Porphyrospora. The records described within this paper, represent the first recognition that the genus Hebeloma, and indeed that members of the ectomycorrhizal Hymenogastraceae, are present on the Malay Peninsula.
Oil palm empty fruit bunches (EFB) were subjected to microbial pre-treatment of lignocellulosic biomass bioconversion to fermentable sugar. Microbial pre-treatment was carried out by inoculating Ganoderma boninense spores through solid state fermentation. The samples were initially treated with Sulphuric acid method prior to reading with UV-Visible Spectrometer. The readings were taken before and after inoculation of EFB with G. boninense. Bioconversion of 20 g EFB via solid state fermentation was done in five different amounts of G. boninense spore namely 0.0 g (control), 0.5 g (T2), 0.7 g (T3), 0.9 g (T4) and 1.1 g (T5) in 7 days. The result shows the highest delignification in sample inoculated with 1.1g of G. boninense spores, in which the spores are successfully reduced by 61.97% of lignin from total EFB biomass in 7 days compared to 60.08% (T4), 58.65% (T3) and 54.85% (T2). Meanwhile, for control the lignin content was reduced by 5.07% in 7 days. The study shows that G. boninense has the ability to remove lignin from EFB whereby longer incubation period and higher number of spores contribute to higher delignification percentage.
The present study aimed to assess the efficiency of silver bio-nanoparticles (Ag-NPs) in inactivating of the Aspergillus fumigatus, A. parasiticus and A. flavus var. columnaris and A. aculeatus spores. The AgNPs were synthesized in secondary metabolic products of Penicillium pedernalens 604 EAN. The inactivation process was optimized by response surface methodology (RSM) as a function of Ag NPs volume (1-10 μL/mL); time (10-120 min); pH (5-8); initial fungal concentrations (log10) (3-6). The artificial neural network (ANN) model was used to understand the behavior of spores for the factors affecting inactivation process. The best conditions to achieved SAL 10-6 of the fungal spores were recorded with 3.46 μl/mL of AgNPs, after 120 min at pH 5 and with 6 log of initial fungal spore concentrations, at which 5.99 vs. 6.09 (SAL 10-6) log reduction was recorded in actual and predicted results respectively with coefficient of 87.00%. The ANN revealed that the timehas major contribution in the inactivation process compare to Ag NPs volume. The fungal spores were totally inactivated (SAL 10-6, 6 log reduction with 99.9999%) after 110 min of the inactivation process, 10 min more was required to insure the irreversible inactivation of the fungal spores. The absence of protease and cellulase enzymes production confirm the total inactivation of the fungal spores. FESEM analysis revealed that the AgNPs which penetrated the fungal spores leading to damage and deform the fungal spore morphology. The AFM analysis confirmed the total spore surface damage. The bands in the range of the Raman spectroscopy from 1300 to 1600 cm-1 in the inactivated spores indicate the presence of CH3, CH2 and the deformation of lipids released outside the spore cytoplasm. These finding indicate that the AgNPs has high potential as a green alternative inactivation process for the airborne fungal spores.
Global trade increases plant introductions, but joint introduction of associated microbes is overlooked. We analyzed the ectomycorrhizal fungi of a Caribbean beach tree, seagrape (Coccoloba uvifera, Polygonacaeae), introduced pantropically to stabilize coastal soils and produce edible fruits. Seagrape displays a limited symbiont diversity in the Caribbean. In five regions of introduction (Brazil, Japan, Malaysia, Réunion and Senegal), molecular barcoding showed that seagrape mostly or exclusively associates with Scleroderma species (Basidiomycota) that were hitherto only known from Caribbean seagrape stands. An unknown Scleroderma species dominates in Brazil, Japan and Malaysia, while Scleroderma bermudense exclusively occurs in Réunion and Senegal. Population genetics analysis of S. bermudense did not detect any demographic bottleneck associated with a possible founder effect, but fungal populations from regions where seagrape is introduced are little differentiated from the Caribbean ones, separated by thousands of kilometers, consistently with relatively recent introduction. Moreover, dry seagrape fruits carry Scleroderma spores, probably because, when drying on beach sand, they aggregate spores from the spore bank accumulated by semi-hypogeous Scleroderma sporocarps. Aggregated spores inoculate seedlings, and their abundance may limit the founder effect after seagrape introduction. This rare pseudo-vertical transmission of mycorrhizal fungi likely contributed to efficient and repeated seagrape/Scleroderma co-introductions.
Hydrophobins are small secreted fungal proteins that play roles in growth and development of filamentous fungi, i.e. in the formation of aerial structures and the attachment of hyphae to hydrophobic surfaces. In Botrytis cinerea, three hydrophobin genes have been identified. Studies by Mosbach et al. (2011) showed that hydrophobins are neither involved in conferring surface hydrophobicity to conidia and aerial hyphae of B. cinerea, nor are they required for virulence. The present study investigated the role of hydrophobins in sclerotium and apothecium development. Expression analysis revealed high expression of the Bhp1 gene during different stages of apothecium development. Two Bhp1 splice variants were detected that differ by an internal stretch of 13 amino acid residues. Seven different mutants in which either a single, two or three hydrophobin genes were knocked out, as well as two wild type strains of opposite mating types, were characterized for sclerotium and apothecium development. No aberrant morphology was observed in sclerotium development when single deletion mutants in hydrophobin genes were analyzed. Sclerotia of double knock out mutant ΔBhp1/ΔBhp3 and the triple knock out mutant, however, showed easily wettable phenotypes. For analyzing apothecium development, a reciprocal crossing scheme was setup. Morphological aberrations were observed in crosses with two hydrophobin mutants. When the double knock out mutant ΔBhp1/ΔBhp2 and the triple knock out mutant were used as the maternal parent (sclerotia), and fertilized with wild type microconidia, the resulting apothecia were swollen, dark brown in color and had a blotched surface. After initially growing upwards toward the light source, the apothecia in many cases collapsed due to loss of structural integrity. Aberrant apothecium development was not observed in the reciprocal cross, when these same mutants were used as the paternal parent (microconidia). These results indicate that the presence of hydrophobins in maternal tissue is important for normal development of apothecia of B. cinerea.
Two new loci found in one strain of Neurospora crassa (P2604) collected in Malaya are related to the meiotic drive system Spore killer Sk-2. Sk-2 was found in Neurospora intermedia and introgressed into N. crassa. P2604 showed high resistance to killing when crossed to Sk-2. This resistance was found to be linked to, but not allelic to, resistance locus r(Sk-2) on LGIIIL. Analysis showed that the high resistance phenotype of P2604 requires resistance alleles at two different loci on LGIIIR. Strains carrying a resistance allele at only the proximal or the distal locus, respectively, were obtained and intercrossed. Highly resistant strains were obtained by rejoining the two genes. The proximal locus alone confers a low level of resistance. This locus was named pr(Sk-2) for partial resistance to Sk-2. The distal locus was named mod(pr) because its only known phenotype is to modify pr(Sk-2).
Aerosampling using Rotorod samplers was conducted in the Institute for Medical Research, Kuala Lumpur, Malaysia, from December 1991 to November 1993. Samples were collected twice a week between 10.00 hours to 12.00 hours. Rods were stained and examined microscopically. A total of 8 and 20 types of pollens and mold spores were collected, respectively. More mold spores were collected than pollens. Grass pollen constituted more than 40 percent of total pollen counts. Gramineae pollen counts peaked in March and September. The most abundant mold spore was Cladosporium followed by Rust, Nigrospora, Curvularia and Smut. Cladosporium counts peaked in February and August. Rust counts peaked in June and December whereas counts for Nigrospora peaked in February and October. Highest counts of Smut were recorded in March and October. Curvularia counts peaked in January, June and September.
Due to problems with chemical control, there is increasing interest in the use of microsporidia for control of lepidopteran pests. However, there have been few studies to evaluate the susceptibility of exotic species to microsporidia from indigenous Lepidoptera.
Many chilli producers in Malaysia are facing a huge problem of anthracnose disease that infect the chilli fruits either at pre- or post-harvest stage. Due to this infection, the chilli fruits become rotten and could be colonized by grey conidial masses which is unfit for human consumption and cannot be sold. This study was aimed to isolate the fungal species associated with anthracnose symptoms collected from several mini-markets near Gong Badak, Terengganu. Pathogenicity test was conducted for all these isolates to evaluate their pathogenic level. Based on the results, five fungal isolates were successfully isolated and morphologically identified as Colletotrichum spp. All these isolates were labelled as UMTT46C, UMTT48C, UMTT50C, UMTT52C and UMTT54C. From these, fungal isolate (UMTT54C) showed the highest pathogenic level with disease severity (DS), 68.9%. Other Colletotrichum spp. recorded percentage of DS ranged from 53.3% to 60%. Identification of fungal isolates and their pathogenic level from the infected chilli is very important to assist in control strategy which can further provide a good protection measure for many chilli producers.
Fungal endophytes are found inside host plants but do not produce any noticeable disease symptoms in their host. In the present study, endophytic Fusarium species were isolated from roots of lawn grass (Axonopus compressus). A total of 51 isolates were recovered from 100 root segments. Two Fusarium species, F. oxysporum (53%) and F. solani (47%), were identified based on macroconidia and conidiogenous cell morphology. The detection of endophytic F. oxysporum and F. solani in the roots of lawn grass contributes to the knowledge of both the distribution of the two Fusarium species and the importance of roots as endophytic niches for Fusarium species.
Routine diagnosis of intestinal microsporidiosis in clinical diagnostic laboratories relies mostly on detection of microsporidial spores via special staining and microscopic techniques. This paper describes the comparative evaluation of Calcofluor White M2R method, with modified Gram-chromotrope Kinyoun method as the reference standard. One hundred and six stool samples were examined for the presence of microsporidial spores. Sensitivity, specificity, positive and negative predictive values of the Calcofluor White M2R method compared to the reference technique were 95.2%, 4.3%, 78.2% and 20.0%, respectively. The positive predictive value (PPV) was 78.2% and the negative predictive value (NPV) was 20.0%. Despite low specificity of the CFW method due to its ability to stain chitinous wall of microorganisms, the presence of distinct deep-blue horizontal or equitorial stripes in microsporidial spores in modified Gram-chromotrope Kinyoun would likely reduce the false positive results obtained in the Calcofluor White M2R. Hence, the simultaneous use of these two methods would give better performance and accuracy for the detection of microsporidial spores in patients with intestinal microsporidiosis.
Anthracnose caused by Colletotrichum species is a common postharvest disease of banana fruit. We investigated and identified Colletotrichum species associated with anthracnose in several local banana cultivars based on morphological characteristics and sequencing of ITS regions and of the β-tubulin gene. Thirty-eight Colletotrichum isolates were encountered in anthracnose lesions of five local banana cultivars, 'berangan', 'mas', 'awak', 'rastali', and 'nangka'. Based on morphological characteristics, 32 isolates were identified as Colletotrichum gloeosporioides and 6 isolates as C. musae. C. gloeosporioides isolates were divided into two morphotypes, with differences in colony color, shape of the conidia and growth rate. Based on ITS regions and β-tubulin sequences, 35 of the isolates were identified as C. gloeosporioides and only 3 isolates as C. musae; the percentage of similarity from BLAST ranged from 95-100% for ITS regions and 97-100% for β-tubulin. C. gloeosporioides isolates were more prevalent compared to C. musae. This is the first record of C. gloeosporioides associated with banana anthracnose in Malaysia. In a phylogenetic analysis of the combined dataset of ITS regions and β-tubulin using a maximum likelihood method, C. gloeosporioides and C. musae isolates were clearly separated into two groups. We concluded that C. gloeosporioides and C. musae isolates are associated with anthracnose in the local banana cultivars and that C. gloeosporioides is more prevalent than C. musae.
The protease activity of different isolates of dermatophytes representing different ecological groups namely geophilic, zoopahilic and anthropophilic, in their vegetative and sporulation growth phases were compared. Unlike their geophilic and zoophilic counterparts, all the isolates of anthropophilic dermatophytes viz. Trichophyton rubrum, T. mentagrophytes, T. tonsurans, T. violaceum and Epidermophyton floccosum recorded reduced protease activity during artificially induced sporulation phase in comparison to their vegetative growth phase. Even among the anthropophilic group, a classical moderation of protease activity was recorded in Trichyphyton rubrum which also correlates to its clinical manifestation. This enzyme moderation could also be an evolutionary adaptation of the anthropization of these species.
Matched MeSH terms: Spores, Fungal/enzymology; Spores, Fungal/growth & development
Morphological features and Inter Simple Sequence Repeat (ISSR) polymorphism were employed to analyse 21 Corynespora cassiicola isolates obtained from a number of Hevea clones grown in rubber plantations in Malaysia. The C. cassiicola isolates used in this study were collected from several states in Malaysia from 1998 to 2005. The morphology of the isolates was characteristic of that previously described for C. cassiicola. Variations in colony and conidial morphology were observed not only among isolates but also within a single isolate with no inclination to either clonal or geographical origin of the isolates. ISSR analysis delineated the isolates into two distinct clusters. The dendrogram created from UPGMA analysis based on Nei and Li's coefficient (calculated from the binary matrix data of 106 amplified DNA bands generated from 8 ISSR primers) showed that cluster 1 encompasses 12 isolates from the states of Johor and Selangor (this cluster was further split into 2 sub clusters (1A, 1B), sub cluster 1B consists of a unique isolate, CKT05D); while cluster 2 comprises of 9 isolates that were obtained from the other states. Detached leaf assay performed on selected Hevea clones showed that the pathogenicity of representative isolates from cluster 1 (with the exception of CKT05D) resembled that of race 1; and isolates in cluster 2 showed pathogenicity similar to race 2 of the fungus that was previously identified in Malaysia. The isolate CKT05D from sub cluster 1B showed pathogenicity dissimilar to either race 1 or race 2.
Matched MeSH terms: Spores, Fungal/genetics; Spores, Fungal/growth & development
Three polycentric rumen fungi, LL, LC2 and Ruminomyces elegans (C2), isolated from the rumen of cattle were grown in six culture media. LL and LC2 were morphologically similar. Their characteristics resembled those of Orpinomyces and Neocallimastix joyonii, and they grew well and produced sporangia after 3-4 d growth in all the media. R. elegans differed morphologically from LL and LC2, but although it also grew well in all media, abundant sporangia occurred only after 2-3 d growth in media containing cellulose. Undifferentiated sporangia were produced by all three isolates; differentiation of the sporangia did not occur in the spent growth media. However, if thalli possessing recently-formed sporangia were transferred to, or flooded with, fresh liquid medium or rumen fluid, zoosporogenesis and liberation of zoospores occurred within 17-20 min for isolates LL and LC2 and 30 min for R. elegans. Procedures for inducing zoosporogenesis by polycentric anaerobic fungi are described.
The potential for using agricultural and industrial by-products as substrate for the production of the edible mushroom, Auricularia polytricha, was evaluated using several formulations of selected palm oil wastes mixed with sawdust and further supplemented with selected nitrogen sources. The best substrate formulations were sawdust (SD) mixed with oil palm frond (OPF; 90:10) added with 15% spent grain (SG) and sawdust mixed with empty fruit bunch (EFB; 50:50) added with 10% spent grain (SG) with mycelia growth rate of 8 mm/day and 7 mm/day respectively. These two substrate formulations were then subjected to different moisture content levels (65%, 75% and 85%). Highest total fresh sporophore yield at 0.43% was obtained on SD+OPF (90:10)+15% SG at 85% moisture content, followed closely by SD+EFB (50:50)+10% SG with 0.40% total yield, also at 85% moisture content. Each of the substrate formulations at 85% moisture content gave the highest biological efficiency (BE) at 288.9% and 260.7%, respectively. Both yield and biological efficiency of A. polytricha on these two formulations were almost three times higher when compared to sawdust substrate alone, thus proving the potential of these formulations to improve yield of this mushroom.
Ganoderma boninense, a phytopathogenic white rot fungus had sought minimal genetic characterizations despite huge biotechnological potentials. Thus, efficient collection of fruiting body, basidiospore and protoplast of G. boninense is described. Matured basidiocarp raised under the glasshouse conditions yielded a total of 8.3 × 104 basidiospores/ml using the low speed centrifugation technique. Mycelium aged 3-day-old treated under an incubation period of 3 h in lysing enzyme from Trichoderma harzianum (10 mg/ml) suspended in osmotic stabilizer (0.6 M potassium chloride and 20 mM dipotassium phosphate buffer) yielded the highest number of viable protoplasts (8.9 × 106 single colonies) among all possible combinations tested (regeneration media, age of mycelium, osmotic stabilizer, digestive enzyme and incubation period).