Molecular identification of endophytic fungi in lawn grass (Axonopus compressus) and their pathogenic ability.

Lawn grass (Axonopus compressus) is a widely distributed grass species from the family Poaceae that is ubiquitous in Malaysia. We isolated endophytic fungi from the leaves of A. compressus and molecularly identified them as Fusarium parceramosum, Colletotrichum siamense, C. gigasporum, C. endophyticum, Curvularia lunata, Stagonospora bicolor, Calonectria gracilis, and Albifimbria verrucari. These fungal endophytes are considered host generalists, as they have been isolated from other plants and have also been reported to be latent plant pathogens. We tested the pathogenicity of selected endophytic fungal isolates on A. compressus leaves, chili (Capsicum annum), and tomato (Solanum lycopersicum), and found that they were pathogenic to wounded A. compressus leaves with low to moderate virulence, and several were pathogenic to wounded and unwounded chili and tomato fruits. This indicated that the endophytes could infect both vegetable fruits with low to very high virulence. Pathogenicity tests demonstrated that endophytic fungi from the leaves of A. compressus can become pathogenic and infect the host and other plant species. The findings also indicated that leaves of A. compressus may harbor pathogens with latent ability that can become active due to changes in environmental conditions, thereby disrupting the balance between host-endophyte antagonism.

Basal Stem Rot of Oil Palm: The Pathogen, Disease Incidence, and Control Methods.

Basal stem rot of oil palm caused by Ganoderma boninense is the most serious disease of oil palm in Malaysia, Indonesia, and other oil-palm-producing countries. Economic losses caused by the disease can be up to USD500 million a year. For many years, basal stem rot was found to infect older palm trees of more than 25 to 30 years in age. Only in the 1950s, the disease began to appear in much younger palm trees, 10 to 15 years old, and, in the last decade or so, palm trees as young as 1 year were infected by the disease. The highest incidence occurs in coastal areas of Southeast Asia but the disease has now infected oil palm in inland areas, mainly oil palm planted in peat soils. Disease incidence is also high in areas previously growing coconut or forest. Basal stem rot infection and spread occur through root-to-root contact, and basidiospores that colonize the roots also play a role. In the early stages of infection by G. boninense, the pathogen behaves as a biotroph and later as a necrotroph, secreting cell-wall-degrading enzymes and triggering host defense responses. Genes, gene products, and metabolic pathways involved in oil palm defense mechanisms against G. boninense have been identified and these metabolites have the potential to be used as markers for early detection of the disease. Integrated disease management used to control basal stem rot includes cultural practices, chemical control, and application of biocontrol agents or fertilizers. Early detection tools have also been developed that could assist in management of basal stem rot infections. Development of resistant or tolerant oil palm is still at an early stage; therefore, the existing integrated disease management practices remain the most appropriate methods for managing basal stem rot of oil palm.

Prevalence, Identification and Mycotoxigenic Potential of Fungi in Common Spices Used in Local Malaysian Cuisines.

Spices are widely used in various cuisines in Malaysia to enhance the flavour and aroma. However, spices are susceptible to fungal infection, leading to mycotoxin contamination if the storage conditions are favourable for fungal growth. Thus, this study aimed to identify fungal species in spices commonly used in local Malaysian cuisines and determine their prevalence and mycotoxigenic potential. A total of 110 spice samples consisting of cumin, fennel, coriander, peppers (black pepper and white pepper), chillies (dried chilli, chilli paste and chilli powder), cinnamon, star anise, cloves, curry powder and korma powder were randomly purchased from retail markets in Penang. The samples were analysed for the total fungal count (ground spices) and the incidence of fungal infection (whole spices). The fungal species isolated from spices were identified based on morphological and molecular approaches, and the mycotoxigenic potential was determined using the Coconut Cream Agar method. The results showed that coriander seeds (ground) recorded the highest total fungal count (ADM 3.08 log CFU/g; DG18 3.14 log CFU/g), while black pepper (whole) recorded the highest incidence of fungal infection (94%). Interestingly, star anise and cloves were free from fungal contamination. The mycotoxigenic fungi of A. flavus and A. niger recorded the highest isolation frequency in ground and whole spices. These findings indicate the risk of mycotoxin exposure to consumers due to the high consumption of spices in local Malaysian cuisine.

Evaluation of the pathogenicity of endophytic fungi isolated from spines of rattan (Calamus castaneus) against other plant hosts.

AIMS: In this study, we sought to determine the pathogenic ability of endophytic fungi recovered from the spines of Calamus castaneus, a common rattan palm growing in the forests of Peninsula Malaysia. METHODS AND RESULTS: Ten endophytic fungal isolates were selected for pathogenicity tests: Colletotrichum boninense, Colletotrichum fructicola, Colletotrichum cliviae, Diaporthe hongkongensis, Diaporthe arengae, Diaporthe cf. nobilis, Neopestalotiopsis saprophytica, Neopestalotiopsis formicarum, Fusarium solani and Fusarium oxysporum. These endophytes were tested against leaves of bertam (Eugeissona sp.), oil palm (Elaeis guineensis) and mango (Mangiferae indica) and the fruits of chilli (Capsicum annum), tomato (Solanum lycopersicum) and banana (Musa acuminata). The fungal isolates showed infectivity against bertam, oil palm and mango leaves with degrees of virulence ranging from low to moderate, whereas infectivity against chilli, tomato and banana ranged from low to very high. CONCLUSIONS: Fungal endophytes isolated from the spines of C. castaneus are pathogenic to different crop plants with differing degrees of virulence or aggressiveness. SIGNIFICANCE AND IMPACT OF THE STUDY: Spines of C. castaneus can harbour fungal pathogens of a number of different crops as endophytes. The ability of the fungal endophytes to colonize and infect different crops demonstrate their importance towards agricultural crops. There is a possibility the endophytes behave as latent pathogen. When conditions become favourable, the fungal endophytes transform to pathogenic form and potentially infect other plants.

Morphology, Phenotype, and Molecular Identification of Clinical and Environmental Fusarium solani Species Complex Isolates from Malaysia.

Fusarium infections in humans (fusariosis) and in economically important plants involve species of several Fusarium species complexes. Species of the Fusarium solani species complex (FSSC) are the most frequent cause of human fusariosis. The FSSC comprises more than 60 closely related species that can be separated into three major clades by multi-locus sequence typing (MLST) using translation elongation factor 1-alpha (TEF1-α) and RNA polymerase II (RPB2) DNA sequences. The MLST nomenclature for clade 3 of the FSSC assigns numbers to species types (e.g., FSSC 2) and lowercase letters to identify unique haplotypes. The aim of this study was to analyse the genotypic and phenotypic characteristics of 15 environmental and 15 clinical FSSC isolates from Malaysia. MLST was used for the genotypic characterisation of FSSC isolates from various locations within Malaysia, which was complemented by their morphological characterisation on potato dextrose and carnation leaf agar. MLST identified eight different FSSC species: thirteen Fusarium keratoplasticum (i.e., FSSC 2), six Fusarium suttonianum (FSSC 20), five Fusarium falciforme (FSSC 3+4), two Fusarium cyanescens (FSSC 27), and one each of Fusarium petroliphilum (FSSC 1), Fusarium waltergamsii (FSSC 7), Fusarium sp. (FSSC 12), and Fusarium striatum (FSSC 21). Consistent with previous reports from Malaysia, most (11 of 15) clinical FSSC isolates were F. keratoplasticum and the majority (9 of 15) of environmental isolates were F. suttonianum (5) or F. falciforme (4) strains. The taxonomic relationships of the isolates were resolved phylogenetically. The eight Fusarium species also showed distinct morphological characteristics, but these were less clearly defined and reached across species boundaries. Although TEF1-α and RPB2 sequences were sufficient for the species identification of most FSSC isolates, a more precise MLST scheme needs to be established to reliably assign individual isolates of the species-rich FSSC to their geographically-, epidemiologically-, and host-associated sub-lineages.

Histopathology of Corn Plants Infected by Endophytic Fungi.

Endophytic fungi inhabiting plant tissues show extensive functional diversity, ranging from mutualism to pathogenicity. The present study evaluated the histological responses of corn plants to colonization by three species of endophytic fungi isolated from corn. Corn seedlings were inoculated with 1 × 106 conidia per mL spore suspensions of endophytic Fusarium verticillioides, Fusarium sacchari, and Penicillium citrinum and observed for 14 days for the emergence of disease symptoms. Histological examination of diseased root, stem, and leaf tissues was conducted using light and transmission electron microscopy. The results indicated that the mean diameters of root phloem, stem vascular bundles, and leaf vascular bundles, of corn plants infected with endophytic P. citrinum (18.91 µm, 146.96 µm, and 107.86 µm, respectively), F. verticillioides (18.75 µm, 85.45 µm, and 118.24 µm, respectively), and F. sacchari (24.15 µm root phloem, and 98.90 µm stem vascular bundle diameters), were significantly lower than the root phloem (33.68 µm), stem vascular bundle (186.77 µm), and leaf vascular bundle (155.88 µm) of the uninfected corn plants (p ≤ 0.05). Endophytic F. verticillioides was the most virulent, resulting in severe degradation and the eventual collapse of infected plant tissues. The study showed that endophytic fungi recovered from corn plants are capable of initiating significant disease responses in infected corn tissues.

Lasiodiplodia theobromae as a causal pathogen of leaf blight, stem canker, and pod rot of Theobroma cacao in Malaysia.

Symptoms of leaf blight, stem canker, and pod rot were observed on T. cacao during a series of samplings conducted in several states of Malaysia from September 2018 to March 2019. The identity of the pathogen that was responsible for the diseases was determined using morphological characteristics, DNA sequences, and phylogenetic analyses of multiple genes, namely, internal transcribed spacer (ITS), elongation translation factor 1-alpha (tef1-α), ß-tubulin (tub2), and RNA polymerase subunit II (rpb2). A total of 57 isolates recovered from diseased leaves of T. cacao (13 isolates), stems (20 isolates), and pods (24 isolates) showed morphological features that resembled Lasiodiplodia sp. The identity of the isolates was further determined up to the species level by comparing DNA sequences and phylogenetic analyses of multiple genes. The phylogenetic analysis of the combined dataset of ITS, tef1-α, tub2, and rpb2 elucidated that all of the isolates obtained were Lasiodiplodia theobromae as supported by 97% bootstrap value. The results of pathogenicity tests revealed L. theobromae as the causal pathogen of leaf blight, stem canker, and pod rot of T. cacao.

Molecular Phylogeny of Endophytic Fungi from Rattan (Calamus castaneus Griff.) Spines and Their Antagonistic Activities against Plant Pathogenic Fungi.

Calamus castaneus is a common rattan palm species in the tropical forests of Peninsular Malaysia and is noticeable by the yellow-based spines that cover the stems. This study aimed to determine the prevalence of fungal endophytes within C. castaneus spines and whether they inhibit the growth of fungal pathogens. Twenty-one genera with 40 species of fungal endophytes were isolated and identified from rattan palm spines. Based on molecular identification, the most common isolates recovered from the spines were Colletotrichum (n = 19) and Diaporthe spp. (n = 18), followed by Phyllosticta spp., Xylaria sp., Trichoderma spp., Helminthosporium spp., Penicillium spp., Fusarium spp., Neopestalotiopsis spp., Arthrinium sp., Cyphellophora sp., Cladosporium spp., Curvularia sp., Bionectria sp., and Acremonium spp. Non-sporulating fungi were also identified, namely Nemania primolutea, Pidoplitchkoviella terricola, Muyocopron laterale, Acrocalymma fici, Acrocalymma medicaginis, and Endomelanconiopsis endophytica. The isolation of these endophytes showed that the spines harbor endophytic fungi. Most of the fungal endophytes inhibited the growth of several plant pathogenic fungi, with 68% of the interactions resulting in mutual inhibition, producing a clear inhibition zone of <2 mm. Our findings demonstrate the potential of the fungal endophytes from C. castaneus spines as biocontrol agents.

Fusarium Species in Mangrove Soil in Northern Peninsular Malaysia and the Soil Physico-Chemical Properties.

Fusarium genus comprises important saprophytic and phytopathogenic fungi and is widespread in nature. The present study reports the occurrence of Fusarium spp. in soils from two mangrove forests in northern Peninsular Malaysia and analyzed physico-chemical properties of the mangrove soil. Based on TEF-1α sequences, nine Fusarium species were identified: Fusarium solani species complex (FSSC) (n = 77), Fusarium verticillioides (n = 20), Fusarium incarnatum (n = 10), Fusarium proliferatum (n = 7), Fusarium lateritium (n = 4), Fusarium oxysporum (n = 3), Fusarium rigidiuscula (n = 2), Fusarium chlamydosporum (n = 1), and Fusarium camptoceras (n = 1); FSSC isolates were the most prevalent. Phylogenetic analysis of the combined TEF-1α and ITS sequences revealed diverse phylogenetic affinities among the FSSC isolates and potentially new phylogenetic clades of FSSC. Soil analysis showed varied carbon content, pH, soil moisture, and salinity, but not nitrogen content, between sampling locations. Regardless of the physico-chemical properties, various Fusarium species were recovered from the mangrove soils. These were likely saprophytes; however, some were well-known plant pathogens and opportunistic human pathogens. Thus, mangrove soils might serve as inoculum sources for plant and human pathogenic Fusarium species. The present study demonstrates the occurrence of various Fusarium species in the extreme environment of mangrove soil, thereby contributing to the knowledge on species diversity in Fusarium.

Diaporthe species causing stem gray blight of red-fleshed dragon fruit (Hylocereus polyrhizus) in Malaysia.

This study aimed to characterize the new fungal disease on the stem of red-fleshed dragon fruit (Hylocereus polyrhizus) in Malaysia, which is known as gray blight through morphological, molecular and pathogenicity analyses. Nine fungal isolates were isolated from nine blighted stems of H. polyrhizus. Based on morphological characteristics, DNA sequences and phylogeny (ITS, TEF1-α, and ß-tubulin), the fungal isolates were identified as Diaporthe arecae, D. eugeniae, D. hongkongensis, D. phaseolorum, and D. tectonendophytica. Six isolates recovered from the Cameron Highlands, Pahang belonged to D. eugeniae (DF1 and DF3), D. hongkongensis (DF9), D. phaseolorum (DF2 and DF12), and D. tectonendophytica (DF7), whereas three isolates from Bukit Kor, Terengganu were recognized as D. arecae (DFP3), D. eugeniae (DFP4), and D. tectonendophytica (DFP2). Diaporthe eugeniae and D. tectonendophytica were found in both Pahang and Terengganu, D. phaseolorum and D. hongkongensis in Pahang, whereas D. arecae only in Terengganu. The role of the Diaporthe isolates in causing stem gray blight of H. polyrhizus was confirmed. To date, only D. phaseolorum has been previously reported on Hylocereus undatus. This is the first report on D. arecae, D. eugeniae, D. hongkongensis, D. phaseolorum, and D. tectonendophytica causing stem gray blight of H. polyrhizus worldwide.

Virulence Pattern of Pyricularia oryzae Pathotypes Towards Blast Monogenic Lines.

Rice blast caused by Pyricularia oryzae (P. oryzae) is one of the most serious diseases infecting rice worldwide. In the present study, virulence pattern of six P. oryzae pathotypes (P0.0, P0.2, P1.0, P3.0, P7.0 and P9.0) identified from the blast pathogen collected in Peninsular Malaysia, were evaluated using a set of 22 IRRI-bred blast resistance lines (IRBL) as well as to determine the resistance genes involved. The information on the virulence of the blast pathotypes and the resistance genes involved is important for breeding of new rice variety for durable resistance against blast disease. The IRBL was established from 22 monogenic lines, harbouring 22 resistance genes [Pia, Pib, Pii, Pit, Pi3, Pi5(t), Pish, Pi1, Pik, Pik-s, Pik-m, Pik-h, Pik-p, Pi7(t), Pi9, Piz, Piz-5, Piz-t, Pi19, Pi20(t), Pita-2, and Pita=Pi4(t)]. Based on the disease severity patterns, the tested pathotypes were avirulence towards seven IRBLs [IRBLi-F5, IRBLk-Ka, IRBLkh-K3, IRBLz-Fu, IRBLsh-S, IRBLPi7 (t) and IRBL9-W] of which these IRBLs harbouring Pii, Pik, Pik-h, Piz, Pish, Pi7(t) and Pi9 resistance genes, respectively. Therefore, the results suggested that the seven IRBLs carrying seven resistance genes [Pii, Pik, Pik-h, Piz, Pish, Pi7(t) and Pi9] would be suitable candidates of resistance genes to be incorporated in new breeding lines to combat the current blast pathotypes in the field.

Karah padi yang disebabkan oleh Pyricularia oryzae merupakan salah satu penyakit paling signifikan yang memberi kesan terhadap tanaman padi di seluruh dunia. Dalam kajian ini, corak kevirulenan enam patotip, P0.0, P0.2, P1.0, P3.0, P7.0 dan P9.0 yang dikenal pasti dari pencilan P. oryzae yang dikumpul dari Semenanjung Malaysia, dinilai menggunakan satu set 22 galur rintangan biakan karah-IRRI (IRBL) serta untuk menganggar gen rintangan yang terlibat. Maklumat mengenai kevirulenan patotaip karah dan gen rintangan terlibat adalah penting untuk pembiakbakaan varieti padi baru untuk kerintangan terhadap penyakit karah. IRBL dibangunkan dari 22 galur monogen, mengandungi 22 gen rintangan [Pia, Pib, Pii, Pit, Pi3, Pi5(t), Pish, Pi1, Pik, Pik-s, Pik-m, Pik-h, Pik-p, Pi7(t), Pi9, Piz, Piz-5, Piz-t, Pi19, Pi20(t), Pita-2, and Pita=Pi4(t)]. Berdasarkan corak tindakbalas penyakit, patotip yang diuji adalah avirulens terhadap tujuh IRBL [IRBLi-F5, IRBLk-Ka, IRBLkh-K3, IRBLz-Fu, IRBLsh-S, IRBLPi7(t) dan IRBL9-W] yang mengandungi gen rintangan Pii, Pik, Pik-h, Piz, Pish, Pi7(t) dan Pi9, masing-masing. Oleh itu, hasil kajian mencadangkan tujuh IRBL yang membawa tujuh gen rintangan [Pii, Pik, Pik-h, Piz, Pish, Pi7(t) dan Pi9] boleh menjadi calon gen rintangan yang sesuai untuk digabungkan ke dalam galur pembiakbakaan baharu untuk memerangi patotip karah di lapangan.

Characterisation and Pathogenicity of Aspergillus tamarii Causing Banana Fruit Rot.

Banana fruit rot is a common postharvest disease of the banana fruit. The appearance of rot symptoms on the surface of the fruits reduces the quality and marketability of banana. From rot lesions on banana fruits, three Aspergillus isolates were isolated. Based on morphological characteristics and sequences of Internal Transcribed Spacer, ß-tubulin and calmodulin, the isolates were identified as A. tamarii. Pathogenicity tests of the isolates, conducted using mycelial plugs with wounded and unwounded treatments, showed A. tamarii as the pathogen of banana fruit rot. Rot symptoms were highly severe on wounded banana fruits compared to unwounded fruits, and therefore, wounded banana fruits are more susceptible to A. tamarii infection. To the best of our knowledge, this is the first report of A. tamarii as a causal pathogen of banana fruit rot. This study indicated A. tamarii is one of postharvest rot pathogens of banana.

Reput buah pisang merupakan penyakit lepas tuai yang lazim pada buah pisang. Kemunculan gejala reput pada permukaan buah mengurangkan kualiti dan kebolehpasaran buah pisang. Dari lesi reput pada buah pisang, tiga pencilan Aspergillus telah dipencilkan. Berdasarkan ciri-ciri morfologi dan jujukan Trankripsi Penjarak Dalaman (ITS), ß-tubulin dan kalmodulin, pencilan tersebut dikenal pasti sebagai A. tamarii. Ujian kepatogenan yang dilakukan menggunakan palam miselium dengan rawatan luka dan tidak luka, menunjukkan A. tamarii merupakan patogen reput buah pisang. Gejala reput amat teruk pada buah pisang yang luka berbanding buah yang tidak luka. Oleh itu, buah pisang yang luka lebih mudah dijangkiti A. tamarii. Pada pengetahuan kami, ini adalah laporan pertama A. tamarii sebagai patogen penyebab reput buah pisang. Kajian ini menunjukkan A. tamarii merupakan salah satu patogen reput pisang lepas tuai.

A 23 bp cyp51A Promoter Deletion Associated With Voriconazole Resistance in Clinical and Environmental Isolates of Neocosmospora keratoplastica.

In the fungal pathogen Aspergillus fumigatus, resistance to azole antifungals is often linked to mutations in CYP51A, a gene that encodes the azole antifungal drug target lanosterol 14α-demethylase. The aim of this study was to investigate whether similar changes could be associated with azole resistance in a Malaysian Fusarium solani species complex (FSSC) isolate collection. Most (11 of 15) clinical FSSC isolates were Neocosmospora keratoplastica and the majority (6 of 10) of environmental isolates were Neocosmospora suttoniana strains. All 25 FSSC isolates had high minimum inhibitory concentrations (MICs) for itraconazole and posaconazole, low MICs for amphotericin B, and various (1 to >32 mg/l) voriconazole susceptibilities. There was a tight association between a 23 bp CYP51A promoter deletion and high (>32 mg/l) voriconazole MICs; of 19 FSSC strains sequenced, nine isolates had voriconazole MICs > 32 mg/l, and they all contained the 23 bp CYP51A promoter deletion, although it was absent in the ten remaining isolates with low (≤12 mg/l) voriconazole MICs. Surprisingly, this association between voriconazole resistance and the 23 bp CYP51A promoter deletion held true across species boundaries. It was randomly distributed within and across species boundaries and both types of FSSC isolates were found among environmental and clinical isolates. Three randomly selected N. keratoplastica isolates with low (≤8 mg/l) voriconazole MICs had significantly lower (1.3-7.5 times) CYP51A mRNA expression levels than three randomly selected N. keratoplastica isolates with high (>32 mg/l) voriconazole MICs. CYP51A expression levels, however, were equally strongly induced (~6,500-fold) by voriconazole in two representative strains reaching levels, after 80 min of induction, that were comparable to those of CYP51B. Our results suggest that FSSC isolates with high voriconazole MICs have a 23 bp CYP51A promoter deletion that provides a potentially useful marker for voriconazole resistance in FSSC isolates. Early detection of possible voriconazole resistance is critical for choosing the correct treatment option for patients with invasive fusariosis.

Identification and Characterization of Macrophomina phaseolina Causing Leaf Blight on White Spider Lilies (Crinum asiaticum and Hymenocallis littoralis) in Malaysia.

Crinum asiaticum and Hymenocallis littoralis, commonly known as spider lilies are bulbous perennial and herbaceous plants that widely planted in Malaysia as ornamental. During 2015-2016, symptom of leaf blight was noticed on the hosts from several locations in Penang. The symptom appeared as irregular brown to reddish lesions surrounded by yellow halos. As the disease progressed, the infected leaves became blighted, dried, and fell off with the presence of black microsclerotia and pycnidia on the lesions parts. The present study was conducted to investigate the causal pathogen of leaf blight on C. asiaticum and H. littoralis. Based on morphological characteristics and DNA sequences of internal transcribed spacer (ITS) region and translation elongation factor 1-alpha (TEF1-α) gene, the causal pathogen was identified as Macrophomina phaseolina. Phylogenetic analysis of combined dataset of ITS and TEF1-α grouped the isolates studied with other isolates of M. phaseolina from GenBank. The grouping of the isolates was supported by 96% bootstrap value. Pathogenicity test proved the role of the fungus in causing leaf blight on both hosts.

Molecular Identification of Endophytic Fungi from Banana Leaves (Musa spp.).

Endophytic fungi are part of microbial community found in various types of plant tissues including the leave, and display a range of symbiotic interactions with the plant host. In this study, endophytic fungi isolated from banana leaves were identified using ITS (Internal Transcribed Spacer region) sequences of which 10 genera comprising 17 species were molecularly identified. Endophytic fungal species identified were Nigrospora oryzae, Nigrospora sphaerica, Colletotrichum gloeosporioides, Colletotrichum siamense, Fusarium equiseti, Fusarium chlamydosporum, Phoma sorghina, Pestalotiopsis oxyanthi, Pestalotiopsis theae, Pestalotiopsis eugeniae, Penicillium steckii, Penicillium purpurogenum, Bipolaris papendorfii, Bipolaris sp., Lasidiodiplodia theobromae, Cochliobolus intermedius dan Aspergillus niger. The present study showed that several endophytic fungal genera/species are common plant pathogen and there is a possibility that these endophytes can become pathogenic. Some of the fungal endophyte might be mutualist or saprophyte.

Characterization and pathogenicity of Fusarium species associated with leaf spot of mango (Mangifera indica L.).

Leaf spot diseases are mainly caused by fungi including Fusarium. In the present study several species of Fusarium were isolated from the leaf spot lesion of mango (Mangifera indica L.) Based on morphological characteristics, TEF-1α sequences and phylogenetic analysis, five species were identified as F. proliferatum, F. semitectum, F. mangiferae, F. solani and F. chlamydosporum. Pathogenicity test indicated that representative isolates of F. proliferatum, F. semitectum and F. chlamydosporum were pathogenic on mango leaves causing leaf spot with low to moderate virulence. Nevertheless, abundance of spots on the leaf can disrupt photosynthesis which in turn reduced growth, and lead to susceptibility to infection by opportunistic pathogens due to weakening of the plant. Fusarium solani and F. mangiferae were non-pathogenic and it is possible that both species are saprophyte which associated with nutrient availability on the surface of the leaf through decaying leave tissues. The occurrence of Fusarium spp. on the leaf spot lesion and the effect from the disease needs to be considered when developing disease management method of mango cultivation as numerous spot on the leaves could effect the photosynthesis process and finally giving low yield and less quality of mango.

Endophytic Diaporthe sp. ED2 Produces a Novel Anti-Candidal Ketone Derivative.

This study aimed to examine the anti-candidal efficacy of a novel ketone derivative isolated from Diaporthe sp. ED2, an endophytic fungus residing in medicinal herb Orthosiphon stamieus Benth. The ethyl acetate extract of the fungal culture was separated by open column and reverse phase high-performance liquid chromatography (HPLC). The eluent at retention time 5.64 min in the HPLC system was the only compound that exhibited anti-candidal activity on Kirby-Bauer assay. The structure of the compound was also elucidated by nuclear magnetic resonance and spectroscopy techniques. The purified anti-candidal compound was obtainedas a colorless solid and characterized as 3-hydroxy-5-methoxyhex-5-ene-2,4-dione. On broth microdilution assay, the compound also exhibited fungicidal activity on a clinical strain of Candida albicans at a minimal inhibitory concentration of 3.1 µg/ml. The killing kinetic analysis also revealed that the compound was fungicidal against C. albicans in a concentration- and time-dependent manner. The compound was heat-stable up to 70°C, but its anti-candidal activity was affected at pH 2.

Saprophytic and Potentially Pathogenic Fusarium Species from Peat Soil in Perak and Pahang.

Isolates of Fusarium were discovered in peat soil samples collected from peat swamp forest, waterlogged peat soil, and peat soil from oil palm plantations. Morphological characteristics were used to tentatively identify the isolates, and species confirmation was based on the sequence of translation elongation factor-1α (TEF-1α) and phylogenetic analysis. Based on the closest match of Basic Local Alignment Search Tool (BLAST) searches against the GenBank and Fusarium-ID databases, five Fusarium species were identified, namely F. oxysporum (60%), F. solani (23%), F. proliferatum (14%), F. semitectum (1%), and F. verticillioides (1%). From a neighbour-joining tree of combined TEF-1α and ß-tubulin sequences, isolates from the same species were clustered in the same clade, though intraspecies variations were observed from the phylogenetic analysis. The Fusarium species isolated in the present study are soil inhabitants and are widely distributed worldwide. These species can act as saprophytes and decomposers as well as plant pathogens. The presence of Fusarium species in peat soils suggested that peat soils could be a reservoir of plant pathogens, as well-known plant pathogenic species such F. oxysporum, F. solani, F. proliferatum, and F. verticillioides were identified. The results of the present study provide knowledge on the survival and distribution of Fusarium species.

Molecular Characterisation of Endophytic Fungi from Roots of Wild Banana (Musa acuminata).

Endophytic fungi inhabit apparently healthy plant tissues and are prevalent in terrestrial plants, especially root tissues, which harbour a wide assemblage of fungal endophytes. Therefore, this study focused on the isolation and characterisation of endophytic fungi from the roots of wild banana (Musa acuminata). A total of 31 isolates of endophytic fungi were isolated from 80 root fragments. The endophytic fungi were initially sorted according to morphological characteristics and identified using the sequences of the translation elongation factor-1α (TEF-1α) gene of Fusarium spp. and the Internal Transcribed Spacer (ITS) regions of other fungi. The most common fungal isolates were species of the genus Fusarium, which were identified as F. proliferatum, Fusarium sp., F. solani species complex, and F. oxysporum. Other isolated endophytic fungi included Curvularia lunata, Trichoderma atroviride, Calonectria gracilis, Rhizoctonia solani, Bionectria ochroleuca, and Stromatoneurospora phoenix (Xylariceae). Several of the fungal genera, such as Fusarium, Trichoderma, Rhizoctonia, and Xylariceae, are among the common fungal endophytes reported in plants. This study showed that the roots of wild banana harbour a diverse group of endophytic fungi.

Effects of fusaric acid treatment on the protocorm-like bodies of Dendrobium sonia-28.

Dendrobium sonia-28 is a popular orchid hybrid due to its flowering recurrence and dense inflorescences. Unfortunately, it is being decimated by fungal diseases, especially those caused by Fusarium proliferatum. In this study, selection of F. proliferatum-tolerant protocorm-like bodies (PLBs) was carried out by assessing the effects of differing concentrations of fusaric acid (FA). PLBs were cultured on Murashige and Skoog (MS) medium supplemented with 0.05 to 0.2 millimolar (mM) concentrations of FA. Higher concentrations of FA increased mortality of PLBs and reduced their growth. The survival rate for 0.05 mM FA was 20 % but only 1 % at the highest dose of 0.2 mM. Additionally, two different size ranges of PLBs were investigated, and growth increased more at lower FA concentrations for larger PLBs, whilst the growth rate of smaller PLBs was inhibited at an FA concentration of 0.2 mM. Histological examination using transmission electron microscopy (TEM) and scanning electron microscopy (SEM) analyses disclosed severe cell wall and organelle damage, as well as stomatal closure in PLBs treated with the high FA concentrations. Reductions in plantlet growth were much greater at the highest concentrations of FA. Some randomly amplified polymorphic DNA (RAPD) markers clearly discriminated between selected and non-selected variants of Dendrobium sonia-28, showing different banding patterns for each FA concentration and specific bands for selected and control plants.