Boonsom Bussaban

Diversity of saprobic microfungi

The data needed to derive an accurate estimate of saprobic microfungi are insufficient, incomplete and contradictory. We therefore address issues that will ultimately reveal whether there are 1.5 million global fungal species, which is the generally accepted working estimate. Our data indicates that large numbers of fungi occur on host families, such as Musaceae, host genera such as Nothofagus and individual host species such as Eucalyptus globulus, and that fungi may be specific or recurrent on different plant groups. Recent studies have shown that fungal numbers on hosts may be larger than originally thought as saprobes are organ-specific/-recurrent and changes in fungal communities occur as substrata decays. Other issues, such as the impact of geography, of methodology and of taxonomy are also addressed. There is evidence that fungi on the same host at different locations also differs; site-specific factors and geographic distance may be more important than host/substrate in shaping fungal assemblages. Methodology impacts on estimates of species diversity with many more taxa observed using indirect isolation protocols as compared to direct isolations from leaves. Our understanding of fungal species numbers in speciose genera is important. In some fungal groups accepted species have been reduced to a few species, while in other groups many cryptic species are being uncovered. While we make a number of generalisations from the studies reported here, this review also highlights some of the limitations mycologists currently have to contend with. A large body of knowledge exists for certain groups of microfungi or for microfungi occurring on certain substrata/hosts. However, it is likely that we are drawing conclusions from data that are somewhat biased toward fungi and host/substrata that are of interest to human endeavours. The discrepancy between the numbers of fungi described from only one economically important genus, Eucalyptus, and all the other members of the Myrtaceae is but one example of this bias. By incorporating the large body of work that is already available and adding appropriate complementary studies, we can accelerate our understanding of microfungal diversity and this will eventually lead us to a realistic estimate of global fungal species numbers.

Metabolites from the mangrove-derived fungus Xylaria cubensis PSU-MA34

Two new succinic acid derivatives, xylacinic acids A (1) and B (2), along with seven known compounds, including one succinic acid derivative (3), three mellein derivatives (4–6), cytochalasin D (7), 2-chloro-5-methoxy-3-methylcyclohexa-2,5-diene-1,4-dione (8) and isosclerone (9), were isolated from the mangrove-derived fungus Xylaria cubensis PSU-MA34. Their structures were established by spectroscopic evidence. They were evaluated for cytotoxicity against KB cells and antibacterial activity against Staphylococcus aureus ATCC 25923 and methicillin-resistant S. aureus.

Molecular and morphological evidence support four new species in the genus Muscodor from northern Thailand

The genus Muscodor comprises fungal endophytes which produce mixtures of volatile compounds (VOCs) with antimicrobial activities. In the present study, four novel species, Muscodor musae, M. oryzae, M. suthepensis and M. equiseti were isolated from Musa acuminata, Oryza rufipogon, Cinnamomum bejolghota and Equisetum debile, respectively; these are medicinal plants of northern Thailand. The new Muscodor species are distinguished based on morphological and physiological characteristics and on molecular analysis of ITS-rDNA. Volatile compound analysis showed that 2-methylpropanoic acid was the main VOCs produced by M. musae, M. suthepensis and M. equiseti. The mixed volatiles from each fungus showed in vitro antimicrobial activity. Muscodor suthepensis had the highest antifungal activity.

Isolation, structure elucidation and biological activity of metabolites from Sch-642305-producing endophytic fungus Phomopsis sp. CMU-LMA.

Eight polyketide compounds were isolated from the cultivation broth of Phomopsis sp. CMU-LMA. We have recently described LMA-P1, a bicyclic 10-membered macrolide, obtained as a bioconversion derivative of Sch-642305, the major compound isolated in this study. Benquinol is the ethyl ester derivative of the 13-dihydroxytetradeca-2,4,8-trienoic acid produced by Valsa ambiens. This compound is concomitantly produced with the 6,13-dihydroxytetradeca-2,4,8-trienoic acid (DHTTA) previously isolated from Mycosphaerellarubella. The absolute configuration of the new compound, (2R,3R,4S,5R)-3-hydroxy-2,4-dimethyl-5-[(S,Z)-3-methylpentenyl]-tetrahydro-pyranone LMA-P2 was confirmed by X-ray crystallography. The δ-lactone 2,3-dihydroxytetradecan-5-olide (DHTO) was previously isolated from Seiridium unicorne. This compound may form through the cyclization of the methyl-2,3,5-trihydroxytridecanoate LMA-P3, a new linear polyketide isolated in this study. Benquoine, a new 14-membered lactone generated from the cyclization of benquinol, is proposed as the key precursor for the biosynthesis of Sch-642305. Antimicrobial activity and cancer cell viability inhibition by the new compounds were investigated. Benquoine exhibits antimicrobial activity against Gram positive bacteria, and cytotoxicity against HCT-116 cancer cell line.

Three new species of Pyricularia are isolated as zingiberaceous endophytes from Thailand

Pyricularia costina and three undescribed Pyricularia species were found as endophytes on wild ginger Amomum siamense and Alpinia malaccensis in Doi Suthep-Pui National Park, Chiang Mai, Thailand. Three new species, Pyricularia kookicola, P. longispora, and P. variabilis are described, illustrated and compared to similar Pyricularia species.

Evaluation of Muscodor suthepensis strain CMU‐Cib462 as a postharvest biofumigant for tangerine fruit rot caused by Penicillium digitatum

BACKGROUND This study investigated both the in vitro and in vivo biofumigant ability of the endophytic fungus Muscodor suthepensis CMU-Cib462 to control Penicillium digitatum, the main cause of tangerine fruit rot. RESULTS Volatile compounds from M. suthepensis inhibited mycelial growth of the pathogen. The most abundant compound was 2-methylpropanoic acid, followed by 3-methylbutan-1-ol. They showed median effective doses (ED50) on P. digitatum growth of 74.91 ± 0.73 and 250.29 ± 0.29 µL L(-1) airspace respectively. Rye grain was found to be a suitable solid medium for M. suthepensis inoculum production. The results indicated that mycofumigation with a 30 g rye grain culture of M. suthepensis for 12 h controlled tangerine fruit rot. The percentage weight loss and soluble solids concentration of fumigated tangerines were similar to those of non-infected and non-fumigated fruits. CONCLUSION Muscodor suthepensis has potential as a biofumigant for controlling postharvest disease of tangerine fruit.

First report of leaf spot disease on oil palm caused by Pestalotiopsis theae in Thailand

A leaf spot on oil palm, caused by Pestalotiopsis theae, was found in a plantation of Elaeis guineensis for the first time in the world in Chiang Mai Province, Thailand. The fungus was isolated from lesions on leaves, and its pathogenicity was confirmed. Pathogenicity tests showed that P. theae could infect E. guineensis, which developed the same symptoms after inoculation as those observed naturally in the field. The fungus was identified based on morphological characteristics and confirmed using comparisons of DNA sequences of internal transcribed spacer (ITS)1, ITS2 and 5.8S rDNA. This report is the first on oil palm leaf spot disease caused by P. theae.

Basidiome formation of an edible wild, putatively ectomycorrhizal fungus, Phlebopus portentosus without host plant

Phlebopus portentosus is a popular wild edible ectomycorrhizal fungus in northern Thailand. In general ectomycorrhizal fungi produce basidiomes when associated with a host plant. In this paper mycelium growth and basidiome production of P. portentosus were examined in pure culture both in vitro and in pot-culture experiments. Five mycelial strains of P. portentosus were isolated from basidiomes and used in the experiments. The mycelia grew fastest on sorghum grains supplemented with fungal-host solution. The mycelia produced sclerotia-like structures after 3 wk incubation in darkness at 30 C. All strains of P. portentosus had the ability to form primordia. The primordia were formed under lowered temperature, high humidity and a 12 h photo-period. They developed to mature basidiomes after 8–12 d in in vitro. In the pot-culture primordia were found after 28–35 d incubation in the greenhouse and mature basidiomes released basidiospores within 6–8 d. Basidiospores were germinated on fungal-host medium and formed mycelial colonies. This fungus showed an ability to produce basidiomes even 2 y after the original isolation from tissues. This research provides valuable information concerning the techniques and protocols for the large scale commercial production of P. portentosus basidiomes in the absence of a host plant.

Biotransformation of natural compounds. Oxido-reduction of Sch-642305 by Aspergillus ochraceus ATCC 1009

Sch-642305 is the major compound produced by the endophytic fungi Phomopsis sp. CMU-LMA. Incubation of Sch-642305 with Aspergillus ochraceus ATCC 1009 resting cells leads to three new derivatives through an oxido-reduction of the six-membered ring of the molecule. Reduction of the double bound leads to compound (1), which subsequently undergoes carbonyl reduction to (2) and ring hydroxylation to (3). According to the previously solved crystal structure of Sch-642305 coupled with (1)H NMR NOE correlation and the crystal structure of compound 1, the absolute configurations of the new derivatives were established. In contrast to the parent compound Sch-642305, compound (1) exhibits antimicrobial activity against gram-negative bacteria. Furthermore, while all derivatives exhibit cytotoxic activity against various cancer cell lines, compound (2) achieved an IC(50) of 4 nM against human myelogenous leukemia K 562, compared to 20 nM for the parent Sch-642305.

First Report of Curvularia lunata Causing Leaf Blight on Mulberry (Morus sp.) in Thailand

Mulberry (Morus L.) is a perennial and heterogeneous outbreeding tree of the Moraceae family. In Thailand, mulberry is cultivated mainly in rural areas where mulberry leaves are used to feed silk worms for silk production. In August 2013, we first observed leaf blight on the mulberry cultivar ‘Burriram 60’ in Chiang Mai province (18.79 N, 98.96 E). The infected leaves exhibited brownish to black lesions, starting from the tips or edges of lamina, mainly on younger leaves. Some lesions extended and coalesced, causing leaves fall as disease progressed. However, this leaf fall did not cause plant mortality. Four diseased mulberry leaves were randomly collected, surface sterilized in 1% NaOCl solution for 3 min, rinsed 3 times with sterile water and incubated in moist chamber to induce sporulation. Single spore isolation (Choi et al. 1999) was performed by preparing conidial suspension in 250 µl of sterile distilled water and dropping on 1.5% (w/v) water agar. After 24 h incubation at room temperature (27 ± 2...