Cryphonectria parasitica, chestnut blight fungus, has a characteristic of decreasing pathogenicity when infected with Cryphonectria hypovirus 1. C. parasitica is known to be one of the most representative model systems used to observe the interaction between viruses, plants and fungi. The mitogen-activated protein kinase (MAPK) pathway, which is well conserved in various organisms ranging from yeast to humans, functions in relaying phosphorylation-dependent signals within MAPK cascades to diverse cellular functions involved in the regulation of pheromone, cell wall integrity, and osmotolerance in filamentous fungi. Several genes in the MAPK pathway were revealed to be regulated by hypovirus, or to be involved in pathogenicity in C. parasitica. Among these pathways, the CWI pathway has aroused interest because CpBck1, an ortholog of yeast Bck1 (a CWI MAPKKK), was previously reported to be involved in cell wall integrity and sectorization. Interestingly, sporadic sectorization was observed in the CpBck1 mutant and sectored phenotypes were stably inherited in the progeny that were successively transferred from sectored mycelia. In this study, we analyzed the biological function of CpSlt2, downstream gene of CpBck1, to confirm whether the sectorization phenomenon occurred in the specific single gene or cell wall integrity (CWI) pathway. As results, the CpSlt2-null mutant exhibited marked changes in colonial growth, near absence of conidiation and aerial hyphae, abnormal pigmentation, CWI-related phenotypic defects, and dramatically impaired virulence. As cultivation of the mutant strains progressed, the majority of the colonies showed sporadic sectorization and mycelia from the sectored area stably maintained the sectored phenotype. These results suggest that the unique sectorization is CWI pathway-specific, though the components in the same CWI pathway have common and specific functions.

Alternaria is a ubiquitous fungal genus, widely distributed in the environment and a range of different habitats. It includes both plant pathogenic and saprophytic species, which can affect crops in the field or cause post-harvest spoilage of plant fruits and kernels. Numerous Alternaria species cause damage to agricultural products including cereal grains, fruits and vegetables, and are responsible for severe economic losses worldwide. Most Alternaria species have the ability to produce a variety of secondary metabolites, which may play important roles in plant pathology as well as food quality and safety. Alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA), tentoxin (TEN) and altenuene (ALT) are considered the main Alternaria compounds thought to pose a risk to human health. However, food-borne Alternaria species are able to produce many additional metabolites, whose toxicity has been tested incompletely or not tested at all. Both alternariols are mutagenic and their presence in cereal grain has been associated with high levels of human esophageal cancer in China. TeA exerts cytotoxic and phytotoxic properties, and is acutely toxic in different animal species, causing hemorrhages in several organs. The possible involvement of TA in the etiology of onyalai, a human hematological disorder occurring in Africa, has been suggested. Altertoxins (ALXs) have been found to be more potent mutagens and acutely toxic to mice than AOH and AME. Other metabolites, such as TEN, are reported to be phytotoxins, and their toxicity on animals has not been demonstrated up to now. Vegetable foods infected by Alternaria rot are obviously not suitable for consumption. Thus, whole fresh fruits are not believed to contribute significantly with Alternaria toxins to human exposure. However, processed vegetable products may introduce considerable amounts of these toxins to the human diet if decayed or moldy fruit is not removed before processing. The taxonomy of the genus is not well defined yet, which makes it difficult to establish an accurate relationship between the contaminant species and their associated mycotoxins. Great efforts have been made to organize taxa into subgeneric taxonomic levels, especially for the small-spored, food associated species, which are closely related and constitute the most relevant food pathogens from this genus. Several crops of agricultural value are susceptible to infection by different Alternaria species and can contribute to the entry of Alternaria mycotoxins in the food chain. The distribution of Alternaria species was studied in different commodities grown in Argentina. These food populations were characterized through a polyphasic approach, with special interest in their secondary metabolite profiles, to understand their full chemical potential. Alternaria species associated with tomato, bell pepper, blueberry, apples and wheat cultivated in Argentina showed a surprisingly high metabolomic and mycotoxigenic potential. The natural occurrence of Alternaria toxins in these foods was also investigated. The results here presented will provide background for discussion on regulations for Alternaria toxins in foods.

Fungal pathogens have huge impact on health and economic wellbeing of human by causing life-threatening mycoses in immune-compromised patients or by destroying crop plants. A key determinant of fungal pathogenesis is their ability to undergo developmental change in response to host or environmental factors. Genetic pathways that regulate such morphological transitions and adaptation are therefore extensively studied during the last few decades. Given that epigenetic as well as genetic components play pivotal roles in development of plants and mammals, contribution of microbial epigenetic counterparts to this morphogenetic process is intriguing yet nearly unappreciated question to date. To bridge this gap in our knowledge, we set out to investigate histone modifications among epigenetic mechanisms that possibly regulate fungal adaptation and processes involved in pathogenesis of a model plant pathogenic fungus, Magnaporthe oryzae. For functional and comparative analysis of histone modifications, a web-based database (dbHiMo) was constructed first to archive and analyze histone modifying enzymes from eukaryotic species whose genome sequences are available. Based on the database entries, we carried out functional analysis of genes encoding histone modifying enzymes. Here I provide examples of such analyses that show how histone acetylation and methylation is implicated in regulating important aspects of fungal pathogenesis. Current analysis of histone modifying enzymes is followed by ChIP-seq and RNA-seq experiments to pinpoint the genes that are controlled by particular histone modifications. We anticipate that our work will provide not only the significant advances in our understanding of epigenetic mechanisms operating in microbial eukaryotes but also basis to expand our perspective on regulation of development in fungal pathogens.

Fungal genome sequencing and assembly have been trivial in these days. Genome analysis relies on high quality of gene prediction and annotation. Automatic fungal genome annotation pipeline is essential for handling genomic sequence data accumulated exponentially. However, building an automatic annotation procedure for fungal genomes is not an easy task. FunGAP (Fungal Genome Annotation Pipeline) is developed for precise and accurate prediction of gene models from any fungal genome assembly. To make high-quality gene models, this pipeline employs multiple gene prediction programs encompassing ab initio, evidence-, and homology-based evaluation. FunGAP aims to evaluate all predicted genes by filtering gene models. To make a successful filtering guide for removal of false-positive genes, we used a scoring function that seeks for a consensus by estimating each gene model based on homology to the known proteins or domains. FunGAP is freely available for non-commercial users at the GitHub site (https://github.com/CompSynBioLab-KoreaUniv/FunGAP).

The production scale of mushroom cultivation in Korea is approximately 600 billion won, which is 1.6% of the Korean gross agricultural output. Annually, ca. 190,000 tons of mushrooms are harvested in Korea. Although the numbers of mushroom farms and cultivators are constantly decreasing, the total mushroom yields are increasing due to the large-scale cultivation facilities and automation. The recent expansion of the well-being trend causes increase in mushroom consumption in Korea: annual per capita consumption of mushroom was 3.9kg ('13) that is a little higher than European's average. Thus the exports of mushrooms, mainly Flammulina velutipes and Pleurotus ostreatus, have been increased since the middle of 2000s. Recently, however, it is slightly reduced. However, Vietnam, Hong Kong, the United States, the Netherlands and continued to export, and the country has increased recently been exported to Australia, Canada, Southeast Asia and so on. Canned foods of Agaricus bisporus was the first exports of the Korean mushroom industry. This business has reached the peak of the sale in 1977-1978. As Korea initiated trade with China in 1980, the international prices of mushrooms were sharply fall that led to shrink the domestic markets. According to the high demand to develop new items to substitute for A. bisporus, oyster mushroom (Pleurotus ostreatus) was received the attention since it seems to suit the taste of Korean consumers. Although log cultivation technique was developed in the early 1970s for oyster mushroom, this method requires a great deal of labor. Thus we developed shelf cultivation technique which is easier to manage and allows the mass production. In this technique, the growing shelf is manly made from fermented rice straw, that is the unique P. ostreatus medium in the world, was used only in South Korea. After then, the use of cotton wastes as an additional material of medium, the productivity. Currently it is developing a standard cultivation techniques and environmental control system that can stably produce mushrooms throughout the year. The increase of oyster mushroom production may activate the domestic market and contribute to the industrial development. In addition, oyster mushroom production technology has a role in forming the basis of the development of bottle cultivation. Developed mushroom cultivation technology using bottles made possible the mass production. In particular, bottle cultivation method using a liquid spawn can be an opportunity to export the F.velutipes and P.eryngii. In addition, the white varieties of F.velutipes were second developed in the world after Japan. We also developed the new A.bisporus cultivar "Sae-ah" that is easy to grown in Korea. To lead the mushroom industry, we will continue to develop the cultivars with an international competitive power and to improve the cultivation techniques. Mushroom research in Korea nowadays focuses on analysis of mushroom genetics in combination with development of new mushroom varieties, mushroom physiology and cultivation. Further studied are environmental factors for cultivation, disease control, development and utilization of mushroom substrate resources, post-harvest management and improvement of marketable traits. Finally, the RDA manages the collection, classification, identification and preservation of mushroom resources. To keep up with the increasing application of biotechnology in agricultural research the genome project of various mushrooms and the draft of the genetic map has just been completed. A broad range of future studies based on this project is anticipated. The mushroom industry in Korea continually grows and its productivity rapidly increases through the development of new mushrooms cultivars and automated plastic bottle cultivation. Consumption of medicinal mushrooms like Ganoderma lucidum and Phellinus linteus is also increasing strongly. Recently, business of edible and medicinal mushrooms was suffering under over-production and problems in distribution. Fortunately, expansion of the mushroom export helped ease the negative effects for the mushroom industry.

Sparassis latifolia is called "Cauliflower Mushroom" and is known as an edible mushroom that has high content of ${\beta}$-glucan. Recently, artificial cultivation of S. latifolia has been done by bottle, plastic bag and wood cultivation in Korea. However it is not widely used because there are low incubation ratio and yield. For the high efficiency of production, we aim to find the superior strains and media for better mycelial and fruit body growth. First, we analyzed the genetic relationship among 31 strains and divided five groups with three kinds of URP primers. And then ten strains were selected from five groups based on the experiment of mycelial growth. The suitability of media for mycelial growth was different according to media type. The suitable solid and liquid media for mycelial growth of S. latifolia isolates were PDA and M2, respectively. In addition, with regard to C/N ratio, the mycelial growth increased even until C/N 160. Second, we investigated the production of fruitbody of the strains by plastic bag cultivation. The substrate was mixed with larch sawdust, corn flour, and wheat flour (8:1:1, v/v). Moisture content of substrate was controlled by about 60% with 10% molasses solution. Out of 31 strains, 19 strains formed primordia. The eight strains produced more than 140g/1kg in fresh weight. Third, molasses culture media was selected for the mycelial growth. And molasses suitable sugar content and input aeration were around 8Brix% and 0.3~0.6vvm, respectively. The longer the incubation period is, the more dried weight of mycelia increased, but medium volume decreased. Therefore, the best incubation period was 9 to 11 days depending on strains. In the future, research project entitled development of culture system and new variety for stable production of S. latifolia will be considered as a new item.

Air-borne plant pathogenic fungus Fusarium graminearum and seed-borne plant pathogenic bacterium Burkholderia glumae are cause similar disease symptoms in rice heads. Here we showed that two pathogens frequently co-isolated in rice heads and F. graminearum is resistant to toxoflavin produced by B. glumae while other fungal genera are sensitive to the toxin. We have tried to clarify the resistant mechanism of F. graminearum against toxoflavin and the ecological reason of co-existence of the two pathogens in rice. We found that F. graminearum carries resistance to toxoflavin as accumulating lipid in fungal cells. Co-cultivation of two pathogens resulted in increased conidia and enhanced chemical attraction and attachment of the bacterial cells to the fungal conidia. Bacteria physically attached to fungal conidia, which protected bacterium cells from UV light and allowed disease dispersal. Chemotaxis analysis showed that bacterial cells moved toward the fungal exudation compared to a control. Even enhanced the production of phytotoxic trichothecene by the fungal under presence of toxoflavin and disease severity on rice heads was significantly increased by co-inoculation rather than single inoculation. This study suggested that the undisclosed potentiality of air-born infection of bacteria using the fungal spores for survival and dispersal.

Arbuscular mycorrhizal fungi (AMF) are one of the most widespread symbionts globally. Owing to their enhanced nutrient absorption capacity, AMF significantly contribute to the survival of individual plants and the ecosystem functioning. Community structures of AMF are affected by many environmental factors Inland wetlands have a different environment from common forest soils, therefore, plants inhabiting wetlands may have characteristic AMF communities. The purpose of this study was to compare the AMF communities in wetlands, among the species of host plants. We sampled the roots of 3 host plant species, Phragmites communis, Miscanthus sacchariflorus, and Trisetum bifidum with rhizospheres from 3 isolated areas in Upo wetland, Korea. We extracted DNA from AMF spores in rhizospheres and the roots of 3 plant species. We amplified 18S rDNA of AMF using AMF specific primer. As a result, we confirmed 9 species from 5 genera in AMF spores, and 5 species from 3 genera in plant roots. Funneliformis caledonium was the most dominant species in field soils, on the other hand, Diversispora aurantia was the most dominant species in plant roots. We confirmed that species diversity and abundance of AMF communities were different among host plant species. These results showed that the AMF community had specific to host plants in the inland wetland.

Mating type of Lentinula edodes is determined by two unlinked genetic loci, A and B. To better understand mating behavior of L. edodes, we investigated variations in mating type genes in129 dikaryotic strains collected from East Asia. Through sequence analysis of A locus, we discovered that hypervariable region spanning N-term of HD2-intergenic region-N-term of HD1 could represent A mating type. Mating and hypervariable region analyses revealed 70 unique A mating types: 27 from 98 cultivated strains, 53 from 31 wild strains, and 10 commonly found. It was also revealed that only a few A mating type alleles such as A1, A4, A5, and A7 were prevalent in cultivated strains. Contrarily, A mating type in wild strains was highly diverse: 23 unique A alleles were discovered in small mountainous area in Korean peninsula, suggesting rapid evolution of A mating type in nature. The B locus was assessed by allelic variations in pheromone (PHB) and pheromone receptor (RCB) pairs which constituted subloci Ba and Bb. Sequence analyses and mating assay revealed 5 alleles of RCB1 with 9 associated PHBs in Ba sublocus and 3 alleles of RCB2 with 5 associated PHBs in Bb sublocus. Each RCB was primarily associated with two PHBs. Each PHB-RCB pair was always discovered as a distinct unit. This allowed us to propose 15 B mating types via combinations of five Ba and three Bb subloci. Further investigation on 129 strains confirmed that the B locus, unlike the A locus, was indeed restricted to 15 mating types. Thus, the total number of mating types became 1,050 in L. edodes through a combination of 70 A and 15 B. This number will further increase because of rapid diversification of A mating type. Our findings provide a comprehensive and practical knowledge on mating behaviors of L. edodes.

There has been an impetus in the development of biocontrol agents (BCAs) with the removal of a number of chemical compounds in the market, especially in the European Union. This has been a major driver in the development of Integrated Pest Management systems (IPM) for both pest and disease control. For control of mycotoxigenic fungi, there is interest in both control of colonization and more importantly toxin contamination of staple food commodities. Thus the relative inoculum potential of biocontrol agent vs the toxigenic specie sis important. The major bottlenecks in the production and development of formulations of biocontrol agents are the resilience of the strains, inoculum quality and formulation with effective field efficacy. It was recently been shown for mycotoxigenic fungi such as Aspergillus flavus, under extreme climate change conditions, growth is not affected although there may be a stimulation of aflatoxin production. Thus, the development of resilient biocontrol strains which can may have conserved control efficacy but have the necessary resilience becomes critical form a food security point of view. Indeed, under predicted climate change scenarios the diversity of pests and fungal diseases are expected to have profound impacts on food security. Thus, when examining the identification of potential biocontrol strains, production and formulation it is critical that the resilience to CC environmental factors are included and quantified. The problems in relation to the physiological competence and the relative humidity range over which efficacy can occur, especially pre-harvest may be increase under climate change conditions. We have examined the efficacy of atoxigenic strains of A. flavus and Clanostachys rosea and other candidates for control of A. flavus and aflatoxin contamination of maize, and for Fusarium verticillioides and fumonisin toxin control. We have also examined the potential use of fluidized-bed drying, nanoparticles/nanospheres and encapsulation approaches to enhance the potential for the production of resilient biocontrol formulations. The objective being the delivery of biocontrol efficacy under extreme interacting climatic conditions. The potential impact of climate change factors on the efficacy of biocontrol of fungal diseases and mycotoxins are discussed.

Fusarium head blight (FHB) caused by infection with Fusarium graminearum leads to enormous losses to crop growers, and may contaminate grains with a number of Fusarium mycotoxins that pose serious risks to human and animal health. Antagonistic bacteria that are used to prevent FHB offer attractive alternatives or supplements to synthetic fungicides for controlling FHB without the negative effects of chemical management. Out of 500 bacterial strains isolated from soil, Bacillus amyloliquefaciens JCK-12 showed strong antifungal activity and was considered a potential source for control strategies to reduce FHB. B. amyloliquefaciens JCK-12 produces several cyclic lipopeptides (CLPs) including iturin A, fengycin, and surfactin. Iturin A inhibits spore germination of F. graminearum. Fengycin or surfactin alone did not display any inhibitory activity against spore germination at concentrations less than 30 ug/ml, but a mixture of iturin A, fengycin, and surfactin showed a remarkable synergistic inhibitory effect on F. graminearum spore germination. The fermentation broth and formulation of B. amyloliquefaciens JCK-12 strain reduced the disease incidence of FHB in wheat. Furthermore, co-application of B. amyloliquefaciens JCK-12 and chemical fungicides resulted in synergistic in vitro antifungal effects and significant disease control efficacy against FHB under greenhouse and field conditions, suggesting that B. amyloliquefaciens JCK-12 has a strong chemosensitizing effect. The synergistic antifungal effect of B. amyloliquefaciens JCK-12 and chemical fungicides in combination may result from the cell wall damage and altered cell membrane permeability in the phytopathogenic fungi caused by the CLP mixtures and subsequent increased sensitivity of F. graminearum to fungicides. In addition, B. amyloliquefaciens JCK-12 showed the potential to reduce trichothecenes mycotoxin production. The results of this study indicate that B. amyloliquefaciens JCK-12 could be used as an available biocontrol agent or as a chemosensitizer to chemical fungicides for controlling FHB disease and as a strategy for preventing the contamination of harvested crops with mycotoxins.

A variety of bioactive secondary metabolites have been reported from plant-associated microorganisms. Halophytes, plants that can only grow in hypersaline area, were reported to host beneficial microorganisms such as plant growth-promoting endophytes. The microorganisms have been reported to show notable mutualistic symbiosis with halophytes to help them survive in high saline condition. Finding out bioactive secondary metabolites as well as elucidation of relationship(s) between microbes and the host halophyte has been paid attention, because of their functional diversity. Novel microbes often have associated with novel natural products. In an effort to investigate natural compounds with interesting structures from fungi, we selected plants from a distinct environmental setting which could be a promising source. Several fungi were isolated from halophyte or medicinal plants. Some strains of the fungi were cultivated on a large scale and extracted with ethyl acetate, which were subjected to a series of chromatographic methods, leading to the isolation of tens of compounds. The isolated compounds were identified by analysis of spectroscopic methods such as 1D-, 2D-NMR, and MS. Details of isolation, structure determination, and biological activities will be discussed.

The application of recombinant DNA technology has been remarkable and nearly replaced commonly used traditional methods. Traditional industrial microbiology long depended on the discovery of valuable strains and mutagenesis of such strains to improve its secretion capacity of enzymes and secondary metabolites on the industrial scale. Commodities included industrial enzymes and biopharmaceuticals. The purpose of genome manipulation by the crossing of different strains or genetic recombination of naked DNA to the genome is of increased production of valuable metabolites. We optimized a transformation method to either for removal of innate genes, introduction of heterologous genes, or combination of both. We have been used selected whole or partial genes to manipulate target fungi toward the development of strains overproducing invaluable proteins. We have also used the whole genome sequence information of fungal genomes in public databases and functional genomics approach to select genes to manipulate and eventually contributing greatly to the development of overproducing industrial strains overproducing proteins or secondary metabolites. I will briefly review 1) filamentous fungi as a host for production of recombinant proteins and secondary metabolites, 2) markets of industrial metabolites, 3) a new approach to manipulate up to five genes at the same time in the system that ProxEnrem uses.