Flammulina velutipes belonging to white rot fungi is one of the commercially important edible mushrooms and is produced in large quantities due to the introduction of a automated and mechanized cultivation system in Korea. Despite the chief item of export among edible mushrooms, Flammulina velutipes has the lowest distribution rate of domestic cultivar, estimated that about 20 percent. As the result that most white cultivars of Flammulina velutipes produced and exported in Korea were introduced from Japan, farmers pay a large amount of royalties. Therefore, we try to develop a new pure domestic cultivars as a substitute for Japanese cultivars. To breed both white and gold superior strains, we selected the crossing mother groups including 10 white strains of ASI 4198 etc. and 7 brown strains of ASI 4049 etc. and mated each of the 17 strains by mon-mon hybridization. 19 white and 14 brown strains were chosen through two selection experiment over 2014 2016. In the third selection experiment this year, we finally selected one white(Fv 16 c 37) and the other gold(Fv 15 a 31) strain. Two selected strains were cultivated in the same environmental conditions. Spawn running period on the sawdust substrate required 30days at 20°C. The cultivation period and optimum temperature were 12±1 days at 14°C for primordia formation, 5 days at 4°C for inhibition phase, and 14±1 days at 7°C for fruiting body development. The length of pilei and stipes in two selected strains and Megumi as a control Japanese cultivar harvested in optimal stage was as follows: 10.5±0.81mm and 139.7±4.23mm in Fv 16 c 37, 10.8±0.43mm and 128..2±7.31mm in Fv 15 a 31, and 10.9±0.41mm and 141.8±4.64mm in Megumi respectively. The Yield of Fv 16 c 37, Fv 15 a 31 and Megumi was 271.2±11.84g, 237.7±9.05g and 270.7±16.87g per 1100ml in bottle cultivation.

Volvariella volvacea is mainly cultivated in subtropics area like South-East Asia. Because it is cultivated in rice straws, it is called a straw mushroom. That mushroom grows well in high temperature about 30~38°C and high humidity. Straw mushroom is a homothallic mushroom, so it is difficult to identify whether the offspring is different from the parents. This study was carried out to investigate RAPD primers that can be used for identification the DNA polymorphism of Volvariella volvacea genetic resources. 9 strains were collected from various countries like China, Vietnam etc. When ITS regions of their DNA were analyzed, they proved to be Volvariella volvacea. A cultivation test was conducted to measure the morphological characteristics of them 2 strains, KMCC04380 and KMCC04382 were selected for breeding resources because the mycelium of them grew well on medium and fruiting bodies were formed quickly. The Universal PCR Fingerprinting kits(UPF primers) were used to confirm the genetic polymorphisms of the 2 strains. As a result of confirming the DNA bands, 2 of 12 primers could be used to genetically distinguish 2 strains. About 50 spores were isolated from their fruiting bodies respectively and they also will be confirmed DNA polymorphisms by using UPF primers.

Tremella fuciformis has about 40 species worldwide, and is widely distributed in the subtropical, tropical, and temperate regions. Tremella fuciformis is known to have been eaten by poppy in Chinese classics and has been a symbol of beauty since ancient times. In fact, it has been known that the extract of the Tremella fuciformis induces inhibition of melanin formation of the cells to have a skin whitening effect and to prevent wrinkles caused by UV stimulation damage, thereby improving wrinkles. Tremella fuciformis was first cultivated in Sichuan province, China, and is now cultivated in China and Southeast Asia. China has succeeded in large-scale artificial cultivation of Tremella fuciformis, but there is no research on mass artificial cultivation in Korea. Therefore, this study was carried out to develop standard cultivation technique for mass production of Tremella fuciformis. In order to select the optimal medium for Tremella fuciformis, five kinds of medium such as PDA, MCM, MEA, YM and CDA were used. Tremella fuciformis showed the best growth on PDA medium, and Annulohypoxylon stygium showed the best growth on MCM. The growth of Tremella fuciformis was good at pH 5 , and Annulohypoxylon stygium showed the best growth at pH 6. The optimum temperature for Tremella fuciformis and Annulohypoxylon stygium was 25 °C. Carbon source and nitrogen source were selected to establish optimal culture conditions.

Cladobotryum mycophilum is the causal agent of cobweb disease of commercial mushrooms. Symptoms were noticed as round, fleshy, yellowish brown lesions on mushroom caps and progressed when the parasitic fungus formed white cobweb circular colonies on dead or damaged pinheads, spread on the surface of the casing. Two gram-positive bacterium were isolated from mushroom media that markedly showed the antagonistic activity against C. mycophilum.. These isolates were identified as Bacillus altitudinis and Bacillus subtilis by analysis of the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacteria is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacteria cell was sufficient for inhibition in vitro for C. mycophilum. Control efficacy of browning disease of Bacillus altitudinis treatment was 78% on Agaricus bisporus. The optimal culture medium was determined as follows: 3% Soluble startch, 10% Soytone, 1% (NH4)2HPO4, 1 mmol KCl, and 0.5% L-asparagin at pH 6.0 at 30°C. Control efficacy of browning disease of Bacillus subtilis treatment was 71% on Agaricus bisporus. The optimal culture medium was determined as follows: 1.5% Xylose, 2% Soytone, 1% NH4H2PO4, 7 mmol CaCl2, and 0.5% Histidine at pH 6.0 at 25°C. Accordingly, the suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by C. mycophilum.

Microsatellite SSR markers were developed and utilized to reveal the genetic diversity of 32 strains of Flammulina velutipes collected in Korea, China, and Japan. From SSR-enriched library, 490 white colonies were randomly selected and sequenced. In the 490 sequenced clones, 85 clones (17.35%) were redundant. Among the remaining 405 unique clones, 201 clones (49.6%) contained microsatellite sequences. As a result, 12 primer pairs produced reproducible polymorphic bands within diverse 4 strains and these selected markers were further characterized in 32 Flammulina velutipes strains. A total of 34 alleles were detected using the 12 markers, with an average of 3.42 alleles and the number of alleles ranged from two to seven per locus. The major allele frequency ranged from 0.42 (GB-FV-127) to 0.98 (GB-FV-166), and values for observed (HO) and expected (HE) heterozygosity ranged from 0.00 to 0.94 (mean = 0.18) and from 0.03 to 0.67 (mean = 0.32), respectively. SSR loci amplified with GB-FV-127 markers gave the highest polymorphism information content (PIC) of 0.61 and mean allele number of five, while for loci amplified with GB-FV-166 markers these values were the lowest, namely 0.03 and two. The mean PIC value (0.29) observed in the present study with average number of alleles (3.42). The genetic relationships among 32 Flammulina velutipes strains based on SSR data were generated by UPGMA cluster analysis. In conclusion, we succeeded in developing 12 polymorphic SSRs markers from SSR-enriched library of Flammulina velutipes. These SSRs are presently being used for phylogenic analysis and evaluation of genetic variations. In future, these SSR markers will be used in clarifying taxonomic relationships among the Flammulina velutipes.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 11,493 M/T in 2014. Several fungus are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Trichoderma harzianum is the causal agent of green mould disease of commercial mushrooms. Early symptoms were noticed as round, fleshy, brown lesions on mushroom caps. Late symptoms spread on the surface of the casing, and covered entirely fruiting bodies. A Gram-positive bacterium was isolated from mushroom media that markedly showed the antagonistic activity against Trichoderma harzianum, the destructive pathogen of cultivated mushrooms. The CH518 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus methylotrophicus by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for Trichoderma harzianum cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Trichoderma harzianum. Control efficacy of browning disease of strain CH518 treatment was 77.7% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 3.0% saccharose, 1.5% Soytone, 1% NH4H2PO4, 10 mM MgSO4, and 2.0% glutamic acid at pH 6.0 at 25°C. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by Trichoderma harzianum.

This study investigated the effect of Charcoal:Dextran Stripped fetal bovine serum (CDS FBS) and heat-inactivated FBS (HI FBS) in embryo culture medium on their ability to support in vitro development of bovine embryos. The developmental ability and quality of bovine embryos were determined by assessing their cell number, lipid content, mitochondrial activity, gene expression, and cryo-tolerance. The percentages of embryos that underwent cleavage and formed a blastocyst were significantly (P<0.05) higher in medium containing CDS FBS than in medium containing HI FBS (42.84 ± 0.78% vs. 36.85 ± 0.89%, respectively). Furthermore, the beneficial effects of CDS FBS on embryos were associated with a significantly reduced intracellular lipid content, as identified by Nile red staining, which increased their cryo-tolerance. The post-thaw survival rate of blastocysts was significantly (P<0.05) higher in the CDS FBS than in the HI FBS group (85.33 ± 4.84% vs. 68.67 ± 1.20%). Quantitative real-time PCR showed that the mRNA levels of acyl-CoA synthetase long-chain family member 3, acyl-coenzyme A dehydrogenase long-chain, hydroxymethylglutaryl-CoA reductase, and insulin-like growth factor 2 receptor were significantly increased upon culture with CDS FBS. Moreover, the mRNA levels of sirtuin 1, superoxide dismutase 2, and anti-apoptotic associated gene B-cell lymphoma 2 in frozen-thawed blastocysts were significantly (P<0.05) higher in the CDS FBS group than in the HI FBS group, however, the mRNA level of the pro-apoptotic gene BCL2-associated X protein was significantly reduced. Taken together, these data suggest that supplementation of medium with CDS FBS improves in vitro bovine embryo developmental competence and cryo-tolerance.

The production scale of mushrooms in Korea is approximately 600 billion won, which is 1.6% of Korea’s gross agricultural output. In Korea, ca. 190,000 tons of mushrooms are harvested annually. Although the numbers of mushroom farms and cultivators are constantly decreasing, total mushroom yields are increasing owing to large-scale cultivation facilities and automation. The recent expansion of the well-being trend has caused an increase in mushroom consumption in Korea: the annual per capita mushroom was 3.9 kg (’13), whichis a little higher than that in Europe. Thus, mushroom export, mainly Flammulina velutipes and Pleurotus ostreatus, has increased since the mid-2000s. Recently, however, it is slightly reduced. Nevertheless, Vietnam, Hong Kong, the United States, and the Netherlands continue to export mushrooms, and Korea has increased its export to Australia, Canada, Southeast Asia, etc. Canned Agaricus bisporus, the first export of the Korean mushroom industry, reached it speak sales in 1977-1978. When Korea initiated trade with China in 1980, the international prices of mushrooms fell sharply, leading to shrinkage of the domestic markets. Spurred by the high demand to develop substitute goods for A. bisporus, the oyster mushroom (P. ostreatus) gained attention since it seemed to suit the taste of Korean consumers. Although the log cultivation technique for oyster mushroom was developed in the early 1970s, it required a great deal of labor. Thus, we developed the shelf cultivation technique, which is easier to manage and allows for mass production. In this technique, the growing shelf is made mafrom fermented rice straw, whichis the only P. ostreatus medium in the world and isused only in South Korea. After then, the use of cotton wastes as an additional material of medium, the productivity. Currently, we are developing a standard cultivation technique and environmental control system that can stably produce mushrooms throughout the year. The increase of oyster mushroom production may boostthe domestic market and contribute to industrial development. In addition, oyster mushroom production technology played a role in forming the basis for the development of bottle cultivation, which made mass production . In particular, bottle cultivation using liquid spawn could allow for the export of F. velutipes and Pleurotus 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 ‘Saeah’, which is easy to grow in Korea. In hopes to advance the mushroom industry, we will continue to develop cultivars with international competitive power and to improve cultivation techniques.

The market distribution channels has changed from single to multi-channels due to the diffusion of digital development products. In addition the new consumption emergence has been applied in showrooming and reverse-showrooming. One of well-known shopping district in South Korea, Dongdaemun, has been introduced Dongdaemun-based fashion brand which use on/offline multi-channel distribution. This study aims to segment multi-channel attributes of Dongdaemun-based fashion brands and analyse the effects of customer equity attributes, customer satisfaction, and re-purchase intention. The goals of this study are as follows: 1) to identify multi-channel attributes of Dongdaemun-based fashion brands, 2) to analyse the influence of multi-channel attributes on customer equity, customer satisfaction, and repurchase intention, 3) to explore the consumer group differences on shopping orientation. This study applied means-end chain theory to conduct in-depth interviews for the purpose of deriving the multi-channel attributes of Dongdaemun-based fashion brands and explore the influence of such attributes on customer equity, customer satisfaction, and re-purchase intention. This study collected 493 samples and structural equation modeling was used for final data analysis. The main findings of this study are as follows. The multi-channel attributes have a significant impact on value equity and brand equity. Relationship equity is influenced by product diversity, entertainment, and informativeness. The all of customer equity drivers have significantly impacts on repurchase intention and only brand equity have significant effect on repurchase intention. Also, customer satisfaction affects re-purchase intention. Brand equity have a significant influence on CLV. The consumers group differ in how consumers recognize multi-channel attributes of Dongdaemun-based fashion brands, based on channel preference. This study shows the importance of customer equity and suggests long-term implications for the development of Dongdaemun-based fashion brands.

Oyster mushroom is one of mushrooms that are cultivated and consumed a lot in Korea. P. ostreatus 'ASI 2504(Suhan)' is a preferable cultivar to mushroom farmers because it has a dark pileus and a thick stipe. But it is very sensitive to environmental conditions, so farmers demand an alternative cultivar of ‘Suhan’ continuously. To develop a new cultivar, parental strains ‘ASI 2504(Suhan)’ and ‘ASI 0665(Heuktari)’ were selected from P. ostreatus’s various collected strains according to morphological characteristics. P. ostreatus ‘Soltari’ was developed by the method of Di-Mono crossing between dikaryotic strain ‘Suhan’ and monokaryotic strain derived from ‘Heuktari’. Analysis of the mitochondrial genetic characteristics was performed for primary selection in 100 crossed strains. The mitochondrial DNA profile of ‘Soltari’ was same as that of ‘Heuktari’, when mitochondrial DNA primer MtPo1 was used. And a nuclear DNA profile of ‘Soltari’ was similar as those of the parental strains, ‘Suhan’ and ‘Heuktari’, when RAPD(Random Amplified Polymorphic DNA) primer URP 1, 3 were used. The optimum temperature for mycelial growth was 30°C for ‘Soltari’. ‘Soltari’ was appropriate for middle high temperature to grow, especially 13~18°C. Fruiting body production per bottle (1,100mL) was about 158.6g. When compared to the control strain ‘Suhan’, the stipe’s length and thickness of ‘Soltari’ were similar to those of ’Suhan’. But the pileus diameter of ‘Soltari’ was a little shorter than that of ‘Suhan’, the former was 42.72mm, while the latter was 51.33mm. And the pileus thickness of ‘Soltari’ and ‘Suhan’ were 18.18mm and 25.46mm, respectively. ‘Soltari’ was more resistant at high CO2 concentration than ‘Suhan’ and the color of pileus of ‘Soltari’ was kept dark gray at high temperature. Therefore, it is suggested that this new cultivar ‘Soltari’ be an alternative of ‘Suhan’ and contribute to energy saving effect in oyster mushroom farms.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 11,493 M/T in 2014. Several fungus are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Cladobotryum mycophilum is the causal agent of cobweb disease of commercial mushrooms. Early symptoms were noticed as round, fleshy, yellowish brown lesions on mushroom caps. Late symptoms progressed when the parasitic fungus formed white cobweb circular colonies on dead or damaged pinheads, spread on the surface of the casing, and covered entirely fruiting bodies. A Gram-positive bacterium was isolated from mushroom media that markedly showed the antagonistic activity against Cladobotryum mycophilum, the most destructive pathogen of cultivated mushrooms. The HC57 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus subtilis by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for Cladobotryum mycophilum cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Cladobotryum mycophilum. Control efficacy of browning disease of strain HC57 treatment was 71% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 1.5% Xylose, 2% Soytone, 1% NH4H2PO4, 7 mmol CaCl2, and 0.5% Histidine at pH 6.0 at 25℃. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by Cladobotryum mycophilum.

A total of 25 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains, we found 9376 SNPs, of which 8178 were non-polymorphic and 1198 were polymorphic. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4120 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr08: 950kb-2650kb, chr09: 500kb-1400kb, chr09: 2800kb-4350kb, and chr11: 2450kb-3500kb regions were identified to be associated with white fruitbody forming.

Matrix Metalloproteinases (both MMP2 and -9) play a pivotal role of the embryos hatching and implantation. Therefore, the objective of this study was carried out to investigate the influence of MMP2 and MMP9 on embryo development potential and subsequent effect at molecular level. There was no significant difference of cleavage rate among the groups. The development competence of blastocyst was significantly higher (P<0.05) in MMP9 treatment (39.81±16.61) than that to the combined treatment of MMP2 and –9 (23.68±0.27), but there was no significant difference among the control vs. MMP2 vs. MMP9 (35.05±2.74 vs. 32.71±6.18 vs. 39.81±16.61, respectively). On the other hand, the hatching rate of blastocysts was significantly lower (P<0.05) in combined group of MMP2 and –9 (12.55±0.09) (Table1). The expression level of MMP2 and MMP9 was significantly lower (P<0.05) in the entire treatment groups than that in the control group. But the expression of MMP9 was significantly higher (P<0.05) when compared in the entire treatment groups. The relative expression embryonic developmental gene, IFNt expression level significantly lower (P < 0.05) in the MMP9 embryos. The placenta establishment genes, PLAC8 and SSLP1, expression were significantly higher (P < 0.05) in the MMP2 embryos compared to other groups. Transcription regulation gene, HNRNPA2B1, was higher (P < 0.05) in the combined group of MMP2+MMP9 than that in the other groups. In conclusion, our results suggest that MMPs to culture medium improves the blastocyst development rate and further impact on target gene expression analysis.

MYC (v-myelocytomatosis viral oncogene homologue) is a regulator gene that encodes for a nuclear phosphoprotein. Porcine MYC gene was mapped on chromosome SSC 4p13 and is associated with a variety of functions such as cell proliferation and cell growth. MYC expression is coupled to a multitude of physiological processes and is regulated by hormones, growth factors, cytokines, lymphokines, nutritional status, development and differentiation. MYC is also involved in myogenesis, muscle hyperplasia and adipogenesis. In this study, we investigated SNPs in MYC gene and their association with economic traits in Duroc, Landrace and Yorkshire populations. We detected a single point mutation in exon 3 of porcine MYC gene as a change of T to C at 906 base (amino acid position 302, nonsynomous mutation of alanine) in MYC-N domain. MYC mutation (T906C) was significantly associated with age at 90 kg in these breeds, signifying that this mutation can serve as a selection marker for growth traits in pigs.

The purpose of this study was attempted to new methods in mammalian embryos vitrification. This method was affected to increase of the embryo vitrification efficiency and it would be applied to the field of embryo transfer to recipient by modified loading method of embryo into 0.25 ml plastic straw. The frozen mouse embryos were carried out warmed from two different cell stages (8-cell and blastocyst, respectively) by attachment of an embryo in the vitrification straw (aV) method. All groups were cultured in M-16 medium to determine the development and survivability for 24 h, respectively. Results shown that, the survivability of two different groups were significantly different (94.8% vs. 70.9%). Total cell number was not significantly different the non-frozen blastocyst (99.7 ± 12.4) compared to the post-thaw blastocyst (94.8 ± 15.1). From the 8-cell embryo, total cell number of frozen blastocysts were significantly lower than others groups (74.7 ± 14.6, p<0.05). In the case of cell death analysis, the blastocysts from non-frozen and frozen-thawed 8-cell group were not different (0.0 ± 0.0 vs. 1.9 ± 3.1, p>0.05). However, the apoptotic nuclei of blastocyst were significantly observed the frozen-thawed group (5.4 ± 4.4) compared to non-frozen group (p<0.05). Therefore, this new method of embryos using in-straw dilution and direct transfer into other species would be more simple procedure of embryo transfer rather than step-wise dilution method and cryopreservation vessels, so we can be applied in animal as well as human embryo cryopreservation in further.

Cobweb disease symptoms were observed in a mushroom farm in Buye, Korea during a disease survey in 2008-2011. Five isolates of Cladobotryum sp. were obtained from the infected caps and stipes. These isolates of Cladobotryum sp. were identified as C. mycophilum based on their morphological, cultural characteristics and analysis of the ITS sequences. Early symptoms were noticed as round, fleshy, yellowish brown lesions on mushroom caps. Late symptoms progressed when the parasitic fungus formed white cobweb circular colonies on dead or damaged pinheads, spread on the surface of the casing, and covered entirely fruiting bodies. Optimal temperature and pH for mycelial growth on MEA is 23℃ and 6.0. Microscopically the spores of the fungus are large and most 2~3 celled produced on vertically branched conidiophores. Mushroom caps turned dark brown and shrunk due to soft rot. Testing of sensitivity to selected fungicides showed that isolate was highly resistance to Mancozeb and Thiophanate-methyl, moderately sensitivity to Iprodione, and highly sensitivity to Benomyl, Prochloraz-Mn and Carbendazim.

This study was focused on improvement of milk production in Mongolia dairy industry by artificial insemination (AI) technology, of which was supported from ODA project of KOICA in Republic of Korea. The study was started from January 2009 to present and 3rd years in this year. So, all data, especially synchronization and pregnancy of dairy cows (Holstein) will be summarized in final result in this year. For synchronization, total 81 dairy cows selected from 4 private farms that were 38, 30, 8 and 5 in Undarmal milk, Onjin (Enkhbayer), Jargalant, and BRM School, respectively. All the dairy cows were injected intramuscular (IM) of 5 ml PGF2α in the vulva and detected estrus 2 to 3 days after PGF2α injection. Total 78 out of 81 dairy cows (96.3%) were detected estrus by only 1 time injection of PGF2α. The dairy cows that were induced estrus, inseminated with 0.5 ml dairy frozen semen by conventional AI techniques. The pregnancy diagnosis of the AI dairy cows was detected by uterus palpation after 60 days of insemination. Total 75 from 78 inseminated dairy cows (90.1%) were diagnosis pregnant. The estrus induction and pregnant rate were very effective using PGF2α injection and conventional AI techniques in Mongolia dairy cow. The results indicated that AI after estrus induction in Mongolia dairy cows could be applied to dairy breeding technology to improve the breeding efficiency and milk production.

Many basidiomycetes, especially mushroom species, thrive on saccharides that they obtain from the decomposition of cellulose, hemi-cellulose and or lignin. Cellulose is degraded by specific enzymes called cellulases. Cellulases play an important role in the biosphere by recycling cellulose, the most abundant carbohydrate produced by plants. Also cellulases have many potential biotechnologic and industrial applications. Detection of cellulose degrading activity is commonly performed on carboxymethylcellulose CMC medium in combination with a stain that reveals the degraded CMC. In order to efficiently screen the F. velutipes knockout library, obtained through Agrobacterium-mediated transformation (ATMT), for those important enzymes we compared two different staining methods i.e. Congo Red and Gram’s Iodine (as reported in R. C. Kasana et al 2008). The latter stain showed strongly enhanced detection in time and intensity, facilitating mass screening of our mutant database. Using Gram’s iodine and square petri dishes containing up to 9 colonies at a time we can now rapidly screen multiple mutants in a short period. We are going to find the genes. Inactivated genes of mutants with altered cellulase degradation activity will be identified and cloned for further analysis.