Bacillus thuringiensis (Bt) is characterized by its ability to synthesize crystal toxins and also able to produce bacteriocins such as thuricin, tochicin, entomocin and bacthuricin. The present work, for the first time, describes the biological activity of bacteriocins from B. thuringiensis subsp. cameroun (Btc). Supernatant which was produced from a liquid culture of Btc had antimicrobial activity against Bacillus cereus, ending up to making a inhibition zone on an agar medium. A significant reduction in antimicrobial activity was observed when the supernatant was exposed to heat at 75~100°C for 15 min. Proteins were separated from the supernatant by a fast protein liquid chromatography (FPLC) given the thermal instability. A group of FPLC fractions had antimicrobial activity against Bt subsp. palmanyolensis, israelensis, 1-3, morrisoni, toguchini and kurstaki, and B. cereus ACTC21768, ATCC14579 and NRRLB-569. Interestingly, when the supernatant was individually incorporated into the liquid cultures of Bt subsp. israelensis (Bti) and mogi (Btm) with mosquitocidal activity, a vegetative cell growth was observed only in the Btm culture 10 h post-incubation. A possible recovery of vegetative Btm cell growth was observed, compared to a control without the supernatant. These results suggest that Btc produced proteinous antimicrobial substances, one of which may be used as a selection marker to separate Btm after possibly conjugating the two mosquitocidal strains.

The genomic structure and phylogenetic relationships of HSP88 genes from P. tenuipes Jocheon-1, P. tenuipes, C. militaris and C. pruinosa are described. The HSP88 genomic DNA from P. tenuipes Jocheon-1, P. tenuipes and C. militaris all contain 5 introns and 6 exons with the length of 13, 62, 32, 1438, 306, 288 bp, encoding 713 amino acid residues. C. pruinosa HSP88 genomic DNA contains 4 introns and 5 exons encoding 713 amino acids. The length of each exon of C. pruinosa HSP88 is 13, 62, 32, 1744, 288 bp and the length of exon 4 is identical to the total length of exon 4 and exon 5 of HSP88 of P. tenuipes Jochoen-1, P. tenuipes, and C. militaris. The deduced amino acid sequence of P. tenuipes Jocheon-1 HSP88 showed 99% identity with the P. tenuipes, 97% identity with the Cordyceps militaris, and 98% identity with the C. pruinosa. Phylogenetic analysis confirmed that the P. tenuipes Jocheon-1, P. tenuipes, C. militaris and C. pruinosa HSP88 are placed together within the ascomycetes group of fungal clade.

In this study, a full-length heat shock protein88 complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening of P. tenuipesJocheon-1 Uni-Zap cDNA library and 5' RACE polymerase chain reaction. The Paecilomyces tenuipes Jocheon-1 heat shock protein88 cDNA contains an open reading frame of 2,139 bp encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipes Jocheon-1 HSP88 cDNA showed 77% identity to N. haematococca HSP88 and 45-76% identity to other fungi HSP88. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade and P. tenuipes Jocheon-1 HSP88 also contains the conserved ATPase domain at the N-terminal. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as a 88 kDa polypeptide in baculovirus-infected insect Sf9 cells. Under different stress conditions, mRNA expression of P. tenuipes Jocheon-1 HSP88 were quantified by real-time PCR and the result showed that heat shock stress affected the mRNA expression levels of P. tenuipes Jocheon-1 HSP88.

Bee venom contains a variety of protein allergens, including serine proteases. Additionally, bee venom has been used in therapeutic application through immunotherapy for bee venom hypersensitivity and venom therapy as an alternative medicine. Here we present a novel view of the application of bee venom through which bee venom serine protease exhibits fibrin(ogen)olytic activity. Compared to honeybee venom, bumblebee venom contains a larger amount of a serine protease as one of its major components. Immunologically, venom serine proteases from bumblebees did not show cross-reactivity with the honeybee venom serine protease. We provide functional evidence indicating that bumblebee (Bombus terrestris) venom serine protease (Bt-VSP) acts as a fibrin(ogen)olytic enzyme. Bt-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles for Bt-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. However, Bt-VSP did not activate plasminogen and the fibrinolytic activity of Bt-VSP is less than plasmin. These findings offer insight into the allergic reaction sequence of bee venom serine protease and its potential usefulness as a clinical agent in the field of hemostasis and thrombosis.

Bacillus thuringiensis (Bt) strain K4 was isolated from fallen leaves which had been collected at a forest stand in Mungyeong city, Republic of Korea. The flagellated vegetative cells of Bt K4 were agglutinated with the H3 reference antiserum among 55 reference H-antisera. In a further test to identify subfactors, 3b and 3d monospecific antisera were reactive to the cells, followed up with introducing a novel serogroup of 3a3b3d, designated as serovar mogi. The strain K4 had mosquitocidal activity against Dipteran larvae, Anopheles sinensis and Culex pipiens pallens, with no Lepidopteran toxicity observed. The SDS-PAGE profile of K4 crystal protein, ovoidal-shaped, included several bands ranging from 30-75 kDa. Four putative peptides, Cry19Ba, Cry40ORF2, Cry27Aa and Cry20Aa were detected from the bands by a nano-LC-ESI-IT MS analysis. Through a thermal asymmetric interlaced PCR, cry19Ba, cry40ORF2 and cry27Aa genes were partially cloned from K4 strain. Three cry genes were further found in the strain by a 454 pyrosequencing, ending up to showing 58%, 39% and 84% homology in amino acids with Cry56Aa, Cry8Ba and Cry39ORF2 toxins, respectively. This novel 3a3b3d type strain, B. thuringiensis subsp. mogi, can be used as a good resource for studying unknown mosquitocidal cry genes.

Aphids feed on host plants by penetrating the stems or leaves with stylets. The feeding behavior of aphids consists of probing, penetration, salivation, and sap ingestion. To assess the effects of sound on feeding behavior, we monitored the stylet activity of the green peach aphid, Myzus persicae (Sulzer), using electrical penetration graph (EPG). The use of EPG was critical for determining the stage, frequency, and duration of feeding in aphids. We played back three acoustic stimuli of sine waves with frequencies of 100, 1000, and 5000 Hz to adult aphids. In the sound treatment group, the frequencies of probing, penetration, and salivation increased, whereas the duration of sap ingestion decreased. The results of EPG revealed that the acoustic stimuli may restrict aphid feeding by inhibiting sap ingestion.

To understand the evolution and speciation of closely related species, a multiple approach encompassing morphological, behavioral, and genetic analyses is necessary. In Korea, three species of Loxoblemmus crickets occur widely. L. campestris and L. equestris are morphologically indistinguishable, whereas males of L. doenitzi are different from the other two species in head morphology. The genetic analyses using the partial mitochondrial COI sequences showed that L. doenitzi diverged off earlier than L. campestris and L. equestris. The analyses of laboratory recordings revealed that distributions of calling song characters generally overlapped among three cricket species. However, the number of pulses in a chirp was two in L. doenitzi and four in L. campestris, but it was greater than or equal to six in L. equestris. Provided that females make mate choice based on this calling song character, the differentiation in this character may lead to premating reproductive isolation and may have evolved during the speciation proccess in these closely related species.

Light brown apple moth, Epiphyas postvittana, is a significant horticultural pest native to Australia, and currently with a limited global distribution. However it can tolerate very heterogeneous climatic and vegetation conditions and has recently invaded California with considerable consequences for US international and domestic trade. A genetic factor that may contribute to its environmental adaptability, and consequently invasive capability, is the phosphoglucose isomerase gene (pgi). This gene codes for a key enzyme in the second step of glycolysis and for which the isozyme composition has been associated with the fitness and dispersal capacity of other moths. As a first step, to determine if this locus is variable within E. postvittana, novel primers were designed enabling access to 957 bp of the coding region across exons 4 to 11 of pgi. Exon-primed intron-crossing (EPIC) primers were then designed to compare sequences of 17 specimens across one laboratory and three wild New Zealand populations from a laditudinal range of ~39-45°S. A total of 70 segregating sites in the exons were found, including 61 synonymous and nine nonsynonymous. Introns 3 to 11 (excluding intron 10) were also sequenced for 13 individuals revealing significant length variation within and between introns and populations. The level of variation revealed here indicates that this could be a useful target gene to assess fitness factors associated with invasibility of E. postvittana.

Light brown apple moth, Epiphyas postvittana, is a significant horticultural pest native to Australia, and currently with a limited global distribution. However it can tolerate very heterogeneous climatic and vegetation conditions and has recently invaded California with considerable consequences for US international and domestic trade. By comparing the climatic conditions of its native (Australia) and long-established (New Zealand) ranges to the rest of the world using CLIMEX, it was suggested that E. postvittana has potential to establish mainly in countries in Central and South America, southern Africa, west Europe and South-east Asia. However, the predicted global distribution of E. postvittana using a new multiple-species-distribution model system suggested that there are additional climatically suitable areas around the world where this species could potentially survive and establish. Our study provides basic but important information for further assessment of the establishment capacity of this species in new habitats, wihch will provide the knowledge required to make science-based decisions in biosecurity.

The toxicity of imperatorin (1) and osthol (2) identified in Cnidium monnieri seed and four structurally related compounds to third instar larvae of insecticidesusceptible (KS-CP strain) and field-collected (DJ-CP colony) of Culex pipiens pallens was examined. Results were compared with those of to conventional mosquito larvicide, fenitrothion and temephos. Based on 24-h LC50 values, imperatorin was 1.9, 3.7, 4.2, 12.4, and 15.1 times more toxic than isopimpinellin, isoimperatorin, osthole, xanthotoxin, and bergapten against KS-CP larvae, respectively. Overall, these compounds were less toxic than either fenitrothion or temephos. However, these compounds did not differ in toxicity against larvae from the two Culex strains, even though the DJ-CP larvae exhibited high levels of resistance α-cypermethrin, deltamethrin, chlorpyrifos, fenthion, and chlorfenapyr (resistance ratio, 94-1179). This finding indicates that the isolated compounds and the pyrethroid, organophosphorus, and pyrrole insecticides do not share a common mode of action or elicit cross-resistance.

본 연구는 국내에서 생산된 주요 동계 사료 작물인 청보리 및 호밀 사일리지를 이용하여 거세 한우의 비육 중기용으로 조제된 TMR의 소 체내 이용성을 배합사료 및 볏짚으로 구성된 관행 사료와 비교하고자 반추위 누관이 장착된 소 3두를 이용하여 3 3 Latin square design 방법으로 실시되었다. 관행사료 급여구(대조구)에는 1일 두당 7kg (비육 중기용 배합 사료 5.6kg 및 볏짚 1.4kg, 건물 기준)을, 그리고 청보리 사일리지 TMR구

The genetic diversity of 16 Ganoderma strains was investigated by rDNA-ITS sequencing. Alignment analysis showed that whole length of internal transcribed spacer(ITS1+ITS2) was 500bp and with 139 variation sites (accounting for 23.3％, ITS1 was 66 and ITS2 was 73), 337 conserved sites (accounting for 72.2％), 59 informative sites (accounting for 9.88％), 86 conversion sites (G-A, C-T), 13 transversion site(C-G, T-A). The ratio of transition and transversion in ITS1 was higher than that in ITS2, and the variable sites of ITS2 were more than those of ITS1. The genetic distance among 16 Ganoderma strains is from 0 to 0.121. The genetic distance between G. lipsiense and F-1 was 0, and the genetic distance between Heizhi 02 and Huizhou, Jingda, G. capense was 0.121, 0.117 and 0.120, respectively. The 16 Ganoderma strains were classed into 4 groups. The biggest group is comprised of 12 strains, including Xinzhou, Huizhou, Jingda, 902, F-1, Xianzhi, Meiluo, Taishan, G. applanatum, 05, G. luteomarginatum. The G. atrum and G. sinense were clustered into one group. The G. capense and Zhongzhi was independent group, respectively. These results showed that there were some genetic difference among groups, and there was lower genetic diversity among strains in same groups.

Recently, farmers of mushroom farms in Korea express the difficulty of shiitake cultivation due to the damage of oak log beds by unknown reason. To elucidate the cause of the damage, we investigated and isolated fungi from inside wood chip of the damaged shiitake log bed. Four of green fungi were obtained and their morphology and molecular properties examined. Trichoderma atroviride, Trichoderma citrinoviride, Hypocrea lixii (Trichoderma harzianum), and Hypocrea lutea (Gliocladiumviride) were identified. Hypocarea lutea is newly described in Korea. These species were able to produce extracellular enzymes such as cellulase, pectinase, and xylanse that might degrade wood components of oak logs. This study provides strong evidence that Trichoderma fungi are involved in the recent damages of shiitake log beds In Korea.

Four Ganoderma lucidum strains, Chizhi 05, Jingda, Huizhou and Xinzhou, were screened out as hybrid parent in order to establish G. lucidum cross breeding system that based on protoplast monokaryogenesis method. Monokaryotic strains of each parental strains were obtained and mating type of each monokaryotic strains were determined. One to three monokaryotic strains that have different mating types were mated, and hybrids were identified by clamp connection observation and antagonist response. The results showed that the number of monokaryon came from Chizhi 05, Jingda, Huizhou and Xinzhou was 9, 14, 40 and 38, respectively. Only one mating type was obtained from Jingda, and two mating types were obtained from the other three strains, Chizhi 05, Huizhou and Xinzhou, respectively. Chi-square test showed that the ratio of two mating types of the three strains was 1:1. Fourteen monokaryotic strains of different mating types from 4 parental strains were select as a cross- breeding materia, and 17 hybrids were obtained, which were identified by clamp connection observation and antagonist response. This study proclaimed that the practicality of the hybridization breeding of G. lucidum by protoplast monokaryogenesis method.

Genomic DNA prepared from fruitbdies of 25 Grifola frondosa strains were amplified with the ITS primers and the PCR reaction products were enzymed. The ITS bands have same electrophoresis patterns but part of them have different restriction enzyme cutting site. These strains were divided into three broad categories. SRAP amplification employing 47 SRAP primer pairs were carried on and 138 polymorphic bands were detected. The phylogram tree showed that: ten strains were differentiated from the others effectively and fifteen strains have no obvious differences between each other. The results implied that the ITS-RFLP and SRAP markers were effective methods for strains identifica tion and germplasm evaluation but other markers were also needed to be used in conjunction.

Phellinus sp. are assigned to the Basidiomycotina, Hymenomycetes, Aphyllophorales and Hymenochaetaceae, and have been shown containing various bioactive substances including triterpenoids, polysaccharides and flavones[1]. Traditional Chinese herbalists believe that Phellinus species are effective in treating many gynecopathic ailments[2] and are also reported to exhibit other pharmacological functions including tumor cell inhibition, antioxidant activity and anti-hepatic fibrosis effects[3]. Polysaccharides of Phellinus sp. has been reported to possess antitumor activities and inhibit tumor recrudescence and metastasis. There are little studies comparing the chemical composition and biological activities difference among polysaccharides from different Phellinus sp and little report about the pure polysaccharide structure analysis. In this study, eight kinds of crude polysaccharides were extracted from Phellinus fruit bodies and their chemical composition and bioactivities were researched. The polysaccharide and protein contents of eight crude polysaccharides had a certain extent differences. Monosaccharide composition and content of amino acids also existed some differences in eight crude polysaccharides. Eight different polysaccharides all showed enhancing splenocyte proliferation effect in vitro. PB-10P and JSHP had high cell proliferation rates with 50㎍/ml concentrations. The results indicated in some extent the immune activity of crude polysaccharides were correlation with the polysaccharide and protein content and composition of each sample. The crude polysaccharides of P. igniarius were further isolated and purified using DEAE Sepharose F. F. and gel-filtration chromatography (Sephacryl S-100-500 ）repeatedly. Five water-soluble homogeneous polysaccharides (P60w1、P60s1、P1SP1、P10SP1and P100SP1) were obtained. Lack of absorption at 280 nm and 260 nm by UV scanning indicated that contained no protein and nucleic acid. HPLC produced a single symmetrical peak, indicated homogeneity and their molecular weigh were 1.71×104 Da、2.07×104 Da、1.48×104 Da、2.20×104 Da and 2.56×104 Da respectively. Structural of P60w-1 were determined using sugar and methylation analysis combined with 1H and 13C NMR spectroscopy, including COSY, TOCSY, NOESY, HSQC and HMBC experiments. The effect of P60w-1 on tumor growth was examined using subcutaneously transplanted H22 and Lewis Lung Carcinoma (LLC) tumor mouse models. Cyclophosphamide or Coriolus versicolor polysaccharopeptide served as positive controls in evaluating tumor response. Results showed that P60w-1 at the most effective dose of 100 mg/kg inhibited the growth of H22 and LLC by 48% and 37%, respectively.

Related research on artificial cultivation of the edible fungi in modern China can be traced back to 100 years ago. The earliest article about the cultivation techniques of the edible fungi was published in 1897 on Agricultural Study Newspaper sponsored by Shanghai Agricultural Society. From the late 1800s to 1940s, the elder generation of scholars including Zou Bingwen, Hu Changzhi, Pan Zhinong, Li Shiyi, Sun Yunwei, and Yu Xiaotie, etc. not only introduced many foreign techniques in edible fungi cultivation a nd disseminated scientific knowledge and cultivation techniques, but also held various edible fungi talents training courses, set up experimental bases, and conduct edible fungi cultivation experiments. Those have laid a preliminary foundation for the modernization of China’s edible fungi cultivation techniques. After the founding of the PRC, the edible fungi cultivation industry has gained more space for development, and has achieved many milestone achievements, mainly including the tremella artificial cultivation technique, the hedgehog hydnum artificial cultivation technique, the Xianggu artificial cultivation with crushed-wood material technique, the white mushroom fine breed selection and breeding and cultivation technique improvement, the black fungus cultivation with crushed-wood material and fine breed selection and breeding, and the golden needle mushroom find breed selection and year-round cultivation technique innovation, as well as the acclimatization of wild edible fungi and the development of new varieties of edible fungi. These inventions and innovations have provided solid technical support to the development of the edible fungi industry in China. The reform and opening-up starting 1978 has provided a favorable policy environment for the development of the edible fungi industry in China. In the period of more than 30 years thereafter, the edible fungi industry in China has been developing rapidly, with the annual yield rocketing from 60,000 tons in 1978 to 20.2 million tons in 2009, and the proportion of the yield against total world yield growing from 5% in the past to more than 70% at present. During this historical period, many research institutions and scientific research staffs have made important contributions to the development of the edible fungi industry in China. Among them, the most important achievements are made by Researcher Chen Meipeng from the Institute of Edible Fungi of Shanghai Academy of Agricultural Sciences, Professor Yang Xinmei from Huazhong Agricultural University, Researcher Huang Nianlai from Fujian Sanming Mycological Institute, and Professor Zhang Shuting from Chinese University of Hong Kong. When we look back, we treasure the outstanding achievements made by the elder generation of scholars on the development of the edible fungi industry in China. When we look into the future, we are geared with enthusiasm and confidence, and we will work hard to achieve higher in the edible fungi industry in China.