Paecilomyces tenuipes로부터 분리한 Acetoxyscirpendiol의 세포사멸작용

한희창,김미정,김종수,김하원 대한약학회 2004 약학회지 Vol.48 No.2

Paecilomyces tenuipes is one of the famous Chinese medicinal entomopathogenic fungi that parasite in the lavae of silkworm. A cytotoxic compound, 4$\beta$-acetoxyscirpendiol (ASD) was isolated from a methanolic extracts of Paecilomyces tenuipes. The ASD compound belongs to scirpenol subfamily of trichothecene mycotoxin. In a continuation of the elucidation of the mechanism of ASD, we report here the evidences of induction of apoptosis by ASD in human Jurkat T cell line. In MTT reduction assay for monitoring cell viability, ASD showed strong toxicity. The 50 percent inhibitory concentrations of ASD against human T lymphoid Jurkat cell was 59.5 ng/$m\ell$. Phosphatidylserine externalization was increased by ASD at 3 and 6 hrs when compared with that of 6 hrs in the cell line showing in a time-dependent manner. When whole lysates of cells treated with ASD were subjected to western blot assay, 113 kDa poly(ADP-ribose) polymerase (PARP) was significantly cleaved to 89 kDa fragment. Time-dependent DNA fragmentation was also observed when Jurkat T cells were treated with ASD at 100 ng/$m\ell$ for 6 hrs and 18 hrs at the ratios of 8.5% and 15.0%, respectively. From these data, Jurkat T lymphocytes treated with ASD from Paecilomyces tenuipes underwent typical cascades of apoptotic cell death.

국내 자생 눈꽃동충하초 균의 명명 고찰

남성희 ( Nam Seong Hee ),이선주 ( Lee Seon Ju ),조세연 ( Cho Sae Yun ) 한국잠사학회 2002 한국잠사곤충학회지 Vol.44 No.1

눈꽃동충하초는 국내에서 몇 가지 학명으로 불려지고 있으나 그 중 오명이 포함되어 있음으로 인해 학자들간에 논란이 되고 있으며, 균학도들에 의해서 잘못 인용되는 사례가 빈번함에 따라 본 시험에서는 균을 재 동정코자 하였다. 수집균은 담황색 분생자병속을 1∼5여개 형성하며 그 끝마디에는 다량의 분생포자가 발생되었다. MA 배지 상에서는 25℃, 14일간 직경 2.1㎝로 더딘 생장을 보이며, colony의 색상은 앞면은 옅은 담황색, 뒷면은 연노랑 색이며, 분생포자는 타원형 혹은 난형으로 2.9∼6.6×1.5∼2.5 ㎛이며 분생포자를 지탱하고 있는 phialide는 부풀고 팽창한 형태로 3∼4개가 형성되며 4.0∼6.6×2.0∼2.2 ㎛이다. 본 균주는 Samson 분류에 의거하여 주요 특성이 일치하여 Paecilomyces tenuipes(Peck) Samson 균과 동일 종으로 인정되었다. Paecilomyces tenuipes (Nunkkot Dongchunghacho) was erroneously treated in three different scientific names: Paecilomyces japonica,k Isaria japonica and Paecilomyces tenuipes. Since the fungus has been widely used in various fields of research and industry, its nomenclatural uncertainty caused many problems. Therefore, morphological studies of korean isolate were carried out and compared with published data on other isolates of P. tenuipes to establish its proper taxonomec position.

눈꽃동충하초(Paecilomyces tenuipes)의 분생자경속 형성

박기범 외 한국버섯학회 2004 한국버섯학회지 Vol.2 No.2

눈꽃동충하초 분생자경속을 형성하는데 액체배양을 이용한 접종원의 배지는 PDB가 가장 우수하였으며, 분생자경속 형성에 유리한 배양온도는 20℃로 나타났다. 조명은 500lx.에서 가장 양호한 성장을 하였으며 광조사 시간은 18hr이 가장 유리한 분생자경형성 조건을 나타내었다. 알맞은 배지 조건은 번데기와 현미를 50 : 50 의 비율에 증류수를 현미에 1 : 1비율로 첨가한 혼합배지에서 높게 나타났다. 번데기배지는 원형상태와 분쇄상태 중에서는 분쇄된 상태에서 번데기배지의 공극률이 높아 균사의 침입이 용이하여 분생자경속 회수율과 균사의 배지내에 형성이 원형상태의 번데기보다 더욱 높게 나타났으며, 균사의 생장도 역시 분쇄한 번데기에서 더욱 빠르고 고르게 나타났다. 종균 접종량은 15ml의 종균을 접종하면 분생자경속이 잘 형성되었다. 눈꽃동충하초 재배에 가장 안정하게 재배하려면 분쇄한 번데기에 현미와 혼합한 배지에 종균 15ml를 접종하여 24 에서 발이를 시킨 후 20℃의 배양실에서 재배하는 것이 가장 좋은 것으로 나타났다. Potato dextrose broth was the best inoculation medium to produce in vitro synnemata of Paecilomyces tenuipes. The optimum temperature for synnemata formation of P. tenuipes was 20℃ under 500 lx of light intensity. Highest synnemata production was observed at 18 hr of light period per day. The medium containing 50 g of each silkworm pupae and brown rice produced highest number of synnemata. The optimum ratio of brown rice to distilled water was found as 1:1. Mycelial growth and synnemata production of P. tenuipes was faster and higher in medium containing grinded pupae as compared to whole pupae. The optimum inoculum amount per bottle of medium was 15ml. The highest synnemata production of P. tenuipes was obtained by incubating rice pupae medium at 24℃ until mycelium grows sufficiently after inoculation and then transferring it to 20℃ chamber till harvest.

Synnema formation of Paecilomyces tenuipes using blue LED lamp

Joo-Hong Yeo,Sang-Hoon Han,Kwang-Gill Lee,HaeYong Kweon,You-Young Jo,Ji-Young Mun,Heui-Sam Lee 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10

To synnema formation and entomopathogenic of Paecilomyces tenuipes using some of media under the blue LED lamp. Lyophilized matured silkworm hemolymph and Bombyx mori powder were used as nutrient source in cultivation of an entomopathogenic fungus, Paecilomyces tenuipes. Matured silkworm (MS, aged seventh days of fifth instars) was in whole lyophilized without any treatment. Hemolymph (HL) powder was prepared by water-soluble extraction from matured silkworms which was silk gland removed and also lyophilized, too. Both of media showed better growth rates and synnemata formations than potato dextrose (PD) medium, and B. mori was controlled under the blue LED lamp conditions. To investigate their effect on mycelial growth of P. tenuipes, each of them were provided as nutrient sources by final concentrations of 2, 3 and 4%, respectively. Total 18 test cultures were cultivated for 14 days with no light. Three PD cultures were used as control group. Each of growth areas was measured with an image analyzer. The result showed that MS and HL medium promoted mycelial growth of P. tenuipes by 287 and 310%, respectively, in comparison with PD medium. The synnema formation and its shape of Paecilomyces tenuipes using B. mori under the blue LED lamp group are not so good than normal conditions. But, how to change of a compositions are not yet. It will be investigation in the future.

누에 품종에 따른 눈꽃동충하초 배양 특성

조유영 ( You Young Jo ),권해용 ( Haeyong Kweon ),이광길 ( Kwang Gill Lee ),김현복 ( Hyun Bok Kim ),김기영 ( Kee Young Kim ) 한국잠사학회 2015 한국잠사곤충학회지 Vol.53 No.2

동충하초는 곤충의 몸속에 들어가 버섯이 발생하는 곤충 기생성 약용 버섯으로 세계적으로 인체에 유용한 활성물질이 확인되었으며, 이를 누에를 이용하여 인공재배할 수 있는 기술이 개발되어 활용되고 있다. 그러나 누에 품종에 따른 동충하초의 생산성에 대한 연구 보고는 없는 실정이다. 본 연구에서는 장려품종 3품종(백옥잠, 금옥잠, 대성잠)과 특수품종 2품종(연록잠, 골든실크)을 이용하여 눈꽃동충하초의 재배조건과 특징을 조사하였다. 눈꽃동충하초 균주(P. tenuipes)에 대한 감염율은 금옥잠이 가장 높았으며, 백옥잠, 대성잠, 골든실크, 연록잠 순으로 감염율이 높았다. 눈꽃동충하초를 재배하기 위한 최적조건은 어두운 장소에서 번데기 당 약 4 cm2의 재배밀도로 22oC ± 1를 유지해 주는 것이었다. 또한 우리는 농가소득 향상을 위해, 동충하초의 수요가 있을 때 마다 수시로 재배할 수 있는 연중재배 기술을 개발하였다. 연중재배 방법은 먼저 동충하초균에 감염된 번데기 체내의 수분함량을 자연건조로 3% 이하로 낮추고, 이를 진공포장 후 4oC 이하에서 보관하는 3단계의 과정이 필수적이다. 이렇게 저온에 보관되어 있는 감염번데기는 언제든지 물에 침지하여 번데기 내의 함수율을 원래대로 복원 시킨 다음 최적 재배조건에서 재배하여 동충하초를 생산할 수 있게 함으로 연중생산이 가능하다. 그러므로 우리는 동충하초 재배농가가 누에 품종 중 금옥잠을 이용하고 연중생산 기술을 활용하여 소득증대를 이룰 수 있을 것으로 기대한다. There is no specific data on Entomogenous fungus (DongChungHaCho) in accordance with the silkworm varieties, despite of very high value industrial use as functional materials at domestic and abroad. In this study, we investigated culture condition and characteristics of Paecilomyces tenuipes in 3 silkworm cultivars and 2 specific silkworm breeds. Infection rate of P. tenuipes for silkworm varieties was the highest Kumokjam, followed by Baegokjam, Daeseongjam, Golden silk, Yeonnokjam in that order. The optimum culture conditions were as follows: dark condition, 22oC ± 1, and about 4 cm2 in planting density per pupa. Also, we have developed annual production technologies of P. tenuipes. First, in order that the moisture content of the infection pupa became 3% or less in the natural condition. Second, the dried pupa must be vacuum-packed and was kept under 4oC. Finally, by restoring the moisture content of the dried pupa and culture in optimal condition, the technique for annual production of the P. tenuipes was achieved. Therefore, we expect that the use of Kumokjam and annual production technique will contribute significantly to farm income.

눈꽃동충하초(Paecilomyces tenuipes)의 분생자경속 형성

박기범,박기병,부샨 쓰레스타,성재모 한국버섯학회 2004 한국버섯학회지 Vol.2 No.2

Potato dextrose broth was the best inoculation medium to produce in vitro synnemata of Paecilomyces tenuipes. The optimum temperature for synnemata formation of P. tenuipes was 20℃ under 500 lx of light intensity. Highest synnemata production was observed at 18 hr of light period per day. The medium containing 50 g of each silkworm pupae and brown rice produced highest number of synnemata. The optimum ratio of brown rice to distilled water was found as 1:1. Mycelial growth and synnemata production of P. tenuipes was faster and higher in medium containing grinded pupae as compared to whole pupae. The optimum inoculum amount per bottle of medium was 15ml. The highest synnemata production of P. tenuipes was obtained by incubating rice pupae medium at 24℃ until mycelium grows sufficiently after inoculation and then transferring it to 20℃ chamber till harvest.

Effects of Paecilomyces tenuipes‐inoculated brown rice combined with fermented oak sawdust on the developmental growth of Protaetia brevitarsis larvae

Kwak Kyu‐Won,Kim Sun Young,Kim Eunsun,Park Kwanho,Nam Sung‐Hee,Kim Yong‐Soon 한국곤충학회 2022 Entomological Research Vol.52 No.7

Protaetia brevitarsis is an edible insect registered as an alternative food ingredient by the Korean Ministry of Food and Drug Safety. Herein, we investigated the effects of Paecilomyces tenuipes on diseases reported in the larvae of edible insects with the aim of improving the productivity of larvae by promoting their development and enhancing their disease resistance. P. tenuipes exhibits several biological and pharmaceutical properties. We developed the feed for P. brevitarsis larvae with fermented oak sawdust combined with P. tenuipes–inoculated brown rice. The larval group that reared on feed combined with P. tenuipes showed a 6.5-fold increase in weight and 5-fold increase in survival rate against Metarhizium anisopliae relative to that seen with the controls. The nutrient composition of this feed formulation was 2.37 times higher in crude protein than in the other feed types, including the control. Our results suggest that P. tenuipes–inoculated brown rice can be used to increase larval weight and promote disease resistance, thereby potentially reducing the prevalence and severity fungal disease caused by M. anisopliae.

동충하초 재배상 내 기형균 및 오염균의 발생현황 및 특성

남성희 ( Nam Seong Hee ),윤철식 ( Yoon Cheol Sik ),정이연 ( Jung I Yeon ),지상덕 ( Ji Sang Duk ),조세연 ( Cho Sae Yun ),한명세 ( Han Myung Sae ) 한국잠사학회 2002 한국잠사곤충학회지 Vol.44 No.1

7개도 22개 재배농가로부터 수집한 시료는 총 529점이었으며, 수집균은 두 가지 유헝으로 구분되며 생육장애에 의한 기형균은 23.2%, 오염균은 50.9%였다. 생육장애는 C-1, C-2, C-3 및 C-4의 4가지 유형이 있으며 이들 분생자병속은 20∼70여 개가 발생되는데 회색, 순백색 또는 등황색을 띤다. 분생자병속 말단은 분지하지 않고 기주 표피에 각각 고유색상을 띠며 PDA 배지상에서 균사생장량은 14일간 43∼75 mm 범위로 눈꽃동충하초균보다 빠른 생장을 보인다. Occurrence and charaberistics of infectious fungi and physiologically abnormal Paecilomyces tenuipes were examined. These unfavorable fungi are recently known to cause yield loss, decreased quality of products in artificial cultivation farms of P. tenuipes. Total 529 fungal isolates were collected from 22 farms of 7 provinces. These isolates were composed of physilolgically abnormal P. genuipes strains and other infectious fungal strains, which constituted 23.2% abd 50.9%,k resoectuvekt. Physiologecally abnormal P. tenuipes strains showed irregular synemata,k and absence of local color. They were divided into 4 type of C-1, C-2, C-3,and C-4. Other infectious fungal strains were idevified to 5 species,Bearveria bassiana, Fusarium sp., P.fumosoroseus, Tricothecium roseum,k Aspergillus parasiticus,k which were 22.1, 13.8, 7.6, 40. and 3.4%, respectively. All of them were hyphomycetous fungi, did not produce synemata, and revealed faster growth rate than that of P. tenuipes.

Genomic Structure of the Cu,Zn Superoxide Dismutase (SOD1) Gene of Paecillomyces tenuipes and Paecilomyces sp.

박남숙,손흥대,진병래,제연호,이광식,이상몽,박은주 한국잠사학회 2005 International Journal of Industrial Entomology Vol.10 No.1

We describe here the complete nucleotide sequence and the exon-intron structure of the Cu,Zn superoxide dismutase (SOD1) gene of Paecilomyces tenuipes and Paecilomyces sp. The SOD1 gene of P. tenuipes spans 966 bp, and consisted of three introns and four exons coding for 154 amino acid residues. Three unambiguous introns in P. tenuipes separate exons of 13, 332, 97, and 20 bp, all exhibiting exon sizes identical to Cordyceps militaris SOD1 gene. The SOD1 gene of Paecilomyces sp. contains 946 bp and consisted of four introns and five exons coding for 154 amino acid residues.Five exons of Paecilomyces sp. SOD1 are composed of 13, 180, 152, 97, and 20 bp. Interestingly, this result showed that the total length of exons 2 (180 bp) and 3 (152 bp) of Paecilomyces sp. SOD1 is same to exon 2 length (332 bp) of C. militaris SOD1 and P.tenuipes SOD1. The deduced amino acid sequence of the P. tenuipes SOD1 showed 95% identity to C. militaris SOD1 and 78% to Paecilomyces sp. SOD1. Phylogenetic analysis confirmed that the C. militaris SOD1, P. tenuipes SOD1 and Paecilomyces sp. SOD1 are placed together within the ascomycetes group of fungal clade.

Molecular Cloning of the cDNA of Heat Shock Protein 88 Gene from the Entomopathogenic Fungus, Paecilomyces tenuipes Jocheon-1

Liu, Ya-Qi,Park, Nam Sook,Kim, Yong Gyun,Kim, Keun Ki,Park, Hyun Chul,Son, Hong Joo,Hong, Chang Ho,Lee, Sang Mong Korean Society of Sericultural Science 2014 International Journal of Industrial Entomology Vol.28 No.2

The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes) Jocheon-1 Uni-Zap cDNA library and performing 5' RACE polymerase chain reaction (PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of 2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88 and 45-76% identity to other fungal homologous HSP88s. 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. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminal region. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton (kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditions for the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1 HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stress induced a higher level of mRNA expression compared to normal growth conditions.