벼 흰잎마름병균(Xanthomonas oryzae pv. oryzae)의 병원성 유전자의 분자유전학적 연구현황 및 비교유전체 분석

강희완,박영진,이병무,Kang, Hee-Wan,Park, Young-Jin,Lee, Byeong-Moo 한국미생물학회 2008 미생물학회지 Vol.44 No.1

본 논문은 벼 흰 잎마름병균인 Xanthomonas oryzae pv. oryzae(Xoo)의 병원성 유전자의 분자유전학적 연구현황을 기술하고자 한다. 또한 국내 고유 벼 흰 잎마름병균 KACC10331의 유전체해독 정보를 기반으로 다른 Xanthomonas의 유전체와 비교 분석함으로써, Xoo의 주요 병원성 유전자의 분자구조를 구명하고자 한다. 이를 통해 Xoo 고유 병원성 유전자 탐색 및 기능 해석을 위한 기초자료를 제공하는데 목적이 있다. Xoo 유전체에는 5개 과(family)에 속하는 9 type의 Insertion sequence(IS)가 611 copy로 존재하고 있으며, 주로 병원성 관련 유전자군 주위에 많이 분포하고 있는 것으로 나타났다. 현재까지 연구가 수행된 주요 병원성 관련 유전자인 hypersensitive response and pathogenicity (hrp) 유전자, extracellular polysaccharide (EPS) 유전자, extracellular enzyme 유전자, lipopolysaccharides (LPS) 유전자, 그리고 avilulence 유전자의 분자유전학적 연구현황을 기술하였다. Xanthomonas oryzae pv. oryzae (Xoo) is the causal agent of bacterial blight on rice. In this paper, current status on molecular genetic study of major virulence genes, hypersensitive response and pathogenicity (hrp), productions of extracellular polysaccharide (EPS), extracellular enzymes and lipopolysaccharides (LPS), avr genes were reviewed. The IS elements with 611 copies including 133 ORF IS were inserted in various regions of the Xoo genome and in expecially regions franking virulence genes. Whole genome sequence of X. oryzae pv. oryzae KACC10331 were used for defining genetic organization of the virulence genes. Futhermore, the virulence genes in Xoo genome were compared to those of other Xanthomonas species in Blast GenBank data base.

총설: Universal Rice Primer (URP)-PCR에 의한 곰팡이 종의 유전적 다양성 검정

강희완 ( Hee Wan Kang ) 한국균학회 2012 韓國菌學會誌 Vol.40 No.2

20mer의 URP primers 가 한국 재래적 미의 반복배열 DNA 염기서열로부터 고안되어 다양한 곰팡이 종의 PCR 다형성검출에 적용 되어 왔다. URP-PCR 방법은 PCR반응중 55˚C이상의 고온에서 annealing반응을 함으로 하여 재생적인 PCR결과를 얻을 수 있으며 효모균류에서 담자 균류의 고등 균류까지 33속, 142 종, 1,489 균주의 다양한 곰팡이 종의 유전체 DNA에 적용되어 그 유용성이 평가 되어 왔다. 본 논문은 URP-PCR의 특성과 지금까지 다양한 곰팡이 종 다양성 검정결과를 종합하여 보고 한다. URP primers that were derived from repetitive DNA sequence of rice weedy rice have been applied for producing PCR polymorphisms in different fungal species. URP-PCR protocol employed stringent PCR with high annealing temperature over 55˚C throughout the thermo-cycling reaction, giving high reproducibility, Under the PCR condition, Each single URP primer produced characteristic fingerprints from diverse genomes of different fungal species, indicating its universal applicability. URP-PCR has been accessed for applicability to various fungal species with 33 genus, 142 species and 1,489 isolates. Numerous related papers have demonstrated that URP-PCR profiles of fungal species are very useful for identifying fungal species at intra and inter species levels. The results were reviewed in this paper.

보문 : 목질진흙버섯 Phellinus Linteus HN1009K의 항산화 활성 과 항염증효과

강희완 ( Hee Wan Kang ),이만휘 ( Man Hyee Lee ),서건식 ( Geon Sik Seo ) 한국균학회 2013 韓國菌學會誌 Vol.41 No.4

Novel Phellinus linteus HN1009K (HK1009) has been characterized on speedy mycelial growth and yearly artificial cultivation. In this study, antioxidant and anti-inflammatory activities of ethanol extract from mycelium and fruiting body of P. linteus HN1009K (HK1009) were compared to those of P. linteus (PL, Korea Sangwhang) and P. baumii (PB, Jangsu Sanghwang) as controls. HK1009 extract showed higher DPPH free radical scavenging activity in both mycelial and fruiting body samples than those of PL and PB. However, in xanthine oxidase assay, antioxidant effect was highly measured on mycelial samples of PL and HK1009. Mycelial extract of HK1009 induced high inhibitory effect on LPS-induced nitric oxide (NO) production in RAW264.7 cells comparing to PL and PB without toxicity on the cells. The results suggest that P. linteus HN1009K possesses superior antioxidant activity and anti-inflammatory effects.

상치유묘에서 Ferritin 유전자의 Cloning 및 발현

박진희(Jin-Hee Park),강희완(Hee-Wan Kang),노일섭(IL-Sup Nou),강권규(Kwon-Kyoo Kang) 한국원예학회 1997 한국원예학회 학술발표요지 Vol.15 No.1

느타리버섯 자실체 생육 및 미세구조에 미치는 배지의 영향

주영철,윤선미,강희완,Ju, Young-Cheoul,Yoon, Seon-Mee,Kang, Hee-Wan 한국생명과학회 2007 생명과학회지 Vol.17 No.9

느타리버섯 자실체의 형성과정에서 생육 및 미세구조에 미치는 배지기재의 영향을 검토한 결과, 배지의 종류에 따른 초발이소요일수는 폐면이 5일로 가장 빨랐고 볏짚은 대조와 같이 6일이었다. 볏짚과 폐면에서 생육한 느타리의 경도는 대조의 5715.4 $g/cm^2$에 비해 1/2정도로 낮게 나타났으나 톱밥 단독배지에서 생육한 느타리의 경도는 17067.3 $g/cm^2$으로 3배이상의 높은 경도를 나타내어 톱방이 느타리버섯의 경도에 큰 영향을 미치는 것으로 생각된다. 배지별 느타리자실체의 갓부위를 관찰한 결과, 톱밥배지의 경우 다른 자실체의 주름으로부터 비산된 담자포자들이 대량으로 떨어져 있는 것이 관찰되었으며, 볏짚배지의 경우 갓낭상체의 발달이 관찰되었으며 폐면에서 생육한 자실체의 갓의 균사와 노화가 다른 처리에 비해 빠르게 진행되어 있는 것이 관찰되었다. 또한, 톱밥 및 폐면에서 생육한 자실체의 갓의 균사의 노화가 다른 처리에 비해 빠르게 진행되어 있는 것이 관찰되었다. 또한, 톱밥 및 폐면에서 생육시킨 느타리자실체의 담자기의 자가분해 및 담자포자 비산이 빠르게 진행되었음을 확인 하였다. 톱밥배지에서 생육시킨 느타리자실체의 대부의 균사의 생육상태는 두꺼운 균사다발이 대부위 전체를 피복한 듯한 형태가 관찰되어 이러한 균사다발층이 대의 경도를 높이는 요인으로 작용했을 것으로 판단되며, 대조군에서는 풍부한 세포질 물질을 갖는 균사들이 관찰된데 반해, 톱밥에서 생육시킨 느타리자실체의 균사는 다른 처리에 비해 착색이 많이 되어 있는 균사와 액포화된 균사의 발달이 두드러짐을 확인하였다. Comparison of fruit bodies of Pleurotus ostreatus cultivar chunchu No .2 grown on the sawdust, rice straw and cotton waste substrates revealed differences in the pattern of differentiation of hyphal compartments. Required period for primordium induction of fruit bodies grown on sawdust substrate was 13 days. Physical structure shown as hardness of stipes grown on the sawdust substrate, fruit bodies were harden than control. Pileocystidia were well developed on the surface of pileus in the fruit body cultivated on rice straw. Microstructures of fruit body grown on the sawdust and cotton wastes substrates shown fast-discharge of basidiospore and sytoms ageing. Hyphae of fruit bodies formed on sawdust substrate had less stainable cytoplasmic material and many more vacuoles than hyphae of fruit bodies formed on synthetic substrate with 50% of pine sawdust, 30% of cotton seed hull and 20 of beet pulp(control).

보문 : URP-PCR핵산지문에 의한 눈꽃동충하초 (Paecilomyces japonica.)와 번데기동충하초(Cordypces militaris) 유전적 다양성분석

김종군 ( Jong Kun Kim ),강희완 ( Hee Wan Kang ) 한국균학회 2011 韓國菌學會誌 Vol.39 No.3

This study was carried out to identify the genetic characteristics among isolates of Paecilomyces spp.and Cordyceps spp. by URP-PCR analysis. Twenty URP (universal rice primer) primers of 20 mer which were designed from repetitive sequence of rice, were used for producing PCR DNA fingerprints of the mushrooms. Of them, 5 URP primers, URP2F, URP2R, URP9F, URP4R, and URP17R amplified genomic DNA of the mushrooms with polymorphic PCR patterns. On isolates of Cordyceps militaris, primers URP1F, URP2R, URP6R and URP17R produced PCR polymorphic bands of 4 types. Isolates of Cordypces sp. that are isolated from different area of Korea were identical to isolate of C. militaris, while other species of Cordypces were different to the PCR profiles. However, the URP primers did not identify the polymorphism of PCR profile on isolates of P. japonica.

버섯수확후배지의 퇴비화에 따른 물리 화학적 변화 및 식물 생육 효과

송지민,김자윤,강대선,유정연,강희완,Song, Ji-Min,Phong, Nguyen Hong,Kim, Ja-Yoon,Kang, Dae-Sun,Yu, Jeong-Yeon,Kang, Hee-Wan 한국버섯학회 2020 한국버섯학회지 Vol.18 No.3

표고(Lentinula edodes), 노루궁뎅이(Hericium erinaceus), 느타리(Pleurotus ostreatus)의 버섯수확후배지(spent mushroom substrate, SMS)를 퇴비화에 활용하였다. 퇴비화 온도변화는 50℃ 이상에서 7일에서 10일 동안 지속되어 30일에 부숙이 완료되었다. 퇴비화 된 LeCSMS, HeCSMS는 130%, 80% 무종자에 대한 종자 발아지수를 보여 부숙 도에 대한 종자 발아지수를 충족하였다. 퇴비화에 따른 물리 화학적 변화를 조사 한 바 pH 범위가 4-5에서 6-7로 증가 되었고 EC는 1-1.4 dS/m로 소폭 감소되었으며 유기질 함량은 LeCSMS에서 36.9%로 SMS에 비하여 60% 이상 가장 큰 폭으로 감소 되었다. LeCSMS를 기준으로 N (1.2%), P(2.3%), K(0.77%)함량이 조사되었으며 중금속은 모든 CSMS에서 기준치 이하였으며 Ca, Mg는 30%에서 60%이상 증가하였다. C/N비는 LeCSMS, HeCSMS에서 15% 낮았으나 PoSMS에서 32%로 높게 나타났다. LeCSMS 처리에 따른 고추 유묘 생육효과는 시판 유기퇴비 처리구 등 대조 구에 비하여 초장이 60%이상 높게 성장했으며 옆폭, 옆장, 옆수에서 다른 대조 구에 비하여 모두 우수한 생육 효과를 보였다. This study aimed to assess the feasibility of composting spent mushroom substrate (SMS) materials of Lentinula edodes (Le), Hericium erinaceus (He), and Pleurotus ostreatus (Po). The different SMSs were composted for 7 to 10 days at high temperatures over 50℃; the composting procedure was completed in 30 days. A maturity test was conducted using the radish seed germination index and CoMMe-100. The composted SMS (CSMS) from Le and He showed gemination indices of 130% and 81%, respectively, that satisfied the criteria of maturity standard (gemination index over 70%) and the CoMMe-100 analysis. The physicochemical changes of CSMSs included an increase in the pH range from 4-5 to 6-7, slight reduction in the EC to 1-1.4 dS/m, and an organic content of 36.9% in LeCSMS. In LeCSMS, the contents of N, P, and K were 1.2%, 2.3%, and 0.77%, respectively, and heavy metals were detected below the standard value in all CSMSs; the Ca and Mg contents in the CSMSs were increased from 30% to 60% when compared to those in the SMSs. The C/N ratio (from 26-33) in LeSMS and HeSMS decreased to 15.3-15.9 in CSMSs. The growth effect of LeCSMS treatment on pepper seedlings was 60% higher than that in the control groups, one of which was treated with commercial organic compost; the former showed a superior growth effect on the leaf width, leaf length, and leaf number compared to other control groups. In conclusion, LeCSMS and HeCSMS could be utilized as compost resources capable of efficient soil amendment and plant growth promotion.

원형질체융합 기법을 이용한 산느타리 계통육성

권희민,이윤혜,김정한,백일선,강희완,최종인,Gwon, Hee-Min,Lee, Yun-Hae,Kim, Jeong-Han,Baek, Il-Sun,Kang, Hee-Wan,Choi, Jong-In 한국버섯학회 2021 한국버섯학회지 Vol.19 No.3

원형질체 융합 기술은 종·속간 유전적 한계를 넘어 육종과 그 소재로 활용하고데 목적이 있다. 본 연구에서는 '흑타리'(P. ostreatus)와 '호산'(P. pulmonarius)의 단핵균사를 이용하여 원형질체를 나출하고 나출된 원형질체를 융합하여 종간 교배 계통을 육성하였다. 육성계통의 균사생장속도는 '호산', '흑타리', PF160313, PF160306 계통 순으로 빠른 편이었다. 균사 밀도는 PF160306 계통이 가장 높았고, 나머지는 중간 수준의 밀도를 나타내었다. 원형질체 융합계통인 PF160306과 PF160313 계통은 '흑타리' 품종보다 배양 기간이 10일, '호산' 품종보다 2일 단축되었다. 자실체 생장 기간은 '흑타리'와 '호산에 비하여 각각 3일, 1일 단축되었다. PF160306 계통의 생산량은 135.9 g/병으로 '호산'에 비하여 높았으나 통계적으로 유의차가 없었다. 자실체 발생기간은 15℃에서 9일, 25℃에서 4.5일로 온도가 높아짐에 빨라졌다. 자실체의 갓색은 21℃ 노란색이 가장 선명하게 발현되었다. URP primer 7을 사용하여 PCR 밴드 패턴을 비교하였을 때, 전체적으로 '호산' 품종과 유사하였다. DPPH radical 소거능과 폴리페놀 함량에 있어 '순정'은 각각 62.5%, 43.5 mg/mL였으며, PF160313 계통은 각각 65.7%, 49.9 mg/mL를 나타내어 계통간 유의차가 있었다. ACE 활성은 '순정' 74%, PF160313 계통 75%로 유사한 수준이었다. In this study, monokaryons of "Heukari" (Pleurotus ostreatus) and "Hosan" (Pleurotus pulmonarius) were separated to remove the cell wall, and a cross-species protoplast fusion was developed through chemical treatment with polyethylene glycol. The protoplast-fused PF160306 and PF160313 strains have a culture period of 10 and 2 days shorter than that of the "Heuktari" and "Hosan" cultivars, respectively. Furthermore, the growth of the strains was faster than that of the existing cultivars. The yield was 135.9 g per bottle, which was approximately 8% higher than that of the commercially available "Hosan" cultivar; however, it was not statistically significant. A growth survey was conducted after treatment at five temperatures (15, 18, 21, 23, and 25℃). The growth of the strains accelerated with the increase in temperature. However, at 21℃, the yellow color of pileus was the brightest. Band pattern, assessed using URP Primer 7, was similar to that of the "Hosan" cultivar. The DPPH radical scavenging capacity and polyphenol content were 62.5% and 43.5 mg/mL, respectively, for "Sunjung" and 65.7% and 49.9 mg/mL, respectively, for PF160313. Furthermore, the antihypertensive activities of the "Sunjung" cultivar and PF160313 were similarly high at 74% and 75%, respectively. In conclusion, cross-species hybridization via the protoplast fusion technique can be used for obtaining primary data for mushroom breeding to develop new varieties. In addition, the protoplast fusion technique might aid in expanding the market for yellow mushrooms.

Protective Effect of Composted Spent Mushroom Substrate (Hericium Erinaceus) on Phytophthora Capsici

응웬홍파옹 ( Nguyen Hong Phong ),강희완 ( Hee-wan Kang ) 한국농공학회 2020 한국농공학회 학술대회초록집 Vol.2020 No.-

Understanding crop water requirements (CWR) is essential for the better irrigation practices. Scheduling and efficient use of water since the water supply through rainfall is limited. The main objectives of the research were estimated crop water requirement and irrigation scheduling for rice crops, in addition, to increase agricultural productivity and improving irrigation efficiency uses. The irrigation schedules and water requirements were simulated using CROPWAT, and the model result was verified with field values in the Maliana Irrigation Scheme, Bobonaro District of East Timor. The results indicated the wider applicability of CROPWAT in calculating water requirement as well as developing irrigation schedules for rice irrespective of the agro-climate conditions. The specific objectives were (i) to estimate reference evapotranspiration (Eto), (ii) to an estimate of the effective rainfall from the Maliana stations, (iii) to estimate of crop water requirements for rice crops, (iv) to estimate irrigation schedule. The average value of reference evapotranspiration of the crop was 4.98mm/day. It ranges from 4.21mm/day to 5.90mm/day. The maximum evapotranspiration of crop was accorded in the month of September and the minimum were January. The maximum effective rainfall of 155. 2mm in the month of January and a minimum of 5.3mm during August. The total irrigation water requirement for the rice crop the season from December to April was 435.3mm. This low water requirement for rice is mainly due to higher effective rainfall in the site of study. The net irrigation applied to the field varies from 90mm to107.8mm with an average net irrigation of 486.9mm. The gross irrigation water requirement ranges from 128.57mm to 154mm with average gross irrigation water of 695. 6mm.

효모세포벽추출물에 의한 고추내 salicylic acid 생성유도

강대선 ( Dae Sun Kang ),조수묵 ( Soo Muk Cho ),강희완 ( Hee Wan Kang ) 한국균학회 2012 韓國菌學會誌 Vol.40 No.4

효모추출물 4 mg/mL를 고추유묘에 경엽 및 뿌리 관주처리 24시간 후에 salicylic acid(SA)함량을 ultra high performance liquid chromatography(UHPLC)분석방법으로 측정한 결과 yeast cell wall extract(YCWE)처리구에서 대조구의 SA처리구와 동일한 peak가 검출되어 SA가생성 유도 되는 것으로 확인 되었다. SA는 YCWE를 경엽처리 48시간 후 20.29 ug/g으로 가장 높게 나타났으며 관주처리보다 약 3.7배의 높은 SA검출함량을 보였으나 처리 후 72시간부터는 점차적으로 SA함량이 감소하는 양상을 보였다. Yeast cell wall extract (YCWE) was treated on leaves and roots of pepper seedlings at the dosage of 4mg/mL and salicylic acid (SA) production in pepper was detected by ultra high performance liquid chromatography (UHPLC). The SA production in pepper stem was induced by YCWE. SA was produced at the highest level of 20.29ug/g after 48 hrs of foliar spray with YCWE, which is 3.7 times higher than that of root perfusion with YCWM. SA production was gradually reduced after 72 hrs of YCWE teatment.