RFLP, PLFA, CLSU를 이용한 폐기물연용토양의 토양미생물 특성 평가 비교

장갑열(Kab-Yeul Jang),원항연(Hang-Yeon Weon),이강효(Kang-Hyo Lee),권순익(Sun-Ik Kwon),공원식(Won-sik Kong),서장선(Jang-sun Suh),성재모(Jae-Mo Sung) 한국토양비료학회 2008 한국토양비료학회지 Vol.42 No.6

The application of sludge wastes into agricultural fields has been increasing annually in Korea. In particular, sewage sludge application has been widely accepted in decades. Sewage sludge application aid in the recycling of essential nutrients and act as a source of organic matter improving the structure and water-holding properties of the soil. The efficient use of sludge wastes, however, requires an individual assessment of waste products. This study assessed the biological characteristics of organic waste-treated lysimeter soils and develop its indicator to assess the soil health of organic waste-treated lysimeter soils. Several analytical techniques more recently developed such as restriction fragment length polymorphism (RFLP), phospholipid fatty acid (PLFA), and community level substrate utilization (CLSU) fingerprints allow for detailed analyses of soil microbial communities. PLFA and RFLP was, therefore, used in the study to characterize the microbial communities in soil without the need to isolate individual fungi and bacteria. PLFA, RFLP and CLSU have been utilized to assess microbial characteristics of the lysimeter soils with four different sludge wastes for eight consecutive years. Each of these methods was analyzed for a different aspect of soil microbial characteristics. The study would disclose those methods yielded highly reproductive results for each soil and allow distinguishing the soils based on the structures of specific geneand PLFA-pools more than CLSU fingerprints. PLFA methods, especially, revealed the same relative similarities of the treated soils based on cluster analysis of the biological characteristics. Pig manure compost-treated soil, however, was only the same relative resemblance among the three methods. These results indicated that PLFA easily assessed the biological soil characterization.

Bacillus subtilis S37-2 균주의 항진균활성 및 식물생육촉진 효과

권장식(Jang-Sik Kwon),원항연(Hang-Yeon Weon),서장선(Jang-Sun Suh),김완규(Wan-Gyu Kim),장갑열(Kab-Yeul Jang),노형준(Hyung-Jun Noh) 한국토양비료학회 2007 한국토양비료학회지 Vol.40 No.6

양송이 액체접종원을 이용한 종균 제조 방법 개발

오연이,장갑열,오민지,임지훈,Oh, Youn-Lee,Jang, Kab-Yeul,Oh, Min Ji,Im, Ji-Hoon 한국버섯학회 2021 한국버섯학회지 Vol.19 No.2

Currently, the spawn of the mushroom Agarcus bisporus is produced by a method developed in the 1980s, and anew spawning method needs to be developed to improve the quality of the spawn. In this study, the condition for a maximum mycelium weight(5.92±0.52 g/L) was shaking culture (24 hours/day) at 24℃ and 120 rpm in CDB (compost dextrose broth). Based on this, the ventilated liquid culture method (2.5 L/min) was cultured for 10 days. This method was appropriate, andwhen the inoculum was cultured at 50 g/mL for about 10 days, it was cultured well without agglomeration and shaking of seed. 현재 양송이 종균은 1980년대 개발된 방법으로 제조되고 있어, 양송이 종균 품질 향상을 위한 새로운 제조 방법을 개발하였다. 그 결과 균사체량이 5.92±0.52 g/L로 가장 많이 배양된 CDB(compost dextrose broth) 배지에서 24℃에서 120 rpm으로 진탕 배양(24시간/일) 하는 통기식(2.5 L/min) 액체배양법으로 배양기간 10일이 액체 접종원 사용으로 양호했으며, 양송이 밀 배지 종균 생산은 밀 배지 50 g당 액체 접종원 1ml을 적정 접종량으로 하여 10일 배양하면 종균의 뭉침과 흔드는 작업없이 적절하게 배양되었음을 보고한다.

양송이 원형질체 분리와 PEG 형질전환법의 최적화

김민식,장갑열,이윤상,오민지,임지훈,오연이,Kim, Minseek,Jang, Kab-yeul,Lee, Yun-Sang,Oh, Min Ji,Im, Ji-Hoon,Oh, Youn-Lee 한국버섯학회 2021 한국버섯학회지 Vol.19 No.3

현재 양송이 품종의 개발은 1980년대에 개발된 방법에 의존하여 진행되고 있다. 유전자가위를 이용한 유전자교정 기술이 다양한 분야에서 각광받고 있고, 이 기술을 버섯 육종에도 적용하기 위하여 진행된 이 연구에서는, CRISPR/Cas9 활용에 필수적인 원형질체 분리 효율을 1.0 × 10⁸/mL까지 안정적으로 끌어올렸고, spermidine을 이용하여 PEG 형질전환의 효율 또한 기존 방법에 비해 100배가량 끌어올렸음을 보고한다. Currently, button mushroom, Agaricus bisporus is one of the most consumed mushroom in the world. However, despite of its importance in food market, molecular genetic modification method for breeding of A. bisporus is not well established. In this study, we optimized yield of A. bisporus protoplast with Lysing enzyme, Chimax-N and cellulase. With this composition, 1.0 × 10⁸/mL of protoplasts were obtained reliably. PEG-mediated transformation with spermidine showed almost 100-fold higher yield than non-spermidine method.

고온성 백색 양송이 신품종 '하담' 육성

오연이,오민지,임지훈,장갑열,Oh, Youn-Lee,Oh, Min Ji,Im, Ji-Hoon,Jang, Kab-Yeul 한국버섯학회 2020 한국버섯학회지 Vol.18 No.3

기후변화에 대비하여 고온성 양송이 백색 품종을 육성한 결과는 다음과 같다. 모본은 KMCC00540, KMCC00591, KMCC00643을 선발하였고, ISSR과 SSR마커로 각각 동핵균주를 선발하여 교잡하였다. 선발 교잡주를 고온 20~25℃, 습도 80% 이상에서 3반복으로 재배하여 농업적 형질과 자실체 특성의 변이가 가장 적은 것은 Abs4-2016-121 계통이었다. 이 계통은 KMCC00000와 KMCC0000를 교잡한 계통이며, 경주와 부여 농가에서 현장실증시험결과 Abs4-2016-121 계통이 자실체 경도가 높으며 고온에서 갓개열이 지연되어 고온성 품종 '하담'으로 육성하였다. We undertook a breeding program to produce a white button mushroom cultivar with high temperature tolerance in preparation for climate change. The results were as follows. The strains KMCC00540, KMCC00591, and KMCC00643 were selected, and homokaryons were selected and hybrids were identified with ISSR and SSR markers, respectively. The selected hybrids were cultivated in three repetitions at a temperature of 20-25 ℃ and a humidity of 80% or higher. The variation in agricultural traits and fruiting body characteristics was least in the Abs4-2016-121 strain. This line is a hybrid of KMCC00591 and KMCC00875, and field experiments at Gyeongju and Buyeo farms found that the Abs4-2016-121 line has high fruiting body with hardness and delayed opening of pileus at high temperatures, so it was bred as a high-temperature cultivar named 'Hadam'.

CAPS 마커를 이용한 국내 개발 양송이 품종 구분

이화용,안혜진,오연이,장갑열,정종욱,Lee, Hwa-Yong,An, Hyejin,Oh, Youn-Lee,Jang, Kab-Yeul,Chung, Jong-Wook 한국버섯학회 2021 한국버섯학회지 Vol.19 No.4

The cleaved amplified polymorphic sequence (CAPS) marker uses a restriction enzyme recognition site resulting from single nucleotide polymorphisms and insertions and deletions on the DNA sequence. This technique does not require expensive equipment, the process is simple, and clear results can be obtained reliably. In this study, Agaricus bisporus cultivars SaeA, SaeDo, SaeHan, SaeYeon, SaeJeong, Dodam, Seolgang, Dahyang, Hogam, and Hadam developed in Korea were discriminated using four CAPS markers. Our results indicated that it is possible to distinguish the ten cultivars and determine the genetic diversity among them. 본 연구에서는 국내에서 개발된 국내에서 개발된 10개의 양송이 품종을 CAPS 마커를 이용하여 구분하였다. An 등(2021)이 개발한 CAPS 마커 AB-gCAPs-017, AB-gCAPs-047, AB-gCAPs-055, AB-gCAPs-071을 이용하여 새아, 새도, 새한, 새연, 새정, 도담, 설강, 다향, 호감, 하담을 구분할 수 있었다. 본 연구의 결과는 국내 개발 양송이 품종에 대하여 품종간의 구별성과 유전적 다양성을 부여하여 품종보호에 대한 분자생물학적 근거를 마련하였다.

보문 : 느타리 버섯에서 원형 품종 특이 SCAR marker 개발

서경인 ( Kyoung In Seo ),장갑열 ( Kab Yeul Jang ),유영복 ( Young Bok Yoo ),박순영 ( Soon Young Park ),김광호 ( Kwang Ho Kim ),공원식 ( Won Sik Kong ) 한국균학회 2011 韓國菌學會誌 Vol.39 No.1

Weonhyeong is one of important commercial strains. It has good characteristics of bundle formation, grey colored pilei and high productivity. We previously reported grouping of 70 strains of Pleurotus ostreatus in which one group contained 35 strains including Weonhyeong. Four strains in that group showed same profiles implicating no variety distinction for mushroom cultivation. Now we developed a specific marker for identification of Weonhyeong. Sequence Characterized Amplified Region (SCAR) marker was developed from the RAPD amplicon. SCAR marker ``S-OPO5`` produced only one band specific to 2183, 2240, 2595 and 2725 strains showing similar banding patterns to Weonhyeong in RAPD-PCR results. The sequence of ``S-OPO5`` marker was unknown when compared with the data in the Genbank using BLASTN. BLASTX results indicated that the marker showed significant alignment with the protein sequences in Tricholoma bakamatsutake reverse transcriptase. The results indicate that this new SCAR marker (``S-OPO5``) will be valuable to distinguish the Weonhyeong similar strains from Pleurotus spp.

보문 : Universal Rice Primer(URP)에 의한 DNA 핵산지문법을 이용한 느타리의 유통 품종간 구분

서경인 ( Kyoung In Seo ),장갑열 ( Kab Yeul Jang ),유영복 ( Young Bok Yoo ),박순영 ( Soon Young Park ),김광호 ( Kwang Ho Kim ),공원식 ( Won Sik Kong ) 한국균학회 2008 韓國菌學會誌 Vol.36 No.2

계절별 온도에 따른 느타리 품종의 재배적 특성

유영복,서경인,공원식,장갑열,신평균,박윤정,Yoo, Young Bok,Seo, Kyoung In,Kong, Won Sik,Jang, Kab Yeul,Shin, Pyung Gyun,Park, Yunjung 한국버섯학회 2009 한국버섯학회지 Vol.7 No.3

최근까지 보급된 느타리버섯류 81개 품종을 URP를 이용하여 PCR에 의한 DNA 패턴으로 유연관계 분석과 자실체 특성조사로 품종간의 유사성을 결정하였고, 계절별로 상자재배 하여 온도 적응성에 대한 그 특성을 분석하였다. 재배품종의 DNA 패턴과 자실체 특성분석으로 품종의 유연관계를 조사한 결과 원형느타리는 1, 수한은 15, 왕흑평은 4, 춘추2호는 6, 신농11호는 1, 삼구01호는 2품종과 유사하거나 구별성이 뚜렷하지 않았다. 또한 수한 근연종으로는 5품종으로 나타났다. 느타리는 버섯 생육 시 산소공급을 위하여 환기가 필수적이므로 재배사내에서 외부기온의 영향을 크게 받는다. 따라서 계절에 따라 알맞은 품종을 선택하여야 에너지 절감과 생산량을 높일 수 있다. 봄~여름 재배 시 높은 온도($16^{\circ}C$ 이상)에 적응성이 강한 품종(내서성이 강함) 으로는 사철느타리종, 여름느타리종, 분홍느타리종의 8개 품종들이 속하였다. 봄~여름 재배 시 높은 온도에 적응성이 다소 강한 품종(내서성이 다소 강함)으로는 느타리종 51개 품종으로 나타났다. 가을~겨울 재배 시 낮은 온도($11^{\circ}C$ 이하)에 적응성이 다소 강한 품종으로는 느타리종 64품종으로 나타났다. 사계절 재배 시 적응성이 다소 강한 품종으로는 49개 품종으로 나타났다. 큰느타리, 아위느타리, 전복느타리종은 균사생장이 느려 오염이 심하여 상자 재배에서는 적합하지 않았다. 이러한 품종 특성 구명은 품종의 계절별 올바른 선택으로 에너지 절감과 생산성 향상에 의한 농가소득에 기여할 것이며, 나아가 농가와 종균배양소간의 분쟁해소, 품종육성자의 본인 개발 품종에 대한 상대적 위치, 육종모본 선택, 농가지도 사업에 유용하게 활용될 수 있을 것이다. We determined the genetic relatedness among 81 commercial varieties of oyster mushrooms by either comparison of fruiting body characteristics or using PCR amplification with URP primers. Also, we assessed their temperature adaptation by using box cultivation. Based on our data, some varieties such as Wonhyung and variety 1, Suhan and variety 20, Wang-hukpyung and variety 4, Chunchu #2 and variety 6, Shinong #11 and variety 1, Samgu #1 and variety 2, showed the close genetic relatedness between each other. It is known that mushroom culture room is affected by outer temperature because mushroom need the adequate aeration for their growth. Therefore, in aspect of saving energy, the cultivation of season favorable mushroom species has been recommended. Eight different commercial varieties including Sa-chul, Yeoreum and Bunhong(Pink) grew well at high temperature ($16^{\circ}C$). Also, about 51 different oyster mushroom strains were good at relatively high temperature. However, other 64 different oyster mushroom varieties were grew well at low temperature ($11^{\circ}C$). Fourty-nine varieties were adapted well in most temperatures. The slow growth rate were observed in some varieties including Kun(P. eryngii ), Awi(P. nebrodensis ), and Cheunbok(P. abalonus ), which are not suitable for box cultivation. We believe that the culture characterization of each mushroom varieties should help many farmers in many aspects such as saving energy, quantity improvement and new variety development etc.

버들송이버섯(Agrocybe cylindracea)유전자원 및 선발계통의 특성

허병수,유영진,서상영,최규환,최영민,권석주,장갑열,Heo, Byong-Soo,Yoo, Young-Jin,Seo, Sang-Young,Choi, Kyu-Hwan,Choi, Young Min,Kwon, Seog-Ju,Jang, Kab-Yeul 한국버섯학회 2019 한국버섯학회지 Vol.17 No.2

주름버섯목(Agricales), 소똥버섯과(Bolbitiaceae), 볏짚버섯속(Agrocybe)에 속하는 버들송이버섯은 국내에서 대부분 병 재배용 품종으로 육성되어 있어 수량 증대를 위한 봉지재배용 품종을 개발하고자 본 연구를 수행하였다. 버들송이버섯 33개 유전자원을 수집하고 유전자원의 봉지재배 특성과 수량을 조사한 후 재배일수와 자실체 모양을 고려하여 5개 모본을 선발하고 3개의 교배조합을 작성하여 7계통을 선발하였다. 선발된 7계통과 대조품종인 '참'을 봉지재배한 결과 재배일수는 'JBAC15-1'이 49일, 'JBAC15-6'이 50일로 대조품종의 재배일수인 53일 대비 각각 4일과 3일이 단축되었으며, 그 외의 계통은 대조품종보다 긴 재배일수를 나타내었다. 선발계통의 발이율은 'JBAC15-1'이 96.1%로 가장 우수하였으며 다음으로 'JBAC15-6'이 94.5%를 나타내어 대조품종의 발이율인 93% 대비 각각 3.1%, 1.5% 우수한 결과를 나타내었다. 봉지 당 수량은 'JBAC15-1'이 176.8 g으로 가장 우수하였고 'JBAC15-6'이 168.7 g으로 대조품종인 '참'의 160.7 g 대비 각각 10%, 5% 증수되었다. 이러한 결과를 토대로 대조품종인 '참' 대비 재배일수가 짧고 수량이 우수한 'JBAC15-1'과 'JBAC15-6'을 최종 선발하였다. 선발계통인 'JBAC15-1', 'JBAC15-6'의 균사생장 최적 배지는 PDA, 최적 온도는 $27.5^{\circ}C$, 최적 pH는 'JBAC15-1'이 pH 5, 'JBAC15-6'이 pH 6으로 나타났다. 자실체 갓의 색은 'JBAC15-1'과 '참'이 진갈색을 나타냈으며 'JBAC15-6'은 연갈색을 나타내었다. 갓 형태는 반구형으로 모두 동일하였으나, 대의 색은 'JBAC15-1'이 연갈색을 나타냈으며 'JBAC15-6'은 흰색을 나타냈고 대조 품종인 '참'은 상아색으로 각각 서로 다른 특성을 나타내었다. Agrocybe spp. belongs to the Agaricales order, Bolbitiaceae family, and Agrocybe genus. In Korea, so far, it has been cultivated through bottle cultivation; therefore, this study was conducted for the development of a new cultivar using the bag cultivation method for quantitative improvement. Thirty-three gene resources of Agrocybe spp. were collected and their quantity and characteristics of bag cultivation were examined. Next, 5 kinds of crossing parents were selected based on the cultivation period and shape of the fruit body. Seven strains were selected through 3 cross combinations. The 7 selected strains and the comparative cultivar 'Cham' were cultivated in a bag. As a result, the cultivation period was 49 days for 'JBAC15-1' and 50 days for 'JBAC15-6' which are 4 days and 3 days less than the cultivation period of the comparative cultivar 'Cham'(53 days), respectively. Cultivation periods of other strains except for 'JBAC15-1' and 'JBAC15-1' were longer than that of the comparative cultivar'Cham'. The best ratio of primordia formation among the selected strains was found to be that of 'JBAC15-1' with 96.1% followed by 'JBAC15-6' with 94.5%. These rates were 3.1% and 1.5% higher than the ratio of primordia formation of the comparative cultivar 'Cham', which is found to be 93.0%. The quantity was maximum in the 'JBAC15-1' cultivar with 176.8 g per bag followed by 'JBAC15-6' with 168.7 g per bag. The quantities were 10% and 5% more than the comparative cultivar 'Cham' with 160.7 g per bag. Based on these results, 'JBAC15-1' and 'JBAC15-6', which had shorter cultivation periods and more quantities than the comparative cultivar 'Cham' were finally selected. For the selected strains of 'JBAC15-1' and 'JBAC15-6', mycelial growth was observed to be optimal on PDA medium and the optimum temperature was $27.5^{\circ}C$. The optimum pH was pH 5 for 'JBAC15-1' and pH 6 for 'JBAC15-6'. The color of the pileus of the fruit body was dark brown in 'JBAC15-1' and 'Cham' and light brown in 'JBAC15-6'. The pileus was hemispherical in shape in both 'JBAC15-1' and 'Cham'. However, the colors of the stem were different - light brown in 'JBAC15-1', white in 'JBAC15-6', and ivory in the comparative cultivar 'Cham'.