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노태환,강미형,서수좌,심형권,최만영,박종철,백채훈,김형무 한국국제농업개발학회 2013 韓國國際農業開發學會誌 Vol.25 No.2
Real Time PCR을 이용한 벼 흰잎마름병균의 밀도를 측정할 수 있는 새로운 방법을 개발하였다. Real Time PCR을 이용하여 벼 흰잎마름병을 예찰하기 위하여 TaqMan MGB probe를 이용한 프라이머인 Xan_PahgeF & Xan_PahgeR primer와 Xan_Pahge FAM MGB probe를 제작하였으며, 높은 특이성이 인정되었다. 병원균 배양액, 벼 흰잎마름병원균의 DNA, 병원세균에 오염된 물에서도 효과적으로 검출이 되었다. 농수로물이나 관개수에서 벼 흰잎마름병균의 밀도를 측정할 때 정량 PCR의 저해인자 제거 후 신속하게 벼 흰잎마름병균의 밀도를 측정할 수 있다. We developed a new measurement method for the density of Xanthomonas oryzae pv. oryzae(Xoo), a casual pathogen of bacterial leaf blight of rice. The new method used real time PCR with the highly characterized and specific Xan_PahgeF & Xan_PahgeR primer and Xan_Pahge FAM MGB probe primer pairs derived from phage-related integrase gene. The method was effective when tested with a variety of templates including isolated Xoo DNA, pure colonies or liquid culture sources, and could also work on contaminating water. A main advantage of this assay is to accurately and rapidly quantitate the population density of Xoo in paddy field water containing inhibitors.
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애멸구 밀도에 따른 벼 생육시기별 벼 줄무늬잎마름병 발병정도
노태환 ( Tae Hwan Noh ),정관호 ( Kwan Ho Jung ),서수좌 ( Su Joa Seo ),심형권 ( Hyeong Kwon Shim ),최만영 ( Man Young Choi ),박종철 ( Jong Chul Park ),이왕휴 ( Wang Hyu Lee ),김형무 ( Hyung Moo Kim ) 전북대학교 농업과학기술연구소 2013 농업생명과학연구 Vol.44 No.1
We have survey on Rice stripe virus (RSV) incidence according to the density of small brown plant hopper (SBPH) in the individual rice plant, both in the nursery box and in the growing stage. A group of viruliferous SBPH was made by inoculation to rice, Dongjin 1 was infected with RSV for one week and reared continuously on the diseased rice in net box. The inoculation density of 8 viruliferous SBPH per rice resulted in 100% infection of the disease. The infection rate were 15, 30 and 60% with the inoculation density of 1, 2 and 4 SBPH per plant, respectively. Virus disease incidence increased two times as the density of SBPH doubled. The SBPH was inoculated with a group of 10, 20, 40, 80 and 160 SBPH per nursery box. Minimum insect vector density required for the infection of the disease was 80 SBPH with the occurrence rate of 0.3% in RSV. The double score of 0.6% was also reached by the double density of 160 SBPH. However, RSV was not occurred with the inoculation density of 10 to 40 SBPH. In seedling stage of susceptible rice varieties in paddy field, the RSV disease index were 70% for Dongjin 1 and 61% for Unkwang. However for Nampyeong and Onnuri which were resistant varieties, the RSV disease index of 1 and 3% observed. In maximum tillering stage of the rice Dongjin 1 and Unkwang infected with the disease 24 and 19% of disease index, but resistance varieties were not infected with the disease.
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연구논문 : 자연과학; 정량 PCR을 이용한 벼 흰잎마름병균 밀도 측정
노태환 ( Tae Hwan Noh ),강미형 ( Mi Hyung Kang ),서수좌 ( Su Joa Seo ),심형권 ( Hyeong Kwon Shim ),최만영 ( Man Young Choi ),박종철 ( Jong Chul Park ),백채훈 ( Chae Hoon Paik ),김형무 ( Hyung Moo Kim ) 한국국제농업개발학회 2013 韓國國際農業開發學會誌 Vol.25 No.2
Real Time PCR을 이용한 벼 흰잎마름병균의 밀도를 측정할 수 있는 새로운 방법을 개발하였다. Real Time PCR을 이용하여 벼 흰잎마름병을 예찰하기 위하여 TaqMan MGB probe를 이용한 프라이머인 Xan_PahgeF & Xan_PahgeR primer와 Xan_Pahge FAM MGB probe를 제작하였으며, 높은 특이성이 인정되었다. 병원균 배양액, 벼 흰잎마름병원균의 DNA, 병원세균에 오염된 물에서도 효과적으로 검출이 되었다. 농수로물이나 관개수에서 벼 흰잎마름병균의 밀도를 측정할 때 정량 PCR의 저해인자 제거 후 신속하게 벼 흰잎마름병균의 밀도를 측정할 수 있다. We developed a new measurement method for the density of Xanthomonas oryzae pv. oryzae(Xoo), a casual pathogen of bacterial leaf blight of rice. The new method used real time PCR with the highly characterized and specific Xan_PahgeF & Xan_PahgeR primer and Xan_Pahge FAM MGB probe primer pairs derived from phage-related integrase gene. The method was effective when tested with a variety of templates including isolated Xoo DNA, pure colonies or liquid culture sources, and could also work on contaminating water. A main advantage of this assay is to accurately and rapidly quantitate the population density of Xoo in paddy field water containing inhibitors.
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노태환 ( Tae Hwan Noh ),이두구 ( Du Ku Lee ),심형권 ( Hyeong Kwon Shim ),서수좌 ( Su Joa Seo ),최만영 ( Man Young Choi ),박종철 ( Jong Chul Park ),백채훈 ( Chae Hoon Paik ),김형무 ( Hyung Moo Kim ) 전북대학교 농업과학기술연구소 2013 농업생명과학연구 Vol.44 No.1
Changes Distribution and Disease Occurrence of Rice Bacterial Leaf Blight on Rice in Korea We have recorded the date when the first symptom of BLB occurred during the seven years from 2006 to 2012 in south-western coast region in Korea including Iksan, Jeongup, Buan, Kimje, Jangheong and Suncheon. The disease occurrence date ranged from early in July to early in August, which was delayed for 10 to 30 days in 2012 than 2006. The BLB suspected area was decreased during the recent seven years compared to 29,485 ha in 2006. The BLB disease occurrence area was 10,175 to 19,605 ha from 2007 to 2011 and 2,345 ha in 2012. A total of 1,078 Xoo strains collected from 2009 to 2011 were grouped into five races in the southern regions in Korea including Chungnam, Jeonbuk, Jeonnam, Kyeongbuk and Kyeongnam provinces by using Korean race differential system. The Xoo race distribution rate was similar among the races with K1 (28.9~31.0%), K2 (24.9 27.1), K3 (36. 0~40.0), K4 (2.2~3.3), and K5 (3.0~4.4) for three years from 2009 to 2011. The K4 and K5 races with more virulence were distributed in south-western coast in Korea including Kunsan, Iksan, Sunchang, Kimje, Buan, Jeongup, Jeonju, Damyang, Yeongkwang, Hampyeong, Jangsung, Naju, Heanam, Yeongam, Kangjin, Jangheong and Suncheon.
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벼 흰잎마름병의 신속하고 간편한 진단을 위한 Recombinase Polymerase Amplification 등온증폭법
김신화(Shinhwa Kim),이봉춘(Bong Choon Lee),김현주(Hyun Ju Kim),최수연(Soo Yeon Choi),서수좌(Su Jwa Seo),김상민(Sang-Min Kim) 한국식물병리학회 2020 식물병연구 Vol.26 No.4
Xanthomonas oryzae pv. oryzae는 벼 흰잎마름병을 일으키는 세균이며, 벼 흰잎마름병은 벼 주요 재배지에서 꾸준하게 발생하고 있다. 본 연구에서는 벼흰잎마름병균을 신속하고 간편하게 검출하기 위해 등온증폭법 중 하나인 recombinase polymerase amplification (RPA)를 적용하여 현장진단 등을 위한 유전자기반 진단법을 개발하였다. RPA법은 짧은 시간의 등온 조건에서 유전자 증폭이 가능하다는 장점이 있다. 본 연구에서개발한 벼 흰잎마름병 RPA 진단법은 39oC에서 5분간만 반응하면 목표유전자의 증폭이 이루어지므로 기존 진단법보다 신속하고 간편하며 우리나라에 존재하는 K1–K3a의 4종 레이스에모두 적용가능하며, DNA 추출 없이 식물체 잎의 즙액으로 증폭반응 수행이 가능하며 기존 Taq 기반 PCR보다 약 10배 검출민감도가 높고 고가의 thermal cycler 없이도 항온수조, 발열블록 혹은 체온을 이용한 증폭반응 수행이 가능하다. 벼흰잎마름병은 벼의 농작물재해보험 대상재해 병충해 중의 하나이므로과학적 진단결과가 요구되나, 일선 농촌지도 현장에서는 유전자기반 진단을 위한 장비, 기술 등의 부족으로 분자진단이 어려웠다. 본 연구에서 개발한 벼 흰잎마름병 RPA 진단을 활용한다면 병징 의심부위 마쇄액을 주형으로, 손으로 5분 동안 반응시키는 것만으로도 신속하고 간편하게 벼 흰잎마름병의 목표유전자 증폭산물을 형성하므로 벼 흰잎마름병 진단의 최일선에서 활용할 수 있을 것이다. Rice bacterial leaf blight (BLB) by Xanthomonas oryzae pv. oryzae (Xoo) is considered to be one of the major rice diseases steadily occurring around the rice-producing countries. In this study, we developed a recombinase polymerase amplification (RPA) assay for the rapid, convenient and specific diagnosis of Xoo by targeting Xoo-specific transposase A gene. As the target gene can be amplified in 10 min without DNA extraction process and special equipment for temperature control, RPA for BLB can be useful and practical component for on-site diagnosis.
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노태환 ( Tae Hwan Noh ),심형권 ( Hyeong Kwon Shim ),최만영 ( Man Young Choi ),이휘종 ( Hwi Jong Lee ),강미형 ( Mihyung Kang ),서수좌 ( Su Joa Seo ),강인정 ( In Jeong Kang ),김형무 ( Hyung Moo Kim ) 전북대학교 농업과학기술연구소 2014 농업생명과학연구 Vol.45 No.2
The natural infected rate and occurrence rate of bakanae disease were investigated among 12 rice varieties depend on growth stages of rice. In case of infection rate, Nampyeong was the highest infected variety among investigated varieties as 70.8 percent. Hopum, Sindongjin, and Honong are also infected highly in natural circumstances. The highest occurrence rates of Boseokchal was investigated the highest in nursery and field both. According to the result which were investigated infection rate on each growth stages of rice, most varieties were infected from 80% to 100%. The average occurrence rates were 49% and 50% at heading stage and milk stage. Even if rice seeds were infected by Bakanae disease pathogen late, bakanae disease occurrence also decreased. Compared with resistant variety, Nampyeong and susceptible, Hwasung, occurrence rate of Nampeong were 38%, 40%, 28% and 20% at heading stage, milk stege, dough ripe stage and yellow ripe stage. However, occurrence rate of Hwasung were 36%, 34%, 18%, 16% at each steges. When pathogen spores were inoculated artificially in field, Hopum was shown the highest occurred as 82%, 78%, 66%, 54% at each stage but Honong was the lowest. In non-disinfection seeds plots, occurrence rate of Boseokchal was the highest.
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