콩모자이크바이러스병(病)의 방제(防除)를 위(爲)하여 본(本) 바이러스병(病)의 몇가지 발생요인조사(發生要因調査) 및 저항성(抵抗性) 검정(檢定) 결과(結果)는 다음과 같다. 우리나라 각지역(各地域)에서 재배(栽培)되는 대두(大豆)의 콩모자이크바이러스 종자전염율(種子傳染率)은 영주외(榮州外) 14개지역(個地域)의 수집종자(蒐集種子)가 30%이상(以上), 남해외(南海外) 22개지역(個地域)의 수집종자(蒐集種子)는 , 완도외(莞島外) 43개지역(個地域)의 수집종자(蒐集種子)는 이었고 연기용호에는 종자전염주(種子傳染株)가 없었다. 포장(圃場)에서 진딧물발생밀도(發生密度)의 증감(增減) 1개월후(個月後)부터 발병율(發病率)에 차이(差異)가 나타났다. 엽모(葉毛)의 길이가 짧고 조밀(稠密)한 품종(品種)에 가장 매개(媒介)진딧물의 부착(附着) 및 흡즙충수가 적었고, 엽모(葉毛)가 길고 조밀(稠密)한 것에는 시간(時間)의 경과(經過)에 따라 부착충수가 적었으며, 엽모(葉毛)가 길고 엉성한 것에 부착(附着) 및 흡즙충수가 가장 많았다. 감염시기(感染時期)에 따른 종자전염율(種子傳染率)은 6월(月) 20일(日)에 감염(感染)된 대두(大豆)에서 높았고, 7월(月) 20일(日)과 8월(月) 20일(日)에 감염(感染)된 대두(大豆)에서는 아주 낮았다. 대두종자부분(大豆種子部分)의 부위별(部位別) 바이러스전염(傳染) 조사(調査)에서는 배(胚)와 배유(胚乳)에서 바이러스소재(所在)가 확인(確認)되었고, 미숙종자(未熟種子)가 완숙종자(完熟種子)보다는 감염율(感染率)이 높았다. 반문(斑紋)이 있는 종자(種子)는 반문(斑紋)이 없는 종자(種子)보다 다소(多少) 높은 종자전염율(種子傳染率)을 보였으나 반문(斑紋)이 심(甚)하여도 종자전염율(種子傳染率)은 높지 않았다. 그리고 발아율(發芽率)에서는 반문(斑紋)의 영향(影響)이 없었다. 저항성품종(抵抗性品種) 검정(檢定)에서 Columbus외(外) 14품종(品種)이 이병성(罹病性), Chief외(外) 14품종(品種)이 중도저항성(中度抵抗性), 장백(長白)콩외(外) 17품종(品種)이 저항성(抵抗性)인 것으로 나타났다.
콩 모자이크 바이러스계통 SMV-G7의 감염에 의하여 모자이크 병징이 나타나는 함안 품종과 괴저병징이 나타나는 광교를 공시하여 (99평)의 포장 중앙 (1평)에 함안을 파종, 접종한 후 주변 시험구(1평씩 99구)에는 광교를 파종하여 SMV의 발생생태를 조사하였다. 광교에서 SMV 이병주율은 7월 13일에 가장 심하였고 황색수반으로 채집한 진딧물 밀도는 6월 22일에 가장 높게 나타났으며 함안과 인접한 시험구의 SMV 이병주율은 평균 , 포장전체의 이병주율은 평균 로 나타났다. 풍향과 바이러스 이병율을 분석한 결과 SMV의 전파는 바람부는 쪽으로 일정한 gradient를 형성하여 SMV 전파에 중요한 요인으로 나타났다
우리 나라 콩 종자의 모자이크 바이러스(Soybean mosaic virus, SMV) 감염상을 조사하기 위하여 국내에서 수집한 8개 품종을 공시해서 Lima와 Purcifull의 면역이중확산법으로 SMV 검정을 한 결과는 다음과 같다. 1. 공시한 7개 품종 중 6개 품종에서 SMV가 검출됨으로서, 우리 나라의 콩 품종에서 SMV의 종자전염성이 혈청학적 방법으로 확인되었다. SMV가 검출된 종자의 종자감염성율은 최저 에서 최고 를 나타냈으며, 전체적으로는 총검정립수 336립 중 18립에서 SMV가 검출됨으로서 약 의 감염율을 보였다. 2. 이병주에서 채집한 갈반립과 무갈반립수의 SMV 감염율은 북해 1호 품종의 경우 각각 와 Clark 품종의 경우 각각 와 , Woodworth 품종의 경우는 갈반립과 무갈반립 모두 의 감염율을 나타냄으로써, 공시한 콩 품종의 갈반립과 무갈반립간엔 SMV 감염율에서 뚜렷한 차이를 발견할 수 없었다. 3. SMV에 감염된 광교품종 중 괴저병징을 나타내는 개체에서 채종한 종자에서는 전혀 SMV가 검출되지 않아 광교품종에서는 SMV-N가 종자전염되지 않는 것으로 보였다.
Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.
Soybean mosaic virus (SMV) is a prevalent pathogen that causes significant yield reduction in soybean production worldwide. SMV belongs to potyvirus and causes typical symptoms such as mild mosaic, mosaic and lethal necrosis. SMV is seed-borne and also transmitted by aphid. Eleven SMV strains, G1 to G7, G5H, G6H, G7H, and G7a were reported in soybean varieties in Korea. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method allowed one-step detection of gene amplification by simple procedure and needed only a simple incubator for isothermal template. This RT-LAMP method allowed direct detection of RNA from virus-infected plants without thermal cycling and gel electrophoresis. In this study, we designed RT-LAMP primers named SML-F3/B3/FIP/BIP from coat protein gene sequence of SMV. After the reaction of RT-LAMP, products were identified by electrophoresis and with the detective fluorescent dye, SYBR Green I. under daylight and UV light. Opmtimal reaction condition was at 58℃ for 60min and the primers of RT-LAMP showed the specificity for nine SMV strains tested in this study.
Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.
In this study, we constructed viral vector for soybean by using Soybean yellow common mosaic virus (SYCMV) infecting both Glycine max and Glycine soja. SYCMV-derived viral vector was tested to use as Virus-induced gene silencing (VIGS) vector for functional analysis of soybean genes and as protein expression vector for foreign protein expression. In vitro transcript with 5’ cap analog m7GpppG from a full-length infectious vector of SYCMV driven by T7 promoter was inoculated to soybean to test infectivity of the clone (pSYCMVT7-full). 5’-capped transcript was able to infect soybean plants. The symptoms observed in soybean plants infected by either the vector or the sap from SYCMV-infected leaves were indistinguishable, suggesting that the vector had an equal biological activity shown by virus itself. To further utilize the vector, an additional DNA-based vector was constructed. The full-length cDNA was inserted into a binary vector flanked by CaMV 35S promoter and the nopaline synthase terminator (pSYCMV35S-full). To test if the vector infects soybean and subsequently induces gene silencing, we prepared two constructs containing fragments of Phytoene desaturase (PDS) gene (pSYCMV35S-PDS1) and small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcS) gene (pSYCMV35S-rbcS2) from soybean plant. Plants infiltrated with the constructs through Agrobacterium-mediated method showed distinct symptoms such as photobleaching in plants infiltrated with pSYCMV-PDS1 and pale green or yellowing in plants infiltrated with pSYCMV-rbcS2. In addition, down-regulations of mRNA levels of two genes were confirmed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). To test if the vector can be used for foreign protein expression in soybean plants, we prepared a construct encoding amino acids 135-160 of VP1 FMDV serotype O1 Campos (O1C) (pSYCMV35S-FMDV). Plants infiltrated with the construct through Agrobacterium-mediated method showed that soybean plant infiltrated with pSYCMV35S-FMDV only was detected by Western blotting using O1C antibody. These results support that SYCMV-derived viral vector can be used as VIGS vector or protein expression vector in soybean plants.
Soybeans X soybeans mosaic virus (SMV) strains interactions affected plant growth and seed transmission. Strain virulence of SMV depended on host cultivars. Kwangankong and Tawonkong were susceptible to G7H and G5 strains, causing mosaic symptoms. The distribution patterns of two SMV strains in soybean plants inoculated with G7H, G5 and G7H/G5 sets were investigated by RT-PCR/RFLP analysis. In the first treatment, two primary leaves in a single plant were infected with both strains by means of one strain per leaf. The leaves of Kwangankong and Tawonkong at V2, V4 and V6 stage were doubly infected with the two strains and the upper leaves than those had only G7H strain. Secondly, the two soybeans were inoculated with G7H, and 24 h after followed by the other strain inoculation. The leaves of V6 and V8 stages in all infected plants showed mosaic symptoms caused by G7H, and there was no detection of G5 strain. In contrast, the reverse treatment with G5 and G7H induced different results. Pre-inoculated G5 strain detected in every stage besides G7H strain. Host X SMV strain compatibility influenced seed coat mottling, yield, plant height, number of pod per plant. G7H had a seed mottling rate of 98.5% in Kwangankong, while G5 had an incidence of seed mottling of 1.4% in the same cultivar. G5 was more virulent to Kwangankong and had a lower affinity for infecting soybean seed mottling. Additional inoculation of G7H protected soybean yield and growth from G5-inducing loss in Kwangankong.
신강은 콩나물 적성이 뛰어나고 농업형질이 우수하나 콩모자이크병에 약한 소원콩을 반복친으로, 콩모자이크병 저항성 유전자 Rsv3을 보유한 L29를 일회친으로 사용하여 육성된 품종이다. 육성기간의 단축을 위하여 분자표지선발법을 사용하여 목표유전자의 도입과 반복친의 회복정도를 신속히 확인함으로써 품종 개발에 소요되는 기간을 7년으로 단축하였다. 신강의 주요 특성을 요약하면 다음과 같다. 1. 유한신육형이며, 꽃색은 자색이고 엽형은 피침형이다. 입형은 구형이고
The coat protein (CP) gene of the G5H and G7H strains of Soybean mosaic virus (SMV) were cloned, sequenced and transformed into the tobacco plant (Nicotiana tabacum. cv. Havana SR1) via Agrobacterium-mediated transformation. Transformation was confirmed b
Soybean mosaic virus (SMV) resistance of Korean recommended soybeans was evaluated naturally and by mechanical inoculation in Suwon. Based on the differential reaction of forty-four soybean genotypes tested to nine different SMV strains, soybeans were classified into twenty-four groups. Myeongjunamulkong and Ilpumgeom-jeongkong showed a high degree of resistance to nine SMV strains, having no symptom. The other cultivars produced various reactions according to inoculation of each SMV strain: symptomless, mosaic or systemic necrosis. Only five cultivars such as Kwangankong, Eunhakong, Tawonkong, Namhaekong, Sobaegnamulkong were totally susceptible to every strain. There was variation in disease incidence. Soybeans, having the highest levels of resistance to G5H and G7H in the greenhouse, showed the lowest levels of SMV incidence in the field of Suwon. Myeong-junamulkong, Ilpumgeomjeongkong, Soyangkong, Pungsannamulkong, Sodamkong, Jangmikong, Geomjeong-kong2, Pureunkong, Sinpaldalkong2, Duyoukong, and Geumgangkong were fairly resistant to SMV. And SMV incidence of Taekwangkong, Saealkong and Baegunkong was over 45% with symptom of bud necrosis. And soybeans, highly resistant to SMV in the field and the greenhouse, were mainly derived from Jangyeobkong and Hwang-keumkong resistant to G1-G7.
콩 재배포장에 정착하는 매개 진딧물 발생소장과 효율적인 살충제 선발을 통해 콩모자이크병 피해 경감 방법을 모색코자 실시한 시험결과를 요약하면 다음과 같다. 1 적기파종에서의 진딧물 발생최성기는 6월 하순과 8월 중순 2회였으나, 조기파종에서의 진딧물 발생최성기는 6월 중순으로 적기파종에 비해 진딧물 발생최성기가 약 10일정도 빨랐다. 2. 공시약제인 이미다클로프리드수화제, 벤즈유제, 아시트수화제 등은 모두 95% 이상의 진딧물 방제가를 보여 약효가 우수하였고 약해도 없어 효과적인 진딧물 방제 약제로 선발되었다. 3. 이미다클로프리드입제를 토양 혼입처리한 시험구는 무처리구에 비해 처리 후 52일까지 진딧물 발생이 억제되었다. 4. 아시트 50% 수화제를 콩 생육단계 V4, V6, V4/V6에 1회 또는 2회 처리하였을 때, 처리구 모두 무처리구에 비해 SMV 발병률이 낮아 방제효과가 인정되었으며, 특히 V4/V6 시기 2회 처리구에서 SMV 발병률이 낮았다.
Reverse transcription and polymerase chain reaction (RT-PCR) assay was used to detect SMV strains. A pair of oligonucleotide primers were designed to include the cylindrical inclusion (CI) coding region between 4,176 to 5,560 nt. Amplification from the total RNA extracted from infected plants with SMV yielded a 1,385 bp DNA fragment. RT-PCR was shown to be 103 times more sensitive than the ELISA assay and it could detect a virus in 10-6 dilution. Restriction enzyme analysis of RT- PCR products using EcoR I showed that SMV isolates were classified into six groups according to the patterns of restriction fragments.
콩모자이크 바이러스(SMV)에 의한 갈반립의 형성율과 SMV의 종자전염율을 조사하기 위하여 대두품종 Clark, Woodworth, 북해1호를 재료로 SMV 이병정도와 갈반립율 결협절위 및 갈반립형성등의 관계를 조사하고 그 갈반립을 통한 SMV의 종자전염율을 면역이중확산법과 유묘감염률로 조사하였다. 1. 갈반립의 형성율은 식물체의 이병정도와 정비례적인 상관(r=0.796)을 보였다. 2. 이병식물체의 갈반립형성은 결협절립 및 협중종실위치와 유의상관이 없었으며 (r=0.34), 동일절위 및 협내에서도 갈반립과 비갈반립이 함께 형성되었다. 3. 면역이중확산법으로 조사한 SMV의 종자전염율은 4.2 -33.3%로 비교적 낮았으며 품종에 따라 현저한 차이가 있었다. 4. 유묘에 나타난 SMV 증상으로 조사한 종자전염율은 면역이중확산법으로 검정한 종자감염율보다 약관 높게 나타났으며 이는 항혈청의 역가 또는 Virus의 체내증식과 관계가 있는 것으로 보였다. 5. 이병직물의 갈반립은 SMV 종자전염율과 일치하지는 않으나 SMV의 포장이병율 추정에 유용한 지표가 될 수 있으며 파종시 갈반립의 제거는 SMV 전염원의 감소효과를 가져올 수 있을 것으로 보였다.