Freesia is one of the most popular flowers over the world including Korea, due to the fragrance and beauty of the plant flower. The first domestic freesia cultivar ‘Shiny Gold’ was developed by NIHHS, RDA, in 2003, which has yellow double and large petals and strong fragrance. Ten years have passed since ‘Shiny Gold’ was cultivated at floral farms, and the deterioration of cut flower quality and yield are reported from the farms. Virus infection causes a reduction in the quantity and quality of the cut freesia flowers and is one of the most serious problems in Korea. Virus detection was carried by reverse transcription polymer chain reaction (RT-PCR) for FreMV, FreSV, BYMV, CMV, and TRV, as known to infect freesia. FreMV, FreSV, BYMV, and TRV were detected single or multiply, and CMV was not found in the freesia leaves collected from the farms. To produce virus-free freesia, meristem culture of ‘Shiny Gold’ was conducted in MS medium added ribavirin at different concentration. As the increased of ribavirin concentration, the growth of ‘Shiny Gold’ plantlets was inhibited in freesia’Shiny Gold’. The plantlets produced by meristem culture in ‘Shiny Gold’ were virus free at the enzyme-linked immunosorbent assay (ELISA) level.

Genetically modified (GM) pepper H15 containing the gene for cucumber mosaic virus (CMV) coat protein (CP) and its control line non-GM pepper P2377 were investigated for their allergic risk. Amino acid sequence of the inserted gene product CMV-CP was compared with those of known allergens. No known allergen had greater than 35% amino acid sequence homology over an 80 amino acid window or more than 8 consecutive identical amino acids. Protein patterns of GM and non-GM pepper extracts were evaluated by SDS-PAGE, which showed similar distribution of protein bands for both GM and non-GM pepper. Antigen-antibody reactions were compared between GM and its non-transgenic parental control. ELISA and immunoblot analysis of sera from allergic patients showed some IgE reactivity; however, no differences were observed between GM pepper H15 and P2377. We therefore conclude that CMV-CP is less likely to be an allergen; the protein composition and allergenicity of the GM pepper H15 is not different from that of P2377 and safe as a commercial host.

Virus disease surveys on barley cultivation areas in Jeonnam ․ Jeonbuk ․ Gyungnam ․ Gyungbuk-do were conducted during 2014-2015. In this study, we detected Barley yellow mosaic virus (BaYMV), Barley mild mosaic virus (BaMMV) and Soil-borne wheat mosaic virus (SBWMV) by RT-PCR from barley leaves. These viruses are of great economic importance for wheat and barley, causing significant quantitative and qualitative losses in yield. The result of investigation showed that the field incidence of BaYMV in Buan, Gimje was more than 90% in 2014. The infection field rate of barley virus including Boseong, Gangjin, Haenam, Jangheung in Jeonnam was ≈ 30%. In 2014, double infections by BaYMV and BaMMV was detected in Boseong, Gangjin, Haenam and Jangheung. Only as a low rate BaYMV occurred in various fields of Jeonbuk in 2015. At the same time high infection field ratio of 70% was observed in Gunsan. Also in Yeonggwang was double infection of BaYMV and BaMMV. BaYMV occurred single infection has been confirmed in all of the study area of Gyungbuk and Gyungnam except for Goseong during the investigation period.

Endocrine disruptors are exogenous chemicals that their endocrine disrupting effects mediated by androgenic signaling plays crucial roles in the control of development and several androgen-related diseases. However, there are no authorized in vitro screening and testing methods to evaluation of (anti-)androgenic activity. To find out a better in vitro cell line model, we have previously reported that 22Rv1 cells, a human prostate cancer cells contained functional Androgen Receptor (AR), might be an appropriate model for the evaluation of (anti-)androgenic endocrine disruptors. Based on this result, we developed a stable 22Rv1/mouse mammary tumor virus (MMTV) cell line to test AR-mediated transcriptional activation (TA). Using 22Rv1/MMTV cells, we established the test protocol and optimized the testing condition for AR-TA assay. In this study, we performed the inter-validation assay by four different laboratories to evaluate the 20 coded chemicals which were selected from the ICCVAM list (ICCVAM, 2003) or academic articles that exhibited exact (anti-) androgenic activity. The statistical analysis of the results of the inter-laboratory validation study revealed that there was reproducibility between the four participating laboratories. In conclusion, 22Rv1/MMTV AR-TA assay might be a quick and relatively inexpensive method, which can be used to screen large numbers of chemicals for their potential to activate or inhibit AR-mediated gene transcription. Furthermore, it will provide mechanistic data relevant to understanding adverse reactions observed in intact organisms.

Tomato spotted wilt virus (TSWV) causes one of the most destructive viral diseases that threaten tomato (Solanum lycopersicum) worldwide. So far, eight TSWV resistance genes, Sw1a, Sw1b, sw2, sw3, sw4, Sw-5b, Sw-6, and Sw-7 have been identified and Sw-5b has been incorporated into tomato for prevention of TSWV. The objectives of this research are first to discover single nucleotide polymorphisms (SNPs) in Sw-5 alleles and then to develop SNP markers to distinguish resistant genotypes against TSWV for marker-assisted breeding in tomato. First, DNA sequences of Sw-5b alleles from both resistant and susceptible cultivars amplified using known Sw-5 gene-based marker was analyzed. The single functional SNP (G→A) was detected as non-synonymous substitution because this SNP causes change of arginine (Arg599) to glutamine (Gln599). Next, the primer pair for high resolution melting analysis (HRM) was designed around this SNP. To determine accuracy of this SNP marker to distinguish resistant Sw-5b genotypes against TSWV, genotypes of 32 commercial tomato cultivars were checked. The newly developed SNP marker could select six cultivars carrying resistant Sw-5b genotype, which was 100% correlated with genotypes based on the gene-based marker. These results indicate that the SNP maker developed in this study could be useful for better tracking resistance to TSWV in tomato breeding.

This study was conducted to develop the early maturing rice lines with genes conferring resistance to bacterial blight and rice stripe virus to enhance the adaptability in plain area. Unkwang carrying Xa3 was used as a recurrent parent and SR30075 carrying Xa4+xa5+Xa21+Stvb-i was used as a donor parent. RL1(Resistant Line, BC1F7), RL2, RL3, RL4, and RL5(BC2F6) were bred through bio-assay of K3a race inoculation and phenotypic selection of agronomic traits. The presence of introduced genes was confirmed by testing the resistance levels against bacterial blight and rice stripe virus and then double-checked by using DNA marker. RL1 has all target genes, Xa3+xa5+Xa21+Stvb-i. RL2, RL3, and RL5 have Xa3+Xa21+Stvb-i whereas RL4 has only Xa21. Rice lines carrying Stvb-i showed resistance reaction to rice stripe virus. The combinations of bacterial blight resistant genes(Xa3+xa5+Xa21 and Xa3+Xa21) were found to be promising, as the rice lines carrying these genes enhanced a strong resistant reaction against 16 bacterial blight isolates. Also, the inoculation of K3a race did not alter the brown rice yield, ripened grain ratio and kernel quality of brown rice compared to control. Although RL1 containing all the target resistance genes showed excellent resistance performance, it is not suitable to cultivate in plain area due to instability to lodging, 80% yield level than Unkwang, and low grain quality. RL5 backcrossed twice with Unkwang was found to be a promising line due to its effective resistance gene combination, Xa3+Xa21+Stvb-i and good agronomic traits such as stability to lodging, higher yield and quality compared to Unkwang.

To analysis of virus free sweetpotato effect, 5 virus free sweetpotato and virus normal sweetpotato varieties were planted in 5 different regions at 2010 year. The average yields of virus free sweetpotato are showed different results according to regions. Sinjami which cultivated at Iksan were increased maximum 68% compare to normal. However, Sinjami which cultivated in Hamyang were decreased yield 11% compare to normal. Analysis of tuber formation ratio of Sinjami, Yenhwangmi, Singeonmi which cultivated in Nonsan were decreased tuber number compare to normal. However, 3 varieties were all increased on Average storage root weight and yield of marketable storage root. In the results of analysis of marketable storage root according to cultivated regions and varieties, all varieties except Sinjami which cultivated in Hamyang were increased yield. Also, quality of virus free sweetpotato were enhanced 7 to 9 compare to virus infected sweet potato which showing average 3. Contents of starch between virus free and virus infected sweetpotato were not affected by virus. Virus free sweetpotato were more increased starch products according to increased total production yield. Also, Brix° (%) was not showing difference between virus free and virus infected sweetpotatoes. In this experiment, Virus free sweetpotato are enhanced production yields and quality. Therefore, we suggested that virus free sweetpotato is one of the methods to reduce damage by sweetpotato virus.

Potato virus Y (PVY) and potato leafroll virus (PLRV) are among the most damaging potato viruses and prevalent in most potato growing areas. In this study, cryopreservation was used to eradicate PVY and PLRV using two cryogenic methods. Potato shoot tips proliferated in vitro were cryopreserved through droplet-vitrification and encapsulation-vitrification using plant vitrification solution 2 (PVS2; 30% glycerol + 15% dimethyl sulfoxide + 15.0% ethylene glycol + 13.7% sucrose) and modified PVS2. Both cryogenic procedures produced similar rates of survival and regrowth, which were lower than those from shoot tip culture alone. The health status of plantlets regenerated from shoot tip culture alone and cryopreservation was checked by reverse transcription-polymerase chain reaction. The frequency of virus-free plants regenerated directly from highly proliferating shoot tips reached 42.3% and 48.6% for PVY and PLRV, respectively. In comparison, the frequency of PVY and PLRV eradication after cryopreservation was 91.3~99.7% following shoot-tip culture. The highest cryopreserved shoot tip regeneration rate was observed when shoot tips were 1.0~1.5 mm in length, but virus eradication rates were very similar (96.4~99.7%), regardless of shoot tip size. This efficient cryotherapy protocol developed to eliminate viruses can also be used to prepare potato material for safe long-term preservation and the production of virus-free plants.

Reactive oxygen species (ROS) are produced in organisms as the natural products of oxidative metabolism by environmental stress and pathogen invasion. ROS, such as superoxide anion and hydrogen peroxide, can be toxic to cells and tissues to cause oxidative stress. Recent study revealed that olive flounder (Paralichthys olivaceus) superoxide dismutase (SOD) has been identified as a partial gene and strongly induced to benzoin[a]pyrene and it was deduced indicator of aquatic oxidative stress responses, but its transcriptional response against viral infection has not been investigated. In the present study, spatial and temporal expression profile was analyzed to investigate the function of Of-SOD in the anti-viral response. Of-SOD transcripts were ubiquitously detected in diverse tissues with variable levels using a real-time PCR. The expression of Of-SOD was significantly higher in the muscle, liver and brain, but extremely low in the stomach and spleen. Following VHSV challenge, the expression of Of-SOD increased within 3 hours and subsequently decreased to the original level at 2 days post-challenge in kidney. Although expression pattern and induction time are slight differences depending on the tissue, the transcript of Of-SOD was consistently increased in acute infection response, but expression is low in the chronic response. Collectively, Of-SOD expressions were inducible after VHSV infection and they were probably involved in the immune response against viral challenge. These results suggest that SODs may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in this flounder.

In plants, eukaryotic translation elongation factor 1B (eEF1B) is composed of three subunits, eEF1Bα, eEF1Bβ and eEF1B γ. Two subunits are nucleotide exchange subunits (eEF1Bα and eEF1Bβ) and one is a structural protein (eEF1Bγ). In the previous study, eEF1B was identified as a common host factor for several RNA viruses. To test which subunit of eEF1B is essential for Potato virus X (PVX) replication, the virus-induced gene silencing (VIGS) for eEF1Bα, β or γ was performed in Nicotiana benthamiana and green fluorescent protein (GFP)-tagged PVX was inoculated. PVX-GFP accumulation was decreased when eEF1Bβ or γ subunit was silenced, whereas eEF1Bα had no effect on PVX-GFP accumulation in inoculated leaves. Targeting induced local lesions in genome (TILLING) was performed using a Capsicum annuum EMS population to test whether mutations in eEF1Bβ subunit affect virus infection in pepper. We obtained 81 eEF1Bβ mutant lines consisted of 16,759 individuals. These mutant lines are being tested to validate the function of eEF1B β in PVX replication.

Tomato yellow leaf curl disease is a devastating disease of tomato (Solanum lycopersicum), which is caused by begomoviruses generally referred to as tomato yellow leaf curl virus (TYLCV). The breeding for TYLCV resistance has been based on the introgression of the Ty-3 resistance locus. Knowledge about the exact location of the Ty-3 on tomato chromosome 6 is needed to understand the genomic organization of the Ty-3 locus. In this study, we conducted a genetic analysis using a segregating population derived from a cross between resistant accession S. lycopersicum “A45” and susceptible accession S. lycopersicum “A39”. The F1 plants showed resistance to TYLCV and F1 was self-pollinated to produce F2 progeny. To screen the TYLCV resistance in 145 F2 plants, a leaf agroinfiltration method was used. F2 plants showed a classical Mendelian seregation (106 resistance : 39 susceptibility) for resistance to TYLCV respectively. SCAR and CAPS markers linked to the Ty-3 were tested for genotyping F2 plants and .genotyping and agroinfiltration results were cosegregated in the newly developed F2 population.

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.

식중독 유발 바이러스인 HAV는 제 1군 감염병원으로 규정되면서 감염 시 원인식품을 빠르게 분석하게 되었으며 그로 인해 정확하면서 빠른 검출기술을 요구하게 되었다. 본 연구에서는 HAV에 오염된 상추에서 신속하게 바이러스를 검사하기 위해서 IMS를 통하여 신속하게 HAV를 순수 분리 및 농축하였고, 형광물질인 quantum dot을 활용하여 형광검출을 실시하였다. 또한 일반적으로 바이러스 농축에 사용되는 PEG 농축방법과 비교하였을 때 검출능은 유사한 결과를 얻었으나, 농축시간 면에서는 IMS를 통한 방법이 효과적이었다. 또한 IMS 방법으로 확보된 항원을 Quantum dot을 활용하여 10분 이내에 바이러스를 검출할 수 있었다. 본 연구에서 제시된 검출기반은 식품 유통 과정 중 다양한 식중독 바이러스로부터 소비자를 보호 할 수 있는 검사방법으로 활용될 수 있다고 기대된다.

본 연구는 벼멸구, 벼흰잎마름병, 벼줄무늬잎마름병에 저항성인 자포니카 복합내병충성 품종을 조기에 확보하고자 약배양을 수행하여 목표 저항성 유전자 조합의 우량 고정계통을 개발하고 육성 과정 중에 발생할 수 있는 문제점을 파악하여 병해충 저항성 육종사업에 반영하고자 수행하였다. 벼흰잎마름병과 벼줄무늬잎마름병에 저항성인 우량 계통 HR26234- 12-1-1과 벼멸구, 벼흰잎마름병, 벼줄무늬잎마름병에 저항성인 SR30071-3-7-23-6-2-1-1을 인공 교배한 F1을 약배양하여 213개 고정계통을 육성하였다. HR26234는 Xa3+xa5, Stvb-i, SR30071은 Bph18, Xa4, Stvb-i를 가지고 있는 것으로 확인되었다. 이들 유래 약배양 계통들은 모두 Stvb-i를 가지고 있었고, Bph18+Xa3, Bph18+Xa4, Bph18+Xa3+xa5, Bph18+Xa4+xa5, bph18+Xa3, bph18+Xa4, bph18+Xa3+ xa5과 bph18+Xa4+xa5의 조합이 확인되었다. 약배양 집단에서 xa5+Bph18(또는 bph18)+Stvb-i 조합이 발생하지 않았고 Bph18 보유 계통이 bph18 보다 적게 발생하는 등 segregation distortion이 발생하였다. Bhp18을 보유하며 벼멸구에 저항성인 계통들이 감수성인 계통들에 비해 간장이 컸다. 선발된 Bph18+Xa4+xa5+Stvb-i 조합의 계통은 단간이면서 벼멸구, 벼흰잎마름병, 벼줄무늬잎마름병에 저항성을 나타내었으나, 낮은 수량성과 일부 불임의 발생, 단간임에도 도복에 안정적이지 못한 특성을 나타냈다. 병해충에 대한 저항성 육종사업에서 약배양을 활용하여 조기에 육종 목표를 달성하고자 할 경우에 segregation distortion이 발생하여 편의 된 변이가 발생할 수 있고, 저항성 유전자 도입 시 linkage drag에 의해 예기치 못한 열악형질 특성이 나타날 수 있음을 고려하여 신중하게 목표에 접근하여야 할 것으로 생각한다.

We report a double primary gastric cancer of different types seen on an endoscopy at the same time. At the first gastroscopy, Lymphoepithelioma-like carcinoma (LELC) appeared to be a benign submucosal tumor (SMT) and gastric adenocarcinoma was limited to the mucosa. Adenocarcinoma was removed by endoscopic submucosal dissection (ESD). However, a followup gastroscopy revealed an enlarged SMT, and another ESD was performed. The patient was additionally diagnosed with gastric LELC. This is the only case that LELC and gastric adenocarcinoma were found at the same time in Korea; therefore, the authors report this case with literature review.

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 Korea, CMV (cucumber mosaic virus) is the most frequently occurring virus with a single infection rate of 45%. However, a total occurrence of CMV by co-infection, either couple or multiple, with BBWV (broad bean wilt virus), PepMoV (pepper mottle virus) and PMMoV (pepper mild mottle virus) covers over 90% in the field cultivation of pepper. The PepMoV is transmitted by several aphid species, and it has been considered the most frequently detected potyvirus when it co-infects with CMV or PMMoV. Since F1 hybrid that resistant to PepMoV has not been developed, we have developed transgenic peppers using Agrobacterium-mediated transformation with a Hc-Pro gene of the PepMoV. A large number of T1 peppers were tested for resistance to the PepMoV, and T1 peppers tolerant of PepMoV were selected. After consequent self-crossing up to T4 generation, highly tolerant peppers to PepMoV were selected. So far, BC3F1 lines have been selected by back-crossing with 4 elite lines through a breeding program. The horticultural differences of the GM line comparing to inbred lines were investigated and no statistical significance between GM and non-GM lines was found. Based on molecular analysis, One of GM lines, 10-2, contained the transgene in the non-coding region indicating that this line would be a GM event.