Under field conditions, we investigated how transgenic Bt cabbage expressing the insecticidal Cry1Ac1 protein affects two target Lepidoptera species — Plutella xylostella (Plutellidae) and Pieris rapae (Pieridae) — as well as the structure of the local non-target arthropod community. When exposed to Bt cabbage line C30, Plutella xylostella and Pieris rapae were significantly less abundant than when in the presence of the non-transgenic control. Multivariate analyses (PerMANOVA and NMDS) showed that composition of the non-target arthropod community was affected by sampling date but not by cabbage genotype. These results suggest that transgenic cabbage expressing Cry1Ac1 protein can be effective in controlling Plutella xylostella and Pieris rapae in the field and that cultivation of this cabbage may not adversely affect such non-target arthropod communities.

Transgenic lines of insect resistant cabbage (Brassica oleracea var. capitata) expressing Cry1Ac1 protein has been developed to control diamondback moth (Plutella xylostella). The potential adverse effects of Bt crops on non-target arthropod herbivores and predators are major concerns. We conducted a tritrophic bioassay to study the ecological impacts of insecticidal transgenic cabbage on the wolf spider (Pardosa astrigera), a non-target generalist predator. First, we measured the levels of Cry1Ac1 proteins in fruit flies that were fed with the transgenic cabbage as well as those levels in the wolf spiders preying on the Bt cabbage-fed fruit flies using enzyme linked immunosorbent assay (ELISA). Cry1Ac1 proteins were detected in the Bt cabbage fed fruit flies and also in the wolf spiders after preying on Bt cabbage-fed fruit flies. Second, we compared the life history characteristics of the wolf spiders preying on the Bt or non-Bt cabbage. Growth, development time and survival of the wolf spiders were not significantly different between Bt and non-Bt cabbage. Although the wolf spiders were exposed to Cry1Ac1 protein via feeding on the preys containing Cry1Ac1 proteins, their growth and survival was not significantly affected.

이 연구는 형질전환 고추로부터 토양 근권 미생물로의 유전자 이동 가능성을 조사하기 위해 수행되었다. 재배중인 오이 모자이크 바이러스 저항성 고추와 대조구의 근권토양으로부터 DNA를 추출한 후 형질전환 고추 도입유전자에 특이적인 프라이머를 사용하여 PCR 분석을 수행하였다. PCR 결과 도입유전자인 NPTII 유전자가 수집된 전 기간의 근권 토양 샘플에서 발견되었다. 발견된 도입유전자가 고추로부터 토양 미생물로의 유전자 이동에 의한 결과인지를 조사하기 위하여 토양 샘플로부터 미생물들을 분리하여 카나마이신이 첨가된 세균 및 진균 배지에 배양하였다. 약 43만개의 세균 코로니와 16만개의 진균 코로니를 조사 한 결과 카나마이신에 저항성을 나타내는 개체는 발견되지 않아 형질전환 고추로부터 근권 미생물로의 유전자이동은 발생하지 않은 것으로 사료되었다.

Anthocyanin is known for positive health beneficial effects that including reduces age related oxidative stress and inflammatory responses. It was produced by vegetable crops and a lettuce is one of the crops. The general pathway of anthocyanin expression is well defined but it is not clear how environments effects on anthocyanin accumulation in a lettuce. Therefore we initiated to study interaction between anthocyanin expression and environment factors. Frist, we applied RGB leaf images in a lettuce to calculate anthocyanin areas in a leaflet with two different cultivars, different developmental stages, and different environments. Later, we attempted to capture RNA expression level with next generation sequence (NGS) RNA sequencing method called RNA-seq. As a result, combined two technologies showed that quantitate phenotypic data help to understand the gene expression of anthocyanin in lettuce cultivars.

The most important factor in breeding program is to obtain the value-added genetic line. Generally, breeders develop genetic sources using several methods such as segregation-breeding, cross-breeding, backcross-breeding, mutation induction, tissue culture and so on. Here, we present one classical way but very valuable method called cell fusion or protoplast fusion to create genetic sources for the breeding practice. The method we developed was the asymmetric somatic-hybridization of protoplast isolated from carrots. This is rather to transfer the nucleus from the high quality F1 hybrid to other mediocre line to produce a new carrot line. Since the breeding a carrot line for higher quality and purity takes a long time, therefore this nuclear transfer technology is very beneficial to generate a new line that could be useful to breed elite varieties. We had obtained around 200 fused carrots (cybrids), 12 cybrids were self pollinated and produced seeds. Selected progenies (C3) have been evaluated for horticultural characteristics and we have found new genetic lines that show better phenotypes.

The PepMoV has been considered the most frequently detected potyvirus. When it co-infects with CMV or PMMoV, it gives severe impact to total pepper harvest in Korea. 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 GM peppers were tested for resistance to the PepMoV, and after consequent self-crossing up to T4 generation, a highly tolerant pepper to PepMoV called T20 was selected. So far, BC4F1 lines have been selected by back-crossing with 4 elite lines through a breeding program. Very recently, based on molecular analysis, we have selected another event, #10-2, which is also resistant to PepMoV. Horticultural difference was investigated for both GM lines, #T20 and 10-2, and no significance was found comparing to non-GM lines.

Next-generation sequencing (NGS) technology and fast improvement in plant genetics have elevated to massive development of molecular genetic markers through fast analysis of huge molecular biological data. Furthermore, in domestic commercial breeding of horticultural crops, the application of marker assisted breeding (MAB) has been introduced recently. For effective improvement of cultivar breeding, in this research, transcriptome analysis and single nucleotide polymorphism (SNP) comparison with high density pepper map in UC-DAVIS were performed using four lines of Capsicum annuum and C. chinense. For rapid analysis of MAB of tolerant pepper lines, 412 Fluidigm probes were newly designed in this study. These designed probes and SSR and COSII markers were applied for background selection through the MAB program. In addition, powdery mildew (PM), tomato spot wilt virus (TSWV) resistance related markers were subjected to foreground selection of BC1, BC2, and BC3 progenies. The MAB system using Fluidigm probes, and trait-related and common markers was introduced into domestic pepper breeding, which will rapidly approach to a new elite line and a commercial tolerant cultivar.

Unfavorable environmental stresses are major limiting factors that affect plant productivity. Plants perceive and respond adaptively to an abiotic stress condition, and the adaptive process is controlled mainly by phytohormone, consequently, changing in gene expression pattern. Transcriptional regulator ABF3 (Abscisic acid response element Binding Factor 3) mediating the ABA-responsive gene expression plays important roles in drought and temperature tolerance. Here, we report an event of drought tolerant GM gourd in which abf3 was inserted in the genome. For drought stress experiment, T0 plants were self-pollinated and back-crossed to select cultivars for drought stress experiment. The drought tolerant GM gourds were selected for last 3 years and have been crossed to get the several BC and T generation consecutively. In this year, BC3T1, F1 (Bak x GM gourd), BC1F1 and control plants were subjected to drought stress, that is no watering for 12 days and rehydration afterwards. The GM gourds showed high tolerance to drought while the non-transformed plants were totally dried. When the plants were subjected to rehydration, the GM bottle gourds were completely revived and recovered from the drought stress. Tolerance levels to drought of BC3T1, F1 (Bak x GM gourd) and BC1F1 were 92.5%, 50.0% and 65.0%, respectively.

2009년에 ‘MS-BGT’와 ‘NHMF’의 고정계통을 교배하여 육성한 특성 및 수량이 우수한 품종으로 ‘티티찰’을 육성하였다. ‘티티찰’의 주요특성은 TMV, TYLCV, TSWV, 잎곰팡이에 내병성으로 7~9월에 정식하는 품종이다. 숙기는 중조생종으로 당도가 평균 9~10 brix로 높으며 식미가 우수하고 평균과중은 23~25 g 내외의 중대과종 대추형 방울토마토이다. 경도가 강하여 저장성이 우수하며 열과 발생이 적고, 초세가 비교적 강하면서 이상줄기 발생이 적어 재배가 용이한 품종이다.

‘MS-CDE’와 ‘VFR’의 고정된 계통을 교배하여 특성 및 수량이 우수한 F1 품종인 ‘TY알토랑’을 육성하였다. ‘TY알토랑’의 주요특성은 TMV, TYLCV, 근부위조병, 잎곰팡이병에 내병성이며, 2~3월, 7~9월에 정식하는 품종으로 숙기는 중조생종으로 평균과중 225 g 내외의 고구형의 완숙 대과종토마토이다. 경도가 강하여 저장성이 우수하고 과형이 정연하며, 착과가 안정되어 수량성이 높다. 또한 초세가 강하면서 이상줄기 발생이 적어 재배가 안정적인 품종이다.

토마토 과실의 숙성기간은 운송기간과 상품성에 상당한 영향을 미친다. 특히, 고온에서는 과실숙성이 빨라지므로, 숙성기간은 열대지방에 수출하는 토마토 품종에서 반드시 고려되어야 한다. 토마토 과실숙성을 효과적으로 차단하는 RIPENING-INHIBITOR (Rin) 유전자는 열성 유전하는 돌연변이체에서 보고되었고, 교잡후대(Rin/rin)에서는 숙성이 느리게 진행되어 저장성이 상당히 향상된다고 알려졌다. 따라서, 본 연구에서는 수출용 토마토 품종의 육성을 위해, 과실 숙성의 저해와 관련한 Rin 유전자 특이적인 분자 표지를 개발하고자 하였다. Rin과 rin유전자의 mRNA와 genomic DNA의 염기서열 정보를 확보하고 다형성을 나타내는 프라이머 조합을 선발하여, 공우성 SCAR 분자표지로 개발하였다. 토마토 육성 라인 24주에서 과실숙성 정도와 PCR밴드를 비교한 결과, 공우성 마커는 동형(Rin/Rin, rin/rin)과 이형(Rin/rin)의 유전자형을 구별하기에 적합하였으며 이를 이용하여 저장성이 좋은 토마토 품종개발에 도움이 될 것으로 기대한다.

Molecular genetic markers have been widely used as powerful tools for analyzing the genome. In particular, SSR markers have practical applications in breeding systems because they can be used in high-throughput analyses for genetic mapping, heritable diversity testing, purity analysis, and marker-assisted selection. Currently, due to technical advances in DNA sequencing, large sequence databases are available for large-scale SSR mining and marker development. Here, we describe an automated method, the SSR Finder program, for SSR discovery in the onion sequence database, and primer design for amplifying the detected SSRs. A total of 1,049 SSR primers were obtained for genetic purity testing, and 100 SSR sets were analyzed in 14 bulb onion breeding lines using clustering analysis. A total of 20 selected markers from screening of all 1,049 SSR primers, were finally applied for genetic purity testing in three breeding lines, NW1, NW9, and NW10. The initial tests showed that 15%, 71%, and 97% of individuals within NW1, NW9, and NW10, respectively, were genetically homogeneous. These markers produced using the SSR Finder will be useful for investigating the genetic purity of onion breeding lines.

ChiVMV is one of the most destructive pepper pathogens in the East Asia. The resistant cultivar against ChiVMV is necessary to control the ChiVMV infection to pepper farm. However, the genetic source resistant ChiVMV was not fully identified yet and until now, the only recessive resistance gene has been recognized. In order to study more on the inheritance of the resistance and to establish a breeding program pertinent to ChiVMV resistance, firstly, we screened about 30 lines from several foreign countries, and found a new resistant line from several inoculation tests. Here, we report a new dominantly resistant chili pepper. Secondly, we found two AFLP fragments linked to the dominant resistance, which was located on the pepper chromosome number 6. The newly discovered dominant markers will help develop a new resistant pepper cultivar to ChiVMV.

CGMMV (cucumber green mottle mosaic virus) infects many cucurbitaceae species causing mosaic symptoms, yellowish leaf, and fruit deterioration. Several isolates of CGMMV from Korea, Israel, Japan, Greece and Spain have been characterized. In fact, CGMMV outbreaks have caused huge losses in the total yields of cucurbitaceous crops in Korea. Unfortunately, no genetic source is available for the resistance against CGMMV infection, and therefore, the breeding management offers no access to a solution yet. Thus, an alternative was to transform the viral gene such as CGMMV-CP into crops to induce tolerance against CGMMV. Here, we present the successful transformation of watermelon rootstock (gongdae) by Agrobacterium-mediated transformation with the CGMMV-CP gene. We obtained 29 independent T0 watermelon rootstock plants from the transformation of 7,887 explants. T0 plants were self-crossed and T1 plants were screened by the virus infection. Those gongdaes tolerant to virus were selected continuously, and BC2T1 and T3 generation were obtained. We have also obtained watermelons of BCF generation by crossing GM gongdae to watermelon line and those BCF watermelones were strongly tolerant to CGMMV.

Bt gene derived from the B. thuringiensis has been used for developing GM crops and those crops are already on the market. The aim of this study is to construct a genetic transformation with peppers using CryIAc1 gene and consequently to develop GM peppers resistant to the oriental tobacco budworm. We have developed transgenic peppers using Agrobacterium-mediated transformation, and obtained 5 T0 peppers. T0 peppers were self-crossed and T1 pepper fruits were exposed to larvae to test the survival rate. The survival rate of larvae that were fed with GM fruits decreased dramatically while with non-GM fruits the rate was not changed much. In order to establish the selection and the culturing of bug through a year, an incubating system for tobacco budworm in chamber was manufactured and a selection system under the media that are mixed with GM green pepper was obtained. Using those system, T3 peppers tolerant to tobacco budworm were selected.

The most important factor in breeding program is to obtain the value-added genetic line. Generally, breeders develop genetic sources using several methods such as segregation-breeding, cross-breeding, backcross-breeding, mutation induction, tissue culture and so on. Here, we present one classical way but very valuable method called cell fusion or protoplast fusion to create genetic sources for the breeding practice. The method we developed was the asymmetric somatic-hybridization of protoplast isolated from carrots. This is rather to transfer the nucleus from the high quality F1 hybrid to other mediocre line to produce a new carrot line. Since the breeding a carrot line for higher quality and purity takes a long time, therefore this nuclear transfer technology is very beneficial to generate a new line that could be useful to breed elite varieties. We had obtained around 200 fused carrots (cybrids), 12 cybrids were self pollinated and produced seeds. Selected progenies are currently being evaluated for horticultural characteristics including self fertility.