Bumkyu Lee

Variation and correlation analysis of phenolic compounds in mungbean (Vigna radiata L.) varieties

Phenolic compounds from a wide collection of mungbean [Vigna radiata (L.) Wilczek] germplasm (56 varieties) were characterised to determine the diversity among these phytochemicals and to analyse the relationships among their contents. The profiles of 25 phenolic compounds identified from the grains were subjected to data-mining processes, including principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), Pearsons correlation analysis, and hierarchical clustering analysis (HCA). The IT212105 and IT104818 varieties separated from the others in the first two principal components of PCA. PLS-DA showed significant separation between extracts of mungbean originating from three countries: China, Japan, and Korea. HCA of these phytochemicals resulted in clusters derived from common or closely related biochemical pathways. Significant positive relationships were observed between coumaric acid and resveratrol (r=0.7195, p<0.0001). Catechin content was positively correlated with rutin (r=0.6291, p<0.0001). The IT104818 variety appears to be a good candidate for future breeding programs, as it contains high levels of phenolic compounds. These results demonstrate the use of metabolic profiling combined with chemometrics as a tool for assessing the quality of food.

Two-year field study shows little evidence that PPO-transgenic rice affects the structure of soil microbial communities

There is global concern about the environmental consequences associated with transgenic crops. Their effects on the soil ecosystem are of special interest when assessing ecological safety and integrity. Although many efforts have been made to develop crops genetically modified to have resistance to protoporphyrin oxidase (PPO)-inhibiting herbicides, little is known about their influence on soil microbial communities. We conducted a 2-year field study and an analysis via terminal restriction fragment length polymorphism (T-RFLP) to assess the impacts of PPO-transgenic rice on bacterial and fungal communities. In the first year we sampled the rhizosphere and surrounding bulk soil, while in the second year we sampled rhizosphere soil only. No differences were observed in the diversity indices and community composition of microbial communities between transgenic rice and its parental non-transgenic counterpart (cultivar Dongjin). Instead, community variation was strongly dependent on growth stage and year. Therefore, we observed no adverse effects by these crops of modified rice on the microbial community composition in paddy soils.

Nucleoside diphosphate kinase-1 regulates hyphal development via the transcriptional regulation of catalase in Neurospora crassa

A nucleoside diphosphate kinase‐1‐disrupted (ndk‐1RIP‐1 ) mutant was observed to be defective in aerial hyphal and conidial development. In this study, two types of hyphae, fine and thick, were observed in wild‐type (Wt) strains. However, only fine‐type hyphae were observed in the ndk‐1RIP‐1 mutants. The ndk‐1RIP‐1 mutants were stimulated by oxidative stress and constitutively expressed an antioxidant enzyme catalase (CAT)‐3. Furthermore the ndk‐1RIP‐1 mutants could form thick hyphae by oxidative stress and a disruption of cat‐3. These results suggest that the loss of thick hyphae in the ndk‐1RIP‐1 mutants may be caused by the over‐expression of cat‐3.

Appearance / Instance of Genetically Modified Maize at Grain Receiving Harbors and Along Transportation Routes in Korea

Genetically modified (GM) crops are not permitted to be cultivated in Korea, but can only be imported as food or feed purposes. The import of GM crops has sharply increased in recent years, thus raising concerns with regard to the unintentional escape of these crops during transport and manufacturing as well as the subsequent contamination of local, non-GM plants. Hence, monitoring of GM crops was studied in or outside of grain receiving ports as well as from feed-processing plants in Korea during July 2008. We observed spilled maize grains and established plants primarily in storage facilities that are exposed around the harbors and near transportation routes of the feed-processing areas. Based on the PCR analyses, a total of 17 GM maize plants and 11 seeds were found among the samples. In most cases, the established maize plants found in this study were at the vegetative stage and thus failed to reach the reproductive stage. This study concludes that, in order to prevent a genetic admixture in the local environment for GM crops or seeds, frequent monitoring work and proper action should be taken.

Detection of gene flow from GM to non-GM watermelon in a field trial

Gene flow from genetically modified (GM) crops to conventional non-GM crops is a serious concern for protection of conventional and organic farming. Gene flow from GM watermelon developed for rootstock use, containing cucumber green mottle mosaic virus (CGMMV)-coat protein (CP) gene, to a non-GM isogenic control variety “Clhalteok” and grafted watermelon “Keumcheon” was investigated in a small scale field trial as a pilot study. Hybrids between GM and non-GM watermelons were screened from 1304 “Chalteok” seeds and 856 “Keumcheon” seeds using the duplex PCR method targeting theCGMMV- CP gene as a marker. Hybrids were found in all pollen recipient plots. The gene flow frequencies were greater for “Chaiteok” than for “KeumcheonD; with 75% outcrossing in the “Chaiteok” plot at the closest distance (0.8 m) to the GM plot. A much larger scale field trial is necessary to identify the isolation distance between GM and non-GM watermelon, as the behaviors of insect pollinators needs to be clarified in Korea.

Assessment of Gene Flow from Genetically Modified Anthracnose-Resistant Chili Pepper (Capsicum annuum L.) to a Conventional Crop

We conducted a 2-year field assessment of the gene flow from genetically modified (GM) chili pepper (Capsicum annuum L.), containing the PepEST (pepper esterase) gene, to a non-GM control line “WT512” and two commercial hybrid cultivars, “Manidda” and “Cheongpung Myeongwol (CM).” After seeds were collected from the pollen-recipient non-GM plants, hybrids between them and the GM peppers were screened by a hygromycin assay. PCR with the targeting hpt gene was performed to confirm the presence of transgenes in hygromycin-resistant seedlings. Out of 7,071 “WT512” seeds and 6,854 “Manidda” seeds collected in 2006, eight and 12 hybrids, respectively, were detected. In 2007, 33 hybrids from 3,456 “WT512” seeds and 50 hybrids from 3,457 “CM” seeds were found. The highest frequency of gene flow, 6.19%, was observed in that 2007 trial. These results suggest that a limited isolation distance would be sufficient to prevent gene flow from GM to conventionally bred chili peppers.

Assessment of the Persistence of DNA in Decomposing Leaves of CMVP0-CP Transgenic Chili Pepper in the Field Conditions

This study was conducted to evaluate the persistence of DNA in the transgenic chili pepper resistant to cucumber mosaic virus (CMV) in the field condition. We analyzed the persistence of genes in the leaf samples obtained from two field conditions, below and above soil. Transgenic and non-transgenic leaf tissues were buried in the soil at a depth of 10 cm or placed on the soil surface. Qualitative and quantitative PCR analysis showed that the amount of transferred genes from the transgenic peppers below and above soil was dropped to 28.3-42.7% one month after buried and it was rapidly reduced to 0.9-3.3% after two months. The transgenes were not detected three to four month after buried. In addition, DNA of the leaves placed below soil decomposed about 8%more than those on soil surface. The gene transfer from decomposing leaves of the transgenic pepper to soil was investigated by PCR analysis with the soil attached to the samples. The PCR result indicated that the gene transfer from the transgenic pepper to soil was not occurred.

The Effect of the Anthracnose Resistant Chili Pepper (PepEST gene) on the Non-target Insects, Green Peach Aphids (Myzus persicae Sulzer, Homoptera)

To assess the environmental risks of transgenic chili pepper with PepEST gene on non-target organisms before it exposes to the agro-ecosystem environments, we conducted the three sets of green peach aphids (Myzus persicas S.) life table experiment under laboratory conditions (Temp. , R.H. 50-70%, Photoperiod L16 : D8) in series during 2005-2006. We measured the net reproductive rate (, the intrinsic rate of increase (), the mean generation time (), , life span, and reproduction period between non-transgenic chili peppers and transgenic chili peppers, respectively. The life span of green peach aphids from three sets was 31, 27, 25 days, and the period of life span was similar to the general average length of green peach aphids, 25-29 days. Although the first reproduction of transgenic pepper was similar to the non transgenic pepper (P>0.05), the fecundity and the net reproductive rate () by using Jackknife method of transgenic pepper were lower than those of non transgenic pepper (P

Investigation into effects of transgenic glufosinate-resistant Zoysia grasses with herbicide application on bacterial communities under field conditions

Zoysia grass (Zoysia japonica Steud.) is a commercially valuable turfgrass that is popularly cultivated for use in sports and recreational environments. A field study was conducted in 2013 to evaluate the effects of transgenic herbicide-resistant Zoysia grasses in combination with the herbicide glufosinate on microbial communities. We investigated population changes in cultivable total bacteria and glufosinateresistant bacteria using the traditional plate-count method. The results showed that total and glufosinate-resistant bacteria counts decreased in September. Significant differences in the abundance of cultivable bacteria were observed between Zoysia grass lines as well as between March and September samples in both glufosinate-treated and nontreated plots. However, there was no significant difference in the abundance of total cultivable bacteria between Zoysia grass lines in March or between Basta-treated and untreated plots in September. To assess the possible horizontal gene transfer (HGT) of the bar gene across microorganisms, total soil DNAs and genomic DNAs of glufosinate-resistant bacteria isolated from soils and water were analyzed by PCR. No positive DNA bands were found, indicating that HGT did not occur during this experimental period. We also investigated changes in taxonomic distribution of the bacterial community using 16S rRNA gene clone libraries constructed from soil samples. Although bacterial diversities increased in September, there were no significant differences in species richness between genetically modified (GM) and non-GM Zoysia grasses as well as between glufosinatetreated and untreated soils.

Development of a chloroplast DNA marker for monitoring of transgene introgression in Brassica napus L.

Chloroplast molecular markers can provide useful information for high-resolution analysis of inter- and intra-specific variation in Brassicaceae and for differentiation between its species. Combining data generated from nuclear and chloroplast markers enables the study of seed and pollen movement, and assists in the assessment of gene-flow from genetically modified (GM) plants through hybridization studies. To develop chloroplast DNA markers for monitoring of transgene introgression in Brassica napus L., we searched for sequence variations in the chloroplast (cp) genome, and developed a simple cpDNA marker that is reliable, time-saving, and easily discriminates among 4 species (B. napus, B. rapa, Raphanus sativus, and Sinapis alba) based on PCR-product length polymorphism. This marker will be useful to identify maternal lineages and to estimate transgene movement of GM canola.