Influence of Genetically Modified Soybean Expressing Epidermal Growth Factor on Arthropod Biodiversity.

The field study was undertaken to examine the potential for adverse effects of transgenic soybean expressing bioactive human epidermal growth factor (with tolerance to the herbicide glufosinate, PPT) on the abundance and diversity of plant-dwelling arthropods by comparing with those of a non-GM parental cultivar, Gwangan soybean. Field surveys of soybean fields were carried out over two consecutive years, 2016 and 2017 at Ochang and Jeonju, Korea. The number of captured individuals associated with either of EGF and Gwangan soybean plants increased in 2017 compared with 2016 in both Ochang and Jeonju. During the survey period, the diversity and richness of the occurred insects and arachnids increased, dominance decreased, and the evenness of the insects remained static. The insects of Hymenoptera Order occurred most often comprised 25.4% of total captured insect pests. On the contrary, natural enemy from Hymenoptera Order and other insects from Diptera Order occurred more frequently (29.9% and 19.0%, respectively) in both the survey regions during the study periods. The score from PROXSCAL multidimensional scaling using combined data showed that the occurrence of insects and arachnids were separated due to their cultivation regions and years, irrespective of soybean cultivars. Consequently, the results indicated that there happened no notable change in the composition of arthropod communities in soybean agroecosystem due to GM event in soybean expressing EGF.

Elucidation of genomic organizations of transgenic soybean plants through de novo genome assembly with short paired-end reads.

Elucidation of the genomic organizations of transgene insertion sites is essential for the genetic studies of transgenic plants. Herein, we establish an analysis pipeline that identifies the transgene insertion sites as well as the presence of vector backbones, through de novo genome assembly with high-throughput sequencing data in two transgenic soybean lines, AtYUCCA6-#5 and 35S-UGT72E3/2-#7. Sequencing data of approximately 28× and 29× genome coverages for each line generated by high-throughput sequencing were de novo assembled. The databases generated from the de novo assembled sequences were used to search contigs that contained putative insertion sites and their flanking sequences (integration sites) of transgene fragments using transgenic vector sequences as queries. The predicted integration site sequences, which are located at three annotated genes that might regulate plant development or confer disease resistance, were then confirmed by local alignment against the soybean reference genome and PCR amplification. As results, we revealed the precise transgene-flanking sequences and sequence rearrangements at insertion sites in both the transgenic lines, as well as the aberrant insertion of a transgene fragment. Consequently, relative to experimental or enrichment technologies, our approach is straightforward and time-effective, providing an alternative method for the identification of insertion sites in transgenic plants.

Compositional differences in hybrids between protoporphyrinogen IX oxidase (PPO)-inhibiting herbicide-resistant transgenic rice and weedy rice accessions.

We characterized the metabolites in grains of transgenic protoporphyrinogen IX oxidase-inhibiting herbicide-resistant rice and weedy accessions using GC-MS and examined whether the chemical composition of their hybrids differed from that of the parents. We found that the metabolite profiles of transgenic rice and weedy rice were clearly separated. Although the metabolite profiles of F2 progeny were partially separated from their parents, zygosity did not affect the profiles. The F2 progeny had similar or intermediate levels of most major nutritional components compared with their parents. However, levels of galactopyranose, trehalose, xylofuranose, mannitol, and benzoic acid were higher in the F2 progeny. Some fatty acids and organic acids also showed prominent quantitative differences between the F2 progeny and the parents. Changes in the metabolite levels of transgenic crop-weed hybrids compared to their parents might influence not only the ecological consequences of the hybrids, but also the nutritional quality and food safety.

Performance of hybrids between abiotic stress-tolerant transgenic rice and its weedy relatives under water-stressed conditions.

Gene transfer from transgenic crops to their weedy relatives may introduce undesired ecological consequences that can increase the fitness and invasiveness of weedy populations. Here, we examined the rate of gene flow from abiotic stress-tolerant transgenic rice that over-express AtCYP78A7, a gene encoding cytochrome P450 protein, to six weedy rice accessions and compared the phenotypic performance and drought tolerance of their hybrids over generations. The rate of transgene flow from AtCYP78A7-overexpressing transgenic to weedy rice varied between 0% and 0.0396%. F1 hybrids containing AtCYP78A7 were significantly taller and heavier, but the percentage of ripened grains, grain numbers and weight per plant were significantly lower than their transgenic and weedy parents. The homozygous and hemizygous F2 progeny showed higher tolerance to drought stress than the nullizygous F2 progeny, as indicated by leaf rolling scores. Shoot growth of nullizygous F3 progeny was significantly greater than weedy rice under water-deficient conditions in a rainout shelter, however, that of homozygous F3 progeny was similar to weedy rice, indicating the cost of continuous expression of transgene. Our findings imply that gene flow from AtCYP78A7-overexpressing transgenic to weedy rice might increase drought tolerance as shown in the pot experiment, however, increased fitness under stressed conditions in the field were not observed for hybrid progeny containing transgenes.

Erratum to: Confirmation of Drought Tolerance of Ectopically Expressed AtABF3 Gene in Soybean.

[This corrects the article on p. 413 in vol. 41.].

Confirmation of Drought Tolerance of Ectopically Expressed AtABF3 Gene in Soybean.

Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from T2 seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than non-transgenic (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants (T2) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.

The influence of host genotype and salt stress on the seed endophytic community of salt-sensitive and salt-tolerant rice cultivars.

BACKGROUND: Inherent characteristics and changes in the physiology of rice as it attains salt tolerance affect the colonizing bacterial endophytic communities of the rice seeds. These transmissible endophytes also serve as a source of the plant's microbial community and concurrently respond to the host and environmental conditions. This study explores the influence of the rice host as well as the impact of soil salinity on the community structure and diversity of seed bacterial endophytes of rice with varying tolerance to salt stress. Endophytic bacterial diversity was studied through culture-dependent technique and Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. RESULTS: Results revealed considerably diverse communities of bacterial endophytes in the interior of rice seeds. The overall endophytic bacterial communities of the indica rice seeds based on 16S rRNA analysis of clones and isolates are dominated by phylum Proteobacteria followed by Actinobacteria and Firmicutes. Community profiles show common ribotypes found in all cultivars of the indica subspecies representing potential core microbiota belonging to Curtobacterium, Flavobacterium, Enterobacter, Xanthomonas, Herbaspirillum, Microbacterium and Stenotrophomonas. Clustering analysis shows that the host genotype mainly influences the seed endophytic community of the different rice cultivars. Under salt stress conditions, endophytic communities of the salt-sensitive and salt-tolerant rice cultivars shift their dominance to bacterial groups belonging to Flavobacterium, Pantoea, Enterobacter, Microbacterium, Kosakonia and Curtobacterium. CONCLUSION: The endophytic communities of rice indica seeds are shaped by the hosts' genotype, their physiological adaptation to salt stress and phylogenetic relatedness. Under salt stress conditions, a few groups of bacterial communities become prominent causing a shift in bacterial diversity and dominance.

Salinity affects metabolomic profiles of different trophic levels in a food chain.

Salinization is one of the most important abiotic stressors in an ecosystem. To examine how exposing a host plant to excess salt affects the consequent performance and metabolism of insects in a food chain, we determined the life history traits and the metabolite profiles in rice (Oryza sativa), the herbivore Sitobion avenae, and its predator Harmonia axyridis. When compared with performance under normal (non-stressed) conditions, exposing plants to 50mM NaCl significantly delayed the timing of development for S. avenae fed on rice and H. axyridis and also reduced the body mass of the latter. Our GC-MS-based analysis revealed clear differences in metabolite profiles between trophic levels or treatment conditions. Salinity apparently increased the levels of main components in rice, but decreased levels of major components in S. avenae and H. axyridis. In addition, 16 metabolites showed salinity-related contrasts in this trophic interaction for our rice-S. avenae-H. axyridis system. Salinity impeded the accumulation of metabolites, especially several sugars, amino acids, organic acids, and fatty acids in both insects, a response that was possibly associated with the negative impacts on their growth and reproduction under stress conditions.

Transgenic Cabbage Expressing Cry1Ac1 Does Not Affect the Survival and Growth of the Wolf Spider, Pardosa astrigera L. Koch (Araneae: Lycosidae).

Both herbivores that consume transgenic crops and their predators can be exposed to insecticidal proteins expressed in those crops. We conducted a tritrophic bioassay to evaluate the ecotoxicological impacts that Bt cabbage (Brassica oleracea var. capitata) expressing Cry1Ac1 protein might have on the wolf spider (Pardosa astrigera), a non-target generalist predator. Enzyme-Linked Immunosorbent Assays indicated that protein levels were 4.61 ng g(-1) dry weight in fruit flies (Drosophila melanogaster) fed with the transgenic cabbage and 1.86 ng g(-1) dry weight in the wolf spiders that preyed upon them. We also compared the life history traits of spiders collected from Bt versus non-Bt cabbage and found no significant differences in their growth, survival, and developmental rates. Because Bt cabbage did not affect the growth of fruit flies, we conclude that any indirect effects that this crop had on the wolf spider were probably not mediated by prey quality. Therefore, exposure to Cry1Ac1 protein when feeding upon prey containing that substance from transgenic cabbage has only a negligible influence on those non-target predatory spiders.

Comparative analysis of chemical compositions between non-transgenic soybean seeds and those from plants over-expressing AtJMT, the gene for jasmonic acid carboxyl methyltransferase.

Transgenic overexpression of the Arabidopsis gene for jasmonic acid carboxyl methyltransferase (AtJMT) is involved in regulating jasmonate-related plant responses. To examine its role in the compositional profile of soybean (Glycine max), we compared the seeds from field-grown plants that over-express AtJMT with those of the non-transgenic, wild-type (WT) counterpart. Our analysis of chemical compositions included proximates, amino acids, fatty acids, isoflavones, and antinutrients. Overexpression of AtJMT in the seeds resulted in decreased amounts of tryptophan, palmitic acid, linolenic acid, and stachyose, but increased levels of gadoleic acid and genistein. In particular, seeds from the transgenic soybeans contained 120.0-130.5% more genistein and 60.5-82.1% less stachyose than the WT. A separate evaluation of ingredient values showed that all were within the reference ranges reported for commercially available soybeans, thereby demonstrating the substantial equivalence of these transgenic and non-transgenic seeds.

Metabolomic changes in grains of well-watered and drought-stressed transgenic rice.

BACKGROUND: Drought induces a number of physiological and biochemical responses in cereals. This study was designed to examine the metabolite changes in grains of drought-tolerant transgenic rice (Oryza sativa L.) that overexpresses AtCYP78A7 encoding cytochrome P450 protein using proton nuclear magnetic resonance ((1)H-NMR) and gas chromatography/mass spectrometry. RESULTS: Principal component analysis showed that the (1)H-NMR-based profile was clearly separated by soil water status of well-watered and water-deficit. A discrimination of metabolites between transgenic and non-transgenic grains appeared under both watering regimes. Variations in the levels of amino acids and sugars led to the discrimination of metabolites among genotypes. In particular, drought significantly enhanced the levels of γ-aminobutyric acid (GABA, 244.6%), fructose (155.7%), glucose (211.0%), glycerol (57.2%), glycine (65.8%) and aminoethanol (192.4%) in the transgenic grains compared with the non-transgenic control grains. CONCLUSION: These changes in amounts of metabolites may assist in improving drought tolerance in transgenic rice by playing crucial roles in stress-responsive pathways including GABA biosynthesis, sucrose metabolism and antioxidant defenses.

Bystander attitudes toward parents? The perceived meaning of filial piety among Koreans in Australia, New Zealand and Korea.

AIM: The objective of this paper was to explore how present-day filial piety is understood among Koreans in geographically different settings. METHODS: Data were collected from qualitative interviews with 61 Korean participants in Australia, New Zealand and Korea and then thematically analysed and evaluated. RESULTS: The findings from this study show that filial piety for Koreans consists of family care and support, along with respect for parents. The norm of filial piety is regarded as an important aspect of intergenerational family relationships in current Korean culture, while its practice is perceived as increasingly eroded within a context of major socio-cultural and economic changes, including migration. CONCLUSION: The results show that the tension between the social and economic aspects of filial piety often creates a 'bystander' attitude toward parents and provides fertile ground for the seeds of family conflict.

Arbuscular mycorrhizal fungi community structure, abundance and species richness changes in soil by different levels of heavy metal and metalloid concentration.

Arbuscular Mycorrhizal Fungi (AMF) play major roles in ecosystem functioning such as carbon sequestration, nutrient cycling, and plant growth promotion. It is important to know how this ecologically important soil microbial player is affected by soil abiotic factors particularly heavy metal and metalloid (HMM). The objective of this study was to understand the impact of soil HMM concentration on AMF abundance and community structure in the contaminated sites of South Korea. Soil samples were collected from the vicinity of an abandoned smelter and the samples were subjected to three complementary methods such as spore morphology, terminal restriction fragment length polymorphism (T-RFLP) and denaturing gradient gel electrophoresis (DGGE) for diversity analysis. Spore density was found to be significantly higher in highly contaminated soil compared to less contaminated soil. Spore morphological study revealed that Glomeraceae family was more abundant followed by Acaulosporaceae and Gigasporaceae in the vicinity of the smelter. T-RFLP and DGGE analysis confirmed the dominance of Funneliformis mosseae and Rhizophagus intraradices in all the study sites. Claroideoglomus claroideum, Funneliformis caledonium, Rhizophagus clarus and Funneliformis constrictum were found to be sensitive to high concentration of soil HMM. Richness and diversity of Glomeraceae family increased with significant increase in soil arsenic, cadmium and zinc concentrations. Our results revealed that the soil HMM has a vital impact on AMF community structure, especially with Glomeraceae family abundance, richness and diversity.

Drought stress-induced compositional changes in tolerant transgenic rice and its wild type.

Comparing well-watered versus deficit conditions, we evaluated the chemical composition of grains harvested from wild-type (WT) and drought-tolerant, transgenic rice (Oryza sativa L.). The latter had been developed by inserting AtCYP78A7, which encodes a cytochrome P450 protein. Two transgenic Lines, '10B-5' and '18A-4', and the 'Hwayoung' WT were grown under a rainout shelter. After the harvested grains were polished, their levels of key components, including proximates, amino acids, fatty acids, minerals and vitamins were analysed to determine the effect of watering system and genotype. Drought treatment significantly influenced the levels of some nutritional components in both transgenic and WT grains. In particular, the amounts of lignoceric acid and copper in the WT decreased by 12.6% and 39.5%, respectively, by drought stress, whereas those of copper and potassium in the transgenics rose by 88.1-113.3% and 10.4-11.9%, respectively, under water-deficit conditions.

Reliability and Validity of Alcohol Use Disorder dentification Test-Korean Revised Version for Screening At-risk Drinking and Alcohol Use Disorders.

BACKGROUND: There needs to be an amendment to the Korean version of the Alcohol Use Disorder Identification Test (AUDIT) with regards to the recent change in percent alcohol by volume (ABV) Korean liquor. This study was performed to suggest a cutoff value, reliability and validity of AUDIT-Korean revised version (AUDIT-KR), which reflect the change of the ABV of Korean alcohol. METHODS: The subjects were 435 peoples (210 males and 225 females), who visited the Chungnam National University Hospital for a comprehensive medical examination. The respondents completed the AUDIT-KR. At-risk drinking and alcohol use disorders had been evaluated by diagnostic interview. The Cronbach's alpha value, the receiver operating characteristic curve, the appropriate cutoff value, sensitivity and specificity of the AUDIT-KR were evaluated. RESULTS: There were 190 at-risk drinkers (111 males and 79 females), and 66 people with alcohol use disorders (48 males and 18 females). The cutoff value of the AUDIT-KR for at-risk drinking was 3 points (sensitivity 93.69% and specificity 78.79%) for males and 3 points (sensitivity 92.40% and specificity 78.08%) for females. The cutoff value for alcohol use disorders was 10 points (sensitivity 100.00% and specificity 89.51%) for males and 8 points (sensitivity 100.00% and specificity 93.71%) for females. Cronbach's alpha of the AUDIT-KR was 0.885. CONCLUSION: The above results suggest that the AUDIT-KR shows a high reliability and validity in identifying at-risk drinking and alcohol use disorders.

Ageing in an inconvenient paradise: the immigrant experiences of older Korean people in New Zealand.

AIM: The purpose of this paper is to explore the immigrant experiences of older Korean people and their intergenerational family relationships in the New Zealand context. METHODS: Data were collected from qualitative interviews with older people, community leaders and professionals in Christchurch and Auckland. Data analysis was conducted using concept mapping techniques in the cross-cultural context where two languages were simultaneously used. RESULTS: The findings of the study show that older Korean people in New Zealand were likely to face multiple challenges due to the combined effects of immigration and ageing in a new country. Some older people experienced difficulties in managing their immigrant lives and intergenerational relationships in the transnational family context in which their family members were dispersed across two or more nations. CONCLUSION: The immigrant experiences of older migrants might be affected by an 'invisible' source of isolation and exclusion at familial, community, societal and transnational levels.

Gene flow from herbicide-tolerant GM rice and the heterosis of GM rice-weed F2 progeny.

Gene flow from genetically modified (GM) crops to non-GM cultivars or weedy relatives may lead to the development of more aggressive weeds. We quantified the amount of gene flow from herbicide-tolerant GM rice (Protox GM, derived from the cultivar Dongjin) to three cultivars (Dongjin, Aranghyangchal and Hwaseong) and a weedy rice line. Gene flow frequency generally decreased with increasing distance from the pollen donor. At the shortest distance (0.5 m), we observed a maximum frequency (0.039%) of gene flow. We found that the cultivar Dongjin received the greatest amount of gene flow, with the second being weedy rice. Heterosis of F2 inbred progeny was also examined between Protox GM and weedy rice. We compared growth and reproduction between F2 progeny (homozygous or hemizygous for the Protox gene) and parental rice lines (GM and weedy rice). Here, transgene-homozygous F2 progeny was significantly taller and produced more seeds than the transgene-hemizygous F2 progeny and parental lines. Although the gene flow frequency was generally low, our results suggest that F2 progeny between GM and weedy relatives may exhibit heterosis.

Temporal dynamics of bacterial and fungal communities in a genetically modified (GM) rice ecosystem.

We assessed the temporal dynamics of bacterial and fungal communities in a soil ecosystem supporting genetically modified (GM) rice (Oryza sativa L., ABC-TPSP; fusion of trehalose-6-phosphate synthase and phosphatase). Using terminal restriction fragment length polymorphism analysis and real-time quantitative PCR, we compared bacterial and fungal communities in the soils underlying GM rice (ABC-TPSP), and its host cultivar (Nakdong) during growing seasons and non-growing seasons. Overall, the soils supporting GM and non-GM rice did not differ significantly in diversity indices, including ribotype numbers, for either bacteria or fungi. The diversity index (H) in both the bacterial and fungal communities was correlated with water content, dissolved organic carbon (DOC), and ammonium nitrogen, and the correlation was stronger in fungi than in bacteria. Multivariate analysis showed no differences in microbial community structures between the two crop genotypes, but such differences did appear in time, with significant changes observed after harvest. Gene copy number was estimated as 10(8)~10(11) and 10(5)~10(7) per gram of soil for bacteria and fungi, respectively. As observed for community structure, the rice genotypes did not differ significantly in either bacterial- or fungal-specific gene copy numbers, although we observed a seasonal change in number. We summarize the results of this study as follows. (1) GM rice did not influence soil bacterial and fungal community structures as compared to non-GM rice in our system, (2) both bacterial and fungal communities changed with the growth stage of either rice genotype, (3) fungal communities were less variable than bacterial communities, and (4) although several environmental factors, including ammonium nitrogen and DOC correlated with shifts in microbial community structure, no single factor stood out.

Fine genetic mapping of the genomic region controlling leaflet shape and number of seeds per pod in the soybean.

Narrow leaflet cultivars tend to have more seeds per pod than broad leaflet cultivars in soybean [Glycine max (L.) Merr.], which suggests that the leaflet-shape trait locus is tightly linked to or cosegregates with the trait locus controlling the number of seeds per pod (NSPP). Here, we attempted to further elucidate the relationship between leaflet shape and NSPP. A BC(3)F(2) population from a cross between the 'Sowon' (narrow leaflets and high NSPP) and 'V94-5152' (broad leaflets and low NSPP) variants was used. The results of the molecular genetic analyses indicated that, although the NSPP characteristic, in particular, the occurrence of 4-seeded pods, is governed by additional modifying genes that are likely present in Sowon, the two traits cosegregate in the BC(3)F(2) population. The mapping results generated using public markers demonstrated that the narrow leaflet-determining gene in Sowon is an allele of the previously highly studied ln gene on chromosome 20. A high-resolution map delimited the genomic region controlling both the leaflet shape and NSPP traits to a sequence length of 66 kb, corresponding to 0.7 cM. Among the three genes annotated in this 66 kb region, Glyma20g25000.1 appeared to be a good candidate for the Ln-encoding gene, owing to its 47.8% homology with the protein encoding for the JAGGED gene that regulates lateral organ development in Arabidopsis. Taken together, our results suggested that phenotypic variations for narrow leaflet and NSPP are predominantly from the pleiotropic effects of the ln gene. Thus, our results should provide a molecular framework for soybean breeding programs with the objective of improving soybean yield.

Effects of soil conservation measures in a partially vegetated area after forest fires.

After forest fires on the east coast of Korea in 2000, some burnt areas were left untreated. Although 80% of the area was reasonably revegetated within 3 months, about 20% of the area was partially vegetated, mainly due to a low density of sprouters and poor growing conditions (eroded soil and steep slopes). Three years after the fires, the effect of soil conservation measures, such as mulching with wood chips, seeding with native plant species and log erosion barriers (LEBs), on runoff and soil erosion were examined using runoff plots. Wood chip mulching greatly reduced runoff and sediment yields and these effects were consistent regardless of the volume of rainfall. Neither seeding nor LEBs reduced runoff and sediment yields. No positive or negative effects of mulching, seeding or LEBs on ground vegetation cover were observed. The ineffectiveness of seeding and LEBs may have been due to the steep slope, the failure of germination and establishment of seeded plants, and the small diameter of logs. Treating hill slopes with mulch should be considered where post-fire regeneration is slow and there is an absence of organic material such as litter.