Choi, Sang Chul;Chung, Yong Suk;Lee, Yun Gyeong;Park, Yun Ji;Kim, Changsoo
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.104-104
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2017
Sorghum (Sorghum bicolor (L.) Moench.) is one of the most important crops for human and animal nutrition. Nonetheless, sorghum has a cyanogenic glucoside compound which can be degraded into hydrogen cyanide, toxic to humans and animals even with tiny amount. In consequence, breeding materials with a low cyanide level has been a top priority in sorghum breeding programs. To fulfill our long-term goal, we are screening sorghum accessions with low cyanide level, which would be an important breeding material for food safety. We collected seeds of various sorghum accessions and analyzed relevant metabolites to find useful breeding materials of sorghum accessions containing low cyanide. Fourteen wild relatives were obtained from the University of Georgia in US, a reference accession BTx623, and three local varieties from National Agrobiodiversity Center of Rural Development Administration in Korea, and one wild species from the Wild Plant Resources Seed Bank of Korea University in Korea. Sorghum plants were grown in plastic greenhouse under natural conditions. After growing, leaf samples were harvested at different developmental stages: seedling phase, vegetative phase (right before flowering), and reproductive phase (ripening). Using collected samples, quantification analysis were performed by an HPLC system for three metabolites (dhurrin, 4-hydroxybenzaldehyde, and 4-hydroxyphenylacetic acid) in sorghum plants. Prior to metabolome analysis, specific experimental condition for HPLC system was set to be able to separate three metabolites simultaneously. Under this condition, these metabolites were quantified in each accession by HPLC system. We observed that the metabolite contents were changed differently by developmental stages and accessions. We clustered these results into five groups as patterns of their contents by developmental stages. Most of accessions showed that 4-hydroxybenzaldehyde content was very high at seedling stage and decreased rapidly at vegetative phase. Interestingly, the patterns of dhurrin content were very different among clusters. However, 4-hydroxyphenylacetic acid content was maintained at low levels by developmental stages in most accessions. The results would demonstrate how dhurrin and alternative degradation pathways are differentiated in each accession.
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.161-161
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2017
Seed endophytes are very remarkable groups of bacteria for their unique abilities of being vertically transmitted and conserved. As plants attain hybrid vigor and heterosis in the process of crossbreeding, this might also lead to the changes in the community structure and diversity of plant endophytes in the hybrid plants ultimately affecting the endophytes of the seeds. It would be interesting to characterize how seed endophyte composition change over time. The objective of this study is to gain insights into the influence of natural crossbreeding and parental lineage in the seed bacterial endophytic communities of two pure inbred lines exploring contributions of the two most important sources of plant endophytes - colonization from external sources and vertical transmission via seeds. Total genomic DNA was isolated from rice seeds and bacterial DNA was selectively amplified by PCR. The diversity of endophytic bacteria was studied through Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Diversity between the original parents and the pure inbred line may show significant differences in terms of richness, evenness and diversity indices. Heat maps reveal astonishing contributions of both or either parents (IR29 ${\times}$ Pokkali and AT401 ${\times}$ IR31868) in the shaping of the bacterial seed endophytes of the hybrid, FL478 and IC32, respectively. Most of the T-RFs of the subsequent pure inbred line could be traced to any or both of the parents. Comparison of common and genotype-specific T-RFs of parents and their offspring reveals that majority of the T-RFs are shared suggesting higher transmission of bacterial communities common to both parents. The parents influence the bacterial community of their offspring. Unique T-RFs of the offspring also suggest external sources of colonization particularly as the seeds are cultivated in different ecogeographical locations. This study showed that host parental lines contributed greatly in the shaping of bacterial seed endophytes of their offspring. It also revealed transmission and potential conservation of core seed bacterial endophytes that generally become the dominant microbiota in the succeeding generations of plant hosts.
A metabolic study has been conducted to investigate the conversion of pentachlorophenol(PCP) to water soluble metabolites in soybean and rice cell suspension cultures as well as in intact rice plants. PCP in plant cells was found to be exclusively transformed into water soluble metabolites. The relative rate of the metabolic conversion of PCP in decreasing order was soybean cultures > rice cultures > rice plants. Also observed was that, the older the cultures grown, the lower the conversion rate was. Primary water soluble metabolites isolated from both the 5 day old soybean and 8 day old rice cells were specifically hydrolyzed only by ${\beta}$-glucosidic linkage specific glucosidase, suggesting that the metabolites are ${\beta}$-glucose conjugates. The amount of glucose conjugates was increased with increasing time of incubation of PCP up to 24 hr in both soybean and rice cultures; Thereafter, it was decreased progressively. Most of the glucose conjugates were further metabolized to more polar conjugates in cells, but a portion of them was excreted into the culture medium.
The advent of available DNA barcoding technology has been extensively adopted to assist in the reference to differentiate the origin of various medicinal plants species. However, this technology is still far behind the curve of technological advances to be applied in a practical manner in the market to authenticate the counterfeit components or detect the contamination in the admixtures of medicinal plant species. Recently, a high resolution melting curve analysis technique was combined with the procedure of DNA barcoding (Bar-HRM) to accomplish this purpose. In this review, we tried to summarize the current development and bottleneck of processing related to the Bar-HRM technology for the practical application of medicinal plant species' differentiation in a viable global market. Although several successful results have been reported, there are still many obstacles to be resolved, such as limited number of DNA barcodes and single nucleotide polymorphisms, in particular, only one DNA barcode, internal transcribed sequence (ITS) of ribosomal DNA has been reported in the available nuclear genome. In addition, too few cases have been reported about the identification of counterfeit or contamination with processed medicinal plant products, in particular specifically the case of technology based infusion, jam and jelly products and components in which it is noted that DNA can be thereby degraded during the processing of these products and components.
Won, Jong Chan;Won, Ok Jae;Ha, Jun;Im, Il-Bin;Kang, Kwang Sik;Pyon, Jong Yeong;Park, Kee Woong;Lee, Jeung Joo
Weed & Turfgrass Science
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v.7
no.2
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pp.166-169
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2018
Repeated use of ACCase inhibiting herbicides for a long time has resulted in increases of resistant Echinochloa oryzicola populations in paddy fields in middle west area of Korea. This study aims to investigate current status of herbicide resistant E. oryzicola in Gyeongsangnam-do, in which there is less information about herbicide resistance. For resistance frequency and dose-response study, seeds from 100 individual plants of E. oryzicola in Gyeongsangnam-do were collected and tested with cyhalofop-butyl. Seven percent of plants from Gyeongsangnam-do was resistant at a recommended rate of cyhalofop-butyl. $GR_{50}$ values (herbicide rates required to reduce plant growth 50%) for one representative resistant populations and five susceptible populations were $738g\;a.i.\;ha^{-1}$ and 66-234 (average 147)$g\;a.i.\;ha^{-1}$, respectively, indicating average 5 times difference in resistance. Although lower rate of frequency of herbicide resistance in Gyeongsangnam-do than in Jeollabuk-do, increases of herbicide resistance are expected in this area because of increases of direct seeded rice fields and increases of dependence on a specific herbicide. Therefore, it is necessary to monitor herbicide resistance regularly and conduct integrated herbicide resistance management in this area.
This study aimed to determinate the effect of fluorescent light and light-emitting diodes on the leaf morphology, growth and antioxidant capacity of Salvia plebeia. The plants were grown for 56 days after transplanting (DAT) under the fluorescent light (FL) and LEDs (White, Red and Blue (R+B, ratio 2:1), Blue, Red LED) under the same light intensity and photoperiod ($130{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, 12 hours). Leaf length, width and number of Salvia plebeia at the 56 DAT were significantly higher under the FL and red LED, and lower in the RB LED and white LED. The highest fresh and dry weights of shoot and leaf area were observed in the red LED, followed by the FL and blue LED, and the lowest in the RB LED and white LED. After 21 DAT, leaf apinasty symptom was appeared in plants grown under red LED and RB LED. The chlorophyll content was lower in the red LED. The specific leaf weight, the ratio of leaf dry weight to area, was higher in the blue LED, and lowest in the FL. No significant difference in DPPH radical scavenging activity of Salvia plebeia under the different light sources. All the integrated results suggest that the FL light is a proper light conditions for a closed cultivation of Salvia plebeia.
In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme dihydroflavonol 4-reductase (DFR) in Matthiola incana R. Br. A heterologous cDNA probe from Zea mays was used to isolate full-size DFR cDNA clone from a corolla-specific cDNA library. Comparison of the coding region of this DFR cDNA sequence including the sequences of Zea mays, Anthirrinum majus, Petunia hybrida, Callistephus chinensis, Dianthus caryophyllus and Rosa hybrida reveals a identity higher than 61% at the nucleotide level. The DFR transcript is G/C rich in monocotyledonous plants show a strong codon bias preferring codons with a G or C in the third position. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants and by in vitro expression yielding an enzymatically active reductase. Genomic southern blot analysis showed the presence of one gene for DFR in Matthiola incana. Northern blot analysis of the DFR wild type and mutant lines showed that the lack of DFR activity in the stable acyanic mutant k17b is clearly by a transcriptional block of the DFR gene.
Lee, Jeongran;Kim, Chang-Seok;Lee, In-Yong;Oh, Hyun-Ju;Kim, Jung Hyun;Kim, Sun Yu
Weed & Turfgrass Science
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v.4
no.1
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pp.26-34
/
2015
Korean Poaceae includes approximately 80 species of the agricultural weeds. Precise species identification is the first step for more effective weed management in the agricultural fields. However, the identification of species in Poaceae is not easy without the assistance of taxonomists or identification experts although they are relatively easy to distinguish from the plants of the other family by the unique characteristics of caryopsis. Thus, DNA barcode was suggested as an alternative powerful technique for species identification by using short sections of DNA from a specific region of the genome. Two standard barcode markers of vascular plants, chloroplast rbcL and matK, and a supplementary nuclear ribosomal Internal Transcribed Spacer (ITS) region were used for barcode of major Korean Poaceae weeds, 403 individuals of 84 taxa. All the barcode markers revealed a good level of sequencing success with the lowest 73.7% for matK and the highest 88.8% for rbcL. The barcode sequences were deposited to the National Center for Biotechnology Information (NCBI) database for public use. Combined matK and ITS showed very high resolving power with 92.9%. Besides the identification of weeds for weed managment, the generated DNA barcode data could be used for many other applications such as rapid biodiversity assessment and conservation prioritization.
A total of 35 phosphate solubilizing bacterial strains were isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam and screened for the production of indole acetic acid (IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 ${\mu}g$$ml^{-1}$ after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 ${\mu}g$$ml^{-1}$) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of $30^{\circ}C$ followed by $25^{\circ}C$. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains' ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same $R_f$ value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.
Quinacrine, a pH-sensitive fluorescence probe, which exists either as an unprotonated fluorescence form or a protonated noufluorescence form, can be used to measure the proton transport activity of $H^+-ATPase$. Quinacrine was used to determine the optimal conditions for measuring the activity of microsomal $H^+-ATPase$ prepared from the roots of tomato plants. The amount of quinacrine fluorescence quenching obtained at $0.43{\mu}g/{\mu}l$ of microsomal protein concentration was 25-26%, which shows that the enzyme activity of 100 nmol/min decreases 10% of quinacrine fluorescence. Maximal fluorescence quenching was obtained at pH 7.0-7.2 and 2 mM $Mg^{2+}$ Because the activity of microsomal $H^+-ATPase$ is also maximal at these conditions, the quinacrine fluorescence well represents the activity of $H^+-ATPase$. Vanadate and $NO_3-$, specific inhibitors of plasma and vacuolar $H^+-ATPases$, respectively, were successfully applied to inhibit the quinacrine fluorescence quenching mediated by the corresponding $H^+-ATPases$. These results imply that quinacrine is a useful tool for measuring the proton transport activities of microsomes obtained from the root tissue of tomato plants.
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