• Journal of Crop Science and Biotechnology
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• v.11 no.4
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• pp.227-232
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• 2008

Drought stress is one of the major abiotic stresses in agriculture worldwide. We report here a proteomic approach to investigate the impact of post-fertilization drought on grain quality in rice seed endosperm (Oryza sativa cv. IR-64). Plants were stressed for 4 days at 3 days before heading. Total proteins of endosperm were extracted and separated by two-dimensional gel electrophoresis. Not many protein spots showed differential accumulation in drought-stressed samples. More than 400 protein spots were reproducibly detected, including three that were up-regulated and five down-regulated. Mass spectrometry analysis and database searching helped us to identify six spots representing different proteins. Functionally, the identified proteins were related to protein synthesis and carbohydrate metabolism, such as Granule-Bound Starch Synthase (GBSS, Wx protein), which is thought to play a very important role in starch biosynthesis and quality, a very crucial factor in determining rice grain quality.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.216-216
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• 2022

The starch-branching enzymes (BEs) are responsible for synthesizing the amylopectin, which plays an important role in determining the structural and physical properties of starch granules. BE has two differently functioning isoforms (BEI and BEIIa/b) based on their difference in the chain-length pattern by the degree of polymerization (DP), which mainly contributes to the amylopectin chain length distribution in starch biosynthesis. In this study, we investigated functional haplotypes and evolutionary analyses of SBE1 in 374 rice accessions (320 Korean bred and 54 wild). The analyses were performed based on the classified subpopulations. Haplotype analysis generates a total of 8 haplotypes, of which only four haplotypes were functional carrying four functional SNPs in four different exons of SBE1 on chromosome 6. Nucleotide diversity analysis showed a highest pi-value in aromatic group (0.0029), while the lowest diversity value was in temperate japonica (0.0002), indicating the signal of this gene evolution origin. Different directional selections could be estimated by negative Tajima's D value of temperate japonica (-1.1285) and positive Tajima's D value of tropical japonica (0.9456), where the selective sweeps were undergone by both positive purifying and balancing selections. Phylogenetic analysis indicates a closer relationship of the wild with most of the cultivated subgroups indicating a common ancestor for SBE1 gene. FST-values indicate distant genetic relationships of temperate japonica from all other classified groups. PCA and population structure analysis show an admixed structure of wild and cultivated subpopulations in some proportions.

• Journal of Plant Biology
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• v.31 no.4
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• pp.277-288
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• 1988

The effects of ammonium ion and glutamate on CO2 fixation abilities and related carbon metabolism were investigated in pea (Pisum sativum L. cv. Sparkle) leaf discs under conditions favoring photorespiration (21% O2, 0.03% CO2) and nonphotorespiration (5% O2, 0.03% CO2). A concentration of more than 10 mM of NH4+ decreased the photosynthetic CO2 fixation and those inhibitory effects were more remarkable in 21% O2 than in 5% O2 conditions. The effect of glutamate on CO2 fixation was found to be independent of the O2 level, as glutamate increased the CO2 fixation under both 21% and 5% O2 conditions. L-methionine-dl-sulfoximine, an irreversible inhibitor of glutamate synthetase, however, inhibited the CO2 fixation markedly under 21% O2, but did not affect it under 5% O2 conditions. The treatment with NH4+ elevated the relative amounts of 14C incorporated into soluble components from 14CO2 with no relation to O2 levels, while glutamate increased 14C into insoluble components and neutral sugars. Glutamate, especially, seemed to stmulate the biosynthesis of starch under 5% O2 condition. These results indicated that NH4+ stimulated the degradation of sugar or starch and this proposal was confirmed by the increasing of pyruvate kinase activity in leaf discs treated with ammonium ion.

• Plant Breeding and Biotechnology
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• v.6 no.4
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• pp.337-353
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• 2018

We used Illumina/HiSeq sequencing for analysis of gene expression profiling among four maize seed types (dent, CM3 and CM6; waxy, CM5 and CM19) at 10 DAP (days after pollination). A total of 88,993,000 (CM3), 103,817,340 (CM6), 103,139,640 (CM5), and 66,978,958 (CM19) sequence reads were generated with read lengths of about 0.9, 1.0, 1.0, and 0.7 billion bp, respectively. We obtained 69.1 (CM3), 71.0 (CM6), 71.2 (CM5), and 71.8% (CM19) high quality reads from the raw data and compared them with reference RNA sequences in a public DB (NCBI). It was revealed that mapped reads were 58%, 63%, 62%, and 62% of the EST reference in CM3, CM6, CM5 and CM19, respectively; and more than 51,000 genes were expressed based on RPKM criteria (over 0.25 value) in each CM3, CM6, CM5, and CM19 inbred line. In differentially expressed gene (DEG) analysis, we found that 3,527 genes were differentially expressed by at least two-fold with 1,709 upregulated in the two waxy inbred lines and 1,818 upregulated in the two dent inbred lines. We also detected genes for the sucrose and starch biosynthesis pathways based on BINs, and different expression patterns between waxy and dent inbred lines were shown for the gene set for starch synthesis, such as sh2, bt2, du1, wx1, and ae1. Although some genes were more expressed in dent lines, most genes for starch synthesis were much expressed in waxy lines. Especially, there was greater expression of the sus2 gene in both waxy lines compared with the dent lines.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.123-129
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• 2007

The trehalose $({\alpha}-D-glucopyranosyl-[1,1]-{\alpha}-D-glucopyranose)$ biosynthesis genes MhMTS and MhMTH, encoding a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively, have been cloned from the hyperthermophilic archaebacterium Metallosphaera hakonesis. The ORF of MhMTS is 2,142 bp long, and encodes 713 amino acid residues constituting a 83.8 kDa protein. MhMTH is 1,677 bp long, and encodes 558 amino acid residues constituting a 63.7 kDa protein. The deduced amino acid sequences of MhMTS and MhMTH contain four regions highly conserved for MTSs and three for MTHs that are known to constitute substrate-binding sites of starch-hydrolyzing enzymes. Recombinant proteins obtained by expressing the MhMTS and MhMTH genes in E. coli catalyzed a sequential reaction converting maltooligosaccharides to produce trehalose. Optimum pH of the MhMTS/MhMTH enzyme reaction was around 5.0 and optimum temperature was around 70 C. Trehalose-producing activity of the MhMTS/ MhMTH was notably stable, retaining 80% of the activity after preincubation of the enzyme mixture at $70^{\circ}C$ for 48 h, but was gradually abolished by incubating at above $85^{\circ}C$. Addition of thermostable $4-{\alpha}-glucanotransferase$ increased the yield of trehalose production from maltopentaose by 10%. The substrate specificity of the MhMTS/MhMTH-catalyzed reaction was extended to soluble starch, the most abundant maltodextrin in nature.

• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.277-283
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• 2007

Embryogenic calluses derived from shoot apical meristem explants of sweetpotato were subjected to particle bombardment to generate transgenic plants for antisense expression of cDNAs encoding two different AGPase small subunit (ibAGP1 and ibAGP2). Plants were generated via somatic embryogenesis. PCR and Southern analysis demonstrated that the incorporation of ibAGP1 and ibAGP2 into the genome in an antisense orientation. Immunoblot analysis confirmed reduced levels of AGPase small subunit in transgenic plant leaves. Plants with both ibAGP1 and ibAGP2 produced a lower level of the protein than plants with ibAGP1 alone. iodine test demonstrated that transgenic plant leaves and storage root accumulated reduced amounts of starch. Iodine staining of leaf tissues indicated that transgenic plants accumulated less amount of starch than control. In accordance with western blot analysis, plants with both ibAGP1 and ibAGP2 accumulated a lower amount of starch than plants with ibAGP1 alone. Both transgenic plants exhibited a severely retarded growth, resulting in bare survival. It is suggested that disrupted expression of the gene encoding AGPase small subunit is lethal to the growth of sweetpotato contrast to other species including potato.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.2
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• pp.84-90
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• 2005

The effects of UV-B radiation on the seedling growth, carbohydrate metabolism and antioxidants activities of rice (Oryza sativa L.) were investigated under environmentally controlled chamber. Supplementary UV­B radiation reduced dry matter as well as leaf area, there­fore, relative growth rates (RGR) of seedlings were decreased by up to half compared to control. Photosynthetic products such as soluble sugars and starch were rapidly and significantly reduced by within 1 day of enhanced UV-B radiation due to the inhibition and degradation of photosynthetic processes and thylakoid membrane integrity. In our study, nonstructural carbohydrate levels were proved to be a main indicator on UV-B­induced stress. The behavior of SOD, CAT, APX and POD activities was monitored in the leaves of rice seedlings subjected to UV-B radiation. Under UV-B treatments, SOD activity was initially increased, whereas CAT and POD activities were slowly and slightly increased. However, APX activity showed no presumable results with an increase of UV-B dose. In leaves of rice seedlings, supplementary UV-B radiation caused an increase in free putrescine and spermidine, however spermine remained unaltered, although 24-hrs UV-B treatment slightly increased. This result presumes that an excess UV-B dose may induce ethylene biosynthesis (senescence) rather than polyamine biosynthesis (defense).

• Microbiology and Biotechnology Letters
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• v.50 no.4
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• pp.522-532
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• 2022

Cheonggukjang is a traditional fermented food in Korea, which is known to exert beneficial effects on health. In this study, we evaluated the effects of cheonggukjang fermented by Bacillus subtilis SCGB 574 (B574) on high fat diet (HFD)-deteriorated large intestinal health. Rats were fed with HFD or HFD supplemented with 10.1% cheonggukjang (B574). Fecal microbiota was analyzed based on 16S rRNA gene sequences, and the fecal and serum metabolome were measured using GC-MS. Our results showed that SCGB574 intake significantly reduced body weight, restored tight junction components, and ameliorated inflammatory cell infiltration. SCGB574 also shifted gut microbiota by increasing the abundance of short chain fatty acid producers such as Alistipes and Flintibacter, although it decreased the abundance of Lactobacillus. Serum and fecal metabolome analyses showed significantly different metabolic profiles between the groups. The top five metabolites increased by SCGB574 were i) arginine biosynthesis, ii) alanine, aspartate, and glutamate metabolism; iii) starch and sucrose metabolism; iv) neomycin, kanamycin, and gentamicin biosynthesis; and v) galactose metabolism. These results showed that cheonggukjang fermented by SCGB574 ameliorates adverse effects of HFD through improving intestinal health.

• KSBB Journal
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• v.10 no.2
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• pp.196-203
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• 1995

The effects of majors nutrients and environmental factors on polyoxins biosysnthesis were examined for the overprodution of antifungal agent polyoxins using Streptomyces sp. 809-11. The cell mass at the exponential growth phase was increased when soluble starch was used, while the polyoxins biosynthesis was increased with the formation of filamentous mycelium when glucose was used as a carbon source. It was, therefore, recommendable to use both soluble starch and glucose simultaneously as carbon sources for the cell growth and polyoxins production. The high concentration of ammonium sulfate (151.4 mM) resulied in the improvement of polyoxins production. The optimal concentration of $K_2HPO_4$for polyoxins production was found to be 0.5mM. By applying fed-batch culture, polyoxins production was improved to about 2-folds compared to the result from the batch culture.

• Journal of Life Science
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• v.9 no.2
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• pp.160-168
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• 1999

Butyrate is one of the short-chain fatty acids that are present in the colon of mammals in millimolar concentration as a result of microbial anaerobic fermentation of dietary fiber, undigested starch, and proteins. In this study, sodium butyrate was examined in HT29 cell, human colonic cancer cell line, on cell viability, alkaline phosphatase activity, PLC-${\gamma}$1 expression and complex sphingolipid biosynthesis. Treatment with butyrate showed that the decrease of cell adhesion and viability was time-dependent. Sodium butyrate also induced to increase the activity of alkaline phosphatase which is a differentiation marker enzyme and decrease the expression of PLC-${\gamma}$1. Biosynthesis of sphingomyelin and galactosylceramide by butyrate treatment were decreased so fast but ceramide was increased 680dpm/mg protein% more than untreated group on first day and then decreased fast. In addition, acid ceramidase and neutral ceramidase activity were inhibited early stage by sodium butyrate. These results suggest that sodium butyrate causes cell differentiation or cell growth arrest of HT29 cell accompanied by early increase of ceramide content and alkaline phosphatase activity and decrease of galactosylceramide content and PLC-r1 expression.