• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.281-291
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• 2003

In this review, we described the catalogues of the rice proteome which were constructed in our program, and functional characterization of some of these proteins was discussed. Mass-spectrometry is the most prevalent technique to rapidly identify a large number of proteome analysis. However, the conventional Western blotting/sequencing technique has been used in many laboratories. As a first step to efficiently construct protein cata-file in proteome analysis of major cereals, we have analyzed the N-terminal sequences of 100 rice embryo proteins and 70 wheat spike proteins separated by two-dimensional electrophoresis. Edman degradation revealed the N-terminal peptide sequences of only 31 rice proteins and 47 wheat proteins, suggesting that the rest of separated protein sports are N-terminally blocked. To efficiently determine the internal sequence of blocked proteins, we have developed a modified Cleveland peptide mapping method. Using this above method, the internal sequences of all blocked rice proteins(i, e., 69 proteins) were determined. Among these 100 rice proteins, thirty were proteins for which homologous sequence in the rice genome database could be identified. However, the rest of the proteins lacked homologous proteins. This appears to be consistent with the fact that about 45% of total rice cDNA have been deposited in the EMBL database. Also, the major proteins involved in the growth and development of rice can be identified using the proteome approach. Some of these proteins, including a calcium-binding protein that tuned out to be calreticulin, gibberellin-binding protein, which is ribulose-1.5-bisphosphate carboxylase/oxygense active in rice, and leginsulin-binding protein in soybean have functions in the signal transduction pathway. Proteomics is well suited not only to determine interaction between pairs of proteins, but also to identify multisubunit complexes. Currently, a protein-protein interaction database for plant proteins(http://genome.c.kanazawa-u.ac.jp/Y2H)could be a very useful tool for the plant research community. Also, the information thus obtained from the plant proteome would be helpful in predicting the function of the unknown proteins and would be useful be in the plant molecular breeding.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.453-459
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• 2000

Three rice (Oryza sativa L. var Dong-Jin) mutants (DTR1, DTR2, DTR3) resistant to S-methyltryptophan (5MT) were selected by mutagenized M3 seeds. The frequency of chlorophyll mutations induced by the EMS (0.2%) treatment performed 2 hours after flowering is clearly higher than that induced by other treatments in M1 generation. Progeny obtained from the self-pollinating of 5MT-resistant lines segregated with 3 : 1 of resistant to sensitive ratio. Furthermore, the ratio of homozygote to heterozygote in 5MT-resistant plants of the M4 generation was 1：2. These results show that 5MT resistance was inherited as a single dominant nuclear gene. The resistance was also expressed in callus derived from seeds. Total free amino acid content in homozygous seeds of DTR1 and DTR2 showed about 1.7 fold-increased compared to the wild-type seeds. In particular, the levels of phenylalanine and Iysine were, respectively, 6.2 and 3.2 times higher than those in the wild-type seeds. However, seeds of DTR3 had lower levels of free amino acid than the wild-type seeds. This result indicate that these mutants as a significant step towards the production of new rice with balanced amino acid content.

• Journal of Plant Biotechnology
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• v.36 no.3
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• pp.281-288
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• 2009

Seed germination is the important stage to express many genes for regulation of energy metabolism, starch degradation and cell division from seed dormancy state. For the functional analysis of seed germination mechanisms, we were analyzed the rice cDNA clones (Oryzasativa cultivar Ilpum) obtained from seed imbibition during 48 hours. Total number of 18,101 Expressed Sequence Tags (ESTs) were clustered using SeqMan program. Among them, 8,836 clones were identified as unique clones. We identified the chitinase gene specifically expressed in seed germination and amylase gene involved to starch degradation from the full length cDNA analysis, and several genes were registered to NCBI GeneBank. To analyzed the commonly expressed genes between inmature seed and germinated seed, 25,66 inmature ESTs and 18,101 germinated ESTs were clustered using SeqMan program and identified 2,514 clones as commonly expressed unigene. Among them, alpha-glubulin and alcohol dehydrogenase I were supposed to LEA genes only expressed in the immature and germinated seed stages. For the clustering of orthologous group genes, we further analyzed the 8,836 EST clones from germinating seeds using NCBI clusters of orthologous groups database. Among the clones, 5,076 clones were categorized into information storage and processing, cellular processes and signaling, metabolism and poorly characterized genes, proportioning 783 (14.29%), 1,484 (27%), 1,363 (24.8%) and 1,869 (34%) clones to the previous four categories, respectively.

• Journal of Plant Biotechnology
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• v.45 no.1
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• pp.63-70
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• 2018

Brazzein is the smallest sweet protein and was isolated from the fruit pulp of Pentadiplandra brazzeana Baillon, native to tropical Africa. From ancient times, the indigenous people used this fruit in their diet to add sweetness to their daily food. Brazzein is 500 to 2000 times sweeter than sucrose on a weight basis and 9500 times sweeter on a molar basis. This unique property has led to increasing interest in this protein. However, it is expensive and difficult to produce brazzein other than in its native growing conditions which limits its availability for use as a food additive. In this study, we report high production yields of, brazzein protein in transgenic rice plants. An ORF region encoding brazzein and driven by the $2{\times}CaMV\;35S$ promoter was introduced into rice genome (Oryza sativa Japonica) via Agrobacterium-mediated transformation. After transformation, 17 regenerated plant lines were obtained and these transgene-containing plants were confirmed by PCR analysis. In addition, the selected plant lines were analyzed by Taqman PCR and results showed that 9 T0 lines were found to have a single copy out of 17 transgenic plants. Moreover, high and genetically stable expression of brazzein was confirmed by western blot analysis. These results demonstrate that recombinant brazzein was efficiently expressed in transgenic rice plants, and that we have developed a new rice variety with a natural sweetener.

• Journal of Plant Biotechnology
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• v.45 no.1
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• pp.17-29
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• 2018

This study was conducted to investigate a morphological trait in 294 rice accessions including Korean breeding lines. We also carried out a genome-wide association study (GWAS) to detect significant single nucleotide polymorphism markers and candidate genes affecting major agronomic traits. A Manhattan plot analysis of GWAS using morphological traits showed that phenotypic and statistical significance was associated with a chromosome in each group. The significance of SNPs that were detected in this study was investigated by comparing them with those found previously studied QTL regions related to agronomic traits. As a result, SNP (S8-19815442), which is significant with regard to leaf angle, was located in the known QTL regions. To observe gene mutations related to leaf angle in a candidate gene, Os08g31950, its sequences were compared with sequences in previously selected rice varieties. In Os08g31950, a single nucleotide mutation occurred in one region. To compare relative RNA expression levels of candidate gene Os08g31950, obtained from GWAS analysis of 294 rice accessions and related to lateral leaf angle, we investigated relative levels by selecting 10 erect leaf angle varieties and 10 horizontal leaf angle varieties and examining real-time PCR. In Os08g31950, a high level of expression and various expression patterns were observed in all tissues. Also, Os08g31950 showed higher expression levels in the erect leaf angle variety group and higher expression rates in the leaf than in the root. The candidate gene detected through GWAS would be useful in developing new rice varieties with improved yield potential through future molecular breeding.

• Korean Journal of Soil Science and Fertilizer
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• v.7 no.2
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• pp.107-112
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• 1974

In order to establish a rational application method of potash to paddy rice(Oryza sativa L. Jinheung, a Japonica variety) field and pot experiments on split application of two rates (6kg and 12kg $K_2O$ per 10a) of potash at a certain critical growing stage have conducted in 1973 and the results were as follows: 1. At the treatment where a rate of 6kg $K_2O$ per 10a(lower dose) of potash was supplied in two split doses, a half at transplanting time and the other half at young panicle formation stage, the yield of paddy rice appeatred to be low due to less number of panicles per hill. (Field experiment) 2. The treatment where potash supply ceased at transplanting time or at effective tillering stage but supplied at the other two stages (including panicle initiation stage) have failed to increase K content in the flag leaf and have shown low maturity and low thousand grain weight, which resulted in low paddy rice yield. (pot experiment) 3. From above two facts, it is concluded that the application of potash at transplanting time is an important practice for the high yielding of paddy rice. 4. The application of lime seemed to stimulate later growth of paddy depressing the unnecessary increase of poor miserable tillers in early stages of growth.

• BMB Reports
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• v.35 no.3
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• pp.313-319
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• 2002

N-Acetyl-$\beta$-D-hexosaminidase ($\beta$-HexNAc'ase) (EC 3.2.1.52) was purified from rice seeds (Oryza sative L. var. Dongjin) using ammonium sulfate (80%) precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex chromatography, sequentially. The activities were separated into 7 fractions($F_1-F_7$) by CM-Sephadex chromatography. Among them, F6 was further purified to homogeneity with a 13.0% yield and 123.3 purification-fold. The molecular mass was estimated to be about 52 kDa on SDS-PAGE and 37.4 kDa on Sephacryl S-300 gel filtration. The enzyme catalyzed the hydrolysis of both p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GlcNAc) and p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GalNAc) as substrates, which are typical properties of $\beta$-HexNAc'ase. The ratio of the pNP-GlcNAc'ase activity to the pNP-GalNAc'ase activity was 4.0. However, it could not hydrolyze chitin, chitosan, pNP-$\beta$-glucopyranoside, or pNP-$\beta$-glucopyranoside. The enzyme showed $K_m$, $V_{max}$ and $K_{cat}$ for pNP-GlcNAc of 1.65 mM, $79.49\;mM\;min^{-1}$, and $4.79{\times}10^6\;min^{-1}$, respectively. The comparison of kinetic values for pNP-GlcNAc and pNP-GalNAc revealed that the two enzyme activities are associated with a single binding site. The purified enzyme exhibited optimum pH and temperature for pNP-GlcNAc of 5.0 and $50^{\circ}C$, respectively. The enzyme activity for pNP-GlcNAc was stable at pH 5.0-5.5 and $20-40^{\circ}C$. The enzyme activity was completely inhibited at a concentration of 0.1 mM $HgCl_$ and $AgNO_3$, suggesting that the intact thiol group is essential for activity. Chloramine T completely inhibited the activity, indicating the possible involvement of methionines in the mechanism of the enzyme.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.4
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• pp.555-563
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• 2016

Purple rice is a strain of glutaneous rice rich in anthocyanins and ${\gamma}$-oryzanol. Both types of compounds are involved in antioxidant and lipid metabolism of mammals. Three experimental diet types were used which consisted approximately by half either of purple rice, white rice or corn. Diets were fed to $3{\times}10$ pigs growing from about 30 to 100 kg. Meat samples were investigated either as raw or cured loin chops or as smoked bacon produced from the belly. Various physicochemical traits were assessed and data were evaluated by analysis of variance. Traits describing water-holding capacity (drip, thaw, and cooking losses) and tenderness (sensory grading, shear force) of the meat were mostly not significantly affected by the diet type. However, purple rice feeding of pigs resulted in lower fat and cholesterol contents of loin and smoked bacon compared to white rice, but not compared to corn feeding except of the fat content of the loin. The shelf life of the raw loin chops was improved by purple rice as well. In detail, the occurrence of thiobarbituric acid reactive substances after 9 days of chilled storage was three to four times higher in the white rice and corn diets than with purple rice. The n-6:n-3 ratio in the raw loin chops was 9:1 with purple rice and clearly higher with 12:1 with the other diets, meat lipids. Level and kind of effect of purple rice found in raw meat was not always recovered in the cured loin chops and the smoked bacon. Still the impression of flavor and color, as well as overall acceptability were best in the smoked bacon from the purple-rice fed pigs, whereas this effect did not occur in the cured loin chops. These findings suggest that purple rice has a certain, useful, bioactivity in pigs concerning meat quality, but some of these effects are of low practical relevance. Further studies have to show ways how transiency and low recovery in meat products of some of the effects can be counteracted.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.12
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• pp.1755-1760
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• 2006

Studies were carried out at the National Institute of Animal Nutrition and Physiology, India to determine the effect of feeding chopped paddy straw (Oryza sativa) on the energy expenditure in crossbred cattle. Four crossbred cattle male, aged 5-6 years and weighing about 450 kg were used for this study. Three experimental trials, one each for the feeding of un-chopped paddy straw offered ad libitum (UCA), chopped paddy straw fed at restricted level (CR) and chopped paddy straw offered ad libitum (CA) were conducted. The quantity of un-chopped paddy straw consumed during UCA was assumed as the voluntary intake by the cattle and the same quantity was offered after chopping during CR. Each trial comprised of 21 d preliminary feeding period and 5 d of observation recording period. Expired gas was collected in Douglas bags using a face-mask and three-way valve at 6 hourly intervals i.e., at 09.30, 15.30, 21.30, and 03.30 h throughout the observation period. Expired gas and ambient air inspired by the animals were analyzed for the oxygen content through paramagnetic oxygen analyzer. Energy expenditure (EE) by the animals was calculated by determining the volume of oxygen consumed per minute (STP) and multiplying by 4.825. Paddy straw used in all the three trials contained (g/kg DM) 90.0 CP, 786 OM, 700 NDF, 489 ADF, 357 Cellulose and 60.0 ADL. Metabolizable energy (ME) was 6.9 MJ/kg DM. Dry matter intake (DMI) both in UCA and CR was about 6.8 kg, except that it was chopped in CR. Chopping has resulted in 32% improvement (9 kg) in DMI of CA as compared to that of UCA. Although ME intake was similar in UCA and CR (47.2 MJ/day), energy expenditure (EE) was higher in UCA (23.3 MJ) when compared to that of CR (19.5 MJ). The ME intake (63.3 MJ) as well as EE (27.1 MJ) was highest in CA. Energy expenditure when expressed as MJ/kg DMI was 3.48, 2.90 and 3.12; whereas as per cent of ME intake it was 50, 41 and 44 in UCA, CR and CA respectively. Our study has unequivocally confirmed that chopping of poor quality roughages like paddy straw has definite advantages not only in terms of improving the intake by decreasing the time taken for ingestion but also in reducing the energy cost of eating.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.59 no.3
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• pp.230-238
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• 2014

The purpose of this study was to develop the cost-effective and efficiency seed longevity prediction method of rice (Oryza sativa L.) germplasm for viability monitoring. To find an optimum predicting method for rice seed longevity at genebank, an accelerated ageing (AA) test, a controlled deterioration (CD) test and a dry-heat treatment (DHT) were conducted to the four groups of rice germplasm based on ecotype, such as Indica, Japonica, Javanica and Tongil type. Among the three artificial aging treatments, the dry-heat treatment of 36 hours at $90^{\circ}C$ is suggested as a routine predictive test method of rice germplasm longevity at a genebank. The distribution of germination rate on 3,066 accessions which conserved 26.5 years at $4^{\circ}C$ showed similar trend with the result of distribution by dry-heat treatment at $90^{\circ}C$ on 36 hours using 106 accessions of rice selected samples which composed four ecotype groups. The results show that the dry-heat treatment affect not only predicting the rice seed longevity but also determining effective interval for monitoring germination of rice germplasm in genebanks.