• Title/Summary/Keyword: Proline accumulation

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Characterization of Salt Tolerant Rice Mutant Lines Derived from Azetidine-2-Carboxylic Acid Resistant Cell Lines Induced by Gamma Ray Irradiation (AZCA 저항성 돌연변이 세포주로부터 선발 육성만 내염성 벼 돌연변이 계통의 특성 검정)

  • Song, Jae-Young;Kim, Dong-Sub;Lee, Geung-Joo;Lee, In-Sok;Kang, Kwon-Kyoo;Yun, Song-Joong;Kang, Si-Yong
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.61-68
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    • 2007
  • To develop rice (Oryza sativa L.) cultivars to be planted on salt-affected sites, cell lines with enhanced proline content and resistance to growth inhibition by Azetidine-2-carboxylic acid (AZCA), a proline analogue, were screened out among calli irradiated with gamma ray of 50, 70, 90, and 120 Gy. The calli had been derived from embryo culture of the cultivar Donganbyeo. Selected AZCA resistant lines that had high proline accumulation were used as sources for selection of NaCl resistant lines. To determine an optimum concentration for selection of NaCl resistant lines, Donganbyeo seeds were initially cultured on the media containing various NaCl concentrations (0 to 2.5%) for 40 days, and 1.5% NaCl concentration was determined as the optimum concentration. One hundred sixteen salt-tolerant (ST) lines were selected from bulked 20,000 seeds of the AZCA resistant $M_{3}$ seeds in the medium containing 1.5% NaCl. The putative 33 lines ($M_{4}$ generation) considered with salt-tolerance were further analyzed for salt tolerance, amino acid and ion contents, and expression patterns of the salt tolerance-related genes. Out of the 33 lines, 7 lines were confirmed to have superior salt tolerance. Based on growth comparison of the entries, the selected mutant lines exhibited greater shoot length with average 1.5 times, root length with 1.3 times, root numbers with 1.1 times, and fresh weight with 1.5 times than control. Proline contents were increased maximum 20%, 100% and 20% in the leaf, seed and callus, respectively, of the selected lines. Compared to control, amino acid contents of the mutants were 24 to 29%, 49 to 143%, 32 to 60% higher in the leaf, seed and callus, respectively. The ratio of $Na^{+}/K^{+}$ for most of the ST-lines were lower than that of control, ranging from 1.0 to 3.8 for the leaf and 11.5 to 28.5 for the root, while the control had 3.5 and 32.9 in the leaf and root, respectively. The transcription patterns for the P5CS and NHXI genes observed by RT-PCR analysis indicated that these genes were actively expressed under salt stress. The selected mutants will be useful for the development of rice cultivar resistant to salt stress.

Quality Changes of Sterilized Soybean Paste during Its Storage (살균 된장의 저장과정 중 품질변화)

  • 오만진;김종생;최성현;이상덕;이규희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.5
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    • pp.1069-1075
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    • 1999
  • The sterilization was attempted to improve the quality deterioration of soybean paste during its storage. For this experiment, soybean paste was sterilized at 80oC for 30 minutes and stored during 6 months at 15oC and 30oC, respectively. The total approximate composition contents were moisture 52.5%, crude protein 11.94%, crude fat 2.0%, amino nitrogen 413.3mg%, sodium chloride 11.61% and ash 15.5%. According to the increase of storage period, pH was decreased gradually because of the increase of organic acids by the metabolism of microorganisms and the acid accumulation by acid forming bacteria, but titratable acidity was increased during storage. Amino nitrogen was rapidly increased for the first one or two month storage period and maintained as the same level for the rest of them. Each amino acid contents of soybean paste, which were glutamic acid, tryptophan, proline, arginine, and aspartic acid, had much higher level than others. In color changes sterilized soybean paste(SSP) was much lower than that of raw ones(RSP). Hunter L and b values on the surface of soybean paste were decreased during storage, and the decreasing levels were higher at 30oC than at 15oC. Hunter a value, however, was increased a little in the initial storage, and thereafter it was decreased. Lactic acid bacteria, yeasts, and molds were disappeared completely by the sterilization. However, the bacteria of aerobes and anaerobes were not disappeared by this processing.

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Protective Effect of Artificially Enhanced Level of L-Ascorbic Acid against Water Deficit-Induced Oxidative Stress in Rice Seedlings

  • Boo, Yong Chool;Cho, Moonjae;Jung, Jin
    • Journal of Applied Biological Chemistry
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    • v.42 no.2
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    • pp.66-70
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    • 1999
  • Effects of the enhanced level of L-ascorbic acid (AA) on the water deficit-induced oxidative damage were studied in rice (Oryza sativa L.) seedlings. The seedlings sprayed with 20 to 80 mM L-galactono-${\gamma}$-lactone (GL), a putative precursor of AA, showed 2 to 5-fold higher levels of AA compared with controls. Pretreatment of the seedlings with GL prior to water stress imposition caused virtually no effect on dehydration of tissues during water deficit but substantially mitigated oxidative injury, as accessed by 2-thiobarbituric acid-reactive substances, ${\alpha}$-tocopherol, chlorophylls and ${\beta}$-carotene. Proline accumulation during water stress was also significantly lowered in the treated seedlings. In a complementary experiment, AA retarded photodegradation of ${\alpha}$-tocopherol in isolated thylakoids far more efficiently than glutathione. GL in itself did not show any noticeable reactivity toward ${\alpha}$-tocopheroxyl radical. The results demonstrate the antioxidative function of AA in rice seedlings encountering water-limited environments, suggesting a critical role of AA as a defense against oxidative stress in plants.

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Morphological, Physiological and Biochemical Responses of Gerbera Cultivars to Heat Stress

  • Chen, Wen;Zhu, Xiaoyun;Han, Weiqing;Wu, Zheng;Lai, Qixian
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.1-14
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    • 2016
  • Heat stress is an agricultural problem for Gerbera jamesonii, and it often causes poor seedling growth, reduced flower yield and undesirable ornamental characteristics of flowers. However, little is known about the effect of heat stress on the morphological, physiological and biochemical characteristics of gerbera plants. Here, the responses of six cultivars of Gerbera jamesonii to heat stress were investigated. Under a 1-d heat treatment at $45^{\circ}C$, the leaves of gerbera cultivars showed yellowing, wilting, drying and death to varying degrees. The heat treatment also resulted in increased electrical conductivity, decreased soluble protein and chlorophyll contents, and the accumulation of malondialdehyde (MDA) and proline in leaves. Moreover, heat tolerance differed among the six tested gerbera cultivars. Our results demonstrated that among the six gerbera cultivars, 'Meihongheixin' is a heat-resistant cultivar, whereas 'Beijixing' is a heat-sensitive one. 'Shijihong' and 'Linglong' are relatively heat-resistant cultivars, and 'Dadifen' and 'Taiyangfengbao' are relatively heat sensitive.

Ecophysiological Characteristics of Chenopodiaceous Plants - An Approach through Inorganic and Organic Solutes - (명아주과 식물의 생리생태학적 특성 - 무기 및 유기용질을 통한 접근 -)

  • Choo, Yeon-Sik;Song, Seung-Dal
    • The Korean Journal of Ecology
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    • v.23 no.5
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    • pp.397-406
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    • 2000
  • In order to clarify the ecophysiological characteristics of Chenopodiaceae which widely distribute on saline and arid habitats, we collected 10 chenopodiaceous plant species, examined their inorganic and organic solute patterns, and confirmed several common physiological characteristics. In spite of high soil Ca/sup 2+/ contents, chenopodiaceous plants had a little water-soluble Ca within cells, but contained high contents of acid-soluble Ca particularly as a result of Ca-oxalate formation. These plant species also showed accumulation of inorganic ions such as K/sup +/, NO₃/sup -/ and Cl/sup -/, and Na/sup +/especially in saline habitats instead of K/sup +/ Meanwhile, with respect to nitrogen metabolism they retained high N contents in leaves, but showed very low amino acid contents. Additionally, they contained very little proline known to act as a cytoplasmic osmolyte. To ascertain whether this physiological characteristics in the field also can be found under controlled conditions, 7 chenopodiaceous plants (Atriplex gmelini, Corispermum stauntonii, Salicornia herbacea, Suaeda aspayagoides, Suaeda japonica, Chenopodium album var. centrorubrum, C. serotinum) were selected and cultivated under salt treatments. As well as field-grown plants, selected plant species showed similar solute pattern in growth experiment. In summary, the family of Chenopodiaceae represents the following physiological properties; high storage capacity for inorganic ions (especially alkali cations, nitrate and chloride), oxalate synthesis to maintain lower soluble Ca contents within cytoplasm, and low contents of amino acids. In addition to some characteristics mentioned above, the physiological plasticities of Chenopodiaceae which can properly regulate their ion and solute pattern according to soil conditions may enable its representative to grow in dry sand dune and salt marsh habitats.

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Overexpression of Ice Recrystallization Inhibition Protein (HvIRIP) from Barley Enhances Cold Tolerance in Transgenic rapeseed plants (HvIRIP 과발현 유채 형질전환체의 내한성 증진)

  • Roh, Kyung Hee;Park, Jong-Sug;Kang, Han-Chul;Kim, Jong-Bum;Jang, Young-Suk;Kim, Kwang-Soo;Yi, Hankuil
    • Journal of Applied Biological Chemistry
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    • v.58 no.4
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    • pp.325-332
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    • 2015
  • Rapeseed (Brassica napus) is now the second largest oilseed crop after soybean. Cold temperature tolerance is an important agronomic trait in winter rapeseed that determines the plant's ability to control below freezing temperatures. To improve cold tolerance of rapeseed plants, an expression vector containing an Barley Ice recrystallization inhibition protein (HvIRIP) cDNA driven by a cauliflower mosaic virus 35S promoter was transferred into rapeseed plants. Transgenic expression of HvIRIP was proved by southern- and northern-blot analyses. The level of freezing tolerance of transgenic $T_3$ plants was found to be significantly greater than that of wild-type rapeseed plants by freezing assay. Proline accumulation during cold stress was also highly induced in the transgenic rapeseed plants. The transgenic plants exhibited considerable tolerance against oxidative damage induced by cold stress. Our results indicated that heterologous HvIRIP expression in transgenic rapeseed plants may induce several oxidative-stress responsive genes to protect from cold stress.

Genetic regulation for the biosynthesis of glutamate family in Corynebacterium glutamicum (Corynebacterium glutamicum에서의 glutamate계 아미노산 생합성의 유전적 조절)

  • Kim In-Ju;Kyung Hee Min;Sae Bae Lee
    • Microbiology and Biotechnology Letters
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    • v.14 no.5
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    • pp.427-432
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    • 1986
  • The regulation of three ammonia assimilatory enzymes, GDH (glutamate dehydrogenase), GS (glutamine synthetase) and GOGAT (glutamate synthase), has been examined in C. glutamicum. Three kinds of arginine auxotrophs blocked in each step of arginine biosynthetic pathway from glutamate were selected as arg 5, arg 6, arg 8. Histidine and tryptophan auxotrophs were also selected because histidine and tryptophan repressed GS biosynthesis in E. coli. These strains were cultured on the media containing nitrogen-excess and limited conditions, to compare the specific activities of ${\alpha}$-ketoglutarate dehydrogenase(${\alpha}-KGDH$), GDH, GS, GOGAT from the cell-free extracts. These results showed that enzyme levels of ${\alpha}-KGDH$ and GDH from 3 kinds of arginine auxotrophs, histidine and tryptophan auxotrophs in nitrogen-excess condition and those of GS and GOGAT in nitrogen limited condition were increased compared with opposite condition. The tryptophan and histidine auxotrophs showed higher level of glutamate and glutamine than parental strains and other mutants. it is assumed that the higher levels of ${\alpha-KGDH}$ and GDH from mutants in nitrogen-excess condition promoted the accumulation of glutamate and glutamine in fermentation broth. The inhibition of GS activities by ADP suggested that GS is regulated by energy charge in C. glutamicum. The results with histidine, tryptophan, glycine, alanine, serine and GMP implied that a system of feedback inhibition were effective. The GDH, GS and GOGAT biosynthesis in culture broth was markedly repressed by the nature and kinds of available nitrogen sources such as tryptophan, proline, glycine, alanine, serine and tyrosine.

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Salt-Responsive Genes in Salt Tolerant Rice Mutants Revealed through Microarray Analysis

  • Song, Jae Young;Kim, Dong Sub;Lee, Myung-Chul;Kang, Si-Yong;Kim, Jin-Baek;Lee, Kyung Jun;Yun, Song Joong
    • Journal of Radiation Industry
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    • v.4 no.4
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    • pp.325-334
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    • 2010
  • Transcriptional regulation in response to salt in mutant lines was investigated using oligonucleotide microarrays. In order to characterize the salt-responsive genes in rice, the expression profiles of transcripts that responded to salt-treatment were monitored using the microarrays. In the microarray analysis, among 37,299 reliable genes, 5,101, 2,758 and 2,277 genes were up-regulated by more than 2-fold using the salt treatment, while the numbers of down-regulated genes were 4,619, 3,234, and 1,878 in the WT, ST-495, and ST-532, respectively. From genotype changes induced by gamma ray mutagenesis, 3,345 and 2,397 genes were up-regulated, while 2,745 and 2,075 genes were down-regulated more than 2-fold in the two untreated mutants lines compared with untreated WT, respectively. A total of 3,108 and 2,731 genes were up-regulated more than 2-fold, while 3,987 and 3,660 genes were down-regulated by more than 2-fold in the salt treatment of the two mutants lines compared with the salt treated WT, respectively. The expressions of membrane transporter genes such as OsAKT1, OsKUP, and OsNAC increased more severely in ST-495 and ST-532 than in the WT. The expressions of the proline accumulation related genes such as OsP5CS and OsP5CR were also markedly increased in the salt tolerant mutants when compared to the WT plant.

Regulation of Chilling Tolerance in Rice Seedlings by Plant Hormones

  • Chu, Chun;Lee, Tse-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.3
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    • pp.288-298
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    • 1992
  • Since the major important factors limiting plant growth and crop productivity are environmental stresses, of which low temperature is the most serious. It has been well known that many physiological processes are alterant in response to the environmental stress. With regard to the relationship between plant hormones and the regulation of chilling tolerance in rice seedlings, the major physiological roles of plant hormones: abscisic acid, ethylene and polyamines are evaluated and discussed in this paper. Rice seedlings were grown in culture solution to examine the effect of such plant hormones on physiological characters related to chilling tolerance and also to compare the different responses among tested cultivars. Intact seedlings about 14 day-old were chilled at conditions of 5$^{\circ}C$ and 80% relative humidity for various period. Cis-(+)-ABA content was measured by the indirect ELISA technique. Polyamine content and ethylene production in leaves were determined by means of HPLC and GC respectively. Chilling damage of seedlings was evaluated by electrolyte leakage, TTC viability assay or servival test. Our experiment results described here demonstrated the physiological functions of ABA, ethylene, and polyamines related to the regulation of chilling tolerance in rice seedlings. Levels of cis-(+)-ABA in leaves or xylem sap of rice seedlings increased rapidly in response to 5$^{\circ}C$ treatment. The tolerant cultivars had significant higher level of endogenous ABA than the sensitive ones. The ($\pm$)-ABA pretreatment for 48 h increased the chilling tolerance of the sensitive indica cultivar. One possible function of abscisic acid is the adjustment of plants to avoid chilling-induced water stress. Accumulation of proline and other compatible solutes is assumed to be another factor in the prevention of chilling injuies by abscisic acid. In addition, the expression of ABA-responsive gene is reported in some plants and may be involving in the acclimation to low temperature. Ethylene and its immediate precusor, 1-amincyclopropane-1-carboxylic acid(ACC) increased significantly after 5$^{\circ}C$ treatment. The activity of ACC synthase which converts S-adenosylmethionine (SAM) to ACC enhanced earlier than the increase of ethylene and ACC. Low temperature increased ACC synthase activity, whereas prolonged chilling treatment damaged the conversion of ACC to ethylene. It was shown that application of Ethphon was beneficial to recovering from chilling injury in rice seedlings. However, the physiological functions of chilling-induced ethylene are still unclear. Polyamines are thought to be a potential plant hormone and may be involving in the regulation of chilling response. Results indicated that chilling treatment induced a remarkable increase of polyamines, especially putrescine content in rice seedlings. The relative higher putrescine content was found in chilling-tolerant cultivar and the maximal level of enhanced putrescine in shoot of chilling cultivar(TNG. 67) was about 8 folds of controls at two days after chilling. The accumulation of polyamines may protect membrane structure or buffer ionic imbalance from chilling damage. Stress physiology is a rapidly expanding field. Plant growth regulators that improve tolerance to low temperature may affect stress protein production. The molecular or gene approaches will help us to elucidate the functions of plant hormones related to the regulation of chilling tolerance in plants in the near future.

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Metabolic engineering of Vit C: Biofortification of potato

  • Upadhyaya, Chandrama P.;Park, Se-Won
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.14-14
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    • 2010
  • Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.

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