• Title/Summary/Keyword: plant enzymes

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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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식물배양세포를 이용한 항산화연구

  • Kim, Gi-Yeon;Lee, Jeong-Eun;An, Yeong-Ok;Gwon, Seok-Yun;Lee, Haeng-Sun;Gwak, Sang-Su
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.65-68
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    • 2000
  • To understand the antioxidative mechanism in plant cell cultures, we investigated the levels of antioxidant enzymes and low molecular antioxidants in 100 cell lines derived from different plant species. SOD and POD activities in plant cell lines were significantly higher than intact plants. The cell lines from sweet potato (Ipomoea batatas) and cassava (Manihot esculeanta) showed the highest POD and SOD activities, respectively, suggesting that the cell cultures of sweet potato and cassava are good biomaterials for the mass production and molecular study of antioxidant enzymes. The average ascorbate content in plant cell lines was several hundred times lower than intact plants, whereas the glutathione content was 2-3 times higher than plants. Interestingly, the ratio of reduced and oxidized ascorbate and glutathione was different from plant species. In conclusion, the results strongly suggest that plant cell cultures are good biomaterials for the study of antioxidative mechanism and the production of useful components including antioxidants.

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Probing Starch Biosynthesis Enzyme Isoforms by Visualization of Conserved Secondary Structure Patterns

  • Vorapreeda, Tayvich;Kittichotirat, Weerayuth;Meechai, Asawin;Bhumiratana, Sakarindr;Cheevadhanarak, Supapon
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.215-220
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    • 2005
  • Generally, enzymes in the starch biosynthesis pathway exist in many isoforms, contributing to the difficulties in the dissection of their specific roles in controlling starch properties. In this study, we present an algorithm as an alternative method to classify isoforms of starch biosynthesis enzymes based on their conserved secondary structures. Analysis of the predicted secondary structure of plant soluble starch synthase I (SSI) and soluble starch synthase II (SSII) demonstrates that these two classes of isoform can be reclassified into three subsets, SS-A, SS-B and SS-C, according to the differences in the secondary structure of the protein at C-terminus. SS-A reveals unique structural features that are conserved only in cereal plants, while those of SS-B are found in all plants and SS-C is restricted to barley. These findings enable us to increase the accuracy in the estimation of evolutionary distance between isoforms of starch synthases. Moreover, it facilitates the elucidation of correlations between the functions of each enzyme isoforms and the properties of starches. Our secondary structure analysis tool can be applicable to study the functions of other plant enzyme isoforms of economical importance.

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Pathogenicity of Bacillus Strains to Cotton Seedlings and Their Effects on Some Biochemical Components of the Infected Seedlings

  • Aly, Aly A.;El-Mahdy, Omima M.;Habeb, Marian M.;Elhakem, Abeer;Asran, Amal A.;Youssef, Maryan M.;Mohamed, Heba I.;Hanafy, Rania S.
    • The Plant Pathology Journal
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    • v.38 no.2
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    • pp.90-101
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    • 2022
  • Pathogenicity of eight Bacillus strains to seedlings of four cotton cultivars was evaluated under greenhouse conditions. Each of the tested cultivars was individually treated with powdered inoculum of each bacterial strain. Untreated seeds were planted as control treatments in autoclaved soil. Effects of the tested strains on levels and activities of some biochemical components of the infected seedlings were also assayed. The biochemical components included total soluble sugars, total soluble proteins, total free amino acids, peroxidase, polyphenol oxidase, phenols, and lipid peroxidation. ANOVA showed that Bacillus strain (B) was a very highly significant source of variation in damping-off and dry weight. Cotton cultivar (V) was a nonsignificant source of variation in damping-off while it was a significant source of variation in dry weight. B × V interaction was a significant source of variation in damping-off and a nonsignificant source of variation in dry weight. Bacillus strain was the most important source of variation as it accounted for 59.36 and 64.99% of the explained (model) variation in damping-off and dry weight, respectively. The lack of significant correlation between levels and activities of the assayed biochemical components and incidence of damping-off clearly demonstrated that these biochemical components were not involved in the pathogenicity of the tested strains. Therefore, it was hypothesized that the pathogenicity of the tested strains could be due to the effect of cell wall degrading enzymes of pathogenic toxins. Based on the results of the present study, Bacillus strains should be considered in studying the etiology of cotton seedling damping-off.

Digestion efficiency differences of restriction enzymes frequently used for genotype-by-sequencing technology

  • Chung, Yong Suk;Jun, Taehwan;Kim, Changsoo
    • Korean Journal of Agricultural Science
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    • v.44 no.3
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    • pp.318-324
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    • 2017
  • With the development of next-generation sequencing (NGS), a cutting-edge technology, genotype-by-sequencing (GBS) became available at a low cost per sample. GBS makes it possible to customize the process of library preparation to obtain high-quality single nucleotide polymorphisms (SNPs) in the most efficient way. However, a GBS library is hard to construct due to fine-tuning of concentration of each reagent and set-up. The major reason for this is the presence of undigested genomic DNA (gDNA) owing to the efficiency of different restriction enzymes for different species with unknown reasons. Therefore, this proof-concept study is to demonstrate the unpredictable patterns of enzyme digestion from various plants in order to make the reader aware of the caution needed when choosing restriction enzymes for their GBS library preparations. Indeed, no pattern was found for the digestibility of gDNA samples and restriction enzymes in the current study. We suggest that more data should be accumulated on this matter to help researchers who want to apply GBS technologies in a variety of genetic approaches.

The role and characterization of .betha.-1, 3-glucanase in biocontrol of fusarium solani by pseudomonas stutzeri YPL-1

  • Lim, Ho-Seong;KiM, Sang-Dal
    • Journal of Microbiology
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    • v.33 no.4
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    • pp.295-301
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    • 1995
  • An antifungal Pseudomonas stutzeri YPL-1 produced extracellular chitinase and .betha.-1, 3-glucanase that were key enzymes in the decomposition of fungal hyphal walls. These lytic extracellular enzymes markedly inhibited mycelial growth of the phytopathogenic fungus Fusarium solani. A chitinase from P. stutzeri YPL-1 inhibited fungal mycelial growth by 87%, whereas a .betha.-1, 3-glucanase from the bacterium inhibited growth by 53%. Furthermore, co-operative action of the enzymes synergistically inhibited 95% of the fungal growth. The lytic enzymes caused absnormal swelling and retreating on the fungal hyphal walls in a dual cultures. Scanning electron microscopy clearly showed hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. In an in vivo pot test, P. stutzeri YPL-1 proved to have biocontrol ability as a powerful agent in controlling plant disease. Planting of kidney bean (Phaseolus vulgaris L.) seedlings with the bacterial suspension in F. solani-infested soil significantly suppressed the development of fusarial root-rot. The characteristics of a crude preparation of .betha.-1, 3-glucanase produced from P. stutzeri YPL-1 were investigated. The bacterium detected after 2 hr of incubation. The enzyme had optimum temperature and pH of 40.deg.C and pH 5.5, respectively. The enzyme was stable in the pH range of 4.5 to 7.0 and at temperatures below 40.deg.C, with a half-life of 40 min at 60.deg.C.

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Effect of Chromium Stress on Antioxidative Enzymes and Malondialdehyde Content Activities in Leaves and Roots of Mangrove Seedlings Kandelia Candel (L.) Druce

  • Rahman, Mohammed Mahabubur;Rahman, Motiur M.;Islam, Kazi Shakila;Chongling, Yan
    • Journal of Forest and Environmental Science
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    • v.26 no.3
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    • pp.171-179
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    • 2010
  • Effect of chromium (Cr) stress on antioxidant enzyme activities and malondialdehyde (MDA) content were investigated in leaves and roots of mangrove (italic (L.) Druce) seedlings. Cr toxicity effects were also assessed on young seedlings. The seedlings were grown in green house condition for three months in nutrient solution with 0, 0.5, 1, 1.5, 2, 2.5, and 3 mg $L^{-1}$ $CrCl_3$. This study showed that Cr led to the change of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) and activities at different concentrations. The activity of antioxidant enzymes in leaves of K. candel seedlings indicates that enzymes engaged in antioxidant defense in certain level especially in low concentration of Cr treatments. The activities of SOD and POD were activated by Cr in the root level, while CAT activity was inhibited. CAT activity decreased in response to high concentrations of Cr. In the present study indicated that SOD in root was active in scavenging the superoxide produced by Cr. Both in roots and leaves, an increase in malondialdehyde (MDA) content was observed with increase in metal concentration and exposure periods. Our finding indicated that the high concentration of excessive Cr supply may interfere with several metabolic processes of seedlings, causing toxicity to plants as exhibited by chlorosis, necrosis, photosynthetic impairing and finally, plant death.

Recent Advances in the Use of Enzymes for Environment-Friendly Swine Diets (환경친화성양돈사료를 위한 효소제 사용의 최근 경향)

  • Hong Jong-Wok;Kim In-Ho
    • Korean Journal of Organic Agriculture
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    • v.9 no.2
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    • pp.55-67
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    • 2001
  • Exogenous enzymes which, for the purpose of this paper, include phytase, $\beta$-glucanase, pentosanase and $\alpha$-galactosidase, are now extensively used throughout the world as aditives in swine diets. The chemical effects of these enzymes are well understand. However, the manner in which their benefits to the swine are brought about is still under debate. Phytase was to increase the availability of plant phytate phosphorus, which reduces phosphorus pollution and allows reductions in the amount of inorganic phosphate used. Also, enzymes have been discovered which have the potential to break down deleterious compounds commonly found in swine rations such as $\beta$-glucanase contained in barley and oats and the soluble pentosans found in rye and wheat thus increasing the digestibility of these non-starch polysaccharides. Future research in these area will allow for more efficient use of the current enzymes, development of more efficient future products and development of more thermotolerant enzymes.

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Biological Control of Bacterial Fruit Blotch of Watermelon Pathogen (Acidovorax citrulli) with Rhizosphere Associated Bacteria

  • Adhikari, Mahesh;Yadav, Dil Raj;Kim, Sang Woo;Um, Young Hyun;Kim, Hyun Seung;Lee, Seong Chan;Song, Jeong Young;Kim, Hong Gi;Lee, Youn Su
    • The Plant Pathology Journal
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    • v.33 no.2
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    • pp.170-183
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    • 2017
  • Bacterial fruit blotch (BFB), which is caused by Acidovorax citrulli, is a serious threat to watermelon growers around the world. The present study was conducted to screen effective rhizobacterial isolates against 35 different A. citrulli isolates and determine their efficacy on BFB and growth parameters of watermelon. Two rhizobacterial isolates viz. Paenibacillus polymyxa (SN-22), Sinomonas atrocyanea (NSB-27) showed high inhibitory activity in the preliminary screening and were further evaluated for their effect on BFB and growth parameters of three different watermelon varieties under greenhouse conditions. The greenhouse experiment result revealed that SN-22 and NSB-27 significantly reduced BFB and had significant stimulatory effect on total chlorophyll content, plant height, total fresh weight and total dry weight compared to uninoculated plants across the tested three watermelon varieties. Analysis of the 16S ribosomal RNA (rRNA) sequences revealed that strains SN-22 belong to P. polymyxa and NSB-27 to S. atrocyanea with the bootstrap value of 99% and 98%, respectively. The isolates SN-22 and NSB-27 were tested for antagonistic and PGP traits. The result showed that the tested isolates produced siderophore, hydrolytic enzymes (protease and cellulose), chitinase, starch hydrolytic enzymes and they showed phosphate as well as zinc solubilizing capacity. This is the first report of P. polymyxa (SN-22) and S. atrocyanea (NSB-27) as biocontrol-plant growth promoting rhizobacteria on watermelon.

Physiological Characterization of Mono-Cotyledonous Model Plant by Ionizing Irradiation (단자엽 모델 식물의 이온화 에너지원에 따른 생리 활성)

  • Song, Mira;Kim, Sun-Hee;Jang, Duk-Soo;Kang, Si-Yong;Kim, Jin-Baek;Kim, Sang Hoon;Ha, Bo-Keun;Park, Yong Dae;Kim, Dong Sub
    • Journal of Radiation Industry
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    • v.5 no.1
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    • pp.7-13
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    • 2011
  • The present study has been performed to compare the physiological analysis of monocot model plant (rice) in response to ionizing irradiations (cosmic-ray, gamma-ray, and Ion beam). Ionizing radiations were implanted into monocot model plant (rice) seed. After irradiation, the seeds were planted in the plastic pots for a growth period of one month. Thereafter, the morphological and physiological characteristics including malondealdehyde (MDA) and chlorophyll content, activities of antioxidant enzymes in irradiation samples were investigated. We are confirmed that the activity level of MDA and chlorophyll content were not changed by ionizing irradiation samples. However, the free radical contents were increased in all irradiated plants. And the activities of SOD, POD, and APX were significantly increased by irradiation compared with non-irradiation plant.