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Over-expression of L-gulono-γ-lactone oxidase (GLOase) gene leads to ascorbate accumulation with enhanced abiotic stress tolerance in tomato
Vitamin C (ascorbic acid, AsA) is an essential component for collagen biosynthesis and also for correct functioning of the cardiovascular system in humans. Plants and several animals can synthesize ascorbic acid, whereas humans lack the gene essential for ascorbic acid biosynthesis and must acquire...
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Published in: | In vitro cellular & developmental biology. Plant 2012-10, Vol.48 (5), p.453-461 |
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description | Vitamin C (ascorbic acid, AsA) is an essential component for collagen biosynthesis and also for correct functioning of the cardiovascular system in humans. Plants and several animals can synthesize ascorbic acid, whereas humans lack the gene essential for ascorbic acid biosynthesis and must acquire the vitamin from their diet. In the present study, we developed a transgenic tomato plant (Lycopersicon esculentum Mill.) over-expressing the rat L-gulono-γ-lactone oxidase (GLOase) gene driven by CaMV35S constitutive promoter. The over-expression of GLOase resulted in a 1.5-fold increase in AsA levels in transgenic tomato fruits, which was positively correlated with increased GLOase activity. The Southern and reverse transcriptase polymerase chain reaction results revealed the stable integration of the transgene in the progeny of the engineered plants. The transgenic plants with increased vitamin C content in the fruit showed enhanced tolerance to abiotic stresses induced by methyl viologen, NaCl, and mannitol as compared with wild-type plants. The leaf disc senescence assay showed better tolerance in transgenic plants by retaining higher chlorophyll content compared with wild-type plants. The T1 plants survived under salt stress (200 mM NaCl) whereas wild-type plants were unable to survive such high salt concentrations. This study suggests that the increased accumulation of AsA could upregulate the antioxidant system which imparts improved tolerance against various abiotic stresses in transgenic tomato plants as compared with wild-type plants. |
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Plants and several animals can synthesize ascorbic acid, whereas humans lack the gene essential for ascorbic acid biosynthesis and must acquire the vitamin from their diet. In the present study, we developed a transgenic tomato plant (Lycopersicon esculentum Mill.) over-expressing the rat L-gulono-γ-lactone oxidase (GLOase) gene driven by CaMV35S constitutive promoter. The over-expression of GLOase resulted in a 1.5-fold increase in AsA levels in transgenic tomato fruits, which was positively correlated with increased GLOase activity. The Southern and reverse transcriptase polymerase chain reaction results revealed the stable integration of the transgene in the progeny of the engineered plants. The transgenic plants with increased vitamin C content in the fruit showed enhanced tolerance to abiotic stresses induced by methyl viologen, NaCl, and mannitol as compared with wild-type plants. The leaf disc senescence assay showed better tolerance in transgenic plants by retaining higher chlorophyll content compared with wild-type plants. The T1 plants survived under salt stress (200 mM NaCl) whereas wild-type plants were unable to survive such high salt concentrations. This study suggests that the increased accumulation of AsA could upregulate the antioxidant system which imparts improved tolerance against various abiotic stresses in transgenic tomato plants as compared with wild-type plants.</description><identifier>ISSN: 1054-5476</identifier><identifier>EISSN: 1475-2689</identifier><identifier>DOI: 10.1007/s11627-012-9461-0</identifier><language>eng</language><publisher>New York: Springer-Verlag</publisher><subject>Antioxidants ; ascorbic acid ; Biomedical and Life Sciences ; biosynthesis ; BIOTECHNOLOGY ; cardiovascular system ; Cell Biology ; chlorophyll ; collagen ; correlation ; Developmental Biology ; Enzymes ; fruits ; gene overexpression ; Genes ; humans ; leaves ; Life Sciences ; Oxidases ; Oxidative stress ; Ozone ; paraquat ; Plant Breeding/Biotechnology ; Plant Genetics and Genomics ; Plant Sciences ; Plants ; Polymerase chain reaction ; progeny ; rats ; reverse transcriptase polymerase chain reaction ; salt concentration ; salt stress ; senescence ; sodium chloride ; Solanum lycopersicum var. lycopersicum ; stress tolerance ; tomatoes ; transgenes ; Transgenic plants ; Vitamin C</subject><ispartof>In vitro cellular & developmental biology. Plant, 2012-10, Vol.48 (5), p.453-461</ispartof><rights>The Society for In Vitro Biology 2012</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c358t-6caa4eed4a3bb3667ab8e5b9a4cdf5763281b5524d53ab948e7737d671493d483</citedby><cites>FETCH-LOGICAL-c358t-6caa4eed4a3bb3667ab8e5b9a4cdf5763281b5524d53ab948e7737d671493d483</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/23326873$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/23326873$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,58238,58471</link.rule.ids></links><search><creatorcontrib>Lim, Mi Young</creatorcontrib><creatorcontrib>Pulla, Rama Krishna</creatorcontrib><creatorcontrib>Park, Jeong Mi</creatorcontrib><creatorcontrib>Harn, Chee Hark</creatorcontrib><creatorcontrib>Jeong, Byoung Ryong</creatorcontrib><title>Over-expression of L-gulono-γ-lactone oxidase (GLOase) gene leads to ascorbate accumulation with enhanced abiotic stress tolerance in tomato</title><title>In vitro cellular & developmental biology. Plant</title><addtitle>In Vitro Cell.Dev.Biol.-Plant</addtitle><description>Vitamin C (ascorbic acid, AsA) is an essential component for collagen biosynthesis and also for correct functioning of the cardiovascular system in humans. Plants and several animals can synthesize ascorbic acid, whereas humans lack the gene essential for ascorbic acid biosynthesis and must acquire the vitamin from their diet. In the present study, we developed a transgenic tomato plant (Lycopersicon esculentum Mill.) over-expressing the rat L-gulono-γ-lactone oxidase (GLOase) gene driven by CaMV35S constitutive promoter. The over-expression of GLOase resulted in a 1.5-fold increase in AsA levels in transgenic tomato fruits, which was positively correlated with increased GLOase activity. The Southern and reverse transcriptase polymerase chain reaction results revealed the stable integration of the transgene in the progeny of the engineered plants. The transgenic plants with increased vitamin C content in the fruit showed enhanced tolerance to abiotic stresses induced by methyl viologen, NaCl, and mannitol as compared with wild-type plants. The leaf disc senescence assay showed better tolerance in transgenic plants by retaining higher chlorophyll content compared with wild-type plants. The T1 plants survived under salt stress (200 mM NaCl) whereas wild-type plants were unable to survive such high salt concentrations. This study suggests that the increased accumulation of AsA could upregulate the antioxidant system which imparts improved tolerance against various abiotic stresses in transgenic tomato plants as compared with wild-type plants.</description><subject>Antioxidants</subject><subject>ascorbic acid</subject><subject>Biomedical and Life Sciences</subject><subject>biosynthesis</subject><subject>BIOTECHNOLOGY</subject><subject>cardiovascular system</subject><subject>Cell Biology</subject><subject>chlorophyll</subject><subject>collagen</subject><subject>correlation</subject><subject>Developmental Biology</subject><subject>Enzymes</subject><subject>fruits</subject><subject>gene overexpression</subject><subject>Genes</subject><subject>humans</subject><subject>leaves</subject><subject>Life Sciences</subject><subject>Oxidases</subject><subject>Oxidative stress</subject><subject>Ozone</subject><subject>paraquat</subject><subject>Plant Breeding/Biotechnology</subject><subject>Plant Genetics and Genomics</subject><subject>Plant Sciences</subject><subject>Plants</subject><subject>Polymerase chain reaction</subject><subject>progeny</subject><subject>rats</subject><subject>reverse transcriptase polymerase chain reaction</subject><subject>salt concentration</subject><subject>salt stress</subject><subject>senescence</subject><subject>sodium chloride</subject><subject>Solanum lycopersicum var. lycopersicum</subject><subject>stress tolerance</subject><subject>tomatoes</subject><subject>transgenes</subject><subject>Transgenic plants</subject><subject>Vitamin C</subject><issn>1054-5476</issn><issn>1475-2689</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><recordid>eNp9kc1O3DAQx6OqSKXAA_RQ1cf2YOpvJ8cK8SWttAfgbE2cyZJVNka2l8JD8DR9jz4TjoI4cpoZ_-c3Y_2nqr5xdsoZs78T50ZYyrigjTKcsk_VIVdWU2Hq5nPJmVZUK2u-VF9T2jLGOOP2sHpZP2Kk-PQQMaUhTCT0ZEU3-zFMgf7_R0fwOUxIwtPQQULy83K1LvEX2WB5HRG6RHIgkHyILWQk4P1-tx8hz8P-Dvme4HQPk8eOQDuEPHiS8rysYCPGWSHDVIod5HBcHfQwJjx5i0fV3cX57dkVXa0vr8_-rKiXus7UeACF2CmQbSuNsdDWqNsGlO96bY0UNW-1FqrTEtpG1WittJ2xXDWyU7U8qvgy18eQUsTePcRhB_HZceZmP93ipyt-utlPxwojFiaV3mmD0W3DPk7lmx9C3xdom3KI71uElOUuVhb9x6L3EBxs4pDc3Y1gXJUDyVo39YcdgkndyFfbWZd2</recordid><startdate>20121001</startdate><enddate>20121001</enddate><creator>Lim, Mi Young</creator><creator>Pulla, Rama Krishna</creator><creator>Park, Jeong Mi</creator><creator>Harn, Chee Hark</creator><creator>Jeong, Byoung Ryong</creator><general>Springer-Verlag</general><general>Springer Science + Business Media</general><scope>FBQ</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>20121001</creationdate><title>Over-expression of L-gulono-γ-lactone oxidase (GLOase) gene leads to ascorbate accumulation with enhanced abiotic stress tolerance in tomato</title><author>Lim, Mi Young ; Pulla, Rama Krishna ; Park, Jeong Mi ; Harn, Chee Hark ; Jeong, Byoung Ryong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c358t-6caa4eed4a3bb3667ab8e5b9a4cdf5763281b5524d53ab948e7737d671493d483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Antioxidants</topic><topic>ascorbic acid</topic><topic>Biomedical and Life Sciences</topic><topic>biosynthesis</topic><topic>BIOTECHNOLOGY</topic><topic>cardiovascular system</topic><topic>Cell Biology</topic><topic>chlorophyll</topic><topic>collagen</topic><topic>correlation</topic><topic>Developmental Biology</topic><topic>Enzymes</topic><topic>fruits</topic><topic>gene overexpression</topic><topic>Genes</topic><topic>humans</topic><topic>leaves</topic><topic>Life Sciences</topic><topic>Oxidases</topic><topic>Oxidative stress</topic><topic>Ozone</topic><topic>paraquat</topic><topic>Plant Breeding/Biotechnology</topic><topic>Plant Genetics and Genomics</topic><topic>Plant Sciences</topic><topic>Plants</topic><topic>Polymerase chain reaction</topic><topic>progeny</topic><topic>rats</topic><topic>reverse transcriptase polymerase chain reaction</topic><topic>salt concentration</topic><topic>salt stress</topic><topic>senescence</topic><topic>sodium chloride</topic><topic>Solanum lycopersicum var. lycopersicum</topic><topic>stress tolerance</topic><topic>tomatoes</topic><topic>transgenes</topic><topic>Transgenic plants</topic><topic>Vitamin C</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lim, Mi Young</creatorcontrib><creatorcontrib>Pulla, Rama Krishna</creatorcontrib><creatorcontrib>Park, Jeong Mi</creatorcontrib><creatorcontrib>Harn, Chee Hark</creatorcontrib><creatorcontrib>Jeong, Byoung Ryong</creatorcontrib><collection>AGRIS</collection><collection>CrossRef</collection><jtitle>In vitro cellular & developmental biology. Plant</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lim, Mi Young</au><au>Pulla, Rama Krishna</au><au>Park, Jeong Mi</au><au>Harn, Chee Hark</au><au>Jeong, Byoung Ryong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Over-expression of L-gulono-γ-lactone oxidase (GLOase) gene leads to ascorbate accumulation with enhanced abiotic stress tolerance in tomato</atitle><jtitle>In vitro cellular & developmental biology. Plant</jtitle><stitle>In Vitro Cell.Dev.Biol.-Plant</stitle><date>2012-10-01</date><risdate>2012</risdate><volume>48</volume><issue>5</issue><spage>453</spage><epage>461</epage><pages>453-461</pages><issn>1054-5476</issn><eissn>1475-2689</eissn><abstract>Vitamin C (ascorbic acid, AsA) is an essential component for collagen biosynthesis and also for correct functioning of the cardiovascular system in humans. Plants and several animals can synthesize ascorbic acid, whereas humans lack the gene essential for ascorbic acid biosynthesis and must acquire the vitamin from their diet. In the present study, we developed a transgenic tomato plant (Lycopersicon esculentum Mill.) over-expressing the rat L-gulono-γ-lactone oxidase (GLOase) gene driven by CaMV35S constitutive promoter. The over-expression of GLOase resulted in a 1.5-fold increase in AsA levels in transgenic tomato fruits, which was positively correlated with increased GLOase activity. The Southern and reverse transcriptase polymerase chain reaction results revealed the stable integration of the transgene in the progeny of the engineered plants. The transgenic plants with increased vitamin C content in the fruit showed enhanced tolerance to abiotic stresses induced by methyl viologen, NaCl, and mannitol as compared with wild-type plants. The leaf disc senescence assay showed better tolerance in transgenic plants by retaining higher chlorophyll content compared with wild-type plants. The T1 plants survived under salt stress (200 mM NaCl) whereas wild-type plants were unable to survive such high salt concentrations. This study suggests that the increased accumulation of AsA could upregulate the antioxidant system which imparts improved tolerance against various abiotic stresses in transgenic tomato plants as compared with wild-type plants.</abstract><cop>New York</cop><pub>Springer-Verlag</pub><doi>10.1007/s11627-012-9461-0</doi><tpages>9</tpages></addata></record> |
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subjects | Antioxidants ascorbic acid Biomedical and Life Sciences biosynthesis BIOTECHNOLOGY cardiovascular system Cell Biology chlorophyll collagen correlation Developmental Biology Enzymes fruits gene overexpression Genes humans leaves Life Sciences Oxidases Oxidative stress Ozone paraquat Plant Breeding/Biotechnology Plant Genetics and Genomics Plant Sciences Plants Polymerase chain reaction progeny rats reverse transcriptase polymerase chain reaction salt concentration salt stress senescence sodium chloride Solanum lycopersicum var. lycopersicum stress tolerance tomatoes transgenes Transgenic plants Vitamin C |
title | Over-expression of L-gulono-γ-lactone oxidase (GLOase) gene leads to ascorbate accumulation with enhanced abiotic stress tolerance in tomato |
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