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Expression of a yeast‐derived invertase in developing cotyledons of Vicia narbonensis alters the carbohydrate state and affects storage functions

Summary In plants the carbohydrate state provides signals to adjust metabolism to specific physiological conditions. Storage‐active sink organs like seeds often contain high levels of sucrose. In order to change the sugar status during seed development a yeast‐derived invertase gene was expressed in...

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Published in:The Plant journal : for cell and molecular biology 1998-10, Vol.16 (2), p.163-172
Main Authors: Weber, Hans, Heim, Ute, Golombek, Sabine, Borisjuk, Ljudmilla, Manteuffel, Renate, Wobus, Ulrich
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description Summary In plants the carbohydrate state provides signals to adjust metabolism to specific physiological conditions. Storage‐active sink organs like seeds often contain high levels of sucrose. In order to change the sugar status during seed development a yeast‐derived invertase gene was expressed in Vicia narbonensis under control of the LeguminB4 promoter. A signal sequence targeted the invertase to the apoplast in maturing embryos. In the cotyledons, sucrose was decreased whereas hexoses strongly accumulated. There was a major reduction of starch whereas proteins were less affected. Vacuoles of cotyledon cells were enlarged and dry seeds wrinkled. Transcripts and enzyme activity of sucrose synthase, the small and large subunit of ADP‐glucose pyrophosphorylase as well as vicilin were downregulated. Sucrose phosphate synthase and legumin‐mRNAs were not affected. Analysing single seeds with different sucrose levels revealed a positive correlation of sucrose concentration to mRNA levels of sucrose synthase and most pronounced to ADP‐glucose pyrophosphorylase‐mRNA levels as well as to starch content. Glucose on the other hand did not show any correlation. After feeding 14C‐sucrose in vitro , the invertase‐expressing cotyledons partitioned less carbon into starch compared to the wild‐type. In the transgenic cotyledons, a relatively higher amount was directed into proteins compared to starch. We conclude that starch accumulation in developing cotyledons could be a function of sucrose concentration. Our results are consistent with a possible sucrose‐mediated induction of storage‐associated differentiation indicated by upregulation of specific genes of the starch synthesis pathway.
doi_str_mv 10.1046/j.1365-313x.1998.00282.x
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Storage‐active sink organs like seeds often contain high levels of sucrose. In order to change the sugar status during seed development a yeast‐derived invertase gene was expressed in Vicia narbonensis under control of the LeguminB4 promoter. A signal sequence targeted the invertase to the apoplast in maturing embryos. In the cotyledons, sucrose was decreased whereas hexoses strongly accumulated. There was a major reduction of starch whereas proteins were less affected. Vacuoles of cotyledon cells were enlarged and dry seeds wrinkled. Transcripts and enzyme activity of sucrose synthase, the small and large subunit of ADP‐glucose pyrophosphorylase as well as vicilin were downregulated. Sucrose phosphate synthase and legumin‐mRNAs were not affected. Analysing single seeds with different sucrose levels revealed a positive correlation of sucrose concentration to mRNA levels of sucrose synthase and most pronounced to ADP‐glucose pyrophosphorylase‐mRNA levels as well as to starch content. Glucose on the other hand did not show any correlation. After feeding 14C‐sucrose in vitro , the invertase‐expressing cotyledons partitioned less carbon into starch compared to the wild‐type. In the transgenic cotyledons, a relatively higher amount was directed into proteins compared to starch. We conclude that starch accumulation in developing cotyledons could be a function of sucrose concentration. Our results are consistent with a possible sucrose‐mediated induction of storage‐associated differentiation indicated by upregulation of specific genes of the starch synthesis pathway.</description><identifier>ISSN: 0960-7412</identifier><identifier>EISSN: 1365-313X</identifier><identifier>DOI: 10.1046/j.1365-313x.1998.00282.x</identifier><identifier>PMID: 9839463</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Science Ltd</publisher><subject>Agronomy. 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Storage‐active sink organs like seeds often contain high levels of sucrose. In order to change the sugar status during seed development a yeast‐derived invertase gene was expressed in Vicia narbonensis under control of the LeguminB4 promoter. A signal sequence targeted the invertase to the apoplast in maturing embryos. In the cotyledons, sucrose was decreased whereas hexoses strongly accumulated. There was a major reduction of starch whereas proteins were less affected. Vacuoles of cotyledon cells were enlarged and dry seeds wrinkled. Transcripts and enzyme activity of sucrose synthase, the small and large subunit of ADP‐glucose pyrophosphorylase as well as vicilin were downregulated. Sucrose phosphate synthase and legumin‐mRNAs were not affected. Analysing single seeds with different sucrose levels revealed a positive correlation of sucrose concentration to mRNA levels of sucrose synthase and most pronounced to ADP‐glucose pyrophosphorylase‐mRNA levels as well as to starch content. Glucose on the other hand did not show any correlation. After feeding 14C‐sucrose in vitro , the invertase‐expressing cotyledons partitioned less carbon into starch compared to the wild‐type. In the transgenic cotyledons, a relatively higher amount was directed into proteins compared to starch. We conclude that starch accumulation in developing cotyledons could be a function of sucrose concentration. Our results are consistent with a possible sucrose‐mediated induction of storage‐associated differentiation indicated by upregulation of specific genes of the starch synthesis pathway.</description><subject>Agronomy. Soil science and plant productions</subject><subject>beta-Fructofuranosidase</subject><subject>Biological and medical sciences</subject><subject>Carbohydrate Metabolism</subject><subject>Cotyledon - enzymology</subject><subject>Economic plant physiology</subject><subject>Enzyme Induction</subject><subject>Fabaceae - enzymology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glycoside Hydrolases - biosynthesis</subject><subject>Glycoside Hydrolases - genetics</subject><subject>Metabolism</subject><subject>Net assimilation, photosynthesis, carbon metabolism. Photorespiration, respiration, fermentation (anoxia, hypoxia)</subject><subject>Nutrition. Photosynthesis. Respiration. Metabolism</subject><subject>Photosynthesis, respiration. 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Soil science and plant productions</topic><topic>beta-Fructofuranosidase</topic><topic>Biological and medical sciences</topic><topic>Carbohydrate Metabolism</topic><topic>Cotyledon - enzymology</topic><topic>Economic plant physiology</topic><topic>Enzyme Induction</topic><topic>Fabaceae - enzymology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glycoside Hydrolases - biosynthesis</topic><topic>Glycoside Hydrolases - genetics</topic><topic>Metabolism</topic><topic>Net assimilation, photosynthesis, carbon metabolism. Photorespiration, respiration, fermentation (anoxia, hypoxia)</topic><topic>Nutrition. Photosynthesis. Respiration. Metabolism</topic><topic>Photosynthesis, respiration. Anabolism, catabolism</topic><topic>Plant physiology and development</topic><topic>Plant Proteins - metabolism</topic><topic>Plants, Medicinal</topic><topic>Protease Inhibitors - metabolism</topic><topic>Protein Sorting Signals - metabolism</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Solanum tuberosum</topic><topic>Starch - analysis</topic><topic>Sucrose - analysis</topic><topic>Transcription, Genetic</topic><topic>Transfection</topic><topic>Vacuoles - enzymology</topic><topic>Vicia narbonensis</topic><topic>Yeasts - enzymology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Weber, Hans</creatorcontrib><creatorcontrib>Heim, Ute</creatorcontrib><creatorcontrib>Golombek, Sabine</creatorcontrib><creatorcontrib>Borisjuk, Ljudmilla</creatorcontrib><creatorcontrib>Manteuffel, Renate</creatorcontrib><creatorcontrib>Wobus, Ulrich</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Plant journal : for cell and molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Weber, Hans</au><au>Heim, Ute</au><au>Golombek, Sabine</au><au>Borisjuk, Ljudmilla</au><au>Manteuffel, Renate</au><au>Wobus, Ulrich</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of a yeast‐derived invertase in developing cotyledons of Vicia narbonensis alters the carbohydrate state and affects storage functions</atitle><jtitle>The Plant journal : for cell and molecular biology</jtitle><addtitle>Plant J</addtitle><date>1998-10</date><risdate>1998</risdate><volume>16</volume><issue>2</issue><spage>163</spage><epage>172</epage><pages>163-172</pages><issn>0960-7412</issn><eissn>1365-313X</eissn><abstract>Summary In plants the carbohydrate state provides signals to adjust metabolism to specific physiological conditions. Storage‐active sink organs like seeds often contain high levels of sucrose. In order to change the sugar status during seed development a yeast‐derived invertase gene was expressed in Vicia narbonensis under control of the LeguminB4 promoter. A signal sequence targeted the invertase to the apoplast in maturing embryos. In the cotyledons, sucrose was decreased whereas hexoses strongly accumulated. There was a major reduction of starch whereas proteins were less affected. Vacuoles of cotyledon cells were enlarged and dry seeds wrinkled. Transcripts and enzyme activity of sucrose synthase, the small and large subunit of ADP‐glucose pyrophosphorylase as well as vicilin were downregulated. Sucrose phosphate synthase and legumin‐mRNAs were not affected. Analysing single seeds with different sucrose levels revealed a positive correlation of sucrose concentration to mRNA levels of sucrose synthase and most pronounced to ADP‐glucose pyrophosphorylase‐mRNA levels as well as to starch content. Glucose on the other hand did not show any correlation. After feeding 14C‐sucrose in vitro , the invertase‐expressing cotyledons partitioned less carbon into starch compared to the wild‐type. In the transgenic cotyledons, a relatively higher amount was directed into proteins compared to starch. We conclude that starch accumulation in developing cotyledons could be a function of sucrose concentration. Our results are consistent with a possible sucrose‐mediated induction of storage‐associated differentiation indicated by upregulation of specific genes of the starch synthesis pathway.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>9839463</pmid><doi>10.1046/j.1365-313x.1998.00282.x</doi><tpages>10</tpages></addata></record>
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identifier ISSN: 0960-7412
ispartof The Plant journal : for cell and molecular biology, 1998-10, Vol.16 (2), p.163-172
issn 0960-7412
1365-313X
language eng
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source Wiley; EZB Electronic Journals Library
subjects Agronomy. Soil science and plant productions
beta-Fructofuranosidase
Biological and medical sciences
Carbohydrate Metabolism
Cotyledon - enzymology
Economic plant physiology
Enzyme Induction
Fabaceae - enzymology
Fundamental and applied biological sciences. Psychology
Glycoside Hydrolases - biosynthesis
Glycoside Hydrolases - genetics
Metabolism
Net assimilation, photosynthesis, carbon metabolism. Photorespiration, respiration, fermentation (anoxia, hypoxia)
Nutrition. Photosynthesis. Respiration. Metabolism
Photosynthesis, respiration. Anabolism, catabolism
Plant physiology and development
Plant Proteins - metabolism
Plants, Medicinal
Protease Inhibitors - metabolism
Protein Sorting Signals - metabolism
Recombinant Fusion Proteins - metabolism
Solanum tuberosum
Starch - analysis
Sucrose - analysis
Transcription, Genetic
Transfection
Vacuoles - enzymology
Vicia narbonensis
Yeasts - enzymology
title Expression of a yeast‐derived invertase in developing cotyledons of Vicia narbonensis alters the carbohydrate state and affects storage functions
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