Post-harvest light treatment increases expression levels of recombinant proteins in transformed plastids of potato tubers
Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non‐photosynthetic plastids are generally discouraging. Here, we report the expression of two thioredoxi...
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Veröffentlicht in: | Biotechnology journal 2015-09, Vol.10 (11), p.1803-1813 |
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creator | Larraya, Luis M. Fernández-San Millán, Alicia Ancín, María Farran, Inmaculada Veramendi, Jon |
description | Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non‐photosynthetic plastids are generally discouraging. Here, we report the expression of two thioredoxin genes (trx f and trx m) from the potato plastid genome to study transgene expression in amyloplasts. As expected, the highest transgene expression was detected in the leaf (up to 4.2% of TSP). The Trx protein content in the tuber was approximately two to three orders of magnitude lower than in the leaf. However, we demonstrate that a simple post‐harvest light treatment of microtubers developed in vitro or soil‐grown tubers induces up to 55 times higher accumulation of the recombinant protein in just seven to ten days. After the applied treatment, the Trx f levels in microtubers and soil‐grown tubers increased to 0.14% and 0.11% of TSP, respectively. Moreover, tubers stored for eight months maintained the capacity of increasing the foreign protein levels after the light treatment. Post‐harvest cold induction (up to five times) at 4°C was also detected in microtubers. We conclude that plastid transformation and post‐harvest light treatment could be an interesting approach for the production of foreign proteins in potato.
Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non‐photosynthetic plastids are generally discouraging. In this article, the authors report the expression of two thioredoxin genes from the potato plastid genome to study transgene expression in amyloplasts. The authors demonstrate that a simple post‐harvest light treatment of tubers induces up to 55 times higher accumulation of the recombinant protein in just 7–10 days. Moreover, tubers stored for 8 months maintain the capacity of increasing the foreign protein levels after the light treatment. |
doi_str_mv | 10.1002/biot.201500028 |
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Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non‐photosynthetic plastids are generally discouraging. In this article, the authors report the expression of two thioredoxin genes from the potato plastid genome to study transgene expression in amyloplasts. The authors demonstrate that a simple post‐harvest light treatment of tubers induces up to 55 times higher accumulation of the recombinant protein in just 7–10 days. Moreover, tubers stored for 8 months maintain the capacity of increasing the foreign protein levels after the light treatment.</description><identifier>ISSN: 1860-6768</identifier><identifier>EISSN: 1860-7314</identifier><identifier>DOI: 10.1002/biot.201500028</identifier><identifier>PMID: 26121393</identifier><language>eng</language><publisher>Weinheim: WILEY-VCH Verlag</publisher><subject>Amyloplast ; Gene Expression Regulation, Plant - radiation effects ; Microtuber ; Plant Tubers - metabolism ; Plants, Genetically Modified - genetics ; Plants, Genetically Modified - metabolism ; Plastid transformation ; Plastids - genetics ; Plastids - metabolism ; Potato ; Recombinant Proteins - analysis ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Solanum tuberosum ; Solanum tuberosum - genetics ; Solanum tuberosum - metabolism ; Thioredoxin ; Thioredoxins - analysis ; Thioredoxins - genetics ; Thioredoxins - metabolism</subject><ispartof>Biotechnology journal, 2015-09, Vol.10 (11), p.1803-1813</ispartof><rights>Copyright © 2015 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5268-fbf10634766f15644e8efa160373632c7afd08146aff152c44f44033d8405c993</citedby><cites>FETCH-LOGICAL-c5268-fbf10634766f15644e8efa160373632c7afd08146aff152c44f44033d8405c993</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fbiot.201500028$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fbiot.201500028$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>315,781,785,1418,27929,27930,45579,45580</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26121393$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Larraya, Luis M.</creatorcontrib><creatorcontrib>Fernández-San Millán, Alicia</creatorcontrib><creatorcontrib>Ancín, María</creatorcontrib><creatorcontrib>Farran, Inmaculada</creatorcontrib><creatorcontrib>Veramendi, Jon</creatorcontrib><title>Post-harvest light treatment increases expression levels of recombinant proteins in transformed plastids of potato tubers</title><title>Biotechnology journal</title><addtitle>Biotechnology Journal</addtitle><description>Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non‐photosynthetic plastids are generally discouraging. Here, we report the expression of two thioredoxin genes (trx f and trx m) from the potato plastid genome to study transgene expression in amyloplasts. As expected, the highest transgene expression was detected in the leaf (up to 4.2% of TSP). The Trx protein content in the tuber was approximately two to three orders of magnitude lower than in the leaf. However, we demonstrate that a simple post‐harvest light treatment of microtubers developed in vitro or soil‐grown tubers induces up to 55 times higher accumulation of the recombinant protein in just seven to ten days. After the applied treatment, the Trx f levels in microtubers and soil‐grown tubers increased to 0.14% and 0.11% of TSP, respectively. Moreover, tubers stored for eight months maintained the capacity of increasing the foreign protein levels after the light treatment. Post‐harvest cold induction (up to five times) at 4°C was also detected in microtubers. We conclude that plastid transformation and post‐harvest light treatment could be an interesting approach for the production of foreign proteins in potato.
Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non‐photosynthetic plastids are generally discouraging. In this article, the authors report the expression of two thioredoxin genes from the potato plastid genome to study transgene expression in amyloplasts. The authors demonstrate that a simple post‐harvest light treatment of tubers induces up to 55 times higher accumulation of the recombinant protein in just 7–10 days. Moreover, tubers stored for 8 months maintain the capacity of increasing the foreign protein levels after the light treatment.</description><subject>Amyloplast</subject><subject>Gene Expression Regulation, Plant - radiation effects</subject><subject>Microtuber</subject><subject>Plant Tubers - metabolism</subject><subject>Plants, Genetically Modified - genetics</subject><subject>Plants, Genetically Modified - metabolism</subject><subject>Plastid transformation</subject><subject>Plastids - genetics</subject><subject>Plastids - metabolism</subject><subject>Potato</subject><subject>Recombinant Proteins - analysis</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Solanum tuberosum</subject><subject>Solanum tuberosum - genetics</subject><subject>Solanum tuberosum - metabolism</subject><subject>Thioredoxin</subject><subject>Thioredoxins - analysis</subject><subject>Thioredoxins - genetics</subject><subject>Thioredoxins - metabolism</subject><issn>1860-6768</issn><issn>1860-7314</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc1v1DAQxS0EakvplSPykUsWO_6Ic4SKlkor2sMijpaTHVNDEgePt3T_-3rZZcUNTh7Lv_c0fo-Q15wtOGP1uy7EvKgZV6zczDNyxo1mVSO4fH6YdaPNKXmJ-J0xqQSTJ-S01rzmohVnZHsXMVf3Lj0AZjqEb_eZ5gQujzBlGqa-zAhI4XFOgBjiRAd4gAFp9DRBH8cuTK6gc4oZwoRFUwzchD6mEdZ0HhzmsP7NzzG7HGnedJDwFXnh3YBwcTjPyZerj6vLT9Xy9vrm8v2y6lWtTeU7z5kWstHac6WlBAPecc1EI7So-8b5NTNcaufLe91L6aVkQqyNZKpvW3FO3u59y4Y_N-WXdgzYwzC4CeIGLS_5KK1MI_4DVbxkLfgOXezRPkXEBN7OKYwubS1ndteM3TVjj80UwZuD96YruRzxP1UUoN0Dv8IA23_Y2Q83t6u_zau9NmCGx6PWpR9WN6JR9uvna8uYWt7JlbGteAJC5asa</recordid><startdate>201509</startdate><enddate>201509</enddate><creator>Larraya, Luis M.</creator><creator>Fernández-San Millán, Alicia</creator><creator>Ancín, María</creator><creator>Farran, Inmaculada</creator><creator>Veramendi, Jon</creator><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>201509</creationdate><title>Post-harvest light treatment increases expression levels of recombinant proteins in transformed plastids of potato tubers</title><author>Larraya, Luis M. ; Fernández-San Millán, Alicia ; Ancín, María ; Farran, Inmaculada ; Veramendi, Jon</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5268-fbf10634766f15644e8efa160373632c7afd08146aff152c44f44033d8405c993</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Amyloplast</topic><topic>Gene Expression Regulation, Plant - radiation effects</topic><topic>Microtuber</topic><topic>Plant Tubers - metabolism</topic><topic>Plants, Genetically Modified - genetics</topic><topic>Plants, Genetically Modified - metabolism</topic><topic>Plastid transformation</topic><topic>Plastids - genetics</topic><topic>Plastids - metabolism</topic><topic>Potato</topic><topic>Recombinant Proteins - analysis</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><topic>Solanum tuberosum</topic><topic>Solanum tuberosum - genetics</topic><topic>Solanum tuberosum - metabolism</topic><topic>Thioredoxin</topic><topic>Thioredoxins - analysis</topic><topic>Thioredoxins - genetics</topic><topic>Thioredoxins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Larraya, Luis M.</creatorcontrib><creatorcontrib>Fernández-San Millán, Alicia</creatorcontrib><creatorcontrib>Ancín, María</creatorcontrib><creatorcontrib>Farran, Inmaculada</creatorcontrib><creatorcontrib>Veramendi, Jon</creatorcontrib><collection>Istex</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>Biotechnology journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Larraya, Luis M.</au><au>Fernández-San Millán, Alicia</au><au>Ancín, María</au><au>Farran, Inmaculada</au><au>Veramendi, Jon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Post-harvest light treatment increases expression levels of recombinant proteins in transformed plastids of potato tubers</atitle><jtitle>Biotechnology journal</jtitle><addtitle>Biotechnology Journal</addtitle><date>2015-09</date><risdate>2015</risdate><volume>10</volume><issue>11</issue><spage>1803</spage><epage>1813</epage><pages>1803-1813</pages><issn>1860-6768</issn><eissn>1860-7314</eissn><abstract>Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non‐photosynthetic plastids are generally discouraging. Here, we report the expression of two thioredoxin genes (trx f and trx m) from the potato plastid genome to study transgene expression in amyloplasts. As expected, the highest transgene expression was detected in the leaf (up to 4.2% of TSP). The Trx protein content in the tuber was approximately two to three orders of magnitude lower than in the leaf. However, we demonstrate that a simple post‐harvest light treatment of microtubers developed in vitro or soil‐grown tubers induces up to 55 times higher accumulation of the recombinant protein in just seven to ten days. After the applied treatment, the Trx f levels in microtubers and soil‐grown tubers increased to 0.14% and 0.11% of TSP, respectively. Moreover, tubers stored for eight months maintained the capacity of increasing the foreign protein levels after the light treatment. Post‐harvest cold induction (up to five times) at 4°C was also detected in microtubers. We conclude that plastid transformation and post‐harvest light treatment could be an interesting approach for the production of foreign proteins in potato.
Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non‐photosynthetic plastids are generally discouraging. In this article, the authors report the expression of two thioredoxin genes from the potato plastid genome to study transgene expression in amyloplasts. The authors demonstrate that a simple post‐harvest light treatment of tubers induces up to 55 times higher accumulation of the recombinant protein in just 7–10 days. Moreover, tubers stored for 8 months maintain the capacity of increasing the foreign protein levels after the light treatment.</abstract><cop>Weinheim</cop><pub>WILEY-VCH Verlag</pub><pmid>26121393</pmid><doi>10.1002/biot.201500028</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Amyloplast Gene Expression Regulation, Plant - radiation effects Microtuber Plant Tubers - metabolism Plants, Genetically Modified - genetics Plants, Genetically Modified - metabolism Plastid transformation Plastids - genetics Plastids - metabolism Potato Recombinant Proteins - analysis Recombinant Proteins - genetics Recombinant Proteins - metabolism Solanum tuberosum Solanum tuberosum - genetics Solanum tuberosum - metabolism Thioredoxin Thioredoxins - analysis Thioredoxins - genetics Thioredoxins - metabolism |
title | Post-harvest light treatment increases expression levels of recombinant proteins in transformed plastids of potato tubers |
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