Agrobacterium tumefaciens mediated transient expression of plant cell wall-degrading enzymes in detached sunflower leaves.
Identifieur interne : 000383 ( Main/Corpus ); précédent : 000382; suivant : 000384Agrobacterium tumefaciens mediated transient expression of plant cell wall-degrading enzymes in detached sunflower leaves.
Auteurs : Sang-Kyu Jung ; Benjamin E. Lindenmuth ; Karen A. Mcdonald ; Hwang Hwang ; Mai Q Nguyen Bui ; Bryce W. Falk ; Sandra L. Uratsu ; My L. Phu ; Abhaya M. DandekarSource :
- Biotechnology progress [ 1520-6033 ]
English descriptors
- KwdEn :
- Agrobacterium tumefaciens (genetics), Agrobacterium tumefaciens (metabolism), Biofuels (MeSH), Cellulase (biosynthesis), Cellulase (genetics), Endo-1,4-beta Xylanases (biosynthesis), Endo-1,4-beta Xylanases (genetics), Gene Expression Regulation, Plant (MeSH), Helianthus (enzymology), Helianthus (genetics), Plant Cells (enzymology), Plant Cells (metabolism), Plant Leaves (enzymology), Plants, Genetically Modified (chemistry), Promoter Regions, Genetic (MeSH).
- MESH :
- chemical , biosynthesis : Cellulase, Endo-1,4-beta Xylanases.
- chemical , genetics : Cellulase, Endo-1,4-beta Xylanases.
- chemical : Biofuels.
- chemistry : Plants, Genetically Modified.
- enzymology : Helianthus, Plant Cells, Plant Leaves.
- genetics : Agrobacterium tumefaciens, Helianthus.
- metabolism : Agrobacterium tumefaciens, Plant Cells.
- Gene Expression Regulation, Plant, Promoter Regions, Genetic.
Abstract
For biofuel applications, synthetic endoglucanase E1 and xylanase (Xyn10A) derived from Acidothermus cellulolyticus were transiently expressed in detached whole sunflower (Helianthus annuus L.) leaves using vacuum infiltration. Three different expression systems were tested, including the constitutive CaMV 35S-driven, CMVar (Cucumber mosaic virus advanced replicating), and TRBO (Tobacco mosaic virus RNA-Based Overexpression Vector) systems. For 6-day leaf incubations, codon-optimized E1 and xylanase driven by the CaMV 35S promoter were successfully expressed in sunflower leaves. The two viral expression vectors, CMVar and TRBO, were not successful although we found high expression in Nicotiana benthamiana leaves previously for other recombinant proteins. To further enhance transient expression, we demonstrated two novel methods: using the plant hormone methyl jasmonic acid in the agroinfiltration buffer and two-phase optimization of the leaf incubation temperature. When methyl jasmonic acid was added to Agrobacterium tumefaciens cell suspensions and infiltrated into plant leaves, the functional enzyme production increased 4.6-fold. Production also increased up to 4.2-fold when the leaf incubation temperature was elevated above the typical temperature, 20C, to 30C in the late incubation phase, presumably due to enhanced rate of protein synthesis in plant cells. Finally, we demonstrated co-expression of E1 and xylanase in detached sunflower leaves. To our knowledge, this is the first report of (co)expression of heterologous plant cell wall-degrading enzymes in sunflower.
DOI: 10.1002/btpr.1888
PubMed: 25180328
Links to Exploration step
pubmed:25180328Le document en format XML
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Mcdonald</name></author><author><name sortKey="Hwang, Hwang" sort="Hwang, Hwang" uniqKey="Hwang H" first="Hwang" last="Hwang">Hwang Hwang</name></author><author><name sortKey="Bui, Mai Q Nguyen" sort="Bui, Mai Q Nguyen" uniqKey="Bui M" first="Mai Q Nguyen" last="Bui">Mai Q Nguyen Bui</name></author><author><name sortKey="Falk, Bryce W" sort="Falk, Bryce W" uniqKey="Falk B" first="Bryce W" last="Falk">Bryce W. Falk</name></author><author><name sortKey="Uratsu, Sandra L" sort="Uratsu, Sandra L" uniqKey="Uratsu S" first="Sandra L" last="Uratsu">Sandra L. Uratsu</name></author><author><name sortKey="Phu, My L" sort="Phu, My L" uniqKey="Phu M" first="My L" last="Phu">My L. Phu</name></author><author><name sortKey="Dandekar, Abhaya M" sort="Dandekar, Abhaya M" uniqKey="Dandekar A" first="Abhaya M" last="Dandekar">Abhaya M. Dandekar</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2014">2014 Jul-Aug</date><idno type="RBID">pubmed:25180328</idno><idno type="pmid">25180328</idno><idno type="doi">10.1002/btpr.1888</idno><idno type="wicri:Area/Main/Corpus">000383</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000383</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Agrobacterium tumefaciens mediated transient expression of plant cell wall-degrading enzymes in detached sunflower leaves.</title><author><name sortKey="Jung, Sang Kyu" sort="Jung, Sang Kyu" uniqKey="Jung S" first="Sang-Kyu" last="Jung">Sang-Kyu Jung</name></author><author><name sortKey="Lindenmuth, Benjamin E" sort="Lindenmuth, Benjamin E" uniqKey="Lindenmuth B" first="Benjamin E" last="Lindenmuth">Benjamin E. Lindenmuth</name></author><author><name sortKey="Mcdonald, Karen A" sort="Mcdonald, Karen A" uniqKey="Mcdonald K" first="Karen A" last="Mcdonald">Karen A. Mcdonald</name></author><author><name sortKey="Hwang, Hwang" sort="Hwang, Hwang" uniqKey="Hwang H" first="Hwang" last="Hwang">Hwang Hwang</name></author><author><name sortKey="Bui, Mai Q Nguyen" sort="Bui, Mai Q Nguyen" uniqKey="Bui M" first="Mai Q Nguyen" last="Bui">Mai Q Nguyen Bui</name></author><author><name sortKey="Falk, Bryce W" sort="Falk, Bryce W" uniqKey="Falk B" first="Bryce W" last="Falk">Bryce W. Falk</name></author><author><name sortKey="Uratsu, Sandra L" sort="Uratsu, Sandra L" uniqKey="Uratsu S" first="Sandra L" last="Uratsu">Sandra L. Uratsu</name></author><author><name sortKey="Phu, My L" sort="Phu, My L" uniqKey="Phu M" first="My L" last="Phu">My L. Phu</name></author><author><name sortKey="Dandekar, Abhaya M" sort="Dandekar, Abhaya M" uniqKey="Dandekar A" first="Abhaya M" last="Dandekar">Abhaya M. Dandekar</name></author></analytic><series><title level="j">Biotechnology progress</title><idno type="eISSN">1520-6033</idno></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens (genetics)</term><term>Agrobacterium tumefaciens (metabolism)</term><term>Biofuels (MeSH)</term><term>Cellulase (biosynthesis)</term><term>Cellulase (genetics)</term><term>Endo-1,4-beta Xylanases (biosynthesis)</term><term>Endo-1,4-beta Xylanases (genetics)</term><term>Gene Expression Regulation, Plant (MeSH)</term><term>Helianthus (enzymology)</term><term>Helianthus (genetics)</term><term>Plant Cells (enzymology)</term><term>Plant Cells (metabolism)</term><term>Plant Leaves (enzymology)</term><term>Plants, Genetically Modified (chemistry)</term><term>Promoter Regions, Genetic (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Cellulase</term><term>Endo-1,4-beta Xylanases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Cellulase</term><term>Endo-1,4-beta Xylanases</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Biofuels</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Plants, Genetically Modified</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Helianthus</term><term>Plant Cells</term><term>Plant Leaves</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Helianthus</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Plant Cells</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Expression Regulation, Plant</term><term>Promoter Regions, Genetic</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">For biofuel applications, synthetic endoglucanase E1 and xylanase (Xyn10A) derived from Acidothermus cellulolyticus were transiently expressed in detached whole sunflower (Helianthus annuus L.) leaves using vacuum infiltration. Three different expression systems were tested, including the constitutive CaMV 35S-driven, CMVar (Cucumber mosaic virus advanced replicating), and TRBO (Tobacco mosaic virus RNA-Based Overexpression Vector) systems. For 6-day leaf incubations, codon-optimized E1 and xylanase driven by the CaMV 35S promoter were successfully expressed in sunflower leaves. The two viral expression vectors, CMVar and TRBO, were not successful although we found high expression in Nicotiana benthamiana leaves previously for other recombinant proteins. To further enhance transient expression, we demonstrated two novel methods: using the plant hormone methyl jasmonic acid in the agroinfiltration buffer and two-phase optimization of the leaf incubation temperature. When methyl jasmonic acid was added to Agrobacterium tumefaciens cell suspensions and infiltrated into plant leaves, the functional enzyme production increased 4.6-fold. Production also increased up to 4.2-fold when the leaf incubation temperature was elevated above the typical temperature, 20C, to 30C in the late incubation phase, presumably due to enhanced rate of protein synthesis in plant cells. Finally, we demonstrated co-expression of E1 and xylanase in detached sunflower leaves. To our knowledge, this is the first report of (co)expression of heterologous plant cell wall-degrading enzymes in sunflower.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">25180328</PMID><DateCompleted><Year>2015</Year><Month>03</Month><Day>30</Day></DateCompleted><DateRevised><Year>2019</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1520-6033</ISSN><JournalIssue CitedMedium="Internet"><Volume>30</Volume><Issue>4</Issue><PubDate><MedlineDate>2014 Jul-Aug</MedlineDate></PubDate></JournalIssue><Title>Biotechnology progress</Title><ISOAbbreviation>Biotechnol Prog</ISOAbbreviation></Journal><ArticleTitle>Agrobacterium tumefaciens mediated transient expression of plant cell wall-degrading enzymes in detached sunflower leaves.</ArticleTitle><Pagination><MedlinePgn>905-15</MedlinePgn></Pagination><Abstract><AbstractText>For biofuel applications, synthetic endoglucanase E1 and xylanase (Xyn10A) derived from Acidothermus cellulolyticus were transiently expressed in detached whole sunflower (Helianthus annuus L.) leaves using vacuum infiltration. Three different expression systems were tested, including the constitutive CaMV 35S-driven, CMVar (Cucumber mosaic virus advanced replicating), and TRBO (Tobacco mosaic virus RNA-Based Overexpression Vector) systems. For 6-day leaf incubations, codon-optimized E1 and xylanase driven by the CaMV 35S promoter were successfully expressed in sunflower leaves. The two viral expression vectors, CMVar and TRBO, were not successful although we found high expression in Nicotiana benthamiana leaves previously for other recombinant proteins. To further enhance transient expression, we demonstrated two novel methods: using the plant hormone methyl jasmonic acid in the agroinfiltration buffer and two-phase optimization of the leaf incubation temperature. When methyl jasmonic acid was added to Agrobacterium tumefaciens cell suspensions and infiltrated into plant leaves, the functional enzyme production increased 4.6-fold. Production also increased up to 4.2-fold when the leaf incubation temperature was elevated above the typical temperature, 20C, to 30C in the late incubation phase, presumably due to enhanced rate of protein synthesis in plant cells. Finally, we demonstrated co-expression of E1 and xylanase in detached sunflower leaves. To our knowledge, this is the first report of (co)expression of heterologous plant cell wall-degrading enzymes in sunflower.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Jung</LastName><ForeName>Sang-Kyu</ForeName><Initials>SK</Initials></Author><Author ValidYN="Y"><LastName>Lindenmuth</LastName><ForeName>Benjamin E</ForeName><Initials>BE</Initials></Author><Author ValidYN="Y"><LastName>McDonald</LastName><ForeName>Karen A</ForeName><Initials>KA</Initials></Author><Author ValidYN="Y"><LastName>Hwang</LastName><ForeName>Hwang</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Bui</LastName><ForeName>Mai Q Nguyen</ForeName><Initials>MQ</Initials></Author><Author ValidYN="Y"><LastName>Falk</LastName><ForeName>Bryce W</ForeName><Initials>BW</Initials></Author><Author ValidYN="Y"><LastName>Uratsu</LastName><ForeName>Sandra L</ForeName><Initials>SL</Initials></Author><Author ValidYN="Y"><LastName>Phu</LastName><ForeName>My L</ForeName><Initials>ML</Initials></Author><Author ValidYN="Y"><LastName>Dandekar</LastName><ForeName>Abhaya M</ForeName><Initials>AM</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Biotechnol Prog</MedlineTA><NlmUniqueID>8506292</NlmUniqueID><ISSNLinking>1520-6033</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D056804">Biofuels</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.4</RegistryNumber><NameOfSubstance UI="D002480">Cellulase</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.8</RegistryNumber><NameOfSubstance UI="D043364">Endo-1,4-beta Xylanases</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D016960" MajorTopicYN="N">Agrobacterium tumefaciens</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D056804" MajorTopicYN="Y">Biofuels</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002480" MajorTopicYN="N">Cellulase</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D043364" MajorTopicYN="N">Endo-1,4-beta Xylanases</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018506" MajorTopicYN="N">Gene Expression Regulation, Plant</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006368" MajorTopicYN="N">Helianthus</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="N">enzymology</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D059828" MajorTopicYN="N">Plant Cells</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="N">enzymology</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018515" MajorTopicYN="N">Plant Leaves</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="N">enzymology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D030821" MajorTopicYN="N">Plants, Genetically Modified</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011401" MajorTopicYN="N">Promoter Regions, Genetic</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2014</Year><Month>9</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2014</Year><Month>9</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2015</Year><Month>3</Month><Day>31</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">25180328</ArticleId><ArticleId IdType="doi">10.1002/btpr.1888</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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