Purification and characterization of a CkTLP protein from Cynanchum komarovii seeds that confers antifungal activity.
Identifieur interne : 000262 ( Main/Curation ); précédent : 000261; suivant : 000263Purification and characterization of a CkTLP protein from Cynanchum komarovii seeds that confers antifungal activity.
Auteurs : Qinghua Wang [République populaire de Chine] ; Fuguang Li ; Xue Zhang ; Yongan Zhang ; Yuxia Hou ; Shengrui Zhang ; Zhixia WuSource :
- PloS one [ 1932-6203 ] ; 2011.
Descripteurs français
- KwdFr :
- ADN complémentaire (analyse), ADN complémentaire (isolement et purification), Antifongiques (analyse), Antifongiques (isolement et purification), Antifongiques (métabolisme), Arabidopsis (génétique), Arabidopsis (immunologie), Arabidopsis (métabolisme), Clonage moléculaire (MeSH), Cynanchum (composition chimique), Cynanchum (génétique), Cynanchum (métabolisme), Extraits de plantes (composition chimique), Extraits de plantes (génétique), Graines (composition chimique), Graines (génétique), Graines (métabolisme), Immunité innée (génétique), Maladies des plantes (génétique), Maladies des plantes (prévention et contrôle), Mycoses (génétique), Mycoses (immunologie), Mycoses (prévention et contrôle), Protéines végétales (analyse), Protéines végétales (génétique), Protéines végétales (isolement et purification), Stress physiologique (génétique), Thérapie génétique (MeSH), Verticillium (physiologie), Végétaux génétiquement modifiés (MeSH).
- MESH :
- analyse : ADN complémentaire, Antifongiques, Protéines végétales.
- composition chimique : Cynanchum, Extraits de plantes, Graines.
- génétique : Arabidopsis, Cynanchum, Extraits de plantes, Graines, Immunité innée, Maladies des plantes, Mycoses, Protéines végétales, Stress physiologique.
- immunologie : Arabidopsis, Mycoses.
- isolement et purification : ADN complémentaire, Antifongiques, Protéines végétales.
- métabolisme : Antifongiques, Arabidopsis, Cynanchum, Graines.
- physiologie : Verticillium.
- prévention et contrôle : Maladies des plantes, Mycoses.
- Clonage moléculaire, Thérapie génétique, Végétaux génétiquement modifiés.
English descriptors
- KwdEn :
- Antifungal Agents (analysis), Antifungal Agents (isolation & purification), Antifungal Agents (metabolism), Arabidopsis (genetics), Arabidopsis (immunology), Arabidopsis (metabolism), Cloning, Molecular (MeSH), Cynanchum (chemistry), Cynanchum (genetics), Cynanchum (metabolism), DNA, Complementary (analysis), DNA, Complementary (isolation & purification), Genetic Therapy (MeSH), Immunity, Innate (genetics), Mycoses (genetics), Mycoses (immunology), Mycoses (prevention & control), Plant Diseases (genetics), Plant Diseases (prevention & control), Plant Extracts (chemistry), Plant Extracts (genetics), Plant Proteins (analysis), Plant Proteins (genetics), Plant Proteins (isolation & purification), Plants, Genetically Modified (MeSH), Seeds (chemistry), Seeds (genetics), Seeds (metabolism), Stress, Physiological (genetics), Verticillium (physiology).
- MESH :
- chemical , analysis : Antifungal Agents, DNA, Complementary, Plant Proteins.
- chemical , chemistry : Plant Extracts.
- chemical , genetics : Plant Extracts, Plant Proteins.
- chemical , isolation & purification : Antifungal Agents, DNA, Complementary, Plant Proteins.
- chemical , metabolism : Antifungal Agents.
- chemistry : Cynanchum, Seeds.
- genetics : Arabidopsis, Cynanchum, Immunity, Innate, Mycoses, Plant Diseases, Seeds, Stress, Physiological.
- immunology : Arabidopsis, Mycoses.
- metabolism : Arabidopsis, Cynanchum, Seeds.
- physiology : Verticillium.
- prevention & control : Mycoses, Plant Diseases.
- Cloning, Molecular, Genetic Therapy, Plants, Genetically Modified.
Abstract
BACKGROUND
Cynanchum komarovii Al Iljinski is a desert plant that has been used as analgesic, anthelminthic and antidiarrheal, but also as a herbal medicine to treat cholecystitis in people. We have found that the protein extractions from C. komarovii seeds have strong antifungal activity. There is strong interest to develop protein medication and antifungal pesticides from C. komarovii for pharmacological or other uses.
METHODOLOGY/PRINCIPAL FINDINGS
An antifungal protein with sequence homology to thaumatin-like proteins (TLPs) was isolated from C. komarovii seeds and named CkTLP. The three-dimensional structure prediction of CkTLP indicated the protein has an acid cleft and a hydrophobic patch. The protein showed antifungal activity against fungal growth of Verticillium dahliae, Fusarium oxysporum, Rhizoctonia solani, Botrytis cinerea and Valsa mali. The full-length cDNA was cloned by RT-PCR and RACE-PCR according to the partial protein sequences obtained by nanoESI-MS/MS. The real-time PCR showed the transcription level of CkTLP had a significant increase under the stress of abscisic acid (ABA), salicylic acid (SA), methyl jasmonate (MeJA), NaCl and drought, which indicates that CkTLP may play an important role in response to abiotic stresses. Histochemical staining showed GUS activity in almost the whole plant, especially in cotyledons, trichomes and vascular tissues of primary root and inflorescences. The CkTLP protein was located in the extracellular space/cell wall by CkTLP::GFP fusion protein in transgenic Arabidopsis. Furthermore, over-expression of CkTLP significantly enhanced the resistance of Arabidopsis against V. dahliae.
CONCLUSIONS/SIGNIFICANCE
The results suggest that the CkTLP is a good candidate protein or gene for contributing to the development of disease-resistant crops.
DOI: 10.1371/journal.pone.0016930
PubMed: 21364945
PubMed Central: PMC3043079
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Diseases</term></keywords><keywords scheme="MESH" qualifier="prévention et contrôle" xml:lang="fr"><term>Maladies des plantes</term><term>Mycoses</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cloning, Molecular</term><term>Genetic Therapy</term><term>Plants, Genetically Modified</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Clonage moléculaire</term><term>Thérapie génétique</term><term>Végétaux génétiquement modifiés</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b></p><p>Cynanchum komarovii Al Iljinski is a desert plant that has been used as analgesic, anthelminthic and antidiarrheal, but also as a herbal medicine to treat cholecystitis in people. We have found that the protein extractions from C. komarovii seeds have strong antifungal activity. There is strong interest to develop protein medication and antifungal pesticides from C. komarovii for pharmacological or other uses.</p></div><div type="abstract" xml:lang="en"><p><b>METHODOLOGY/PRINCIPAL FINDINGS</b></p><p>An antifungal protein with sequence homology to thaumatin-like proteins (TLPs) was isolated from C. komarovii seeds and named CkTLP. The three-dimensional structure prediction of CkTLP indicated the protein has an acid cleft and a hydrophobic patch. The protein showed antifungal activity against fungal growth of Verticillium dahliae, Fusarium oxysporum, Rhizoctonia solani, Botrytis cinerea and Valsa mali. The full-length cDNA was cloned by RT-PCR and RACE-PCR according to the partial protein sequences obtained by nanoESI-MS/MS. The real-time PCR showed the transcription level of CkTLP had a significant increase under the stress of abscisic acid (ABA), salicylic acid (SA), methyl jasmonate (MeJA), NaCl and drought, which indicates that CkTLP may play an important role in response to abiotic stresses. Histochemical staining showed GUS activity in almost the whole plant, especially in cotyledons, trichomes and vascular tissues of primary root and inflorescences. The CkTLP protein was located in the extracellular space/cell wall by CkTLP::GFP fusion protein in transgenic Arabidopsis. Furthermore, over-expression of CkTLP significantly enhanced the resistance of Arabidopsis against V. dahliae.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSIONS/SIGNIFICANCE</b></p><p>The results suggest that the CkTLP is a good candidate protein or gene for contributing to the development of disease-resistant crops.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">21364945</PMID><DateCompleted><Year>2011</Year><Month>09</Month><Day>01</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1932-6203</ISSN><JournalIssue CitedMedium="Internet"><Volume>6</Volume><Issue>2</Issue><PubDate><Year>2011</Year><Month>Feb</Month><Day>22</Day></PubDate></JournalIssue><Title>PloS one</Title><ISOAbbreviation>PLoS One</ISOAbbreviation></Journal><ArticleTitle>Purification and characterization of a CkTLP protein from Cynanchum komarovii seeds that confers antifungal activity.</ArticleTitle><Pagination><MedlinePgn>e16930</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1371/journal.pone.0016930</ELocationID><Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">Cynanchum komarovii Al Iljinski is a desert plant that has been used as analgesic, anthelminthic and antidiarrheal, but also as a herbal medicine to treat cholecystitis in people. We have found that the protein extractions from C. komarovii seeds have strong antifungal activity. There is strong interest to develop protein medication and antifungal pesticides from C. komarovii for pharmacological or other uses.</AbstractText><AbstractText Label="METHODOLOGY/PRINCIPAL FINDINGS" NlmCategory="RESULTS">An antifungal protein with sequence homology to thaumatin-like proteins (TLPs) was isolated from C. komarovii seeds and named CkTLP. The three-dimensional structure prediction of CkTLP indicated the protein has an acid cleft and a hydrophobic patch. The protein showed antifungal activity against fungal growth of Verticillium dahliae, Fusarium oxysporum, Rhizoctonia solani, Botrytis cinerea and Valsa mali. The full-length cDNA was cloned by RT-PCR and RACE-PCR according to the partial protein sequences obtained by nanoESI-MS/MS. The real-time PCR showed the transcription level of CkTLP had a significant increase under the stress of abscisic acid (ABA), salicylic acid (SA), methyl jasmonate (MeJA), NaCl and drought, which indicates that CkTLP may play an important role in response to abiotic stresses. Histochemical staining showed GUS activity in almost the whole plant, especially in cotyledons, trichomes and vascular tissues of primary root and inflorescences. The CkTLP protein was located in the extracellular space/cell wall by CkTLP::GFP fusion protein in transgenic Arabidopsis. Furthermore, over-expression of CkTLP significantly enhanced the resistance of Arabidopsis against V. dahliae.</AbstractText><AbstractText Label="CONCLUSIONS/SIGNIFICANCE" NlmCategory="CONCLUSIONS">The results suggest that the CkTLP is a good candidate protein or gene for contributing to the development of disease-resistant crops.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Qinghua</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>College of Science, China Agricultural University, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Fuguang</ForeName><Initials>F</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Xue</ForeName><Initials>X</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Yongan</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Hou</LastName><ForeName>Yuxia</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Shengrui</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Wu</LastName><ForeName>Zhixia</ForeName><Initials>Z</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D023362">Evaluation Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2011</Year><Month>02</Month><Day>22</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>PLoS One</MedlineTA><NlmUniqueID>101285081</NlmUniqueID><ISSNLinking>1932-6203</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000935">Antifungal Agents</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018076">DNA, Complementary</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010936">Plant Extracts</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010940">Plant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>53850-34-3</RegistryNumber><NameOfSubstance UI="C003427">thaumatin protein, plant</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000935" MajorTopicYN="N">Antifungal Agents</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D017360" MajorTopicYN="N">Arabidopsis</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D003001" MajorTopicYN="N">Cloning, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D031105" MajorTopicYN="N">Cynanchum</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018076" MajorTopicYN="N">DNA, Complementary</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015316" MajorTopicYN="N">Genetic Therapy</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007113" MajorTopicYN="N">Immunity, Innate</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009181" MajorTopicYN="N">Mycoses</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName><QualifierName UI="Q000517" MajorTopicYN="Y">prevention & control</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010935" MajorTopicYN="N">Plant Diseases</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000517" MajorTopicYN="Y">prevention & control</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010936" MajorTopicYN="N">Plant Extracts</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" 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