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Article: Overexpression of Brassica juncea wild-type and mutant HMG-CoA synthase 1 in Arabidopsis up-regulates genes in sterol biosynthesis and enhances sterol production and stress tolerance

TitleOverexpression of Brassica juncea wild-type and mutant HMG-CoA synthase 1 in Arabidopsis up-regulates genes in sterol biosynthesis and enhances sterol production and stress tolerance
Authors
KeywordsBrassica juncea
HMG-CoA synthase
Isoprenoid
Mevalonate
Sterol
Issue Date2012
PublisherBlackwell Publishing Ltd. The Journal's web site is located at http://www.blackwellpublishing.com/journals/PBI
Citation
Plant Biotechnology Journal, 2012, v. 10 n. 1, p. 31-42 How to Cite?
AbstractBrassica juncea 3-hydroxy-3-methylglutaryl-CoA synthase (HMGS) is encoded by four isogenes (BjHMGS1-BjHMGS4). In vitro enzyme assays had indicated that the recombinant BjHMGS1 H188N mutant lacked substrate inhibition by acetoacetyl-CoA (AcAc-CoA) and showed 8-fold decreased enzyme activity. The S359A mutant demonstrated 10-fold higher activity, while the H188N/S359A double mutant displayed a 10-fold increased enzyme activity and lacked inhibition by AcAc-CoA. Here, wild-type and mutant BjHMGS1 were overexpressed in Arabidopsis to examine their effects in planta. The expression of selected genes in isoprenoid biosynthesis, isoprenoid content, seed germination and stress tolerance was analysed in HMGS overexpressors (OEs). Those mRNAs encoding enzymes 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), sterol methyltransferase 2 (SMT2), delta-24 sterol reductase (DWF1), C-22 sterol desaturase (CYP710A1) and brassinosteroid-6-oxidase 2 (BR6OX2) were up-regulated in HMGS-OEs. The total sterol content in leaves and seedlings of OE-wtBjHMGS1, OE-S359A and OE-H188N/S359A was significantly higher than OE-H188N. HMGS-OE seeds germinated earlier than wild-type and vector-transformed controls. HMGS-OEs further displayed reduced hydrogen peroxide (H 2O 2)-induced cell death and constitutive expression of salicylic acid (SA)-dependent pathogenesis-related genes (PR1, PR2 and PR5), resulting in an increased resistance to Botrytis cinerea, with OE-S359A showing the highest and OE-H188N the lowest tolerance. These results suggest that overexpression of HMGS up-regulates HMGR, SMT2, DWF1, CYP710A1 and BR6OX2, leading to enhanced sterol content and stress tolerance in Arabidopsis. © 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.
Persistent Identifierhttp://hdl.handle.net/10722/144733
ISSN
2015 Impact Factor: 6.09
2015 SCImago Journal Rankings: 2.591
ISI Accession Number ID
Funding AgencyGrant Number
University of Hong Kong10208034
10400945
France-Hong Kong Joint Research SchemeF-HKA04/00
Funding Information:

This work was supported by the University of Hong Kong (Grants 10208034 and 10400945, and a Postgraduate Studentship to W.H.) and the France-Hong Kong Joint Research Scheme F-HKA04/00 (to TBJ and MLC). We thank Dr. Hubert Schaller (CNRS-IBMP, Strasbourg) for instructions on sterol extraction and quantitative analysis by the use of the instrumentation installed at the IBMP's metabolomics platform and Prof. S. F. Chen (The University of Hong Kong) for providing the conductivity meter.

References

 

DC FieldValueLanguage
dc.contributor.authorWang, Hen_HK
dc.contributor.authorNagegowda, DAen_HK
dc.contributor.authorRawat, Ren_HK
dc.contributor.authorBouvierNavé, Pen_HK
dc.contributor.authorGuo, Den_HK
dc.contributor.authorBach, TJen_HK
dc.contributor.authorChye, MLen_HK
dc.date.accessioned2012-02-03T06:20:12Z-
dc.date.available2012-02-03T06:20:12Z-
dc.date.issued2012en_HK
dc.identifier.citationPlant Biotechnology Journal, 2012, v. 10 n. 1, p. 31-42en_HK
dc.identifier.issn1467-7644en_HK
dc.identifier.urihttp://hdl.handle.net/10722/144733-
dc.description.abstractBrassica juncea 3-hydroxy-3-methylglutaryl-CoA synthase (HMGS) is encoded by four isogenes (BjHMGS1-BjHMGS4). In vitro enzyme assays had indicated that the recombinant BjHMGS1 H188N mutant lacked substrate inhibition by acetoacetyl-CoA (AcAc-CoA) and showed 8-fold decreased enzyme activity. The S359A mutant demonstrated 10-fold higher activity, while the H188N/S359A double mutant displayed a 10-fold increased enzyme activity and lacked inhibition by AcAc-CoA. Here, wild-type and mutant BjHMGS1 were overexpressed in Arabidopsis to examine their effects in planta. The expression of selected genes in isoprenoid biosynthesis, isoprenoid content, seed germination and stress tolerance was analysed in HMGS overexpressors (OEs). Those mRNAs encoding enzymes 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), sterol methyltransferase 2 (SMT2), delta-24 sterol reductase (DWF1), C-22 sterol desaturase (CYP710A1) and brassinosteroid-6-oxidase 2 (BR6OX2) were up-regulated in HMGS-OEs. The total sterol content in leaves and seedlings of OE-wtBjHMGS1, OE-S359A and OE-H188N/S359A was significantly higher than OE-H188N. HMGS-OE seeds germinated earlier than wild-type and vector-transformed controls. HMGS-OEs further displayed reduced hydrogen peroxide (H 2O 2)-induced cell death and constitutive expression of salicylic acid (SA)-dependent pathogenesis-related genes (PR1, PR2 and PR5), resulting in an increased resistance to Botrytis cinerea, with OE-S359A showing the highest and OE-H188N the lowest tolerance. These results suggest that overexpression of HMGS up-regulates HMGR, SMT2, DWF1, CYP710A1 and BR6OX2, leading to enhanced sterol content and stress tolerance in Arabidopsis. © 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.en_HK
dc.languageengen_US
dc.publisherBlackwell Publishing Ltd. The Journal's web site is located at http://www.blackwellpublishing.com/journals/PBIen_HK
dc.relation.ispartofPlant Biotechnology Journalen_HK
dc.rightsThe definitive version is available at www.blackwell-synergy.comen_US
dc.subjectBrassica junceaen_HK
dc.subjectHMG-CoA synthaseen_HK
dc.subjectIsoprenoiden_HK
dc.subjectMevalonateen_HK
dc.subjectSterolen_HK
dc.titleOverexpression of Brassica juncea wild-type and mutant HMG-CoA synthase 1 in Arabidopsis up-regulates genes in sterol biosynthesis and enhances sterol production and stress toleranceen_HK
dc.typeArticleen_HK
dc.identifier.emailChye, ML: mlchye@hkucc.hku.hken_HK
dc.identifier.authorityChye, ML=rp00687en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1111/j.1467-7652.2011.00631.xen_HK
dc.identifier.pmid21645203-
dc.identifier.scopuseid_2-s2.0-83055173108en_HK
dc.identifier.hkuros198598en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-83055173108&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume10en_HK
dc.identifier.issue1en_HK
dc.identifier.spage31en_HK
dc.identifier.epage42en_HK
dc.identifier.isiWOS:000298094800005-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridWang, H=54797121000en_HK
dc.identifier.scopusauthoridNagegowda, DA=8709830800en_HK
dc.identifier.scopusauthoridRawat, R=8525546500en_HK
dc.identifier.scopusauthoridBouvierNavé, P=6603021106en_HK
dc.identifier.scopusauthoridGuo, D=36184025800en_HK
dc.identifier.scopusauthoridBach, TJ=24786011400en_HK
dc.identifier.scopusauthoridChye, ML=7003905460en_HK
dc.identifier.citeulike11814394-

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