|
|
|
Overexpression of a Cytosolic Ascorbate Peroxidase Gene, OsAPX2, Increases Salt Tolerance in Transgenic Alfalfa |
ZHANG Qian, MA Cui, XUE Xin, XU Ming, LI Jing , WU Jin-xia |
1、Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
2、College of Pastoral Agricultural Science and Technology, Lanzhou University, Lanzhou 730000, P.R.China |
|
|
摘要 Alfalfa (Medicago sativa L.) is an important forage crop in the world and it is of great significance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene (OsAPX2) was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T1 transgenic alfalfa were identified by Southern hybridization. Three independent T2 transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H2O2, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species.
Abstract Alfalfa (Medicago sativa L.) is an important forage crop in the world and it is of great significance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene (OsAPX2) was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T1 transgenic alfalfa were identified by Southern hybridization. Three independent T2 transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H2O2, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species.
|
Received: 01 August 2013
Accepted:
|
Fund: This work was supported by the National 973 Program of China (2014CB138700). |
Corresponding Authors:
WU Jin-xia, Tel: +86-10-82106133, Fax: +86-10-82106132, E-mail: wujinxia@caas.cn
E-mail: wujinxia@caas.cn
|
About author: ZHANG Qian, E-mail: springzhang55@163.com |
Cite this article:
ZHANG Qian, MA Cui, XUE Xin, XU Ming, LI Jing , WU Jin-xia.
2014.
Overexpression of a Cytosolic Ascorbate Peroxidase Gene, OsAPX2, Increases Salt Tolerance in Transgenic Alfalfa. Journal of Integrative Agriculture, 13(11): 2500-2507.
|
Agrawal G K, Jwa N S, Iwahashi H, Rakwal R. 2003.Importance of ascorbate peroxidases OsAPX1 andOsAPX2 in the rice pathogen response pathways andgrowth and reproduction revealed by their transcriptionalprofiling. Gene, 322, 93-103Asada K 1999. The water-water cycle in chloroplasts:Scavenging of active oxygens and dissipation of excessphotons. Annual Review of Plant Physiology and PlantMolecular Biology, 50, 601-639Bates L, Walderen R, Taere I. 1973. Rapid determination offree proline for water stress studies. Plant and Soil, 39,205-207Chew O, Whelan J, Millar A H. 2003. Molecular definitionof the ascorbate-glutathione cycle in Arabidopsismitochondria reveals dual targeting of antioxidant defensesin plants. The Journal of Biological Chemistry, 278,46869-46877Davletova S, Rizhsky L, Liang H, Shengqiang Z, OliverD J, Coutu J, Shulaev V, Schlauch K, Mittler R. 2005.Cytosolic ascorbate peroxidase 1 is a central componentof the reactive oxygen gene network of Arabidopsis. ThePlant Cell, 17, 268-281Hare P, Cress W, van Staden J. 1998. Dissecting the rolesof osmolyte accumulation during stress. Plant Cell andEnvironment, 21, 535-553Hodges D, de Long J, Forney C, Prange R. 1999. Improving thethiobarbituric acid-reactive-substances assay for estimatinglipid peroxidation in plant tissues containing anthocyaninand other interfering compounds. Planta, 207, 604-611Jain M, Mathur G, Koul S, Sarin N. 2001. Ameliorative effectof proline on the salt stress induced lipid peroxidation incell lines of groundnut (Arachis hypogea L.). Plant CellReports, 20, 463-468Karpinski S, Escobar C, Karpinska B, Creissen G, MullineauxP M. 1997. Photosynthetic electron transport regulatesthe expression of cytosolic ascorbate peroxidase genesin Arabidopsis during excess light stress. The Plant Cell,9, 627-640Koussevitzky S, Suzuki N, Huntington S, Armijo L, ShaW, Cortes D, Shulaev V, Mittler R. 2008. Ascorbateperoxidase 1 plays a key role in the response of Arabidopsisthaliana to stress combination. The Journal of BiologicalChemistry, 283, 34197-34203Lu Z, Liu D, Liu S. 2007. Two rice cytosolic ascorbateperoxidases differentially improve salt tolerance intransgenic Arabidopsis. Plant Cell Reports, 26, 1909-1917Maruta T, Tanouchi A, Tamoi M, Yabuta Y, Yoshimura K,Ishikawa T, Shigeoka S. 2010. Arabidopsis chloroplasticascorbate peroxidase isoenzymes play a dual role inphotoprotection and gene regulation under photooxidativestress. Plant and Cell Physiology, 51, 190-200Mckersie B D, Bowley S R, Harjanto E, Leprince O. 1996.Water-deficit tolerance and field performance of transgenicalfalfa overexpressing superoxide dismutase. PlantPhysiology, 111, 1177-1181 Miller G, Suzuki N, Rizhsky L, Hegie A, Koussevitzky S,Mittler R. 2007. Double mutants deficient in cytosolicand thylakoid ascorbate peroxidase reveal a complexmode of interaction between reactive oxygen species,plant development, and response to abiotic stresses. PlantPhysiology, 144, 1777-1785Mittler R, Vanderauwera S, Gollery M, van Breusegem F.2004. Reactive oxygen gene network of plants. Trends inPlant Science, 9, 490-498Murashige T, Skoog F. 1962. A revised medium for rapidgrowth and bioassays with tobacco tissue culture. PhysiolPlantarum, 15, 473-497Murray M, Thompson W. 1980. Rapid isolation of highmolecular weight plant DNA. Nucleic Acids Research,8, 4321-4325Noctor G, Foyer C H. 1998. Ascorbate and glutathione:Keeping active oxygen under control. Annual Reviewof Plant Physiology and Plant Molecular Biology, 49,249-279Panchuk I I, Zentgraf U, Volkov R A. 2005. Expression of theApx gene family during leaf senescence of Arabidopsisthaliana. Planta, 222, 926-932Rizhsky L, Hallak-Herr E, van Breusegem F, RachmilevitchS, Barr J E, Rodermel S, Inze D, Mittler R. 2002. Doubleantisense plants lacking ascorbate peroxidase and catalaseare less sensitive to oxidative stress than single antisenseplants lacking ascorbate peroxidase or catalase. PlantJournal, 32, 329-342Rizhsky L, Liang H, Shuman J, Shulaev V, Davletova S,Mittler R. 2004. When defense pathways collide, theresponse of Arabidopsis to a combination of drought andheat stress. Plant Physiology, 134, 1683-1696Rosa S B, Caverzan A, Teixeira F K, Lazzarotto F, SilveiraJ A, Ferreira-Silva S L, Abreu-Neto J, Margis R, Margis-Pinheiro M. 2010. Cytosolic APx knockdown indicatesan ambiguous redox responses in rice. Phytochemistry,71, 548-558Sato Y, Masuta Y, Saito K, Murayama S, Ozawa K. 2011.Enhanced chilling tolerance at the booting stage in riceby transgenic overexpression of the ascorbate peroxidasegene, OsAPXa. Plant Cell Reports, 30, 399-406Scandalios J G. 2002. The rise of ROS. Trends in BiochemicalSciences, 27, 483-486Shigeoka S, Ishikawa T, Tamoi M, Miyagawa Y, Takeda T,Yabuta Y, Yoshimura K. 2002. Regulation and function ofascorbate peroxidase isoenzymes. Journal of ExperimentalBotany, 53, 1305-1319Suarez R, Calderon C, Iturriaga G. 2009. Enhanced tolerance tomultiple abiotic stresses in transgenic alfalfa accumulatingtrehalose. Crop Science, 49, 1791-1799Teixeira F K, Menezes-Benavente L, Galvao V C, Margis R,Margis-Pinheiro M. 2006. Rice ascorbate peroxidase genefamily encodes functionally diverse isoforms localized indifferent subcellular compartments. Planta 224, 300-314Trinh T H, Ratet P, Kondorosi E, Durand P, Kamaté K, BauerP, Kondorosi A. 1998. Rapid and efficient transformationof diploid medicago truncatula and Medicago sativa ssp.falcata lines improved in somatic embryogenesis. PlantCell Reports, 17, 345-355Winicov I. 2000. Alfin1 transcription factor overexpressionenhances plant root growth under normal and salineconditions and improves salt tolerance in alfalfa. Planta210, 416-422Zhang J Y, Broeckling C D, Blancaflor E B, Sledge M K,Sumner L W, Wang Z Y. 2005. Overexpression of WXP1,a putative Medicago truncatula AP2 domain-containingtranscription factor gene, increases cuticular waxaccumulation and enhances drought tolerance in transgenicalfalfa (Medicago sativa). Plant Journal, 42, 689-707Zhang Z, Zhang Q, Wu J, Zheng X, Zheng S, Sun X, Qiu Q,Lu T. 2013. Gene knockout study reveals that cytosolicascorbate peroxidase 2 (OsAPX2) plays a critical role ingrowth and reproduction in rice under drought, salt andcold stresses. PLOS ONE, 8, e57472. |
No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
|
Shared |
|
|
|
|
|
Discussed |
|
|
|
|