Assessing the role of root plasma membrane and tonoplast Na+/H+ exchangers in salinity tolerance in wheat: in planta quantification methods
ABSTRACT This work investigates the role of cytosolic Na+ exclusion in roots as a means of salinity tolerance in wheat, and offers in planta methods for the functional assessment of major transporters contributing to this trait. An electrophysiological protocol was developed to quantify the activity...
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| Veröffentlicht in: | Plant, cell and environment cell and environment, 2011-06, Vol.34 (6), p.947-961 |
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| creator | CUIN, TRACEY A. BOSE, JAYAKUMAR STEFANO, GIOVANNI JHA, DEEPA TESTER, MARK MANCUSO, STEFANO SHABALA, SERGEY |
| description | ABSTRACT
This work investigates the role of cytosolic Na+ exclusion in roots as a means of salinity tolerance in wheat, and offers in planta methods for the functional assessment of major transporters contributing to this trait. An electrophysiological protocol was developed to quantify the activity of plasma membrane Na+ efflux systems in roots, using the microelectrode ion flux estimation (MIFE) technique. We show that active efflux of Na+ from wheat root epidermal cells is mediated by a SOS1‐like homolog, energized by the plasma membrane H+‐ATPase. SOS1‐like efflux activity was highest in Kharchia 65, a salt‐tolerant bread wheat cultivar. Kharchia 65 also had an enhanced ability to sequester large quantities of Na+ into the vacuoles of root cells, as revealed by confocal microscopy using Sodium Green. These findings were consistent with the highest level of expression of both SOS1 and NHX1 transcripts in plant roots in this variety. In the sensitive wheat varieties, a greater proportion of Na+ was located in the root cell cytosol. Overall, our findings suggest a critical role of cytosolic Na+ exclusion for salinity tolerance in wheat and offer convenient protocols to quantify the contribution of the major transporters conferring this trait, to screen plants for salinity tolerance. |
| doi_str_mv | 10.1111/j.1365-3040.2011.02296.x |
| format | Article |
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This work investigates the role of cytosolic Na+ exclusion in roots as a means of salinity tolerance in wheat, and offers in planta methods for the functional assessment of major transporters contributing to this trait. An electrophysiological protocol was developed to quantify the activity of plasma membrane Na+ efflux systems in roots, using the microelectrode ion flux estimation (MIFE) technique. We show that active efflux of Na+ from wheat root epidermal cells is mediated by a SOS1‐like homolog, energized by the plasma membrane H+‐ATPase. SOS1‐like efflux activity was highest in Kharchia 65, a salt‐tolerant bread wheat cultivar. Kharchia 65 also had an enhanced ability to sequester large quantities of Na+ into the vacuoles of root cells, as revealed by confocal microscopy using Sodium Green. These findings were consistent with the highest level of expression of both SOS1 and NHX1 transcripts in plant roots in this variety. In the sensitive wheat varieties, a greater proportion of Na+ was located in the root cell cytosol. Overall, our findings suggest a critical role of cytosolic Na+ exclusion for salinity tolerance in wheat and offer convenient protocols to quantify the contribution of the major transporters conferring this trait, to screen plants for salinity tolerance.</description><identifier>ISSN: 0140-7791</identifier><identifier>EISSN: 1365-3040</identifier><identifier>DOI: 10.1111/j.1365-3040.2011.02296.x</identifier><identifier>PMID: 21342209</identifier><identifier>CODEN: PLCEDV</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Arabidopsis - drug effects ; Arabidopsis - metabolism ; Biochemistry - methods ; Biological and medical sciences ; Bread ; Cell Membrane - metabolism ; cytosol ; Cytosol - drug effects ; Cytosol - metabolism ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation, Plant - drug effects ; Genotype ; Kinetics ; Membranes ; Mutation - genetics ; Osmotic Pressure - drug effects ; Plant Proteins - genetics ; Plant Proteins - metabolism ; Plant Roots - cytology ; Plant Roots - drug effects ; Plant Roots - metabolism ; Potassium - metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; salinity tolerance ; Salt-Tolerance - drug effects ; Salt-Tolerance - genetics ; Seedlings - drug effects ; Seedlings - metabolism ; sodium ; Sodium - metabolism ; Sodium Chloride - pharmacology ; Sodium-Hydrogen Exchangers - metabolism ; Triticum - drug effects ; Triticum - genetics ; Triticum - metabolism ; Triticum aestivum ; vacuolar sequestration ; Vacuoles - metabolism ; wheat</subject><ispartof>Plant, cell and environment, 2011-06, Vol.34 (6), p.947-961</ispartof><rights>2011 Blackwell Publishing Ltd</rights><rights>2015 INIST-CNRS</rights><rights>2011 Blackwell Publishing Ltd.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4966-5205ee56bfc1c346d1fee526bc7f6b1575db4da111e5c27ac34f8e8144eec20c3</citedby><cites>FETCH-LOGICAL-c4966-5205ee56bfc1c346d1fee526bc7f6b1575db4da111e5c27ac34f8e8144eec20c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1365-3040.2011.02296.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1365-3040.2011.02296.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,777,781,1412,1428,27905,27906,45555,45556,46390,46814</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24170756$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21342209$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>CUIN, TRACEY A.</creatorcontrib><creatorcontrib>BOSE, JAYAKUMAR</creatorcontrib><creatorcontrib>STEFANO, GIOVANNI</creatorcontrib><creatorcontrib>JHA, DEEPA</creatorcontrib><creatorcontrib>TESTER, MARK</creatorcontrib><creatorcontrib>MANCUSO, STEFANO</creatorcontrib><creatorcontrib>SHABALA, SERGEY</creatorcontrib><title>Assessing the role of root plasma membrane and tonoplast Na+/H+ exchangers in salinity tolerance in wheat: in planta quantification methods</title><title>Plant, cell and environment</title><addtitle>Plant Cell Environ</addtitle><description>ABSTRACT
This work investigates the role of cytosolic Na+ exclusion in roots as a means of salinity tolerance in wheat, and offers in planta methods for the functional assessment of major transporters contributing to this trait. An electrophysiological protocol was developed to quantify the activity of plasma membrane Na+ efflux systems in roots, using the microelectrode ion flux estimation (MIFE) technique. We show that active efflux of Na+ from wheat root epidermal cells is mediated by a SOS1‐like homolog, energized by the plasma membrane H+‐ATPase. SOS1‐like efflux activity was highest in Kharchia 65, a salt‐tolerant bread wheat cultivar. Kharchia 65 also had an enhanced ability to sequester large quantities of Na+ into the vacuoles of root cells, as revealed by confocal microscopy using Sodium Green. These findings were consistent with the highest level of expression of both SOS1 and NHX1 transcripts in plant roots in this variety. In the sensitive wheat varieties, a greater proportion of Na+ was located in the root cell cytosol. Overall, our findings suggest a critical role of cytosolic Na+ exclusion for salinity tolerance in wheat and offer convenient protocols to quantify the contribution of the major transporters conferring this trait, to screen plants for salinity tolerance.</description><subject>Arabidopsis - drug effects</subject><subject>Arabidopsis - metabolism</subject><subject>Biochemistry - methods</subject><subject>Biological and medical sciences</subject><subject>Bread</subject><subject>Cell Membrane - metabolism</subject><subject>cytosol</subject><subject>Cytosol - drug effects</subject><subject>Cytosol - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation, Plant - drug effects</subject><subject>Genotype</subject><subject>Kinetics</subject><subject>Membranes</subject><subject>Mutation - genetics</subject><subject>Osmotic Pressure - drug effects</subject><subject>Plant Proteins - genetics</subject><subject>Plant Proteins - metabolism</subject><subject>Plant Roots - cytology</subject><subject>Plant Roots - drug effects</subject><subject>Plant Roots - metabolism</subject><subject>Potassium - metabolism</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>salinity tolerance</subject><subject>Salt-Tolerance - drug effects</subject><subject>Salt-Tolerance - genetics</subject><subject>Seedlings - drug effects</subject><subject>Seedlings - metabolism</subject><subject>sodium</subject><subject>Sodium - metabolism</subject><subject>Sodium Chloride - pharmacology</subject><subject>Sodium-Hydrogen Exchangers - metabolism</subject><subject>Triticum - drug effects</subject><subject>Triticum - genetics</subject><subject>Triticum - metabolism</subject><subject>Triticum aestivum</subject><subject>vacuolar sequestration</subject><subject>Vacuoles - metabolism</subject><subject>wheat</subject><issn>0140-7791</issn><issn>1365-3040</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc9u1DAQxi0EokvpKyBfEIcqqe34T4LEoVoVilQBB3q2HGfS9SqJt7FX3X2GvjQ2u7TH4suMx7_PM5oPIUxJSdO5WJe0kqKoCCclI5SWhLFGlrtXaPH08BotCOWkUKqhJ-hdCGtCUkE1b9EJoxVnjDQL9HgZAoTgpjscV4BnPwD2fYo-4s1gwmjwCGM7mwmwmToc_eRzPeIf5vzi-hzDzq7MdAdzwG7CwQxucnGfuAGSyEKuPqzAxM85S9IpGny_TcH1zpro_JQ6xJXvwnv0pjdDgLNjPEW3X69-L6-Lm5_fvi8vbwrLGykLwYgAELLtLbUVlx3t05XJ1qpetlQo0bW8M2lPICxTJjF9DTXlHMAyYqtT9Onw72b291sIUY8uWBjSbOC3QTe0JrxWtXiRrKVkglaKJbI-kHb2IczQ683sRjPvNSU6e6bXOlujszU6e6b_eqZ3Sfrh2GTbjtA9Cf-ZlICPR8AEa4Y-79WFZ45TRZSQifty4B7cAPv_HkD_Wl7lrPoDa1uz2Q</recordid><startdate>201106</startdate><enddate>201106</enddate><creator>CUIN, TRACEY A.</creator><creator>BOSE, JAYAKUMAR</creator><creator>STEFANO, GIOVANNI</creator><creator>JHA, DEEPA</creator><creator>TESTER, MARK</creator><creator>MANCUSO, STEFANO</creator><creator>SHABALA, SERGEY</creator><general>Blackwell Publishing Ltd</general><general>Blackwell</general><scope>IQODW</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>7X8</scope><scope>7ST</scope><scope>C1K</scope><scope>SOI</scope></search><sort><creationdate>201106</creationdate><title>Assessing the role of root plasma membrane and tonoplast Na+/H+ exchangers in salinity tolerance in wheat: in planta quantification methods</title><author>CUIN, TRACEY A. ; BOSE, JAYAKUMAR ; STEFANO, GIOVANNI ; JHA, DEEPA ; TESTER, MARK ; MANCUSO, STEFANO ; SHABALA, SERGEY</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4966-5205ee56bfc1c346d1fee526bc7f6b1575db4da111e5c27ac34f8e8144eec20c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Arabidopsis - drug effects</topic><topic>Arabidopsis - metabolism</topic><topic>Biochemistry - methods</topic><topic>Biological and medical sciences</topic><topic>Bread</topic><topic>Cell Membrane - metabolism</topic><topic>cytosol</topic><topic>Cytosol - drug effects</topic><topic>Cytosol - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation, Plant - drug effects</topic><topic>Genotype</topic><topic>Kinetics</topic><topic>Membranes</topic><topic>Mutation - genetics</topic><topic>Osmotic Pressure - drug effects</topic><topic>Plant Proteins - genetics</topic><topic>Plant Proteins - metabolism</topic><topic>Plant Roots - cytology</topic><topic>Plant Roots - drug effects</topic><topic>Plant Roots - metabolism</topic><topic>Potassium - metabolism</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>salinity tolerance</topic><topic>Salt-Tolerance - drug effects</topic><topic>Salt-Tolerance - genetics</topic><topic>Seedlings - drug effects</topic><topic>Seedlings - metabolism</topic><topic>sodium</topic><topic>Sodium - metabolism</topic><topic>Sodium Chloride - pharmacology</topic><topic>Sodium-Hydrogen Exchangers - metabolism</topic><topic>Triticum - drug effects</topic><topic>Triticum - genetics</topic><topic>Triticum - metabolism</topic><topic>Triticum aestivum</topic><topic>vacuolar sequestration</topic><topic>Vacuoles - metabolism</topic><topic>wheat</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>CUIN, TRACEY A.</creatorcontrib><creatorcontrib>BOSE, JAYAKUMAR</creatorcontrib><creatorcontrib>STEFANO, GIOVANNI</creatorcontrib><creatorcontrib>JHA, DEEPA</creatorcontrib><creatorcontrib>TESTER, MARK</creatorcontrib><creatorcontrib>MANCUSO, STEFANO</creatorcontrib><creatorcontrib>SHABALA, SERGEY</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Environment Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Environment Abstracts</collection><jtitle>Plant, cell and environment</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>CUIN, TRACEY A.</au><au>BOSE, JAYAKUMAR</au><au>STEFANO, GIOVANNI</au><au>JHA, DEEPA</au><au>TESTER, MARK</au><au>MANCUSO, STEFANO</au><au>SHABALA, SERGEY</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Assessing the role of root plasma membrane and tonoplast Na+/H+ exchangers in salinity tolerance in wheat: in planta quantification methods</atitle><jtitle>Plant, cell and environment</jtitle><addtitle>Plant Cell Environ</addtitle><date>2011-06</date><risdate>2011</risdate><volume>34</volume><issue>6</issue><spage>947</spage><epage>961</epage><pages>947-961</pages><issn>0140-7791</issn><eissn>1365-3040</eissn><coden>PLCEDV</coden><abstract>ABSTRACT
This work investigates the role of cytosolic Na+ exclusion in roots as a means of salinity tolerance in wheat, and offers in planta methods for the functional assessment of major transporters contributing to this trait. An electrophysiological protocol was developed to quantify the activity of plasma membrane Na+ efflux systems in roots, using the microelectrode ion flux estimation (MIFE) technique. We show that active efflux of Na+ from wheat root epidermal cells is mediated by a SOS1‐like homolog, energized by the plasma membrane H+‐ATPase. SOS1‐like efflux activity was highest in Kharchia 65, a salt‐tolerant bread wheat cultivar. Kharchia 65 also had an enhanced ability to sequester large quantities of Na+ into the vacuoles of root cells, as revealed by confocal microscopy using Sodium Green. These findings were consistent with the highest level of expression of both SOS1 and NHX1 transcripts in plant roots in this variety. In the sensitive wheat varieties, a greater proportion of Na+ was located in the root cell cytosol. Overall, our findings suggest a critical role of cytosolic Na+ exclusion for salinity tolerance in wheat and offer convenient protocols to quantify the contribution of the major transporters conferring this trait, to screen plants for salinity tolerance.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>21342209</pmid><doi>10.1111/j.1365-3040.2011.02296.x</doi><tpages>15</tpages></addata></record> |
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| subjects | Arabidopsis - drug effects Arabidopsis - metabolism Biochemistry - methods Biological and medical sciences Bread Cell Membrane - metabolism cytosol Cytosol - drug effects Cytosol - metabolism Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Plant - drug effects Genotype Kinetics Membranes Mutation - genetics Osmotic Pressure - drug effects Plant Proteins - genetics Plant Proteins - metabolism Plant Roots - cytology Plant Roots - drug effects Plant Roots - metabolism Potassium - metabolism Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger - genetics RNA, Messenger - metabolism salinity tolerance Salt-Tolerance - drug effects Salt-Tolerance - genetics Seedlings - drug effects Seedlings - metabolism sodium Sodium - metabolism Sodium Chloride - pharmacology Sodium-Hydrogen Exchangers - metabolism Triticum - drug effects Triticum - genetics Triticum - metabolism Triticum aestivum vacuolar sequestration Vacuoles - metabolism wheat |
| title | Assessing the role of root plasma membrane and tonoplast Na+/H+ exchangers in salinity tolerance in wheat: in planta quantification methods |
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