Xylem sap collection and extraction methodologies to determine <it>in vivo </it>concentrations of ABA and its bound forms by gas chromatography-mass spectrometry (GC-MS)
<p>Abstract</p> <p>Background</p> <p>Accurate quantification of xylem sap ABA concentrations is important to underpin models of root-to-shoot ABA signalling to predict the physiological effects of soil drying. Growing tomato plants in a whole plant pressure chamber allo...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-03-01
|
| Series: | Plant Methods |
| Subjects: | |
| Online Access: | http://www.plantmethods.com/content/8/1/11 |
| _version_ | 1818369555349635072 |
|---|---|
| author | Netting Andrew G Theobald Julian C Dodd Ian C |
| author_facet | Netting Andrew G Theobald Julian C Dodd Ian C |
| author_sort | Netting Andrew G |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Accurate quantification of xylem sap ABA concentrations is important to underpin models of root-to-shoot ABA signalling to predict the physiological effects of soil drying. Growing tomato plants in a whole plant pressure chamber allowed sequential xylem sap collection from a detached leaf, the petiole stub of an otherwise intact plant and finally the de-topped root system of the same plant, to determine the impact of xylem sap sampling methodology on xylem ABA concentration. Since xylem sap can contain bound forms of ABA, a novel gas chromatography-mass spectrometry (GC-MS) procedure was developed to chemically separate free ABA from two <it>in planta </it>bound ABA forms known as Adducts I and II and ABA-glucose-ester (ABA-GE).</p> <p>Results</p> <p>Xylem sap ABA concentrations were highly dependent on the sampling methodology used: the highest concentrations were detected in sap collected by applying an overpressure to detached leaves following the measurement of leaf water potential. Irrespective of xylem sap source, the wild-type cultivars Ailsa Craig and Rheinlands Ruhm had higher free ABA concentrations than a range of ABA-deficient mutants (<it>notabilis, flacca </it>and <it>sitiens</it>). However, in the mutants, concentrations of bound forms of ABA were similar to wild-type plants, and similar to free ABA concentrations.</p> <p>Conclusions</p> <p>Although xylem concentrations of these bound ABA forms and ABA-GE suggest they have a limited physiological impact on ABA homeostasis in tomato, the methods developed here will allow a more complete understanding of ABA biochemistry and root-to-shoot signalling in species known to have higher concentrations of these compounds.</p> |
| first_indexed | 2024-12-13T23:25:42Z |
| format | Article |
| id | doaj.art-fc6e86681ee748b48ff066c80d7eac84 |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-12-13T23:25:42Z |
| publishDate | 2012-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-fc6e86681ee748b48ff066c80d7eac842022-12-21T23:27:33ZengBMCPlant Methods1746-48112012-03-01811110.1186/1746-4811-8-11Xylem sap collection and extraction methodologies to determine <it>in vivo </it>concentrations of ABA and its bound forms by gas chromatography-mass spectrometry (GC-MS)Netting Andrew GTheobald Julian CDodd Ian C<p>Abstract</p> <p>Background</p> <p>Accurate quantification of xylem sap ABA concentrations is important to underpin models of root-to-shoot ABA signalling to predict the physiological effects of soil drying. Growing tomato plants in a whole plant pressure chamber allowed sequential xylem sap collection from a detached leaf, the petiole stub of an otherwise intact plant and finally the de-topped root system of the same plant, to determine the impact of xylem sap sampling methodology on xylem ABA concentration. Since xylem sap can contain bound forms of ABA, a novel gas chromatography-mass spectrometry (GC-MS) procedure was developed to chemically separate free ABA from two <it>in planta </it>bound ABA forms known as Adducts I and II and ABA-glucose-ester (ABA-GE).</p> <p>Results</p> <p>Xylem sap ABA concentrations were highly dependent on the sampling methodology used: the highest concentrations were detected in sap collected by applying an overpressure to detached leaves following the measurement of leaf water potential. Irrespective of xylem sap source, the wild-type cultivars Ailsa Craig and Rheinlands Ruhm had higher free ABA concentrations than a range of ABA-deficient mutants (<it>notabilis, flacca </it>and <it>sitiens</it>). However, in the mutants, concentrations of bound forms of ABA were similar to wild-type plants, and similar to free ABA concentrations.</p> <p>Conclusions</p> <p>Although xylem concentrations of these bound ABA forms and ABA-GE suggest they have a limited physiological impact on ABA homeostasis in tomato, the methods developed here will allow a more complete understanding of ABA biochemistry and root-to-shoot signalling in species known to have higher concentrations of these compounds.</p>http://www.plantmethods.com/content/8/1/11ABAABA-GE<it>flacca</it>GC-MS<it>notabilis</it>Mutant<it>sitiens</it>TomatoXylem sap |
| spellingShingle | Netting Andrew G Theobald Julian C Dodd Ian C Xylem sap collection and extraction methodologies to determine <it>in vivo </it>concentrations of ABA and its bound forms by gas chromatography-mass spectrometry (GC-MS) Plant Methods ABA ABA-GE <it>flacca</it> GC-MS <it>notabilis</it> Mutant <it>sitiens</it> Tomato Xylem sap |
| title | Xylem sap collection and extraction methodologies to determine <it>in vivo </it>concentrations of ABA and its bound forms by gas chromatography-mass spectrometry (GC-MS) |
| title_full | Xylem sap collection and extraction methodologies to determine <it>in vivo </it>concentrations of ABA and its bound forms by gas chromatography-mass spectrometry (GC-MS) |
| title_fullStr | Xylem sap collection and extraction methodologies to determine <it>in vivo </it>concentrations of ABA and its bound forms by gas chromatography-mass spectrometry (GC-MS) |
| title_full_unstemmed | Xylem sap collection and extraction methodologies to determine <it>in vivo </it>concentrations of ABA and its bound forms by gas chromatography-mass spectrometry (GC-MS) |
| title_short | Xylem sap collection and extraction methodologies to determine <it>in vivo </it>concentrations of ABA and its bound forms by gas chromatography-mass spectrometry (GC-MS) |
| title_sort | xylem sap collection and extraction methodologies to determine it in vivo it concentrations of aba and its bound forms by gas chromatography mass spectrometry gc ms |
| topic | ABA ABA-GE <it>flacca</it> GC-MS <it>notabilis</it> Mutant <it>sitiens</it> Tomato Xylem sap |
| url | http://www.plantmethods.com/content/8/1/11 |
| work_keys_str_mv | AT nettingandrewg xylemsapcollectionandextractionmethodologiestodetermineitinvivoitconcentrationsofabaanditsboundformsbygaschromatographymassspectrometrygcms AT theobaldjulianc xylemsapcollectionandextractionmethodologiestodetermineitinvivoitconcentrationsofabaanditsboundformsbygaschromatographymassspectrometrygcms AT doddianc xylemsapcollectionandextractionmethodologiestodetermineitinvivoitconcentrationsofabaanditsboundformsbygaschromatographymassspectrometrygcms |