A Monoclonal Antibody to (S)-abscisic Acid: Its Characterisation and Use in a Radioimmunoassay for Measuring Abscisic Acid in Crude Extracts of Cereal and Lupin Leaves

Overview

Journal Planta

Specialty Biology

Date 2013 Nov 15

PMID 24226540

Citations 66

Authors

S A Quarrie

P N Whitford

N E Appleford

T L Wang

S K Cook

I E Henson

B R Loveys

Affiliations

Soon will be listed here.

Abstract

A monoclonal antibody produced to abscisic acid (ABA) has been characterised and the development of a radioimmunoassay (RIA) for ABA using the antibody is described. The antibody had a high selectivity for the free acid of (S)-cis, trans-ABA. Using the antibody, ABA could be assayed reliably in the RIA over a range from 100 to 4000 pg (0.4 to 15 pmol) ABA per assay vial. As methanol and acetone affected ABA-antibody binding, water was used to extract ABA from leaves. Water was as effective as aqueous methanol and acetone in extracting the ABA present. Crude aqueous extracts of wheat, maize and lupin leaves could be analysed without serious interference from other immunoreactive material. This was shown by measuring the distribution of immunoreactivity in crude extracts separated by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), or by comparing the assay with physicochemical methods of analysis. Analysis of crude extracts by RIA and either, after TLC purification, by gas chromatography using an electron-capture detector or, after HPLC purification, by combined gas chromatography-mass spectrometry (GC-MS) gave very similar ABA concentrations in the initial leaf samples. However, RIA analysis of crude aqueous extracts of pea seeds resulted in considerable overestimation of the amount of ABA present. Determinations of ABA content by GC-MS and RIA were similar after pea seed extracts had been purified by HPLC. Although the RIA could not be used to analyse ABA in crude extracts of pea seeds, it is likely that crude extracts of leaves of several other species may be assayed successfully.

Citing Articles

Silicon improves root functioning and water management as well as alleviates oxidative stress in oilseed rape under drought conditions.

Saja-Garbarz D, Libik-Konieczny M, Janowiak F Front Plant Sci. 2024; 15:1359747.

PMID: 38450404 PMC: 10915341. DOI: 10.3389/fpls.2024.1359747.

Species-specific variation of photosynthesis and mesophyll conductance to ozone and drought in three Mediterranean oaks.

Hoshika Y, Paoletti E, Centritto M, Gomes M, Puertolas J, Haworth M Physiol Plant. 2022; 174(1):e13639.

PMID: 35092611 PMC: 9303399. DOI: 10.1111/ppl.13639.

Isohydricity of Two Different Citrus Species under Deficit Irrigation and Reclaimed Water Conditions.

Romero-Trigueros C, Gambin J, Nortes Tortosa P, Cabanero J, Nicolas Nicolas E Plants (Basel). 2021; 10(10).

PMID: 34685931 PMC: 8538605. DOI: 10.3390/plants10102121.

Changes in Ethylene, ABA and Sugars Regulate Freezing Tolerance under Low-Temperature Waterlogging in .

Jurczyk B, Pociecha E, Janowiak F, Dziurka M, Koscik I, Rapacz M Int J Mol Sci. 2021; 22(13).

PMID: 34206693 PMC: 8268127. DOI: 10.3390/ijms22136700.

Accumulation of Silicon and Changes in Water Balance under Drought Stress in var. L.

Saja-Garbarz D, Ostrowska A, Kaczanowska K, Janowiak F Plants (Basel). 2021; 10(2).

PMID: 33535676 PMC: 7912841. DOI: 10.3390/plants10020280.

References

Quarrie S . A rapid and sensitive assay for abscisic acid using ethyl abscisate as an internal standard. Anal Biochem. 1978; 87(1):148-56. DOI: 10.1016/0003-2697(78)90579-1. View

Ciha A, Brenner M, Brun W . Rapid separation and quantification of abscisic Acid from plant tissues using high performance liquid chromatography. Plant Physiol. 1977; 59(5):821-6. PMC: 543303. DOI: 10.1104/pp.59.5.821. View

Little C, Heald J, Browning G . Identification and measurement of indoleacetic and abscisic acids in the cambial region of Picea sitchensis (Bong.) Carr. by combined gas chromatography-mass spectrometry. Planta. 2014; 139(2):133-8. DOI: 10.1007/BF00387138. View

Daie J, Wyse R . Adaptation of the enzyme-linked immunosorbent assay (ELISA) to the quantitative analysis of abscisic acid. Anal Biochem. 1982; 119(2):365-71. DOI: 10.1016/0003-2697(82)90599-1. View

Rosher P, Jones H, Hedden P . Validation of a radioimmunoassay for (+)-abscisic acid in extracts of apple and sweet-pepper tissue using high-pressure liquid chromatography and combined gas chromatography-mass spectrometry. Planta. 2013; 165(1):91-9. DOI: 10.1007/BF00392216. View

Fuchs Y, Mayak S, Fuchs S . Detection and quantitative determination of abscisic acid by immunological assay. Planta. 2014; 103(2):117-25. DOI: 10.1007/BF00387362. View

Quarrie S, Galfre G . Use of different hapten-protein conjugates immobilized on nitrocellulose to screen monoclonal antibodies to abscisic acid. Anal Biochem. 1985; 151(2):389-99. DOI: 10.1016/0003-2697(85)90193-9. View

Mertens R, Eberle J, Arnscheidt A, Ledebur A, Weiler E . Monoclonal antibodies to plant growth regulators. II. Indole-3-acetic acid. Planta. 2013; 166(3):389-93. DOI: 10.1007/BF00401177. View

Walton D, Dashek W, Galson E . A radioimmunoassay for abscisic acid. Planta. 2013; 146(2):139-45. DOI: 10.1007/BF00388224. View

10.

Weiler E . Radioimmunoassays for the differential and direct analysis of free and conjugated abscisic acid in plant extracts. Planta. 2013; 148(3):262-72. DOI: 10.1007/BF00380037. View

11.

Sweetser P, Vatvars A . High-performance liquid chromatographic analysis of abscisic acid in plant extracts. Anal Biochem. 1976; 71(1):68-78. DOI: 10.1016/0003-2697(76)90012-9. View

12.

Weiler E . Radioimmunoassay for the determination of free and conjugated abscisic acid. Planta. 2014; 144(3):255-63. DOI: 10.1007/BF00388767. View