Quantitative Determination of the Gibberellins GA3 and GA1 in Spruce (Picea abies [L.]) Needles by a Highly Specific Enzyme Immunoassay

doi:10.1016/S0176-1617(11)80613-9

Journal of Plant Physiology

Volume 139, Issue 2, December 1991, Pages 229-234

https://doi.org/10.1016/S0176-1617(11)80613-9 Get rights and content

Summary

The gibberellins GA₁ and GA₃ were quantitatively determined in needle extracts from spruce (Picea abies [L.]). The phytohormones were purified by several extraction procedures and by HPLC, and detected by a highly sensitive enzyme-linked immunosorbent assay (ELISA). The performance of the EIA was validated by monitoring the parallelism of the logit-log transformed curves of standard and extract dilutions, and by spiking the samples with known amounts of GA. The concentrations of GA₃ were higher than GA₁ in all samples tested so far. Furthermore, GA₃ levels in needles of strongly damaged trees with yellowing symptoms exceeded those of healthy trees.

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Cited by (3)

Identification and expression analysis of Sorghum bicolor gibberellin oxidase genes with varied gibberellin levels involved in regulation of stem biomass
2020, Industrial Crops and Products
Citation Excerpt :
The extracts were collected after centrifugation at 10.000 × g for 15 min and passed through a C-18 Sep-Pak device (MA, USA), then dried in N2. The residue was dissolved in phosphate-buffer solution (0.01 mol L−1, pH 7.4) to determine the levels of GA1 and GA4 using the enzyme-linked immunosorbent assay (ELISA) technique as described by Schneider et al. (1991). The concentrations of GA1 and GA4 in young leaves and stems were determined and expressed as picogram content of GA1 or GA4 per milligram fresh weight (pg/mg FW).
Sweet sorghum (Sorghum bicolor (L.) Moench) is globally evaluated as a key feedstock for bioethanol production due to its high biomass yield. In the present study, the stem biomass characteristics of the sweet sorghum Yajin 1 were revealed and compared with the grain sorghum Aikang 8 at different growth stages. Dynamic analysis of lignocellulosic composition in stems of Yajin 1 and Aikang 8 showed that cellulose content increased and lignin content decreased in the sweet sorghum over development and thus led to a higher saccharification efficiency, which was opposite in the grain sorghum. These varied biomass-associated traits were suggested to be related to changes of bioactive gibberellins GA1 and GA4 levels in young leaves and stems at different growth stages. To investigate the role of gibberellin (GA) metabolism involved in the regulation of stem biomass accumulation and composition in sorghums, a large scale spatio-temporal expression analysis was carried out on the 9 sorghum gibberellin oxidase (SbGAox) genes that encode three classes of key enzymes in the GA biosynthesis. Most of the SbGAox displayed cultivar-, organ-, or stage-preferential expression patterns. Among them, SbGA20ox1 was more specifically expressed in the stem of the sweet sorghum, which was positively related to the stem biomass accumulation and composition, and bioactive GA levels. SbGA2ox1 was more specifically expressed in the grain sorghum than in the sweet sorghum, which was negatively related to the stem biomass accumulation and composition, and bioactive GA levels. As such, these two SbGAox genes may play opposite regulating roles in sorghum stem biomass accumulation and composition by controlling the bioactive GA levels. This study provides a comprehensive understanding of the SbGAox gene family in sorghums, offering a valuable resource to develop strategies for genetic improvement of sorghum biomass traits.
Comparison of sample pretreatment methods for the determination of multiple phytohormones in plant samples by liquid chromatography-electrospray ionization-tandem mass spectrometry
2015, Microchemical Journal
Citation Excerpt :
In the work by Izumi [27], a mixed-mode SPE consisting of Qasis MCX-HLB and Qasis MAX was used for the determination of phytohormones in plant material samples. The recoveries of the target phytohormones ranged from 8.00 ± 1 and 61.0 ± 11% [30]. Concerning LLE, DCM LLE resulted in recoveries ranging from 57.2% (ABA) to 101% (IAA).
Sample preparation is often the bottleneck in the analysis of phytohormones in plant samples. In the present study, sample pretreatment involving extraction by acidified and non-acidified organic solvents and extract purification using solid-phase extraction (SPE) and/or liquid–liquid extraction (LLE) has been optimized and compared for the determination of multiple phytohormones, including auxins, cytokinins (CKs), abscisic acid (ABA), gibberellins (GAs), jasmonic acid (JA) and salicylic acid (SA) in biological samples by liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS) under multiple-reaction monitoring (MRM). A novel and efficient sample pretreatment method for multiple phytohormones in acidified acetonitrile extracts of plant samples using polymer-based mixed-mode cation-exchange solid phase extraction (SPE) with Oasis MCX cartridges combined with ethyl acetate LLE has been developed. The method detection limits (MDLs) for the target phytohormones ranged from 0.0013 to 0.021 ng·mL^− 1, and the recoveries were obtained by spiking the target phytohormones in the two-month-old leaves of oilseed rape (Brassica napus L.) at concentrations of 0.20, 2.0 and 10 ng·g^− 1, which ranged from 75.1 to 111%, 79.6 to 113% and 89.2 to 111%, respectively. The intra-day precisions were in the range of 1.15 to 10.2%, and the inter-day precisions ranged from 2.92 to 12.4%. Additionally, matrix effects were substantially decreased. The proposed sample pretreatment method has been successfully applied to the analysis of multiple phytohormones in the leaf samples of oilseed rape (B. napus L.).
Chapter 3 Immunological methods in plant hormone research
1999, New Comprehensive Biochemistry

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Journal of Plant Physiology

Summary

References (16)

High performance liquid chromatography of gibberellins

J. Chromatogr.

Partition coefficients of 27 gibberellins

Phytochemistry

Auxin distribution in spruce needles

J. Plant Physiol.

Reversed-phase C₁₈ high-performance liquid chromatography of acidic and conjugated gibberellins

J. Chromatogr.

Influence of ozone and acid mist on the contents of Gibberellic Acid (GA₃) in spruce needles (Picea abies) (L.) Karst

Environmental Pollution

Determination of cytokinins in needles of spruce (Picea abies [L.] Karst.) by an indirect enzyme-linked immunosorbent assay

J. Plant Physiol.

Optimum conditions for bonding of plant phenols to insoluble polyvinylpyrrolidone

Phytochemistry

The immunoassay of gibberellins; II. Quantification of GA₃, GA₄ and GA₇ by ultra-sensitive solid phase enzyme immunoassays

Planta

Cited by (3)

Identification and expression analysis of Sorghum bicolor gibberellin oxidase genes with varied gibberellin levels involved in regulation of stem biomass

Comparison of sample pretreatment methods for the determination of multiple phytohormones in plant samples by liquid chromatography-electrospray ionization-tandem mass spectrometry

Chapter 3 Immunological methods in plant hormone research

Quantitative Determination of the Gibberellins GA3 and GA1 in Spruce (Picea abies [L.]) Needles by a Highly Specific Enzyme Immunoassay

Summary

J. Chromatogr.

Phytochemistry

J. Plant Physiol.

J. Chromatogr.

Environmental Pollution

J. Plant Physiol.

Optimum conditions for bonding of plant phenols to insoluble polyvinylpyrrolidone

Phytochemistry

The immunoassay of gibberellins; II. Quantification of GA3, GA4 and GA7 by ultra-sensitive solid phase enzyme immunoassays

Planta

Quantitative Determination of the Gibberellins GA₃ and GA₁ in Spruce (Picea abies [L.]) Needles by a Highly Specific Enzyme Immunoassay

The immunoassay of gibberellins; II. Quantification of GA₃, GA₄ and GA₇ by ultra-sensitive solid phase enzyme immunoassays