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Journal of Integrative Agriculture  2020, Vol. 19 Issue (3): 700-708    DOI: 10.1016/S2095-3119(19)62640-7
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Detection of seven phytohormones in peanut tissues by ultra-high-performance liquid chromatography-triple quadrupole tandem mass spectrometry
WANG Hai-xia1, 2, WANG Ming-lun2, WANG Xiu-zhong2, DING Yu-long1, 3   
1 College of Biology and the Environment, Nanjing Forestry University, Nanjing 210037, P.R.China
2 College of Chemistry and Pharmaceutical Sciences, Qingdao Agricultural University, Qingdao 266109, P.R.China
3 Co-innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, P.R.China
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Abstract  
Development of highly sensitive and reliable method for detection of phytohormones is of great significance to study plant hormones and agricultural production.  In this study, an ultra-high-performance liquid chromatography-mass spectrometry/mass spectrometry method was established for separation and quantification of trans-zeatin, trans-zeatin riboside, gibberellin A3, indol-3-acetic acid, salicylic acid, abscisic acid, and jasmonic acid (JA) without any label.  The separation was performed on an Agilent Explus Plus C18 column by using methanol and water as mobile phases with gradient elution.  The target compounds were confirmed and quantified by mass spectrum via positive electrospray ionization for trans-zeatin, trans-zeatin riboside, indole-3-acetic acid, and via negative electrospray ionization for gibberellin3, salicylic acid, abscisic acid, and JA.  The limits of detection ranged from 0.0127 ng L–1 for gibberellin A3 (GA3) to 33.26 ng L–1 for JA and were lower than the currently reported values in literature.  The proposed method was applied for qualitative and quantitative analyses of phytohormones in peanut gynophores and pods.  The recoveries of the spiked phytohormones ranged from 80.20 to 102.56%.  The contents of seven endogenous hormones varied specifically in different development stages of peanuts.  This study provides a highly sensitive and selective detection method for hormones and elucidates the growth and development of the gynophore and peanut fruit, which are controlled by seven endogenous hormones.
 
Keywords:  phytohormones        simultaneous detection        UHPLC-MS/MS        peanut  
Received: 23 November 2018   Accepted:
Fund: This work was supported by the earmarked fund for China Agriculture Research System (CARS-14) and partly supported by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China (RAPD). We also thank the Natural Science Foundation of Shandong Province, China (ZR2018MB030).
Corresponding Authors:  Correspondence WANG Xiu-zhong, Tel: +86-532-58957442, E-mail: xzwang@qau.edu.cn; DING Yu-long, E-mail: ylding@vip.163.com   

Cite this article: 

WANG Hai-xia, WANG Ming-lun, WANG Xiu-zhong, DING Yu-long . 2020. Detection of seven phytohormones in peanut tissues by ultra-high-performance liquid chromatography-triple quadrupole tandem mass spectrometry. Journal of Integrative Agriculture, 19(3): 700-708.

Balcke G U, Handrick V, Bergau N, Fichtner M, Henning A, Stellmach H, Tissier A, Hause B, Frolov A. 2012. An UPLC-MS/MS method for highly sensitive high-throughput analysis of phytohormones in plant tissues. Plant Methods, 8, 47–57.
Beilby M J, Turi C E, Baker T C, Tymm F J M, Murch S J. 2015. Circadian changes in endogenous concentrations of indole-3-acetic acid, melatonin, serotonin, abscisic acid and jasmonic acid in Characeae (Chara australis Brown). Plant Signaling & Behavior, 10, e1082697.
Boerjan W, Genetello C, Van Montagu M, Inzé D. 1992. A new bioassay for auxins and cytokinins. Plant Physiology, 99, 1090–1098.
Bosco R, Caser M, Ghione G G, Mansuino A, Giovannini A, Scariot V. 2015. Dynamics of abscisic acid and indole-3-acetic acid during the early-middle stage of seed development in Rosa hybrida. Plant Growth Regulation, 75, 265–270.
Cao Z Y, Sun L H, Mou R X, Zhang L P, Lin X Y, Zhu Z W, Chen M X. 2016. Profiling of phytohormones and their major metabolites in rice using binary solid-phase extraction and liquid chromatography-triple quadrupole mass spectrometry. Journal of Chromatography, 1451, 67–74.
Chiwocha S D S, Abrams S R, Ambrose, S J, Cutler A J, Loewen M, Ross A R S, Kermode A R. 2003. A method for profiling classes of plant hormones and their metabolites using liquid chromatography-electrospray ionization tandem mass spectrometry: an analysis of hormone regulation of thermodormancy of lettuce (Lactuca sativa L.) seeds. The Plant Journal, 35, 405–417.
Creelman R A, Mullet J E. 1995. Jasmonic acid distribution and action in plants: Regulation during development and response to biotic and abiotic stress. Proceedings of the National Academy of Sciences of the United States of America, 92, 4114–4119.
Daie J, Wyse R. 1982. Adaptation of the enzyme-linked immunosorbent assay (ELISA) to the quantitative analysis of abscisic acid. Analytical Biochemistry, 119, 365–371.
Fletcher A T, Mader J C. 2007. Hormone profiling by LC-QToF-MS/MS in dormant macadamia integrifolia: Correlations with abnormal vertical growth. Journal of Plant Growth Regulation, 26, 351–361.
Fu J H, Sun X H, Wang J D, Chu J F, Yan C Y. 2011. Progress in quantitative analysis of plant hormones. Chinese Science Bulletin, 56, 355–366.
Giannarelli S, Muscatello B, Bogani P, Spiriti M M, Buiatti M, Fuoco R. 2010. Comparative determination of some phytohormones in wild-type and genetically modified plants by gas chromatography-mass spectrometry and high-performance liquid chromatography-tandem mass spectrometry. Analytical Biochemistry, 398, 60–68.
Gunes A, Inal A, Alpaslan M, Eraslan F, Bagci E G, Cicek N. 2007. Salicylic acid induced changes on some physiological parameters symptomatic for oxidative stress and mineral nutrition in maize (Zea mays L.) grown under salinity. Journal of Plant Physiology, 164, 728–736.
Hu X H, Jiang Y P, Miao H R, Shi Y Q, Chen J. 2014. Study on the improvement of HPLC method for analyzing endogenous hormone in peanut. Journal of Peanut Science, 43, 50–53. (in Chinese)
Kojima M, Kamada-Nobusada T, Komatsu H, Takei K, Kuroha T, Mizutani M, Ashikari M, Ueguchi-Tanaka M, Matsuoka M, Suzuki K, Sakakibara H. 2009. Highly sensitive and high-throughput analysis of plant hormones using MS-probe modification and liquid chromatography-tandem mass spectrometry: An application for hormone profiling in Oryza sativa. Plant and Cell Physiology, 50, 1201–1214.
Liu H Y, Guo T C, Liu S B, Zhu Y J, Yue C F, Wang C Y, Yang W P, Ma D Y, Wang H H. 2006. Effect of spraying salicylic acid (SA) at anthesis on grain starch properties and yield of winter wheat cultivars with different end-use qualities. Journal of Triticeae Crops, 26, 123–127. (in Chinese)
Luo B, Liu F Z, Wan Y S, Zhang K, Zhao W X. 2013. Dynamic changes of endogenous hormones content and dry matter accumulation of pods and kernels in different varieties (lines) of peanut (Arachis hypogaea L.). Acta Agronomica Sinica, 39, 2083–2093. (in Chinese)
Luo X T, Cai B D, Chen X, Feng Y Q. 2017. Improved methodology for analysis of multiple phytohormones using sequential magnetic solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry. Analytica Chimica Acta, 983, 112–120.
Ma C, Feng Y L, Zhang J, Wang H Z, Yuan J L, L Y J. 2017. Effects of exogenous methyl jasmonate on endogenous hormones and yield formation in wheat after anthesis under drought stress. Plant Physiology Journal, 53, 1051–1058.
Maksimov I V, Yarullina L G. 2007. Salicylic acid and local resistance to pathogens. Salicylic Acid: A Plant Hormone, 11, 323–334.
Van Meulebroek L, Bussche J V, Steppe K, Vanhaecke L. 2012. Ultra-high performance liquid chromatography coupled to high resolution Orbitrap mass spectrometry for metabolomic profiling of the endogenous phytohormonal status of the tomato plant. Journal of Chromatography (A), 1260, 67–80.
Oklestkova J, Tarkowska D, Eyer L, Elbert T, Marek A, Smrzova Z, Novak O, Franek M, Zhabinskii V N, Strnad M. 2017. Immunoaffinity chromatography combined with tandem mass spectrometry: A new tool for the selective capture and analysis of brassinosteroid plant hormones. Talanta, 170, 432–440.
Pan X, Welti R, Wang X. 2008. Simultaneous quantification of major phytohormones and related compounds in crude plant extracts by liquid chromatography-electrospray tandem mass spectrometry. Phytochemistry, 69, 1773–1781.
Pan X, Welti R, Wang X. 2010. Quantitative analysis of major plant hormones in crude plant extracts by high-performance liquid chromatography-mass spectrometry. Nature Protocols, 5, 986–992.
Santner A, Calderon-Villalobos L I A, Estelle M. 2009. Plant hormones are versatile chemical regulators of plant growth. Nature Chemical Biology, 5, 301–307.
Santner A, Estelle M. 2009. Recent advances and emerging trends in plant hormone signalling. Nature, 459, 1071–1078.
Shlamovitz N, Ziv M, Zamski E. 1995. Light dark and growth regulator involvement in groundnut (Arachis hypogaea L.) pod development. Plant Growth Regulation, 16, 37–42.
Shushu D D, Cutter E G. 1990. Growth of the gynophore of the peanut Arachis hypogaea. L. Intact and decapitated gynophores. Canadian Journal of Botany, 68, 955–964.
Šimura J, Antoniadi I, Široká J, Tarkowska D, Strnad M, Ljung K, Novak O. 2018. Plant hormonomics: Multiple phytohormone profiling by targeted metabolomics. Plant Physiology, 177, 476–489.
Stamm P, Kumar P P. 2010. The phytohormone signal network regulating elongation growth during shade avoidance. Journal of Experimental Botany, 61, 2889–2903.
Witek S, Wozniak B, Matraszek-Zuchowska I, Posyniak A. 2018. Analysis of thyreostats in bovine feces using ultra high performance liquid chromatography with tandem mass spectrometry. Journal of Separation Science, 41, 1083–1090.
Yang J C, Peng S B, Gu S L, Visperas R M, Zhu Q S. 2001. Changes in zeatin and zeatin riboside content in rice grains and roots during grain filling and relationship to grain plumpness. Acta Agronomica Sinica, 27, 35–42. (in Chinese)
Yang W B, W Z, Yin Y P, Li W Y, Li Y, Chen X G, Wang P, Chen E Y, Guo J X, Cai T, Ni Y L. 2011. Effects of spraying exogenous ABA or GA on the endogenous hormones concentration and filling of wheat grains. Scientia Agricultura Sinica, 44, 2673–2682. (in Chinese)
Yuan Y, Zhou R, Li D L, Luo C, Li G Y. 2018. Simultaneous quantitative assessment of nine glycosides in tobacco by liquid chromatography-tandem mass spectrometry. Journal of Separation Science, 41, 1009–1016.
Zhang J G, Huang X Y, Ma Y B, Zhang X M, Chen J J, Geng C A. 2018. Dereplication-guided isolation of a new indole alkaloid triglycoside from the hooks of Uncaria rhynchophylla by LC with ion trap time-of-flight MS. Journal of Separation Science, 41, 1532–1538.
 
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