Quantification of six types of cytokinins: Integration of an ultra-performance liquid chromatographic-electrospray tandem mass spectrometric method with antibody based immunoaffinity columns equally recognizing cytokinins in free base and nucleoside forms

被引：2

作者：

Zhang, Man ^{[1
]}

Chen, Xiaojiao ^{[2
]}

Zhao, Yajie ^{[1
]}

Zhang, Jiaqi ^{[1
]}

He, Qingqing ^{[1
]}

Qian, Jingqi ^{[1
]}

Tan, Guiyu ^{[1
]}

Liu, Wei ^{[1
]}

Yang, Xiaoling ^{[1
]}

Wang, Baomin ^{[1
]}

机构：

[1] China Agr Univ, Coll Agron, Beijing 100193, Peoples R China

[2] Ningbo Univ, Sch Marine Sci, Key Lab Marine Biotechnol Zhejiang Prov, Ningbo 315211, Peoples R China

来源：

JOURNAL OF CHROMATOGRAPHY A | 2022年 / 1682卷

基金：

中国国家自然科学基金;

关键词：

Cytokinins; Monoclonal antibody; Immunoaffinity cleanup; UPLC-ESI-MS; MS; Gossypium hirsutum L; MONOCLONAL-ANTIBODIES; ISOPRENOID CYTOKININS; PLANT CYTOKININ; ABSCISIC-ACID; PURIFICATION; ARABIDOPSIS; AUXIN; GROWTH;

D O I：

10.1016/j.chroma.2022.463497

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Cytokinins (CTKs) exist in various types in plants. The accurate quantification of free base and nucleo-side types of cytokinins are helpful for better understanding their physiological role. In the present study, antibodies against trans-zeatin riboside (tZR) and N6-isopentenyladenine riboside (iPR) antibodies with equal recognition to free base and nucleoside cytokinins were developed. The cross-reactivity of tZR mAb 3G101G7 with tZR, trans-zeatin (tZ), dihydrozeatin riboside (DHZR), dihydrozeatin (DHZ), iPR, and N6- isopentenyladenine (iP) was 100.0%, 95.7%, 19.1%, 18.0%, 1.1%, and 0.7%, and that of iPR mAb 5C82F1 with above-mentioned 6 types of cytokinins was 1.5%, 1.4%, 5.7%, 3.1%, 100.0% and 92.6%, respectively. The ob-tained antibodies were used to prepare two immunoaffinity columns (IAC). The elution efficiencies of tZR 3G101G7-IAC for tZ and tZR, DHZ and DHZR and of iPR 5C82F1-IAC for iP and iPR were almost no dif-ference with the same loading amount on their corresponding IACs. Subsequently, six types of cytokinins in mepiquat chloride (MC)-treated cotton ( Gossypium hirsutum L. ) roots were determined by IACs com-bined with ultra-performance liquid chromatography-electrospray tandem mass spectrometry (UPLC-ESI-MS/MS). The contents of tZR, iPR and DHZR were increased by 9.3-38.5%, 6.6-23.5%, and 30.1-110.0%, respectively, whereas those of tZ and iP were reduced by 5.3-20.0% and 27.7-32.1%, respectively. The decreased tZ and iP levels led to the ratio of auxin-to-active cytokinins increase to promote lateral root initiation in MC-treated cotton seeding. Integration of the IACs equally recognizing cytokinins in their free base and nucleoside forms with UPLC-ESI-MS/MS can accurately quantify different cytokinins in plant tis-sues.(c) 2022 Elsevier B.V. All rights reserved.

引用

页数：10