Single-cell-type transcriptomic analysis reveals distinct gene expression profiles in wheat guard cells in response to abscisic acid
Junbin Wang A B # , Yang Li A # , Tianwen Wu A # , Chen Miao C , Meijuan Xie A , Bo Ding A , Ming Li A , Shuguang Bao A , Xiaoqiang Chen A , Zhaorong Hu D and Xiaodong Xie![https://orcid.org/0000-0001-8434-8423](/media/client/orcid_16x16.png)
A International Joint Center for the Mechanismic Dissection and Genetic Improvement of Crop Stress Tolerance, College of Agriculture & Resources and Environmental Sciences, Tianjin Agricultural University, Tianjin 300392, China.
B College of Basic Sciences, Tianjin Agricultural University, Tianjin 300392, China.
C State Key Laboratory of Crop Stress Adaptation and Improvement, School of Life Sciences, Henan University, Kaifeng 475001, China.
D State Key Laboratory for Agrobiotechnology, Key Laboratory of Crop Heterosis Utilization (MOE) and Key Laboratory of Crop Genetic Improvement, China Agricultural University, Beijing 100193, China.
Handling Editor: Sergey Shabala
Functional Plant Biology 48(11) 1087-1099 https://doi.org/10.1071/FP20368
Submitted: 28 November 2020 Accepted: 5 July 2021 Published: 23 September 2021
© 2021 The Author(s) (or their employer(s)). Published by CSIRO Publishing
Abstract
Stomatal closure, driven by shrinking guard cells in response to the accumulation of abscisic acid (ABA) under drought stress, has a great impact on plant growth and environmental acclimation. However, the molecular regulatory mechanism underlying the turgor alteration of guard cells remains elusive, especially in cereal grasses. Here, we develop a modified enzyme digestion-based approach for the isolation of wheat (Triticum aestivum L.) guard cells. With this approach, we can remove mesophyll, pavement cells and subsidiary cells successively from the epidermis of the trichomeless coleoptile in wheat and preserve guard cells on the cuticle layers in an intact and physiologically active conditions. Using a robust single-cell-type RNA sequencing analysis, we discovered 9829 differentially expressed genes (DEGs) as significantly up- or down-regulated in guard cells in response to ABA treatment. Transcriptome analysis revealed a large percent of DEGs encoding multiple phytohormone signalling pathways, transporters, calcium signalling components, protein kinases and other ABA signalling-related proteins, which are primarily involved in key signalling pathways in ABA-regulated stomatal control and stress response. Our findings provide valuable resource for investigating the transcriptional regulatory mechanism underlying wheat guard cells in response to ABA.
Keywords: abscisic acid, gene expression, guard cells isolation, phytohormone signaling pathways, RNA sequencing, stomatal signalling pathways, stress response, Triticum aestivum L.
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