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利用基于水母发光蛋白的钙成像系统筛选保卫细胞对CO/HCO的响应突变体。

guard cell CO/HCO response mutant screening by an aequorin-based calcium imaging system.

作者信息

Tang Mengmeng, Zhao Xiaowei, Hu Yinling, Zeng Miaomiao, Wang Kai, Dong Nannan, Ma Xiaonan, Bai Ling, Song Chun-Peng

机构信息

1State Key Laboratory of Crop Stress Adaptation and Improvement, School of Life Sciences, Henan University, Kaifeng, 475004 China.

Biocover Technology Development Co., Ltd, Shixing Street, Shijingshan District, Beijing, 100195 China.

出版信息

Plant Methods. 2020 Apr 29;16:59. doi: 10.1186/s13007-020-00600-w. eCollection 2020.

DOI:10.1186/s13007-020-00600-w
PMID:32368249
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7189508/
Abstract

BACKGROUND

The increase in atmospheric CO is causing a number of changes in plant growth such as increases in leaf area and number, branching, plant size and biomass, and growth rate. Despite the importance of stomatal responses to CO, little is known about the genetic and molecular mechanisms that mediate stomatal development and movement in response to CO levels. Deciphering the mechanisms that sense changes in CO and/or HCO concentration is critical for unraveling the role of CO in stomatal development movement. In , CO-induced stomatal closure is strongly Ca-dependent. To further dissect this signaling pathway and identify new components in the CO response pathway, we recorded [Ca] changes in mutagenized leaves and screened for mutants with abnormal guard cell behavior in response to CO/HCO .

RESULTS

We observed that 1 mM HCO induces [Ca] transient changes in guard cells and stomatal closure both in light and darkness. The changes in [Ca] induced by HCO could be detected by an aequorin-based calcium imaging system. Using this system, we identified a number of mutants defective in both [Ca] changes and the stomatal response to CO/HCO .

CONCLUSIONS

We provide a sensitive method for isolating stomatal CO/HCO response genes that function early in stomatal closure and that have a role in regulating [Ca]. This method will be helpful in elucidating the Ca-dependent regulation of guard cell behavior in response to CO/HCO .

摘要

背景

大气中二氧化碳浓度的增加正在引起植物生长的一系列变化，例如叶面积、叶片数量、分枝、植株大小和生物量以及生长速率的增加。尽管气孔对二氧化碳的响应很重要，但对于介导气孔发育和响应二氧化碳水平而运动的遗传和分子机制却知之甚少。阐明感知二氧化碳和/或碳酸氢根离子浓度变化的机制对于揭示二氧化碳在气孔发育运动中的作用至关重要。在拟南芥中，二氧化碳诱导的气孔关闭强烈依赖于钙离子。为了进一步剖析这一信号通路并鉴定二氧化碳响应通路中的新组分，我们记录了诱变拟南芥叶片中的钙离子变化，并筛选对二氧化碳/碳酸氢根离子有异常保卫细胞行为的突变体。

结果

我们观察到1 mM碳酸氢根离子在光照和黑暗条件下均能诱导保卫细胞中钙离子的瞬时变化和气孔关闭。基于水母发光蛋白的钙成像系统可以检测到由碳酸氢根离子诱导的钙离子变化。利用该系统，我们鉴定出了一些在钙离子变化和对二氧化碳/碳酸氢根离子的气孔响应方面均有缺陷的拟南芥突变体。

结论

我们提供了一种灵敏的方法来分离在气孔关闭早期起作用且在调节钙离子方面有作用的气孔二氧化碳/碳酸氢根离子响应基因。该方法将有助于阐明保卫细胞对二氧化碳/碳酸氢根离子响应的钙离子依赖性调节机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/f0146d13fa7d/13007_2020_600_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/213c32ad9ea1/13007_2020_600_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/375c44e7341d/13007_2020_600_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/9edd739d1f68/13007_2020_600_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/f0146d13fa7d/13007_2020_600_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/213c32ad9ea1/13007_2020_600_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/375c44e7341d/13007_2020_600_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/9edd739d1f68/13007_2020_600_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8da1/7189508/f0146d13fa7d/13007_2020_600_Fig4_HTML.jpg

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