Shandong Provincial Key Laboratory of Plant Stress, College of Life Sciences, Shandong Normal University, Wenhua East Road 88, Jinan 250014, China.
Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI 53211, USA.
J Integr Plant Biol. 2018 Jul;60(7):591-607. doi: 10.1111/jipb.12649. Epub 2018 Jun 5.
Hydrogen peroxide (H O ) is generated in many metabolic processes. As a signaling molecule, H O plays important roles in plant growth and development, as well as environmental stress response. In Arabidopsis, there are three catalase genes, CAT1, CAT2, and CAT3. The encoded catalases are predominately peroxisomal proteins and are critical for scavenging H O . Since CAT1 and CAT3 are linked on chromosome 1, it has been almost impossible to generate cat1/3 and cat1/2/3 mutants by traditional genetic tools. In this study, we constructed cat1/3 double mutants and cat1/2/3 triple mutants by CRISPR/Cas9 to investigate the role of catalases. The cat1/2/3 triple mutants displayed severe redox disturbance and growth defects under physiological conditions compared with wild-type and the cat2/3 double mutants. Transcriptome analysis showed a more profound transcriptional response in the cat1/2/3 triple mutants compared to the cat2/3 mutants. These differentially expressed genes are involved in plant growth regulation as well as abiotic and biotic stress responses. In addition, expression of OXI1 (OXIDATIVE SIGNAL INDUCIBLE 1) and several MAPK cascade genes were changed dramatically in the catalase triple mutant, suggesting that H O produced in peroxisomes could serve as a peroxisomal retrograde signal.
过氧化氢(H2O2)在许多代谢过程中产生。作为一种信号分子,H2O2 在植物生长发育以及环境胁迫响应中发挥着重要作用。在拟南芥中,有三个过氧化氢酶基因,CAT1、CAT2 和 CAT3。编码的过氧化氢酶主要是过氧化物酶体蛋白,对于清除 H2O2 至关重要。由于 CAT1 和 CAT3 位于染色体 1 上,因此几乎不可能通过传统的遗传工具生成 cat1/3 和 cat1/2/3 突变体。在这项研究中,我们通过 CRISPR/Cas9 构建了 cat1/3 双突变体和 cat1/2/3 三突变体,以研究过氧化氢酶的作用。与野生型和 cat2/3 双突变体相比,cat1/2/3 三突变体在生理条件下表现出严重的氧化还原失调和生长缺陷。转录组分析表明,cat1/2/3 三突变体的转录响应比 cat2/3 突变体更为深刻。这些差异表达的基因参与植物生长调控以及非生物和生物胁迫响应。此外,在过氧化氢酶三突变体中,OXI1(OXIDATIVE SIGNAL INDUCIBLE 1)和几个 MAPK 级联基因的表达发生了显著变化,表明过氧化物酶体中产生的 H2O2 可以作为过氧化物酶体逆行信号。