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拟南芥 2C 蛋白磷酸酶家族的功能筛选发现 PP2C15 通过靶向 BR 受体激酶 1 负调控植物免疫。

Functional screening of the Arabidopsis 2C protein phosphatases family identifies PP2C15 as a negative regulator of plant immunity by targeting BRI1-associated receptor kinase 1.

机构信息

Department of Biotechnology and Applied Biology, College of Life Sciences, Sichuan Agricultural University, Ya'an, Sichuan, China.

Chengdu Lusyno Biotechnology Co., Ltd., Chengdu, China.

出版信息

Mol Plant Pathol. 2024 Apr;25(4):e13447. doi: 10.1111/mpp.13447.

Abstract

Genetic engineering using negative regulators of plant immunity has the potential to provide a huge impetus in agricultural biotechnology to achieve a higher degree of disease resistance without reducing yield. Type 2C protein phosphatases (PP2Cs) represent the largest group of protein phosphatases in plants, with a high potential for negative regulatory functions by blocking the transmission of defence signals through dephosphorylation. Here, we established a PP2C functional protoplast screen using pFRK1::luciferase as a reporter and found that 14 of 56 PP2Cs significantly inhibited the immune response induced by flg22. To verify the reliability of the system, a previously reported MAPK3/4/6-interacting protein phosphatase, PP2C5, was used; it was confirmed to be a negative regulator of PAMP-triggered immunity (PTI). We further identified PP2C15 as an interacting partner of BRI1-associated receptor kinase 1 (BAK1), which is the most well-known co-receptor of plasma membrane-localized pattern recognition receptors (PRRs), and a central component of PTI. PP2C15 dephosphorylates BAK1 and negatively regulates BAK1-mediated PTI responses such as MAPK3/4/6 activation, defence gene expression, reactive oxygen species bursts, stomatal immunity, callose deposition, and pathogen resistance. Although plant growth and 1000-seed weight of pp2c15 mutants were reduced compared to those of wild-type plants, pp2c5 mutants did not show any adverse effects. Thus, our findings strengthen the understanding of the mechanism by which PP2C family members negatively regulate plant immunity at multiple levels and indicate a possible approach to enhance plant resistance by eliminating specific PP2Cs without affecting plant growth and yield.

摘要

利用植物免疫负调控因子的基因工程有可能在农业生物技术中提供巨大的推动力,在不降低产量的情况下实现更高程度的抗病性。2C 型蛋白磷酸酶(PP2Cs)是植物中最大的蛋白磷酸酶家族之一,通过去磷酸化阻断防御信号的传递,具有很强的负调控功能。在这里,我们使用 pFRK1::luciferase 作为报告基因建立了一个 PP2C 功能原生质体筛选系统,发现 56 个 PP2C 中的 14 个显著抑制了 flg22 诱导的免疫反应。为了验证该系统的可靠性,我们使用了之前报道的 MAPK3/4/6 相互作用蛋白磷酸酶 PP2C5,证实其是 PAMP 触发免疫(PTI)的负调控因子。我们进一步鉴定出 PP2C15 是 BRASSINOSTEROID-INSENSITIVE 1 ASSOCIATED RECEPTOR KINASE 1(BAK1)的相互作用伙伴,BAK1 是质膜定位模式识别受体(PRRs)最著名的共受体,也是 PTI 的核心组成部分。PP2C15 使 BAK1 去磷酸化,负调控 BAK1 介导的 PTI 反应,如 MAPK3/4/6 激活、防御基因表达、活性氧爆发、气孔免疫、胼胝质沉积和病原体抗性。尽管与野生型植物相比,pp2c15 突变体的植物生长和千粒重降低,但 pp2c5 突变体没有表现出任何不良影响。因此,我们的研究结果加强了对 PP2C 家族成员在多个水平上负调控植物免疫的机制的理解,并表明通过消除特定的 PP2C 而不影响植物生长和产量来增强植物抗性的一种可能方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1da/10984862/4d72cdd9d616/MPP-25-e13447-g005.jpg

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