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Abstract
The involvement of plant immunophilins in multiple essential processes such as development, various ways of adapting to biotic and abiotic stresses, and photosynthesis has already been established. Previously, research has demonstrated the involvement of three immunophilin genes (AtCYP19-1/ROC3, AtFKBP65/ROF2, and AtCYP57) in the control of plant response to invasion by various pathogens. Current research attempts to identify host target proteins for each of the selected immunophilins. As a result, candidate interactors have been determined and confirmed using a yeast 2-hybrid (Y2H) system for protein–protein interaction assays. The generation of mutant isoforms of ROC3 and AtCYP57 harboring substituted amino acids in the in silico-predicted active sites became essential to achieving significant binding to its target partners. This data shows that ROF2 targets calcium-dependent lipid-binding domain-containing protein (At1g70790; AT1) and putative protein phosphatase (At2g30020; АТ2), whereas ROC3 interacts with GTP-binding protein (At1g30580; ENGD-1) and RmlC-like cupin (At5g39120). The immunophilin AtCYP57 binds to putative pyruvate decarboxylase-1 (Pdc1) and clathrin adaptor complex-related protein (At5g05010). Identified interactors confirm our previous findings that immunophilins ROC3, ROF2, and AtCYP57 are directly involved with stress response control. Further, these findings extend our understanding of the molecular functional pathways of these immunophilins.
Highlights
In the course of evolution, plants have developed various ways to adapt to and counter many biotic and abiotic stresses using physiological and molecular mechanisms
The main molecular tools utilized by plants to resist stress are the activation of specific ion channels; accumulation of reactive oxygen species (ROS) [1,2]; activation of protective kinase cascades [3]; and changes in the synthesis of phytohormones such as abscisic acid (ABA), salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) [4]
Of the 175 clones, 42 were able to grow on Quadruple dropout medium (QDO) medium after replating
Summary
In the course of evolution, plants have developed various ways to adapt to and counter many biotic and abiotic stresses using physiological and molecular mechanisms. Almost all abiotic stresses in plant cells temporarily increase the concentration of free cytosolic Ca2+ in the first seconds of the response [9,10]. It has been shown that if large concentrations of ROS lead to cell death, lower amounts regulate the cellular response processes under stressful conditions [11,12,13]. This is one of the main functions of ROS: to serve as signaling molecules in cells [14,15]
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