Abstract
Plant cytoskeleton regulation has been studied using a new approach based on both (1) pharmacological analysis of tubulin and actin inhibitors and (2) mechanical stimulation achieved by using a slow-rotating (2 rpm) clinostat in combination with transcriptional analysis of genes encoding TUA6, ACT2, MAP65-1, CLASP, PLDδ, FH4 and FH1 proteins in Arabidopsis thaliana seedling roots. The obtained data suggest feedback between the organization of microtubule (MT) and actin filament (AF) networks and the expression of the ACT2, TUA6, MAP65-1, CLASP and FH1/FH4 genes. Different regulation of feedback between MT/AF organization and TUA6, ACT2, MAP65-1, CLASP, FH4 and FH1 gene expression was noted during slow clinorotation, possibly due to altered mechanical impact on the cortical cytoskeleton. For the first time, the expression of the tubulin-associated gene MAP65-1 was shown to be dependent upon the organization of AFs. TUA6, MAP65-1, CLASP, FH1 and FH4 likely participate in mechanical signal transduction. Our work demonstrated that slow clinorotation is able to cause mechanical stress.
Highlights
In interphase plant cells, arrays of cortical microtubules, which are composed of microtubule (MT) bundles and separate MTs, are located beneath the plasma membrane (PM) and arranged transversely to the cell axis
Taking the abovementioned into account, we investigated the expression of the ACT2, TUA6, cytoplasmic linker associated protein (CLASP), MAP651, phospholipase D delta (PLDd), FH1 and FH4 genes in clinorotated A. thaliana plants treated with either an inhibitor of tubulin polymerization (ORY) or an inhibitor of actin polymerization (CD)
Our experiments have shown that the organization of cortical microtubules (cMTs) and actin filament (AF) is able to regulate the expression of the ACT2, TUA6, MAP65-1, CLASP, FH1 and FH4 genes
Summary
Arrays of cortical microtubules (cMTs), which are composed of microtubule (MT) bundles and separate MTs, are located beneath the plasma membrane (PM) and arranged transversely to the cell axis. They are highly dynamic structures able to reorganize fast upon receiving internal or external stimuli (Yuan et al 1994). Since the cortical cell area is the first to respond to environmental stimuli, the cell wall– PM–cytoskeleton continuum is considered to be a susceptive structure, where environmental stimuli are perceived and transduced.
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