Abstract
In this study, when germinated <em>Triticum aestivum</em> L. seeds were treated with 0, 2, 4 and 6 mM ethyl glycol tetraacetic acid (EGTA), root growth was suppressed and the mitotic index decreased. These inhibitory effects were positively correlated with EGTA concentration. RT-PCR analysis revealed that the expression of several gene markers related to the G1/S transition of the cell cycle were significantly downregulated. Confocal microscopy of Fluo-3/AM-stained roots showed chelation of nearly all of the Ca<sup>2+</sup> within the root meristematic regions. Both random amplified polymorphic DNA (RAPD) and coupled restriction enzyme digestion-random amplification (CRED-RA) techniques showed significant increases in the levels of genomic DNA polymorphisms and degree of DNA methylation. The study provides information concerning the impact of Ca<sup>2+</sup> chelator, EGTA, on the growth, expression of cell cycle transition marker genes, and changes in DNA structure and methylation in the wheat roots.
Highlights
Plant growth, development, and differentiation rely on coordination of the cell cycle which can be divided into four consecutive phases: G1, S, G2, and M
When the radicles emerged from the seed coat to a length of 1 mm, germinated seeds were treated with different concentrations of ethyl glycol tetraacetic acid (EGTA) (Solarbio, Beijing, China, #E8050) (2, 4, and 6 mM in distilled water) in darkness at 25°C for 24 h
The data demonstrate that EGTA inhibits roots growth and cell division and that the inhibitory effects are positively correlated with its concentrations
Summary
Development, and differentiation rely on coordination of the cell cycle which can be divided into four consecutive phases: G1, S, G2, and M. To ensure the faithful duplication and passage of genetic information during cell division, the transitions between different phases of the cell cycle are precisely coordinated and controlled by the cyclin-dependent kinases (CDKs) [1]. Cyclin D-CDK4/6 complexes phosphorylate the retinoblastoma protein (pRB) to dissociate it from E2F family of transcription factors, which is a crucial step in the coordination of the G1/S transition [2,3]. During early G1 phase, the pRB protein is hypophosphorylated and binds strongly to E2F transcription factors, preventing transcription of E2F target genes. Hyperphosphorylation of pRB releases E2F, which transactivates target genes that promote Sphase entry [6].
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