Investigation of the Nature of CgCDPK and CgbHLH001 Interaction and the Function of bHLH Transcription Factor in Stress Tolerance in Chenopodium glaucum

Abstract

Calcium-dependent protein kinase (CDPK) and its substrates play important roles in plant response to stress. So far, the documentation on the characterization of the CDPK and downstream interaction components (especially transcription factors, TFs) is limited. In the present study, an interaction between CgCDPK (protein kinase) (accession no. MW26306) and CgbHLH001 (TF) (accession no. MT797813) from a halophyte Chenopodium glaucum was further dissected. Firstly, we revealed that the probable nature between the CgCDPK and CgbHLH001 interaction was the phosphorylation, and the N-terminus of CgbHLH001, especially the 96th serine (the potential phosphorylation site) within it, was essential for the interaction, whereas the mutation of 96Ser to alanine did not change its nuclear localization, which was determined by the N-terminus and bHLH domain together. Furthermore, we verified the function of CgbHLH001 gene in response to stress by ectopic overexpression in tobacco; the transgenic lines presented enhanced stress tolerance probably by improving physiological performance and stress-related gene expression. In conclusion, we characterized the biological significance of the interaction between CDPK and bHLH in C. glaucum and verified the positive function of CgbHLH001 in stress tolerance, which may supply more evidence in better understanding of the CDPK signaling pathway in response to adversity.

Overexpression of the Apple (Malus × domestica) MdERF100 in Arabidopsis Increases Resistance to Powdery Mildew

“…For example, heterologous expression of the cotton ( Gossypium hirsutum ) ERF gene GhB301 in Nicotiana benthamiana enhanced its resistance against fusarium wilt [ 8 ]. The basic helix–loop–helix (bHLH) superfamily is considered as the second largest class of plant transcription factors [ 9 ]. A bHLH domain contains 50–60 amino acids and two functionally distinct regions: the N-terminal basic region, which is involved in the binding to the E-box (CANNTG) DNA sequence, and the HLH domain, which mediates domain dimerization [ 10 , 11 ].…”

iTRAQ based protein profile analysis revealed key proteins involved in regulation of drought-tolerance during seed germination in Adzuki bean

“…The new developping leaves that was uninoculated with 10 d water withholding were harvested for analyzing the activity of SOD, POD, content of proline and MDA, and water loss as described by Zhou et al 53 . Data analysis were conducted by Microsoft Excel 2016 and the SPSS 16.0.…”

Regulatory Mechanisms of bHLH Transcription Factors in Plant Adaptive Responses to Various Abiotic Stresses

“…Chenopodium glaucum CgbHLH001 can positively regulate plant stress tolerance via clearing excessive ROS and accumulating transcripts of stress-related genes (Wang et al, 2017). A recent study indicates that CgbHLH001 activity might be affected by phosphorylation through interaction with CgCDPK (Zhou et al, 2021). Ubiquitination can likewise directly regulate bHLH activity via the 26S proteasome protein degradation pathway (Figure 1).…”