Role Of Transcription Factor AtCPC Regulating Actin Depolymerizing Factor AtADF11 During Root Hair Growth And Development In Arabidopsis Thaliana

Root hairs significantly increase the surface area of the root system and promotes plants the absorption of water and nutrients.In the meantime,as a single cell,the root hair also provides a ideal model for studying the development,differentiation and elongation in plant cells.Therefore,studying the mechanism of root hair growth and development has important scientific and theoretical significance.Actin binding proteins and transcription factors are two key factors that regulate the growth and development of root hairs.However,no research has been reported on the molecular mechanisms involved in the growth and development of root hairs.Early laboratory tests found that the Atadf11 mutant exhibited a long and dense root hair phenotype,therefore,this experiment investigate the mechanism of regulating the actin depolymerizing factor At ADF11 during root hair growth and development,and revealed the role of transcription factor At CPC in regulating At ADF11 during root hair growth and development.The experimental research results are as follows:(1)Atcpc homozygous mutants were identified.q RT-PCR was used to detect the relative expression of At ADF11 gene in the root of Atcpc homozygous mutants.The results showed that the relative expression of At ADF11 gene was significantly up-regulated after the At CPC gene was deleted,indicating that At CPC negatively regulates the expression of At ADF11.(2)Construction of At ADF11 gene expression vector,genetic transformation and screening of over-expressed homozygous lines.Identify the Atadf11-1 homozygous mutants and At ADF11-OE #21 plants.Detection the hybrid materials of Atcpc and At ADF11 genetic plants : Atcpc Atadf11-1 and Atcpc At ADF11-OE.Observe the root hair phenotype of WT,Atcpc,Atadf11-1,At ADF11-OE #21,Atcpc Atadf11-1 and Atcpc At ADF11-OE plants grown for 6 days and statistics of root hair length and number.The results found : the Atcpc has short and few root hairs;The Atadf11-1 exhibits a long and dense root hair phenotype,and the overexpression plant At ADF11-OE #21 has the opposite root hair phenotype as the Atadf11-1;Atcpc Atadf11-1 shows the root hair phenotype of Atadf11-1,Atcpc At ADF11-OE shows the At ADF11-OE #21 phenotype.The above results indicate that At CPC negatively regulates the role of At ADF11 during the growth and development of Arabidopsis thaliana root hairs.(3)The At CPC-p GEX-4T1 prokaryotic expression vector was constructed,and the At CPC-GST tag fusion protein was successfully induced and purified.An At CPC binding site was analyzed at 96 bp upstream of ATG in the At ADF11 promoter sequence on the Plant CARE website,and design specific primer for cloning probes.The EMSA experimentalresults prove that At CPC binds to At ADF11 gene promoter probes in vitro which contains specific binding sites,indicating that At ADF11 is a target gene of At CPC.Based on the results of the above experiments,this study confirmed that the transcription factor At CPC directly bind to the promoter region of At ADF11,regulate its transcription,and participate in the formation and growth of root hair development,elucidates the role of the transcription factor At CPC in regulating actin depolymerizing factor At ADF11 during root hair growth and development.The first to establish the molecular mechanism between transcription factor and microfilament skeleton in the regulation mechanism of root hair growth and development,providing a new scientific theoretical basis for related research.