| When plants are under drought,light,high temperature,saline,cold,pests aind other stress,the cells will be damaged at different degrees.Caragana intermedia is drought,saline and extreme temperature tolerant,and was used widely in sand fixation,soil and water preservation.NACs are one of the largest plant-specific transcription factor families.Members in this family play important roles in growth and abiotic stress responses.In this study,CiNAC1 was cloned from C.intermedia and its primary function was analyzed.The concrete conclusions are as following:1.The full-length cDNA sequence of CiNAC1 from C.intermedia was cloned by PCR technique.It was 1066 bp in length.Bioinformatics analysis shows that CiNAC1 has an open reading frame of 921 bp,which encoding 306 amino acids.The calculated molecular weight of CiNAC1 is 34.57 kD and the isoionic point is 8.35.The CiNAC1 protein is hydrophilic has a conserved NAM domain in the N-terminus.It contains 26 phosphorylation sites and 7 glycosylation sites.2.Real-time quantitative PCR analysis showed that CiNAC1 was induced by drought and saline.pH and dehydration.These results indicated that CiANAC1 might be involved in response of C.intermedia to abiotic stresses.3.Fusion construct CiNACl-GFP was transformed into Arabidopsis thaliana protoplasts.Fluorescence observation under confocal laser-scanning microscopy confirmed that CiNAC1 localized in the nuclei,consistent with its role as transcription factor.4.Binary vector CiNAC1-HA was transformed into A.thaliana to get permanent transgenic lines.Real-time quantitative RT-PCR showed that the target gene was expressed in different transgenic homozygous lines.5.Overexpression of CiNAC1 promoted the lateral root formation and length of the primary root.Under NaCl and mannitol treatment,the seed germination rate of CiNAC1 overexpressing lines was lower than that of wild type.After treatment with ethylene,overexpression of CiNAC1 in Arabidopsis resulted in early senescence. |
PDF Full Text Request