Cloning And Expression Analysis Of Wheat Stress-responsive Transcription Fcator TaSNAC1

Plants are often subjected to various biotic and abiotic stresses during the growth process. Study the molecular mechanisms of plants perceive, resist and tolerate various stresses is not only has theoretical value, but also has important significance on crop breeding and genetic improvement. NAC is widely distributed in the terrestrial plant, is the transcription factor family specific in the plants, play an extremely important role in plant development and stress responses. This study used EST splicing as a foundation, cloned the NAC transcription factor gene of wheat drought and salt stress response, and analysis its gene expression pattern. The results as following:the results are as following:1. We used rice stress response transcription factor gene SNAC1as the basis,designed primes based on the result of wheat ETS sequence splicing, using wheatZhengyin#1as material, PCR and RT-PCR amplification of wheat TaSNAC1genomeDNA (gDNA) and cDNA sequence, amplified fragment are1222bp and1076bp,respectively. The cDNA sequence contains990bp coding sequence (CDS), gDNAincludes a length of146bp intron, and both sides of the intron contains typically"GT/AG" shear signal. The gDNA sequence has been submitted to the GenBank, and theaccession number is JN621240.2. Real-Time PCR analysis the expression of TaSNAC1, and found that TaSNAC1has low expression in both of roots, stems, leaves and seeds, the expression in leaves and roots are higher than the expression in stems and seeds. We choose the wheat after two weeksof water culture, stress treated of20%PEG6000and250mmol NaCl solutions, use wheatbeta-actin gene primers for reference, analysis the gene expression pattern of TaSNAC1, the results showed that: under the presence of salt stress, the expression of TaSNAC1in leaves and roots are up-regulated and have the same expression pattern, but more remarkablein the roots; under the condition of PEG stress, the expression of TaSNAC1were relatively fast and intense in the root, and slower and weaker in the leaves.3. Constructed the transient expression vector pTF486-TaSNAC1(P35S: TaSNAC1-GFP), use PEG mediated method transient transfect the plasmids intoarabidopsis mesophyll cell, fluorescence microscopy observed the fusion proteinTaSNAC1-GFP specifically localized in the nucleus.Our study successfully cloned the wheat TaSNAC1gene, and demonstrated that it isinvolved in abiotic stress responses of wheat, and also established the foundation ofTsSNAC1in the genetic improvement of wheat drought and salt resistance.