Cloning And Functional Analysis Of MYBAS1-like Gene Of Puccinellia Tenuiflora

MYB(v-myb avian myeloblastosis viral oncogene homolog)transcription factor is one of the members of the transcription factor family.Its structural features include the MYB domain.It participates in plants responding to biological,abiotic response reactions in nature.The study of PutMYBAS1-like,a member of Puccinellia tenuiflora transcription factor,,The function of Puccinellia tenuiflora transcription factor PutMYBAS1-like and its expression characteristics under abiotic stress were studied in depth.1.In this study,PutSnRK2 was used as a "bait" protein to screen the protein interacting with it by yeast two-hybrid technique.The MYB gene fragment was cloned from the cDNA library of Puccinellia tenuiflora under NaHCO3 stress The full length of PutMYBAS1-like was cloned by RT-PCR and RACE.Bioinformatics showed that there were 522 bp open reading frame in the gene sequence,encoding 173 amino acids.The molecular weight of the predicted protein was 19.68 kDa,and its N-terminal contained a MYB characteristic domain of about 45 amino acids.(Brachypodium distachyon)MYBAS1-like gene homology is higher,named PutMYBAS1-like(Alternative splicing MYB transcription factor).2.In order to understand the expression pattern of PutMYBAS1-like gene in Astragalus mongolicus,we used real-time fluorescent quantitative RT-PCR to analyze the expression of PutMYBAS1-like gene under NaHCO3,NaCl and PEG.The results showed that the PutMYBAS1-like gene was significantly induced in leaves and roots under these stress conditions.In the leaves,have shown a downward trend;in the roots,have shown an upward trend.Indicating that PutMYBAS1-like gene and adversity stress have a certain response relationship.3.The transcriptional activity was verified by yeast single hybridization and the transcriptional activation region was analyzed.4.PBS-PutMYBAS1-like-GFP was introduced into the onion epidermal cells by the method of particle bombardment.The results showed that PutMYBAS1-like was located in the nucleus.5.At the same time,the pBI121-PutMYBAS1-like plant overexpression vector was constructed and the PutMYBAS1-like overexpression of Arabidopsis thaliana was obtained.The phenotypic analysis of Arabidopsis thaliana under abiotic stress(NaCl,NaHCO3,PEG)The results showed that overexpression of PutMYBAS1-like Arabidopsis thaliana could enhance the tolerance to salt,alkali and drought under NaCl,NaHCO3 and PEG stress.