Functional Analysis Of NACL Transgenic Arabidopsis

The NAC is the largest family of plant transcription factors.It exists only in terrestrial plants and has multiple functions.It not only plays an important role in plant growth and development,hormone regulation,but also can participate in abiotic stress and biotic stress response.In this paper,a NAC gene was cloned and identified from Arabidopsis thaliana and named NACL.A preliminary study on the expression and function of this gene was performed,and the transcriptome changes of AtNACL transgenic plants at 30 days were further analyzed.The results of the study are as follows:1.The transcription activity analysis of AtNACLThe pGBKT7-AtNACL recombinant plasmid was constructed and transferred to competent cells of the yeast cells AH 109 and Y187,obtaining positive colonies.Striation detection was performed on the medium of the two deficiencies(SD/-Trp/-His,SD/-Trp/-Ade),and do X-gal color reaction.The results indicated that AtNACL is a transcriptional repressor.2.The expression pattern analysis of AtNACLThe expression of AtNACL in roots,stems,leaves,flowers and silicle of wild type was detected.The results showed that the expression level of AtNACL in flowers was higher.By promoter activity analysis in transgenic Arabidopsis,AtNACL promoter is dominant expression in flowers.3.The phenotypic analysis in AtNACL overexpressing transgenic ArabidopsisWe hydroponically cultivated WT and overexpressing AtNACL plants.The results showed that the transgenic Arabidopsis of overexpressing AtNACL has three phenotypes:the first one can normally flower and fruit,the size of which was basically the same as of WT,and the second one was not blooming until 100d,which was still in vegetative stage;the third one can flower,but the flowering time is delayed.The latter two are accompanied by dwarfing,anthocyanin accumulation.4.AtNACL and PHR1,flowering and anthocyanin-related gene expression analysisBy observing the phenotype of overexpressing transgenic Arabidopsis,we detected the expression levels of PHR1,flowering genes,and related genes involved in anthocyanin synthesis of overexpressing transgenic Arabidopsis about 60 days and 75 days.The result showed that the expression of PHR1 and FLC was significantly up-regulated,and LFY was down-regulated.The anthocyanin synthesis related genes MYB4,MYB75 and MYB90 were also significantly up-regulated,and the sugar synthesis related genes HXK1 and HXK2 were significantly up-regulated.It is initially assumed that NAC plays an important role in the conversion from vegetative growth to reproductive growth.5.Transcriptome analysis of overexpressing NACL transgenic Arabidopsis thalianaWe performed transcriptome analysis of the aerial parts of AtNACL transgenic plants,focusing on the transcriptome data of WT and NACL-16,18 of no-flowering phenotypes.Significant enrichment analysis of GO function and Pathway enriched analysis was performed on differentially expressed genes.The analysis and induction of differentially expressed genes were performed about three aspects,such as flowering,anthocyanin synthesis,and senescence.