Analysis Of Arabidopsis AtOxR Gene Promoter And Identification Of Upstream Interacting Protein Screening

Light is an important environmental factor affecting plants.It not only provides energy for photosynthesis,but also is one of the key environmental factors that regulate various processes of plant growth and development,including seed germination,seedling de-yellowing,stem elongation,flowering,shade avoidance response and biological clock.Light,as an important environmental signal,not only affects the survival of all organisms on earth,but plants can also receive different wavelengths of light through photoreceptors and transmit light signals to cells,triggering a series of intracellular responses to achieve their effects on growth and development.Our group has shown that Arabidopsis AtOxR is a light-signal-responsive gene,and its expression is significantly up-regulated by light induction.To further investigate the mechanism of AtOxR involvement in light response,this study was conducted to screen and identify proteins interacting with the AtOxR gene promoter by yeast single hybridization.The main findings are as follows.1,The AtOxR gene promoter contains a variety of cis-acting elements,including biotic stress,abiotic stress,growth and development,and light signaling response elements,further indicating that the AtOxR gene is closely related to light signaling response.2,31 genes co-expressed with AtOxR were screened by weighted co-expression network analysis,among which 14 genes were involved in light response.RT-q PCR was further used to verify the strong response of these 14 genes with light signal.3,The interaction between the AtOxR promoter and 14 light-responsive genes was further analyzed by yeast single hybridization,and a total of two interacting proteins with the AtOxR promoter,ascorbate peroxidase Att APX and unknown protein AtL1D2,were obtained,and the interacting elements were identified to be present within the pAbAi-AtOxR-M5 fragment by analysis of AtOxR promoter fragments of different lengths,and the interactions were verified by a dual luciferase reporter system.4,The subcellular localization of AtL1D2 protein was analyzed by transient expression in tobacco and onion epidermis,and the results showed that it was localized in the nucleus and cell membrane,which was consistent with the bioinformatic predictions.Further,pure mutants with deletion of AtL1D2 gene were identified and successfully obtained by molecular level.