The Interaction Between NO₃^- Signaling Components And ABA Signaling Components Is Involved In Water Transport In Arabidopsis

Water plays a vital role in the growth and development of plants.Water balance is necessary for advanced plants to live in a fixation way and to resist adverse circumstances in various environment.It is known that water permeability of plant cells and water transmission within plants are closely related to the response to abiotic stresses.Therefore,it is feasible to control adaptation of plants through regulating water transmission.Currently,most signaling molecules found in plants can be used to regulate hydraulic characteristics,where the nitrate?NO₃^-?and plant hormone?ABA?are very important for optimizing the plant water relation both in theory and practice.However,whether the two signaling molecules interact with each other in regulating water uptake and whether there is a mechanism enabling such interaction do not know yet.Therefore,exploring the interaction between ABA signal and NO₃^-signal for regulating transmembrane plant water transmission is of great importance for us to deeply discover the fine control of hydraulic characteristics.1? Our reseach uses Arabidopsis as materias.Based on the available Protein interaction software,we predicted the protein phosphatase At ABI2 and aquaporin At PIP1.1 can interact with At CIPK8,meanwhile aquaporin At PIP1.3 can interact with nitrate transporters At NRT1.2.This laid the foundation for further identification of their interactions through biochemical methods.2? Using the c DNA of Arabidopsis Col-0 as template,the genes of At ABI2,At CIPK8,At PIP1.1,At PIP1.3 and At NRT1.2 were obtained by PCR.Subsequently these genes were conneted to the Bi FC expression vector,in order to acquire the recombinants of At ABI2-SPYCE,At CIPK8-SPYNE,At PIP1.1-SPYCE,At PIP1.3-SPYCE and At NRT1.2-SPYNE,which were transferred into Agrobacterium strain EHA105 to inject the leaves of Nicotiana benthamiana with different combinations.By doing this,the interactions between varies proteins were observed.The results suggested that aquaporin At PIP1.1 as well as At PIP1.3 can interact with At CIPK8 and At NRT1.2,respectively.Besides,the protein kinase At CIPK8 can also interact with At ABI2.These interactions were located at cell membrane by confocal microscopy observation.3? The novel split-ubiquitin two hybrid system was used to investigate Protein-Protein interactions of membrane Proteins.The recombinants between proteins of Arabidopsis?At PIP1.1,At PIP1.3 and At ABI2?and p BT3-N were set as bait,the recombinants between proteins?At NRT1.2 and At CIPK8?and p PR3-N were set as prey for interaction analysis after co-transformation of yeast strains.Furthermore the colony growth of three transformation groups on four deficiency medium were observed.These results revealed that the protein At PIP1.1 and At ABI2 can interact with At CIPK8,while At PIP1.3 can also interact with At NRT1.2 each other,which laid the foundation of the mechansim of the interaction between NO₃^-signaling and ABA signaling involved in water transport for Arabidopsis.