Interaction Of VAMP With Plant Oil Synthesis Related Proteins And Aquaporins

China is the world's largest producer and consumer of edible vegetable oils,but most of the rapeseed varieties in our country have low oil content.Cultivating rapeseed with high oil content has become the breeding goal of rapeseed in China.The model plant Arabidopsis and oilseed rape belong to the Brassicaceae family,so based on the genetic information of Arabidopsis thaliana,the oil-related regulation mechanism was studied.It can lay a foundation for genetic transformation to increase oil content in rape.Plant oil is mainly accumulated in seeds,and multiple genes are involved in the regulation of metabolic processes.Lysophosphatidic acid acyltransferase(LPAT)is a key enzyme in the triacylglycerol synthesis process.It is located on the endoplasmic reticulum membrane and plays an important role in lipid synthesis,seed development and biofilm synthesis.RIF1 is related to plastid metabolism,participates in proteins involved in lipid synthesis and metabolism,and is related to chloroplast ribosomal function.Aquaporins(AQPs)are the transport channels of water and neutral small molecules across biomembrane.Among them,tonoplast intrinsic proteins(TIPS)are very important for vacuole related physiological activities,plant growth and development,especially seed development.The enzymes involved in lipid metabolism play a catalytic role and are closely related to their correct subcellular localization and protein conformation.Soluble N-ethyl-maleimide sensitive factor attachment protein receptor(SNARE)protein forms a SNARE complex to mediate membrane fusion and participates in intracellular membrane vesicle transport in plants.It plays a role in cytokinesis,gravity,response to biotic and abiotic stress,and plant hormone signal transduction.Protein interactions are the basis of cell physiological activity.The preliminary work screening of this experimental group found that SNARE protein interacts with cytosolic enzyme protein.Whether SNARE protein can participate in regulating lipid metabolism through protein interaction is worth studying.Therefore,in this study,we used the mating based split ubiquitin assay(mb SUS)to study whether RIF1,LPAT2,the key enzyme genes of oil content,and tonoplast intrinsic proteins TIP,which affects seed development and water transport,interact directly with the VAMP7 family protein in R-SNARE protein.This study found that VAMP712,VAMP721 and TIP2;1,TIP1;1,TIP1;2,LPAT2 have strong interactions,VAMP723 and TIP2;1,TIP1;1,TIP1;2have interactions,VAMP724 There is interaction between TIP1;1 and LPAT2,and there is interaction between VAMP726 and TIP3;1.The important interaction results were verified by ratio biometric molecular fluorescence complementation(r Bi FC).This study helps to reveal the effects of membrane transport on plant cell volume growth,seed development,and lipid metabolism,and can lay a foundation for future exploration of the regulatory mechanism of these proteins based on membrane transport,and provide a certain basis for future cultivation of high-yield rape varieties reference value.