Analysis Of The Response Of Sugar Beet BvM14-nsLTP Gene To Abiotic Stress

The beet M14 line is a monomer addition line obtained by crossing and backcrossing beet(diploid)and wild white-flowered beet(tetraploid).It has many advantages of parents and is a good material for discovering high-quality genes.The laboratory has constructed a transcriptome database of sugar beet M14 strains under salt stress.The study found that the BvM14-nsLTP gene,which encodes non-specific lipid-transfer protein(nsLTP),can be found in leaves and roots under salt stress.Both up-regulated expression.In this experiment,the BvM14-nsLTP gene was cloned from the sugar beet M14strain,and bio-information analysis,protein subcellular localization,and specific analysis of the BvM14-nsLTP gene in root and leaf tissue were carried out.Four kinds of abiotic stress response analysis to explore the response mode of the sugar beet BvM14-nsLTP gene to different abiotic stress,laying the foundation for the research of the function of BvM14-nsLTP gene in response to abiotic stress.Through the sequence open reading frame analysis of BvM14-nsLTP,it was found that the ORF of BvM14-nsLTP was 357 bp,encoding 118 Aa;through multiple sequence alignment of beet BvM14-nsLTP and other plant LTP protein sequences,it was found that beet BvM14-nsLTP It has the typical conserved domains of nsLTP protein,including8 highly conserved cysteine residue backbones,a signal peptide sequence near the N-terminus,a Ca M binding site near the C-terminus,and a CGV lipid binding site,belonging to nsLTP I Subfamily members;phylogenetic tree construction was carried out,and it was found that BvM14-nsLTP was closely related to the nsLTP protein of spinach(Spinacia oleracea L.)and quinoa(Chenopodium quinoa L.);by analyzing the functional elements of the beet BvM14-nsLTP sequence promoter,Found that the promoter contains functional elements involved in stress regulation and.The prediction of subcellular localization indicated that BvM14-nsLTP was localized outside the cell.The 35S::pCAMBIA1300-BvM14-nsLTP-eYFP expression vector was constructed and transformed into tobacco.The subcellular localization of BvM14-nsLTP was verified,and it was found that BvM14-nsLTP was localized in the cell membrane.Tissue-specific expression analysis found that the expression level of BvM14-nsLTP in roots was 15.1times that of leaves.Using real-time fluorescent quantitative PCR technology,the beet M14 strains were subjected to salt stress(0,200,400 mM NaCl),alkali stress(0,50 mM alkaline salt mixture,and a mixture of Na₂CO₃ and Na HCO₃ with a molar ratio of 2:1.p H value 9.2),drought stress(0,15%PEG6000)and low temperature stress(room temperature,4?)treatments for 0 h,3 h,6 h,9 h,12 h,the effect of BvM14-nsLTP gene in leaves and roots.The response analysis showed that the expression pattern of BvM14-nsLTP gene was the same under salt,alkali,drought and low temperature stress,all of which increased first and then decreased,and all were induced to be expressed.This study shows that the sugar beet BvM14-nsLTP gene has the ability to respond to different abiotic stresses,which lays a good foundation for perfecting the molecular mechanism of the BvM14-nsLTP gene in response to different abiotic stresses.