Prediction Of The Structure And Research Of The Activity Of Promoter Glycerol-3-pHosphate Acyltransferases Gene In Lilium Spp.

An important and forefront of plant genetic engineering is gene expression and regulation,while promoters play an important role in the regulation of transcription,which has become a hot spot in molecular biology.Inducible promoters can induce the expression of promoter genes under stress and reduce their own injury and avoid energy waste.Liliums has the food,medicinal and ornamental value,but Liliums is susceptible to diseases and adverse environmental impact,and the structure and function characteristics of the inducible promoter can improve Liliums resistance.Based on the research results of the research group,we selected Lilium regale and Lilium dauricum as the test material,sequenced the GPAT genes of two Liliums species using bioinformatics techniques,analyzed the expression of GPAT gene by fluorescence quantitative PCR,The deletion fragment of the promoter of Liliums GPAT gene was then transformed into tobacco.The main results are as follows:1.The amino acids encoded by the GPAT genes of Lilium dauricum and Lilium regale were homologized and their homology was 96.34%.Through searching and comparing the conserved domains of Arabidopsis thaliana ATS1 protein,various other GPAT proteins and two Liliums GPAT proteins,the two Liliums GPAT proteins and other plant GPAT proteins also have a conserved acyltransferase activity domain: motifsI,II,III and IV.The results of using online software SMART analysis shows that the proteins encoded by the 2 kinds of Liliums GPAT genes had a PlsC acylase domain at positions 176-322 and 169-315 indicating that the GPAT protein belongs to acyltransferase family members,with acyltransferase activity.The results of TMHMM Server 2.0 amino acid sequence analysis shows that the proteins of the 2 kinds of Liliums GPAT without transmembrane region.The results of using online software SignalIP 4.0 analysis shows that 2 kinds of Liliums GPAT amino acid sequences have no signal peptide.Through sequence analyses in PLACE and PlantCARE,except for the core promoter elements such as TATA box and CAAT-box and five inducible responsive elements:MYC?TC-rich repeats?MYB? MBS and DRE.2.Using real-time quantitative PCR to research GPAT gene expression by ABA-induced,the results shows that the expression of the different varieties Liliums GPAT gene increased.3.The LpGPAT full-length promoter sequence was truncated from the 5' end to dif ferent length sequences of 1466 bp,884 bp,566 bp,and 412 bp,which were named GPAT F1,GPAT F2,GPAT F3,and GPAT F4,respectively,to replace the promoter of plant expression vector PCAMBIA1303.4.Agrobacterium-mediated leaf injection was used for tobacco transformation.The deletion analysis and GUS histochemical staining revealed that the 844 bp promoter of the LpGPAT promoter and the 384 bp fragment of the LrGPAT promoter of the Lilium davidii could drive the inducible expression of GUS in tobacco.Therefore,that the fragment was the smallest in the other four GPAT gene promoter.