| Spirulina morphology is an important factor to measure the quality of Spirulina,but in the training process is easily affected by external environment,morphological changes,and accompanied by a series of physiology,nutrition,corresponding to the change of genetics and proteomics etc.,not only the algae yield decreased greatly,but also brought difficulties for the identification of species.So as to realize the improvement and regulation of Spirulina morphological changes,it is necessary to build analytical form of the molecular mechanism of Spirulina.This study originated from single filament culture occurred morphological differentiation of straight and spiral individual objects,using iTRAQ proteomics and mass spectrometry,and investigated the difference of protein,and Spirulina morphogenesis related proteins,and combined with the difference of screening to learn information technology proteins from biological processes,involved in cellular localization,three molecular function were analyzed by real-time fluorescence quantitative PCR technology,verified from the transcriptome of protein obtained data,to find key genes regulating Spirulina form built,reveal the molecular mechanism of Spirulina morphogenesis and generation The results are as follows:1.The filaments of length,straight and helical screw pitch,diameter of Spirulina,spiral number and growth curve,phycocyanin,allophycocyanin,chlorophyll a,carotenoids and other morphological indexes and physiological indexes were measured,grasp the morphological and physiological differences between different forms of Spirulina,provide sufficient the basis for further research on the selected object.2.The proteins were identified by iTRAQ proteomics and mass spectrometry.To FDR? 1%,remove the anti-library data and the ratio of the blank value of the invalid,as a trusted protein screening criteria;protein abundance change greater than 2 or less than 0.5,as a differential protein screening criteria.ITRAQ-LC-MS / MS analysis was matched to185123 maps with 37.6% of the spectrum,removal of the less reliable peptides,and identified a total of 30,508 specific peptides,according to the criteria for screening the trusted protein Redundant results were eventually identified to 2156 proteins.TJSD2/TJSD3 protein group.The group showed a total of 167 differentially expressed proteins in the TJBC4-1/TJBC4-2 group,and there were about 35 differentially expressed proteins between the two groups.3.The bioinformatics analysis of the differential proteins was carried out.The differential proteins were identified in the glycolytic TCA cycle,photosynthesis,photosynthesis-antenna protein,starch and sucrose metabolism,lipopolysaccharide biosynthesis,photosynthetic carbon fixation,porphyrin,Chlorophyll metabolism and other metabolic regulation.Based on the analysis of metabolic pathways,the prediction model of Spirulina morphology was established,which was used to analyze the mechanism of Spirulina morphology.And the protein data in the model were verified by real-time fluorescence quantitative PCR.On the basis of proteomics and transcriptome analysis of proteins with the same trend of change,we constructed the energy metabolism network diagram of the Spirulina morphogenesis center,and analyzed the protein interaction network.4.The pgm gene,which plays a key role in the morphology of Spirulina,was verified by prokaryotic expression of two different proteins in Spirulina platensis.Construction of pgm gene fusion expression vector,and successfully transferred to E.coli for expression.The morphological changes of Escherichia coli transformed into the pgm gene of Spirulina platensis showed that the pgm gene of Spirulina platensis could be regulated by E.coli,which provided the theoretical basis for the functional verification of pgm gene in Spirulina. |
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