Transcriptome Analysis Of Glycyrrhiza Inflata Batalin Callus Under Salt Stress

Plants will specifically express some genes under stress conditions,regulate plant physiological and biochemical characteristics to adapt to the stress environment,and have very complex molecular regulation mechanisms.Therefore,the study of the expression pattern of plants responding to stress at the transcriptome level is more conducive to elucidating the molecular mechanisms of stress tolerance in plants.In this study,the high-throughput sequencing technology Illumina 2000 was used to perform transcriptome sequencing of normal callus of Glycyrrhiza inflata Batalin and callus treated with salt.A comprehensive analysis of the differentially expressed genes under salt stress of licorice callus reveals the possible salt-tolerance pathways and molecular mechanisms of licorice callus.It is of great significance to excavate the anti-salt genes of licorice and to use molecular biology to cultivate new varieties of licorice in the future.At the same time,the lupinol synthase related to glycyrrhizic acid synthesis pathway,the key enzymes of chalcone synthase and chalcone isomerase in the pathway of flavone synthesis were selected for further study.The main findings are as follows:1.The transcriptome of Glycyrrhiza samples obtain 46735469 Short read,and the raw sequencing data was processed and spliced to generate 155072 Transcript and 93263 Unigene.Compared with NR,GO,KO,Swiss Prot,PFAM,KOG,and NT databases,the percentages of annotations were 55.76%,42.61%,21.2%,41.98%,41.48%,15.09%,and59.53%,respectively.1913 differentially expressed genes were identified,of which 1109 were up-regulated and 804 were down-regulated.GO enrichment analysis revealed that these differentially expressed genes are mainly involved in catalysis,oxidoreductase activity,and oxidation-reduction process.KEGG metabolic pathway enrichment analysis found that the most enriched metabolic pathways were phenylpropane biosynthetic pathways,phenylalanine metabolism and flavonoid biosynthesis pathways.2.Based on the transcriptome analysis of the callus of Glycyrrhiza inflata Batalin under salt stress,it was speculated that the salt-tolerant network map.The related genes in the SOS signal pathway,MAPK signaling pathway,transcription factors,hormone metabolism and others all interact to regulate the expression of the target gene,maintain the ion balance,oxidation-reduction balance and osmotic balance,and increase the salt tolerance of cells.3.The plant expression vector p BI121 was modified.The cloned ampicillin resistance gene was inserted between multiple cloning site Xba I and Bam H I,and the plant expression vector p BI121-Ampr was successfully constructed,which provides the basis for the construction of subsequent gene plant expression vectors and gene function identification.4.In this study,a 310 bp lupinol synthase gene fragment was selected as the target gene,and some gus fragments were selected as introns.Fusion fragments were obtained by overlap PCR,and a cloning vector was constructed,and an antisense fragment of the target gene was also constructed cloning vector.The sequencing result is correct.This will lay the foundation for the subsequent construction of RNAi expression vector.5.In this study,chalcone synthase and chalcone isomerase,the key enzymes of the flavone synthesis pathway,were selected as the research object.The cloning vector of the chalcone synthase gene and the chalcone isomerase gene was constructed and the sequencing result was correct,which provided the basis for the subsequent plant expression vectors construction and function identification.