| Zanthoxylum bungeanum Maxim.,originated in China,is an economic tree with important economic value and medicinal value.It belongs to a small deciduous tree of the family ruta and Zanthoxylum.It is widely distributed in Sichuan,Gansu,Shaanxi,Jiangsu,etc.The whole body of Z.bungeanum Maxim is treasure.Its fruits,leaves and stems are widely used as spices and oil plants.Z.bungeanum has a pungent and fragrant fruit and is used as a seasoning in food and cooking.Its stem and bark are traditional Chinese medicine,which have the functions of relieving cough,resolving phlegm and eliminating food.The fragrant leaves are used to repel pests in food preservation.Hanyuan prickly ash is a kind of prickly ash,known as the "king of Sichuan prickly ash".Because of its color and quality,it was used as a tribute in ancient times.The planting and processing of Hanyuan Z.schinifolium Sieb.et Zucc.is regarded as an important economic pillar in Hanyuan.Therefore,it is of great theoretical and practical significance to study the transcriptome analysis and key genes identification of Z.schinifolium Sieb.et Zucc.in the process of grain development.In this study,Hanyuan Z.schinifolium Sieb.et Zucc was selected as the research object,and its transcriptome during the key expansion stage of 5 days,10 days and 15 days after pollination was analyzed and the key expression differential genes were verified.The following experimental results are obtained:1.The growth of the early stage of grain development is faster than that of the later stage.The results showed that the growth of Z.schinifolium Sieb.et Zucc.in 5-10 days after pollination was faster than that in 10-15 days after pollination.2.High quality transcriptome sequencing data were obtained.The mRNA was extracted 5 days,10 days and 15 days after pollination in Hanyuan Z.schinifolium Sieb.et Zucc.Each period was repeated three times independently,and the quality analysis was carried out.After sequencing,sequence assembly,sequence alignment,result annotation,expression analysis at different pollination stages,47831201 sequences were measured on average for each sample;the average number of bases of each sample was 7094430816 base pairs,and the average length of each sequencing was 148 base pairs.The accuracy of sequencing was 97.4%,and the average GC%content of the sequence was 44.99%.These results indicate that the quality of transcriptome sequencing is high and can be used for further analysis.3.High quality transcriptome assembly analysis data were obtained.Since the genome of Z.bungeanum has not been published,the sequences measured by transcriptome are assembled from scratch to form overlapping groups and single sequences.After Trinity software analysis,a total of383489 transcripts were obtained and 188038 unigenes were obtained.The average length of these transcripts is about 900 bp,and the average length of unigene is 653 bp.The sequencing length of144831 transcripts and 94540 unigenes ranged from 201-400 bp,accounting for 37.76% of the total transcripts and 50.28% of the single gene respectively;the length of 71493 transcripts and 38001 unigene splices ranged from 401-600 bp,accounting for 18.64% of the total transcripts and 21.21%of the single gene respectively;38259 transcripts and 17998 unigenes was 601-800 bp,accounting for 9.98% of the total transcripts and 9.57% of the single factor,respectively.The proportion of other length transcripts and spliced unigene was 33.72% and 18.94%,respectively.4.Go enrichment analysis showed that in terms of gene function and protein function,these genes were attributed to biological process,cell composition and molecular function.It was found that the most enriched genes were cell processes,with a total of 41836 genes.It was found that the most enriched genes were involved in cell metabolism,with a total of 36655 genes.It was found that the most enriched genes were molecular binding,with a total of 36052 genes Because of genes.These results showed that the development process of Z.bungeanum seeds was complex and involved many genes.5.The analysis of the homologous cluster of the protein showed that these genes have a variety of functions,and there are 25 functional categories.Among them,there are 3311 proteins with replication,recombination and repair at most,2704 proteins predicted by general functional proteins,and 2557 proteins involved in cell signal transduction mechanism.These analysis showed that there were relatively few proteins with specific functions in Z.bungeanum,which reflected the uniqueness of Z.schinifolium Sieb.et Zucc.6.The analysis of KEGG metabolic pathway showed that the main genes involved in 20 signaling pathways in the seeds of 5,10 and 15 days after pollination.Unigene analysis showed that there were more genes involved in carbohydrate metabolism and cell signal transduction pathway.7.The differentially expressed genes in different pollination stages of Z.schinifolium Sieb.et Zucc.were analyzed.Among them,there were significant differences in 2002 genes between 5 days after pollination and 10 days after pollination.174 genes were significantly different between 10 and15 days after pollination.Compared with 15 days after pollination,there were 3751 differential genes.These genes are distributed in different biological processes and have different functions.8.The real time quantitative analysis of the key differential genes showed that the key differential genes were basically consistent with the transcription data.In a word,the growth indexes of three stages of Z.schinifolium Sieb.et Zucc.seeds were measured,the transcriptome of five days,ten days and fifteen days after pollination was analyzed,and the key differential genes were verified by real time fluorescence quantitative analysis.These results showed that Z.schinifolium Sieb.et Zucc.seeds development process was complex and unique.At different developmental stages,4357 differential genes were analyzed.These results laid a foundation for further study on the accumulation process of the unique substances of Z.bungeanum. |
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