| Rosa roxburghii Tratt Also known as golden fruit,Ci pear,and rosa roxburghii tratt,it is a perennial woody plant of Rosaceae Rosaceae.It is a kind of emerging fruit tree native to China.It is mainly distributed in Guizhou province,Sichuan,Yunnan and other provinces in southwestern China.With the widest distribution area in Guizhou,the output is also the highest.Rosa roxburghii Tratt fruit is rich in a variety of natural active ingredients and has high practical medicinal value.Among them,the content of vitamin C is extremely high,and it is called "king of vitamin C".In recent years,the research on Rosa roxburghii Tratt has mainly focused on the determination and quantitative description of the nutritional functional components of rosa roxburghii tratt.Although there have been relevant studies that have revealed the accumulation mechanism of vitamin C of Rosa roxburghii Tratt from the molecular level,no one has ever studied proteomics.Explain the accumulation of vitamin C in Rosa roxburghii Tratt Therefore,this research uses thorn pear fruit as raw material,through the measurement of related physical and chemical indicators,starting from the changes of vitamin C in different development stages of Rosa roxburghii Tratt fruit,using transcriptome and proteomics technology as the core means,screening thorn pear to regulate vitamin C changes The key gene,study the key protein in the process of vitamin C accumulation of Rosa roxburghii Tratt,through GOSlim,KEGG and other bioinformatics analysis methods to clearly regulate the starting gene,protein and regulatory pathways for regulating the accumulation of Rosa roxburghii Tratt vitamin C,to explore the high content accumulation of Rosa roxburghii Tratt Vitamin C formation mechanism.The main research contents and results are as follows:1.The content of vitamin C in the fruit of Rosa roxburghii Tratt showed a linear growth trend throughout the development process.In the early stage of fruit development(7-42 days),the growth of vitamin C in the fruit of Rosa roxburghii Tratt was relatively slow,and the vitamin C in the whole stage was at a low level Concentration,the average concentration is 155.85(mg ·(100g FW)-1.By the middle and late period of flowering(49-84 days),the Vc of Rosa roxburghii Tratt began to show a rapid growth trend,of which 56-70 after flowering Day is the period with the highest accumulation rate.Vitamin C accumulated to nearly half of the total content in 15 days.Vitamin C reached 2274.60(mg ·(100g FW)-1 when the fruit was ripe.2.Through constructing m RNA library and transcriptome sequencing,6 samples obtained a total of 281979432 raw reads,and after filtering with BUSCO software,273744384 clean reads were obtained.The ratio of clean reads to raw reads in all 6 samples was above 97.08%.The average GC content in clean reads is 46.18% ～ 46.41%.Analysis of the gene expression levels of Rosa roxburghii Tratt fruit at three different developmental stages revealed that there were 2,732 differentially expressed genes in the development of Pear pear A-vs-B,including 1,088 differentially expressed genes and 1,644 differentially expressed genes;A-vs-During the C development period,3971 differentially expressed genes were obtained,1857 differentially expressed genes were upregulated,and 2114 differentially expressed genes were down-regulated;during the B-vs-C development period,2626 differential genes were obtained,1553 differentially expressed genes were up-regulated,and 1053 differentially expressed genes were down-regulated.Pcs.3.GO annotation and KEGG enrichment of the differential genes in three different developmental stages of Rosa roxburghii Tratt fruit.It was found that the differential genes related to the accumulation of Vitamin C in Rosa roxburghii Tratt are mainly enriched in ascorbate and aldarate metabolism and Fructose and mannose metabolism(Fructose and mannose metabolism)two metabolic pathways,which annotated 50 genes in the ascorbic acid and alginate metabolism pathway,108 genes in the metabolism of fructose and mannose.The related genes encoding ascorbic acid and alginate metabolism GDP-D-mannose 3 ’,5’-epimerase(GDP-D-mannose 3’,5’-epimerase GME),significantly up-regulated expression.It indicates that the synthetic pathway of vitamin C of Rosa roxburghii Tratt may be L-galactose pathway.In addition,monodehydroascorbate reductase,a related gene encoding ascorbic acid and alginate metabolism,was significantly up-regulated during the growth and development of Rosa roxburghii Tratt)fruit.It shows that the metabolic cycle of vitamin C is also the main reason for the high accumulation of vitamin C in rosa roxburghii tratt.4.Detection by mass spectrometry.A total of 1302 proteins were identified,of which 1020 were identified in group A,with 159(12.2%)specific expression;900 in group B,with 130(10%)specific expression,and 924 in group C,with 69 specific expression(5.3%),of which,there are 598(45.9%)common proteins.Volcanogram analysis was used to compare the differential proteins in groups A,B,and C.The results showed that compared with group A,213 proteins were up-regulated and 333 proteins were down-regulated in group B;152 proteins were up-regulated and 248 proteins were down-regulated in group C;compared with group B,243 proteins were up-regulated Expression,219 proteins were down-regulated;83 proteins were up-regulated in groups B and C,and 159 proteins were down-regulated in groups B and C.5.Comparing the enrichment of A-vs-B,A-vs-C,B-vs-C differentially expressed protein KEGG in three different developmental stages of Rosa roxburghii Tratt fruit,we obtained 106,97,100 metabolic pathways in Among the many metabolic pathways,ascorbate and aldarate metabolism,Asparabate and aldarate metabolism,Fructose and mannose metabolism,and two metabolic pathways have a strong correlation with the accumulation of vitamin C in Rosa roxburghii Tratt)fruits.Through KEGG screening,a total of 32 proteins were annotated.Among them,during B / A development,2 proteins were up-regulated(Cluster-181.6074,Cluster-181.6074)and 6 proteins were down-regulated(Cluster-181.11079,Cluster-181.14656,Cluster-181.6701,Cluster-181.20344,Cluster-181.9654,Cluster-7743.0);during the C / A development period,2 proteins were up-regulated(Cluster-181.3266,Cluster-181.6074),and 7 were down-regulated(Cluster-181.14656,Cluster-181.20344,Cluster-181.9935,Cluster-181.9560,Cluster-181.9654,Cluster-7743.0,Cluster-181.16070);C / B development period,up-regulated expression protein is 3 Cluster-181.11079,Cluster-181.3266,Cluster-181.6701),protein expression is down-regulated 5(Cluster-181.10621,Cluster-181.9935,Cluster-181.9560,Cluster-181.16070,Cluster-181.10970). |
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