| As an important Alpine medicinal plant,the contents of active components of R.crenulata are affected by many factors.Recently,endophytic fungi were found that they were essential part of plant growth,development and metabolism,which can change host gene expression and metabolic pathway through transverse gene transfer,induction effect and biotransformation,and make an impact on plant growth and development ultimately.In this paper,the effects of endophytic fungi,Phialocephala fortinii,on R.crenulata were studied based on transcriptome and metabolomics techniques,which would provide help for understand of plant-microbe interaction theoretical and application of chemical composition regulation,biological stress and other aspects.The results in this stydy were listed as follows:(1)In this study,they were sequenced that the R.crenulata plantlets inoculated with endophytic fungi P.fortinii after 2,4,6,8,10,12 days.After assembly and removal of redundancy,53039 Unigenes were obtained at last.The total length,the average length,the length of N50 and the GC content were 48866 366 bp,912.2520 457 bp,1396 bp,and 47.38%,respectively.These Unigenes were annotated to five functional databases of NR,GO,KEGG,egg NOG and Swis-Prot,so that each gene was annotated to its corresponding function.The number of annotated genes in each database was 53039(100%),30419(57.35%),5591(10.54%),51 252(96.63%)and 41831(78.87%),respectively.From the transcriptome data,we found that the number of differentially expressed genes(DEGs)changed with the interaction time and there were 379(636),517(267),4233(17),3942(56),3048(152)and 2587(205)up-regulated(down-regulated)DEGs on the 2nd,4th,6th,8th,10 th and 12 th days,respectively.Sequencing results showed that these DEGs were involved in many metabolic pathways of R.crenulata,such as carbohydrate metabolism,lipid metabolism,amino acid metabolism,secondary metabolite synthesis and other metabolism.Studies showed that endophytic fungi played an extremely important role in plant metabolism.(2)In this study,the response of R.crenulata to endophytic fungi,P.fortinii,was studied by using liquid chromatography tandem spray ion source four stage time of flight mass spectrometry(UPLC-Q-TOF-MS)technique.27 differently metabolites were picked by multivariate statistical analysis,of which 2 were not identified and others were identified as Acetaldehyde,Cyclopropylamine,Naphthalene,Ethylene sulfide,Homocysteine,4-Methoxybenzyl O-(2-sulfoglucoside),3-Sulfomuconate,Fusaroskyrin,S-Propyl-Lcysteine,Botryplodin,Dimethic,1,3-Budiene,C-diesteine.Cycloobrassinin sulfoxide,Halistanol sulfate,Diacetyl,1-Butanol,Dichlorobenzene and 1-Pyrrolinium,respectively.However,only the relative contents of 4-Methoxybenzyl O-(2-sulfoglucoside)and Halistol sulfate reached their maximum value on the 4th day.3-Sulfomuconate and Cycloobrassinin sulfoxide showed a trend of increasing at first,then decreasing,and increasing at last,of which the relative contents were the lowest on 8th and the highest on 12 th.Among other differential metabolites,the content of metabolite 1-Pyrrolinium changed irregularly,while the relative contents of other metabolites increased firstly,then decreased and the greatly change of metabolites happened the sixth day.The analysis of the metabolome revealed that these metabolites involved in many metabolic pathways,such as sugar metabolism,secondary metabolite metabolism,amino acid metabolism,lipid metabolism and so on.The results showed that endophytic fungi affected not the metabolites but the metabolic pathways of R.crenulata.Studies suggested that endophytic fungi had a great impact on the metabolism of R.crenulata.In sum,the endophytic fungi affected the development and metabolism from molecular and metabolitic levels based on the results of transcriptome data and metabolome data,the influenced metabolic pathway distributed in sugar metabolism,lipid metabolism,amino acid metabolism and secondary metabolite synthesis,et al. |
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