| Baphicacanthus cusia is one of the geoherbs of Fujian,studies on the accumulation of effective conponents;especially the indican biosynthesis and accumulation are limited due to insufficiently available datasets.In addition,the genomic information of this herb is also unavailable.In this study,we investigated the accumulations of inorganic element and effective constituent in the root,stem and leaf,analyzed the transcriptome profiles leaf and root were by Illumina RNA-Seq,annotated the differentially expressed genes(DEGs),especially DEGs related the accumulations of inorganic element and effective constituent,The results from be helpful in illuminating the biosynthetic pathway for tissue-specific synthesis of secondary products in non-model plant organisms,as well as in developing strategies for enhanced biosynthesis of indican through biotechnological approaches and metabolism regulation.It will provide understanding of the biosynthesis and accumulation of effective constituent of Baphicacanthus cusia at the molecular level.1.In this study,contents of fifteen elements(Fe,Ca,Mg,Al,Cu,Ba,Zn,K,Mn,Ni,Na,Cr,As,Pd,Cd)in the root,stem,leaf and soil of Baphicacanthus cusia were determine by Inductively coupled plasma-atomic emission spectrometry(ICP-AES),the results showed that the K element content in the root is highest than in stem and leaf,the Ca element content in the stem is highest than in leaf and root,the Mg element content in the leaf is highest than in root and stem,the Fe element contents in the Baphicacanthus cusia were root<stem<leaf,the Cu,Zn,Ni element contents in the Baphicacanthus cusia were root>stem>leaf,the Mn and Al element contents in the Baphicacanthus cusia were leaf>root>stem,the Na element contents in the Baphicacanthus cusia were root>leaf>stem,the Ba element contents in the Baphicacanthus cusia were stem>root>leaf,by reference to Green Industry Standards of Medicinal Plants and Preparations for the Import and Export,the content of Cu,Pb,As met the minimum performance,while the content of Cd exceeded the standard.2.The spectrum and chromatography fingerprints were established by Fourier transform infrared spectroscopy(FTIR)and high performance liquid chromatography(HPLC).The results showed that the FTIR fingerprints of roots,stems and leaves from ten kinds of habitats are very similar,the FTIR fingerprints of roots and stems had a absorption peak at 1260cm-1,the FTIR fingerprints of leaves were not obvious.Seven common peaks were identified in The leaf HPLC fingerprint of Baphicacanthus cusia,similarities of the leaf HPLC fingerprint were all above 0.98,it showed that ten batches leaf HPLC fingerprints are similar.3.Quantification of indigo,indirubin,adenosine and indican was conducted using HPLC-DAD.The volatile components were extracted with distillation and extraction apparatus and analyzed by gas chromatography mass spectrometry(GC-MS).Indigo and indirubin,as well as adenosine and indican,were compared among the leaves,stems and roots of Baphicacanthus cusia.Leaves showed significantly higher indigo,indirubin,adenosine and indican contents than roots in Baphicacanthus cusia from medicinal garden.contents of indigo,indirubin,adenosine and indican in tender leaf from Fuzhou were 595.04?g·g-1,681.40?g·g-1,4675.36?g·g-1,16062.77?g·g-1,contents of indigo,indirubin,adenosine and indican in tender leaf from Putian were 380.66?g·g-1,315.15?g·g-1,4323.15?g·g-1,20978.26?g·g-1,contents of indigo,indirubin,adenosine and indican in root from Fuzhou were l0.26?g·g-1,4.55?g·g-1,227.52?g·g-1,8515.44?g·g-1,contents of indigo,indirubin,adenosine and indican in root from Putian were 9.31?g·g-1,7.82?g·g-1,197.48?g·g-1,170.45?g·g-1,respectively.The contents of indigo,indirubin(1906.55?g·g-1,1575.48?g·g-1)in leaves collected at July were highest,contents of indigo,indirubin,adenosine and indican in tender leaf from cultivation base were 861.03?g·g-1,271.49?g·g-1,1224.12?g·g-1,3637.46?g·g-1,contents of indigo,indirubin,adenosine and indican in root from cultivation base were 24.33p?g·g-1,10.58?g·g-1,111.94?g·g-1,8901.90?g·g-1.Root showed significantly higher indican contents than leaf in Baphicacanthus cusia from cultivation base.29 chemical components were identified preliminary from essential oil of root,stem and leaf,among them 11 chemical components were common peak.13 chemical components were identified preliminary from essential oil of root,24 chemical components were identified preliminary from essential oil of stem,and nineteen chemical components were identified preliminary from essential oil of leaf.4.At the same time,we performed de novo transcriptome sequencing for the species leaves and roots.Sequencing yielded 43,037,897 and 40,189,081 clean reads from leaves and roots,respectively,which were assembled into 187,765 transcripts and 71,071 unique sequences.A total of 35,046 unigenes could be annotated using the Nr,Swiss-Prot,KEGG,COG,and GO databases.Furthermore,a total of 5,597simple sequence repeats(SSRs),70720 coding aequences and 1332589 SNP were identified from 71,071 unigenes.Based on this resource;two DGEs libraries were established to capture the unigenes differences between the leaf and root libraries.Finally,a set of 6,580(2,751 up-regulated and 3,829down-regulated)differentially expressed genes was screened,and 5,661 unigenes were annotated by Nr,Swiss-Prot,KEGQ COQ and GO database.From 5661 unigenes annotated,we identified four DEGs related to nitrogen metabolism,42 DEGs related to indole and its derivative.A total of 87 CYP450 DEGs were identified,thirty-nine UGT DEGs were identified,a total of 38 glucosidase DEGs were identified,four tryptophan synthase DEGs were identified,three tryptophan amino transferases DEGs were identified.The DEGs related to the contents of mineral nutrition analysis result showed that The DEGs expression and the contents of mineral nutrition element have basical positive correlation.Terpenoid biosynthesis passway analysis result showed that terpenoid biosynthesis passway of Baphicacanthus cusia was DXP.34 DEGs related to terpenoid biosynthesis were identified,such as phosphomevalonate kinase.27 DEGs related to phenylpropanoid biosynthesis were identified,such as flavonoid 3'-monooxygenase. |
PDF Full Text Request