[Synthesis of triterpenoid saponins from Aesculus chinensis based on transcriptome data].

Yi-Ding Wei, Chao Xiong, Tian-Yuan Zhang, Han Gao, Qing-Gang Yin, Hui Yao, Wei Sun, Zhi-Gang Hu, Shi-Lin Chen
Author Information
  1. Yi-Ding Wei: School of Pharmacy,Hubei University of Chinese Medicine Wuhan 430065,China Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.
  2. Chao Xiong: School of Pharmacy,Hubei University of Chinese Medicine Wuhan 430065,China Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.
  3. Tian-Yuan Zhang: Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.
  4. Han Gao: Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.
  5. Qing-Gang Yin: Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.
  6. Hui Yao: Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100193,China.
  7. Wei Sun: Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.
  8. Zhi-Gang Hu: School of Pharmacy,Hubei University of Chinese Medicine Wuhan 430065,China.
  9. Shi-Lin Chen: School of Pharmacy,Hubei University of Chinese Medicine Wuhan 430065,China Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.

Abstract

Aesculus chinensis belongs to Hippocastanaceae family,bears medicinal and ornamental values. The oleanane type triterpenoid saponin aescin is regarded as active ingredient and accumulated in seed. In order to understand its molecular basis of the triterpenoid biosynthesis,we used high-throughput sequencing under Illumina Hi Seq 2000 platform to obtain the transcriptome data of seed and flower from A. chinensis to further mine the genes involved in its metabolic pathway. Unigene's de novo splicing was performed using Trinity software; the transcriptome results were annotated with KEGG database to predict the specific pathways of the aescin triterpenoid metabolism. terpenoid and triterpenoid pathways were found from transcriptome data,and forty seven and twenty seven corresponding genes were uncovered respectively. It was found that there are eight kinds of enzymes related to the terpenoid metabolism pathway precursors and three kinds of enzymes related to the triterpenoid metabolism pathway. In this study,five genes corresponding to triterpene cyclase were analyzed in A. chinensis for the first time,which may participate in the synthesis of triterpenoid. It' s revealed that there were thirty three differential genes associated with the ko00900 and ko00909 pathways by analysis on the difference in transcriptome expression between seeds and flowers; seventeen unigenes were up-regulated and sixteen unigenes were down-regulated in the seeds relative to flowers. In this study, qRT-PCR experiments were used to verify the expression of three key enzyme genes of SQE( Unigene25806),HMGS( Unigene36710),and β-AS( Unigene33291). The results of qRT-PCR were consistent with the transcriptome data. The candidate genes related to triterpenoid saponin aescin synthesis in A. chinensis found in this study can provide theoretical basis for the metabolism synthesis and regulation of aescin.

Keywords

MeSH Term

Aesculus
Flowers
Gene Expression Profiling
Saponins
Transcriptome
Triterpenes

Chemicals

Saponins
Triterpenes

Word Cloud

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