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Verbascoside biosynthesis pathway and verbascoside biosynthesis enzyme related genes

A technology of verbascoside and biosynthesis, which is applied in the direction of plant gene improvement, genetic engineering, biochemical equipment and methods, etc., and can solve the problems of genes to be studied

Inactive Publication Date: 2017-03-15
HENAN NORMAL UNIV
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Problems solved by technology

However, more unknown genes involved in verbascoside biosynthesis remain to be studied

Method used

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  • Verbascoside biosynthesis pathway and verbascoside biosynthesis enzyme related genes
  • Verbascoside biosynthesis pathway and verbascoside biosynthesis enzyme related genes
  • Verbascoside biosynthesis pathway and verbascoside biosynthesis enzyme related genes

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Embodiment 1

[0026] (1) Search for the previously speculated verbascoside biosynthesis pathway

[0027] Retrieval of relevant literature on the website http: / / isiknowledge.com found that some scientists previously speculated that the biosynthetic pathway of verbascoside ( figure 1 ), and some researchers chemically synthesized verbascoside (Synthesis of verbascoside: a dihydroxyphenylethylglycoside with diverse bioactivity. Eur J Org Chem 1999,10: 2623–2632) according to this pathway. However, the true verbascoside enzymatic biosynthetic pathway is unknown, and the enzymes involved in the pathway and their corresponding genes are poorly understood;

[0028] (2) Transcriptome sequencing: Take an appropriate amount of mixed roots of 6 different developmental stages of Rehmannia glutinosa 85-5, and extract total RNA with a plant total RNA extraction kit. Quantity ≥ 6μg, OD260 / 280 in the range of 1.8-2.1, rRNA Ratio (28S:18S) ≥ 1.5: 1, RIN ≥ 7 (RNA integrity number)), and the integrity of tot...

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Abstract

The invention discloses a verbascoside biosynthesis pathway and verbascoside biosynthesis enzyme related genes. Tuberous roots of rehmannia at six different development stages serve as materials, overall RNA (ribonucleic acid) is extracted and detected, combined transcriptome sequencing is performed by an Illumina HiSeq2500 platform to obtain 149.8 million reads, and head assembly is performed to obtain 96961 Unigenes sequences. The Unigenes sequences are compared with a KEGG (Kyoto encyclopedia of genes and genomes) database to perform metabolism pathway analysis and obtain 280 KEGG pathways, a metabolism pathway is compared with a supposed verbascoside biosynthesis pathway, and 5 candidate KEGG pathways are screened. The supposed verbascoside biosynthesis pathway is compared with the screened 5 candidate KEGG pathways, 19 gene coding enzyme catalysis verbascoside biosynthesis pathways are built, the Unigenes sequences and the database are subjected to Blast X comparison, and ESTscan software functions are added to obtain 13 candidate verbascoside biosynthesis enzyme related genes. A foundation is laid for illuminating the complete verbascoside biosynthesis pathway and producing verbascoside by secondary metabolism engineering or synthetic biology.

Description

technical field [0001] The invention belongs to the technical field of verbascoside biosynthesis, and in particular relates to a biosynthesis pathway of verbascoside and a synthetase-related gene thereof. Background technique [0002] Actascoside is a phenylethanol glycoside compound, which is distributed in the roots, stems, leaves and flowers of more than 200 plants of 23 families such as Mullein, Rehmannia glutinosa and Cistanche. It has anti-oxidation, anti-inflammatory, anti-tumor, wound healing, liver protection, Neuroprotective and immunomodulatory effects. Relevant researchers have deduced the verbascoside biosynthetic pathway based on stable isotope-labeled precursor feeding experiments (such as figure 1 shown), and verbascoside was chemically synthesized accordingly (Synthesis of verbascoside: adihydroxyphenylethyl glycoside with diverse bioactivity. Eur J Org Chem 1999,10: 2623–2632). However, the actual verbascoside biosynthetic pathway is unknown, and the enzy...

Claims

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Application Information

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IPC IPC(8): C12P19/44C12N15/60C12N15/54C12N15/53C12N15/52
CPCC12N9/0022C12N9/0073C12N9/1029C12N9/1096C12N9/88C12N9/93C12P19/44C12Y104/03021C12Y114/13011C12Y114/13036C12Y203/01133C12Y206/01001C12Y401/01025C12Y402/01091C12Y602/01012
Inventor 周延清王向楠段红英张亮张丹丹杨柯
Owner HENAN NORMAL UNIV
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