Abstract
To improve the production of cephalotaxine and explore its biosynthesis pathway in the suspension cells of Cephalotaxus mannii, the cell suspension cultures were separately treated with elicitors, including coronatine (0.5, 1.0, 2.0 μmol/L) and methyl jasmonate (100 μmol/L), and then analyzed their transcriptome differences. The results showed that coronatine could improve the content of cephalotaxine in suspension cells of C. mannii. The highest product (6.75 mg/L) appeared in the coronatine treatment of 1.0 μmol/L, which was higher than that of methyl jasmonate treatment (4.29 mg/L) and control (3.14 mg/L). The key enzyme of the shikimic acid pathway, 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase (DS) activity increased after 24 h of adding elicitors above. Transcriptome analysis indicated that the phenylpropane pathway, shikimic acid pathway, glucose metabolism process, phenylalanine, and tyrosine synthesis pathway were involved in the synthesis of cephalotaxine. In addition, glucose-6-phosphate dehydrogenase (G6PDH), hexokinase, phenylalanine ammonialyase (PAL), and Cytochrome P450, etc. were found out to be closely related to the cephalotaxine accumulation. It could be concluded that coronatine promoted the synthesis of cephalotaxine by affecting the expression of some enzymes, especially DS in the synthesis pathway of cephalotaxine.
Key message
Coronatine (1.0 μmol/L) led a yield of 6.75 mg/L cephalotaxine, which was 2.15 times of the control. The transcriptome analysis indicated DS played an important role in cephalotaxine biosynthesis.
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Abbreviations
- CDS:
-
Coding sequence
- COR:
-
Coronatine
- CT:
-
Cycle threshold
- DEGs:
-
Differential expression genes
- DS:
-
3-Deoxy-d-arabino-heptulosonate-7-phosphate synthase
- E4P:
-
Erythrite 4-phosphate
- FW:
-
Fresh weight
- G6P-2Na:
-
d-Glucose-6-Phosphate 2Na
- G6PDH:
-
Glucose-6-phosphate dehydrogenase
- KT:
-
Kinetin
- MJ:
-
Methyl jasmonate
- NAA:
-
1-Naphthylacetic acid
- NADP:
-
Nicotinamide adenine dinucleotide phosphate
- PEP:
-
Phosphoenolpyruvate
- PVP:
-
Polyvinyl pyrrolidone
- TCA:
-
Trichloroacetic acid
- TTC:
-
Triphenyltetrazolium chloride
- VB1:
-
Vitamin B1
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This work was financially supported by the National Natural Science Foundation of China (Grant No. 21766006).
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This research was conceived and designed by LW and YL. The specific experiments were conducted by LW and ZW. The data analysis and writing were accompanied by LW and QZ. The manuscript was finally revised by YL.
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Wang, LY., Zhang, Q., Wang, ZQ. et al. Effect of coronatine on synthesis of cephalotaxine in suspension cells of Cephalotaxus mannii and its transcriptome analysis. Plant Cell Tiss Organ Cult 147, 209–220 (2021). https://doi.org/10.1007/s11240-021-02117-z
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DOI: https://doi.org/10.1007/s11240-021-02117-z