[Determination of huperzine A and semi-quantitative RT-PCR analysis of lysine decarboxylase gene in different parts of Huperzia serrata from western Hunan].

Zhong Yao Cai

College of Biology and Environmental Sciences, Jishou University/Key Laboratory of Plant Resource Conservation and Utilization, College of Hunan Province, Jishou 416000, China.

Published: March 2013

Objective: To determine the content of huperzine A in the root, stem and leaf of Huperzia serrata from western Hunan, and analyze the relationship between the distribution of huperzine A and the expression of lysine decarboxylase gene in different parts of huperzia serrata.

Methods: The content of huperzine A in the root, stem and leaf of Huperzia serrata was determinated by HPLC, of which the chromatographic column was Diamonsil C18 (250 mm x 4.6 mm,5 microm), mobile phase was methanol-0.08 mol/L CH3COONH4 with flow rate at 1.0 mL/min, column temperature at 25 degrees C and detection wavelength at 308 nm. The expression of lysine decarboxylase gene in different parts of Huperzia serrata was analyzed by semi-quantitative RT-PCR, the beta-actin gene was used as internal reference.

Results: Huperzine A had good linear relationships within the range of 0.5 - 10.0 microg/mL, with the recovery rate of 102% and RSD 0.32%. The content of huperzine A in the root,stem and leaf of huperzia serrata was (118.35 +/- 0.77) microg/g and (411.09 +/- 2. 47) microg/g, (562.15 +/- 2.86) microg/g, respectively. Semi-quantitative RT-PCR results showed that lysine decarboxylase gene had nearly identical expression in the root, stem and leaf of Huperzia serrate.

Conclusion: The content of huperzine A changes with different parts of Huperzia serrata, lysine decarboxylase might be not the key enzyme to regulate the biosynthesis of huperzine A.

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