Assessment of genetic and epigenetic variation during long-term Taxus cell culture.

Plant Cell Reports
Chunhua FuLongjiang Yu

Abstract

Gradual loss of secondary metabolite production is a common obstacle in the development of a large-scale plant cell production system. In this study, cell morphology, paclitaxel (Taxol®) biosynthetic ability, and genetic and epigenetic variations in the long-term culture of Taxus media cv Hicksii cells were assessed over a 5-year period to evaluate the mechanisms of the loss of secondary metabolites biosynthesis capacity in Taxus cell. The results revealed that morphological variations, gradual loss of paclitaxel yield and decreased transcriptional level of paclitaxel biosynthesis key genes occurred during long-term subculture. Genetic and epigenetic variations in these cultures were also studied at different times during culture using amplified fragment-length polymorphism (AFLP), methylation-sensitive amplified polymorphism (MSAP), and high-performance liquid chromatography (HPLC) analyses. A total of 32 primer combinations were used in AFLP amplification, and none of the AFLP loci were found to be polymorphic, thus no major genetic rearrangements were detected in any of the tested samples. However, results from both MSAP and HPLC indicated that there was a higher level of DNA methylation in the low-paclitaxel yielding cell l...Continue Reading

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Citations

Sep 6, 2015·Journal of Chromatographic Science·Xiao-li LiLong-jiang Yu
Sep 4, 2015·Physiologia Plantarum·Yan-Shan DongLong-Jiang Yu
Aug 11, 2016·Journal of Biological Research·Renata OrłowskaPiotr Tomasz Bednarek
May 18, 2019·International Journal of Molecular Sciences·Abbas KhojastehJavier Palazon

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