A single base deletion in the MbiPTS is highly associated with failure to produce parthenolide, a promising anti-cancer drug, in Magnolia biondii

Abstract

Parthenolide (PTL) is a bioactive small molecule that selectively targets cancer stem cells. This study quantified the PTL content of 967 Magnolia biondii root bark samples, which ranged from 0.00 to 8.68% of dry weight, with an average of 4.06%. Individuals with > 6% and ~ 0% PTL were selected for genomic resequencing. Genome-wide association analysis identified MbiPTS as being strongly associated with the absence of PTL production in M. biondii. Sequence alignment revealed that the MbiPTS gene in PTL-absent plants contains a single-base deletion, resulting in a truncated, non-functional protein. MbiPTS encodes a monooxygenase that catalyzes the C4–C5 epoxidation of costunolide to form PTL. Consistent with this function, chromatographic assays showed an accumulation of costunolide in PTL-absent plants, indicating blockage of the conversion from costunolide to PTL. A kompetitive allele-specific PCR (KASP) marker was developed to target the base deletion in MbiPTS. Failure to synthesize PTL in homozygous mutants was validated by KASP genotyping in independent samples. MbiPTS function was further verified through virus-induced gene silencing and Agrobacterium rhizogenes-mediated gene overexpression. Collectively, these results elucidate the molecular basis of PTL absence in M. biondii and provide a molecular marker to facilitate selection of plants capable of PTL synthesis in future breeding programs.

Keywords

Parthenolide / Anti-cancer molecule / Magnolia biondii / Genome-wide association study / Protein truncation

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Northeast Forestry University