ABSTRACT
Pueraria mirifica-derived tuberous powder has been long-term consumed in Thailand as female hormone-replacement traditional remedies. The protein profiles of tubers collected in different seasons were evaluated. Phenol extraction, 2D-PAGE, and mass spectrometry were employed for tuberous proteome analysis. Out of the 322 proteins detected, over 59% were functionally classified as being involved in metabolism. The rest proteins were involved in defense, protein synthesis, cell structure, transportation, stress, storage, and also unidentified function. The proteins were found to be differentially expressed with respect to harvest season. Importantly, chalcone isomerase, isoflavone synthase, cytochrome p450, UDP-glycosyltransferase, and isoflavone reductase, which are all involved in the biosynthesis pathway of bioactive isoflavonoids, were most abundantly expressed in the summer-collected tubers. This is the first report on the proteomic patterns in P. mirifica tubers in relevant with seasonal variation. The study enlights the understanding of variance isoflavonoid production in P. mirifica tubers.
Subject(s)
Gene Expression Regulation, Plant , Phytoestrogens/isolation & purification , Plant Proteins/isolation & purification , Plant Tubers/chemistry , Proteome/isolation & purification , Pueraria/chemistry , Cytochrome P-450 Enzyme System/genetics , Cytochrome P-450 Enzyme System/metabolism , Flavonoids/biosynthesis , Gene Expression Profiling , Glucuronosyltransferase/genetics , Glucuronosyltransferase/metabolism , Intramolecular Lyases/genetics , Intramolecular Lyases/metabolism , Liquid-Liquid Extraction , Molecular Sequence Annotation , Oxidoreductases Acting on CH-CH Group Donors/genetics , Oxidoreductases Acting on CH-CH Group Donors/metabolism , Oxygenases/genetics , Oxygenases/metabolism , Phenol/chemistry , Phytoestrogens/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Tubers/genetics , Plant Tubers/metabolism , Proteome/genetics , Proteome/metabolism , Pueraria/genetics , Pueraria/metabolism , SeasonsABSTRACT
We investigated the major leaf isoflavonoid contents of Pueraria mirifica from three different cultivars (PM-III, PM-IV, and PM-V) using reverse RP-HPLC analysis. The proportions and net levels of puerarin, daidzin, genistin, and daidzein in P. mirifica leaves were found to depend on the plant cultivar and to correlate with cultivation temperature and rainfall amount. The crude leaf-extracts were tested using the Yeast Estrogen Screen (YES) assay with both human estrogen receptors (hERα and hERß). Their estrogenic activity was higher when determined by the YES system containing hERß than that with hERα and was also higher when the Δsnq2 than the wildtype yeast was employed. The results open the possibility of selecting and cultivating certain P. mirifica cultivars at a farm scale to produce a sufficient supply of leaf material to act as a starting source for the commercial scale extraction of these major isoflavonoids.
