Flavonoid Compounds and Photosynthesis in Passiflora Plant Leaves under Varying Light Intensities.

Functional constituents in the leaves of Passiflora plants contain antidepressant and antianxiety effects which are beneficial to human health and fitness. The objective of this study was to investigate leaf growth, physiological parameters, and secondary metabolite contents of Tainung No. 1 variety (P. edulis × P. edulis f. flavicarpa.) and P. suberosa in response to three light intensity conditions, including 100% light intensity (LI-100), 50% light intensity (LI-50), and 15% light intensity (LI-15) for 2 months. The leaf number, length, width, area, dry weight (DW), minimal fluorescence (Fo), maximal fluorescence (Fm), maximum photochemical efficiency of photosystem II, and soil-plant analysis development (SPAD) values of all tested plants increased with a decreasing light intensity, except for the leaf number and DW of P. suberosa plants. Low values of the net photosynthetic rate, transpiration rate, and stomatal conductance of Tainung No. 1 leaves in the LI-15 treatment showed the acclimation capacity of these plants. These observations together with high values of leaf growth traits of Fo, Fm, SPAD, and the intercellular-to-atmospheric CO2 concentration ratio indicate their physiological plasticity, which is of fundamental importance when cultivating plants in environments with different light availabilities. Wide variations occurred in total phenol (TP), total flavonoid (TF), orientin (OR), and isovitexin (IV) contents of the two Passiflora varieties, and P. suberosa contained higher TP and TF contents than did Tainung No. 1 in each light treatment but IV content of P. suberosa was lower than that of Tainung No. 1 in the LI-15 treatment. Moreover, increases in TF, OR, and IV contents of Tainung No. 1 and P. suberosa were clear in the LI-50 and LI-100 treatments, respectively, compared to LI-15 treatment. Leaf growth, physiological parameters, and secondary metabolite accumulations in Passiflora species can be optimized for commercial production via lighting control technologies, and this approach may also be applicable to leafy vegetables to produce a stable industrial supply of high leaf yields and metabolite contents.

4-Hydroxybenzoic acid serves as an endogenous ring precursor for antroquinonol biosynthesis in Antrodia cinnamomea.

Antrodia cinnamomea, an endemic fungus species of Taiwan, has long been used as a luxurious dietary supplement to enhance liver functions and as a remedy for various cancers. Antroquinonol (AQ), identified from the mycelium of A. cinnamomea, is currently in phase II clinical trials in the USA and Taiwan for the treatment of non-small-cell lung cancer. In the previous studies, we have demonstrated that AQ and 4-acetylantroquinonol B (4-AAQB) utilize orsellinic acid, via polyketide pathway, as the ring precursor, and their biosynthetic sequences are similar to those of coenzyme Q. In order to test 4-hydroxybenzoic acid (4-HBA), synthesized via shikimate pathway, is the ring precursor of AQ analogs, the strategy of metabolic labeling with stable isotopes was applied in this study. Here we have confirmed that 4-HBA serves as the ring precursor for AQ but not a precursor of 4-AAQB. Experimental results indicated that A. cinnamomea preferentially utilizes endogenous 4-HBA via shikimate pathway for AQ biosynthesis. Exogenous tyrosine and phenylalanine can be utilized for AQ biosynthesis when shikimate pathway is blocked by glyphosate. The benzoquinone ring of 4-AAQB is synthesized only via polyketide pathway, but that of AQ is synthesized via both polyketide pathway and shikimate pathway. The precursor-products relationships diagram of AQ and 4-AAQB in A. cinnamomea are proposed based on the experimental findings.

Biosynthesis of Antroquinonol and 4-Acetylantroquinonol B via a Polyketide Pathway Using Orsellinic Acid as a Ring Precursor in Antrodia cinnamomea.

Antroquinonol (AQ) and 4-acetylantroquinonol B (4-AAQB), isolated from the mycelium of Antrodia cinnamomea, have a similar chemical backbone to coenzyme Q (CoQ). Based on the postulation that biosynthesis of both AQ and 4-AAQB in A. cinnamomea starts from the polyketide pathway, we cultivated this fungus in a culture medium containing [U-13C]oleic acid, and then we analyzed the crude extracts of the mycelium using UHPLC-MS. We found that AQ and 4-AAQB follow similar biosynthetic sequences as CoQ. Obvious [13C2] fragments on the ring backbone were detected in the mass spectrum for [13C2]AQ, [13C2]4-AAQB, and their [13C2] intermediates found in this study. The orsellinic acid, formed from acetyl-CoA and malonyl-CoA via the polyketide pathway, was found to be a novel benzoquinone ring precursor for AQ and 4-AAQB. The identification of endogenously synthesized farnesylated intermediates allows us to postulate the routes of AQ and 4-AAQB biosynthesis in A. cinnamomea.

Enhancement of 4-acetylantroquinonol B production by supplementation of its precursor during submerged fermentation of Antrodia cinnamomea.

The antiproliferation activity of the ethanol extract of A. cinnamomea mycelium on hepatocellular cancer cells HepG2 was found to be associated with aroma intensity of the broth during fermentation. We hypothesized that some of the volatile compounds are the precursors of the key bioactive component 4-acetylantroquinonol B of this fungus. The major volatile compounds of A. cinnamomea were identified by GC/MS, and they are oct-1-en-3-ol, linalool, methyl phenylacetate, nerolidol, γ-cadinene and 2,4,5-trimethoxybenzaldehyde (TMBA). TMBA and nerolidol were further selected and used as supplements during fermentation. It was found that both of them could increase the production of 4-acetylantroquinonol B and enhance the antiproliferation activity of the fungus. In addition, the TMBA was identified as the most promising supplement for increasing the bioactivity of A. cinnamomea during cultivation.

The effects of Davallic acid from Davallia divaricata Blume on apoptosis induction in A549 lung cancer cells.

Traditional or folk medicinal herbs continue to be prescribed in the treatment of various diseases and conditions in many cultures. Recent scientific efforts have focused on the potential roles of extracts of traditional herbs as alternative and complementary medications for cancer treatment. In Taiwan, Davallia divaricata Blume has been traditionally employed in folk medicine for therapy of lung cancer, davallic acid being the major active compound of D. divaricata Blume. In this study, we investigated the inhibitory activity of davallic acid on the proliferation of A549 lung cancer cells. Davallic acid was extracted from D. divaricata Blume, and its effects on cell viability, cell cycle distribution, ROS level, and apoptotic protein expression in A549 cells were determined. Davallic acid significantly induced reactive oxygen species (ROS) generation as well as caspase-3, -8, and -9 activation, thereby repressing A549 cell growth and elevating apoptotic activity. Since lung cancer has a high incidence of recurrence, these results indicate that davallic acid may have the potential to be a natural anti-lung cancer compound, and may provide a basis for further study of its use in combating cancer.