In order to explore the function of the Pq3-O-UGT2 promoter, chromosome walking technology was used to isolate the 1399 bp sequence upstream of the ATG initiation codon of Pq3-O-UGT2 from Panax quinquefolium L. Bioinformatics analysis shows that the nucleic acid sequence contains a large number of typical structures unique to eukaryotic promoters and many other important cis-acting regulatory elements, including light responsive elements, hormone-responsive elements and stress-responsive elements, etc. Seven fragments including the full-length promoter and six 5′ terminal series deleted fragments were fused with the GUS reporter gene to test their activities. The results of histochemical staining show that a strong GUS activity were observed in owers, siliques, leaves, stems and roots of transgenic Arabidopsis containing the full length Pq3-O-UGT2 promoter. Different GUS activity were also observed in the seedlings of transgenic Arabidopsis containing the full length promoter and six 5′ terminal series deleted fragments. Fluorometric assays show that seven fragments were found to drive GUS expression, and the highest enzyme activity is the full-length fragment with 4370 pmol 4-MU/min/mg protein, which is 80.01% of the CaMV35S promoter. Followed by P-801::GUS with 2162 pmol 4-MU/min/mg protein, and the shortest promoter containing P-198::GUS with 45 pmol 4-MU/min/mg protein was su cient to activate GUS expression. In addition, extended light, low temperatures, Methyl jasmonate(MeJA), Abscisic acid(ABA), NAA and GA3 were selected to investigate the Pq3-O-UGT2 promoter in response to abiotic stress and hormone treatment. The promoter activity of the full length can be enhanced much more than the other six 5′ terminal series deleted fragments, and the most signi cant change was detected in MeJA treatment with 2.12 times increased. Furthermore, it was found that the promoter activity of P-998::GUS can be enhanced by ABA with 1.47 times. Above results show that the GUS activity of different promoter fragments had different response to different environmental factors. This article provides a great understanding of complex regulatory mechanisms of Pq3-O-UGT2 and the molecular mechanisms of triterpene biosynthesis. Key MessageThe Pq3-O-UGT2 promoter was successfully cloned. Response to hormones and abiotic stress treatments of seven fragments including the full-length promoter and six 5′ terminal series deleted fragments were studied.

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Introduction: Lycopodiella cernua is a widely distributed species recognized for several uses in traditional medicine. Although relatively common in the wild, relevant pharmacological data on Philippine L. cernua population seems scanty despite its use in traditional medicine among some of the country's tribal groups. Objectives: The study aims to validate the medicinal potential as anti-inflammatory and antioxidant to strengthen traditional claims of L. cernua. Materials and Methods: Crude methanolic extraction was done for aerial and ground parts of Lycopodiella cernua. The extracts were measured for total phenolic content (TPC) and total flavonoid content (TFC) with Folin-Ciocalteu and aluminum chloride methods, respectively. Antioxidant activity was also determined based on DPPH radical scavenging activity relative to ascorbic acid. Antiinflammatory activity was detected by enzyme cyclooxygenase (COX-2 and COX-1) inhibition assays. Moreover, cytotoxicity tests of extracts against neonatal human epidermal keratinocytes (HEKn) and lung adenocarcinoma (A549) cell line were done using MTT assays. Results and Discussion: Phenolics (aerial: 33.04 ± 2.82 mg GAE/g; ground: 4.91 ± 0.1 mg GAE/g) and flavonoids contents (aerial: 11.46 ± 7.19 µg QE/g; ground: 5.82 ± 5.09 µg QE/g) were relatively low. DPPH radical scavenging activity was also relatively low (aerial: 19.13 ± 0.88 %; ground: 12.21 ± 0.9 %). However, COX-2 inhibition activity of the ground parts was 74.78 ± 18.61%. Moreover, the COX-2: COX-1 ratio of 1.29 of the aerial parts suggests selectivity for COX-2. Both extracts were found nontoxic against HEKn normal cell lines and A549 cancer cell lines. Conclusion: L. cernua is nontoxic and has anti-inflammatory and antioxidant activities.

Diplazium esculentum, Drynaria quercifolia, and Asplenium nidus are among the fern species found in Bukidnon, Philippines which are used as traditional herbal medicines.  The HPTLC profile and the anti-inflammatory properties against cyclooxygenase (COX) and 15-lipoxygenase (15-LOX) of the frond ethanolic extracts of D. esculentum, D. quercifolia, and A. nidus were determined.  The High-Performance Thin Layer Chromatography (HPTLC) profile was obtained using ethyl acetate: formic acid: water (16:2:2) as mobile phase and Natural Products (NP) as derivatizing reagent.  The HPTLC profile of the D. esculentum, D. quercifolia, and A. nidus extracts showed 10 (Rf = 0.02-0.97), 13 (Rf = 0.03-0.90), and 14 (Rf = 0.02-0.99) bands, respectively. The profiles for each fern species may be used as marker for quality evaluation and standardization of herbal formulations containing these plants.  For the anti-inflammatory properties, D. esculentum and D. quercifolia extracts which inhibited more than 50% of the COX enzymes showed significantly higher activity than A. nidus and were considered active against COX-2 and COX-1. D. esculentum, however, gave a selectivity ratio (COX-2/COX-1) of 1.03 making its inhibitory activity selective against COX-2.  The percent 15-LOX inhibitory activity of D. quercifolia (58.62%) is significantly higher than that of A. nidus (38.70%) but statistically comparable to D. esculentum (51.19%).  Among the extracts, D. quercifolia and D. esculentum which inhibited more than 50% of the 15-LOX were considered active. D. esculentum and D. quercifolia can therefore be potential sources of anti-inflammatory lead compounds for future drug development.