C. S. Tee

Induction of in vitro flowering in the orchid Dendrobium Sonia 17

In this study, Dendrobium Sonia 17 plantlets were used to induce in vitro flowering. Inflorescences were induced and rooting was inhibited in the half-strength Murashige and Skoog medium containing 20 µM N6-benzyladenine (BA). The medium with high P and low N contents was effective to induce inflorescences while the medium with low P and high N contents was only effective to promote forming of shoots. In addition, the induced in vitro inflorescences were able to multiply and maintain without exhibiting a distinctive vegetative phase. Different morphologies of in vitro flowers such as incomplete flower structures, abnormal and unresupinated in vitro flowers were observed.

Selection of co-transformed Dendrobium Sonia 17 using hygromycin and green fluorescent protein

Dendrobium Sonia 17 calluses were used for co-transformation study using particle bombardment. The bombarded transformed callus tissues were selected using half-strength MS medium containing 25 mg dm−3 hygromycin. Expression of green fluorescent protein (GFP) was observed in the callus and protocorm-like body (PLBs) tissues survived on the selection medium. The presence of green fluorescence protein (sgfp), hygromycin-B-phosphotransferase (hptII) and β-glucuronidase (uidA) genes in the transformed tissues were verified using PCR, Southern blot and dot blot analyses. Based on the results from PCR and expression of sgfp and uidA genes in the calluses and PLBs survived from hygromycin selection, we reported the co-transformation of sgfp, hptII and uidA genes into Dendrobium Sonia 17. GFP-expressing tissues were also observed in the regenerated transformed plantlets.

Alpha-glucosidase inhibitory and antioxidant potential of antidiabetic herb Alternanthera sessilis : Comparative analyses of leaf and callus solvent fractions

Background: Alternanthera sessilis is a medicinal herb which is consumed as vegetable and used as traditional remedies of various ailments in Asia and Africa. Objective: This study aimed to investigate the antiglucosidase and antioxidant activity of solvent fractions of A. sessilis leaf and callus. Materials and Methods: Leaf and callus methanol extracts were fractionated to produce hexane, chloroform, ethyl acetate, butanol, and water fractions. Antiglucosidase and 1,1-diphenyl-2-picrylhydrazyl scavenging activities as well as total phenolic (TP), total flavonoid (TF), and total coumarin (TC) contents were evaluated. Lineweaver–Burk plot analysis was performed on leaf and callus fractions with the strongest antiglucosidase activity. Results: Leaf ethyl acetate fraction (LEF) had the strongest antiglucosidase (EC50 0.55 mg/mL) and radical scavenging (EC50 10.81 μg/mL) activity among leaf fractions. Callus ethyl acetate fraction (CEF) and chloroform fraction had the highest antiglucosidase (EC50 0.25 mg/mL) and radical scavenging (EC50 34.12 μg/mL) activity, respectively, among callus fractions. LEF and CEF were identified as noncompetitive and competitive α-glucosidase inhibitors, respectively. LEF and CEF had greater antiglucosidase activity than acarbose. Leaf fractions had higher phytochemical contents than callus fractions. LEF had the highest TP, TF, and TC contents. Antiglucosidase and antioxidant activities of leaf fractions correlated with phytochemical contents. Conclusion: LEF had potent antiglucosidase activity and concurrent antioxidant activity. CEF had the highest antiglucosidase activity among all fractions. Callus culture is a promising tool for enhancing production of potent α-glucosidase inhibitors. Abbreviations used: LHF: Leaf hexane fraction, LCF: Leaf chloroform fraction, LEF: Leaf ethyl acetate fraction, LBF: Leaf butanol fraction, LWF: Leaf water fraction, CHF: Callus hexane fraction, CCF: Callus chloroform fraction, CEF: Callus ethyl acetate fraction, CBF: Callus butanol fraction, CWF: Callus water fraction, TP: Total phenolic, TF: Total flavonoid, TC: Total coumarin.