This first-attempt study used microtome-based methods to generate a thin cell layer culture for the micropropagation of Phal. Hwafeng Redjewel × Phal. New Cinderella. Protocorms were embedded in various agarose concentrations (8–12%, w/v) and dried from 1 to 8 h before sectioning with a microtome. Optimal conditions for slicing sections of 100 to 300 μm were achieved when the protocorms were embedded at 10% (w/v) agarose and dried for 4 h under laminar flow, and the hardness of the agarose block under these conditions reached 641.8 ± 9.5 g·cm⁻². The sectioned protocorms that were cultured on an MS medium supplemented with 1.2 mg·L⁻¹ 6-benzylaminopurine and 0.1 mg·L⁻¹ α-naphthaleneacetic acid were capable of growth and differentiated through the neoformation of protocorm-like bodies (PLBs) and/or callus before subsequent regeneration into plantlets and development into healthy plants in a nursery environment. The results of this study demonstrate that microtome-based tTCL is a reliable and promising approach for mass propagation and possible virus-free propagation objectives for Phalaenopsis.

Background: Adiantum reniforme var. sinense is a pteridophyte with considerable ornamental importance and medicinal value. However, the wild population of A. reniforme var. sinense has gradually become scarce due to sparse distribution and human activities. Green Globular Bodies (GGBs) are specific tissue which use to propagate ferns in vitro. The study uses GGBs induced from sporophytic embryos to preserve and to proliferate this pteridophyte. Methods: The present study was carried out during year 2018-2020 at National Ilan University. The developing sporophyte of Adiantum reniforme var. sinense were used to induce GGBs. The optimal medium for GGBs proliferation and differentiation in terms of basal salts, plant growth regulators, sugar concentrations and gelling agents was examined. The structure of GGBs and the embryos were analyzed by paraffin section as well. Result: The result indicates the 100% GGBs induction rate from sporophytic embryos for the first time could be obtained under BA (0.1 mg L-1) and NAA (0.15 mg L-1). For subsequent proliferation of GGBs required a higher concentration of NAA (10 mg L-1). Cytokinins such BA, KT and TDZ, inhibited GGBs proliferation. In vitro optimal amount of basal salts, plant growth regulators, sugar concentrations and gelling agents for GGB proliferation were obtained as 2 g L-1 Hyponex, 10 mg L-1 NAA, 15 g L-1 sucrose and 4 g L-1 gelrite, respectively. Paraffin sections revealed histological characteristics of the sporophytic embryo and GGB.