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.
Heritage trees carry both botanical and historical value for a city’s resilience and sustainability and hence are precious and unique. Their transplant is costly and very rare due to tremendous cost and 100% survival requirement by law. Rootless transplant is even more detrimental to the heritage tree due to removal of roots infected by brown root rot (BRR) before transplanting. This study examined the adventitious roots (AR) induction ability of the Ficus elastica Roxb. heritage tree infected with BRR. The experimental design considered three factors: root diameter (RD), wounding method (WM), and auxin solution on aerial roots under fractional factorial experiment in completely randomized design (CRD). There were four RD groups: RDI (RD < 2 cm), RDII (2 ≤ RD ≤ 4.3 cm), RDIII (4.3 < RD ≤ 22), and RDIV (RD > 22); three WMs: cutting off (CF), girdling (GD), and rectangular shape peeling (RP) of aerial roots; and three auxin solutions: 2000 mg·L−1 IBA(Indole-3-butyric acid) (2B), 2000 mg·L−1 IBA + 2000 mg·L−1 NAA(1-Naphthaleneacetic acid) (2NB), and 4000 mg·L−1 IBA (4B) plus water as control (C). The number of rooting wounds, number of roots, and the mean length of the three longest adventitious roots in each wound were recorded to evaluate the AR rooting performance. Twenty four treatment combinations including 328 wounds were tested. The results showed that rooting ability was significantly correlated with RD and WM. Smaller RDs had better rooting and declined with increased RDs. CF had the best rooting followed by GD and then RP. Auxin solution did not significantly affect the rooting ability. It may be due to the abundant endogenous auxin in the heritage tree, which mitigated the effect of exogenous auxin for AR induction. We conclude that cutting off small-diameter aerial roots is the best approach to induce ARs from rootless F. elastica heritage trees to enhance transplantation success.
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