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    Vitro rooting in liquid cultures

    The physical properties of the medium have significant consequences for micropropagation protocols. Vegetative propagation of most commercially important crops, such as vegetables, medicinal plants and fruit trees, can be done using tissue culture. Micropropagation is labor-intensive and time-consuming depending on the species. Micropropagation is often done using semi-solid media. This medium provides support for the plants, but liquid culture is better for some species. It can also reduce subculturing and allows for more automation. Semi-solid media is not recommended for micropropagation. The gel must be removed from the roots by hand to prevent microbial growth in the growing facility. This is not only tedious but can also cause damage to the roots and increase the risk of infection. Micropropagation is not commercially viable due to its high cost and low efficiency. It cannot be used for high-value plants or other unique applications.

    Liquid medium provides

    A liquid medium provides several advantages over a semi-solid medium. Liquid media is easier to use, more convenient to replenish, has high nutrient diffusion rates and lowers oxidative stress on the plant. Liquid media are generally more productive than traditional gel-based micropropagation methods. For a variety of species, liquid has been shown to be more effective than semi-solid media in comparison. Because gel is not required to be removed from roots, rooted in liquid medium is more cost-effective and advantageous. Semi-solid culture is advantageous during rooting because it supports the roots to prevent auxins from reaching the base. This is essential to promote healthy foliar development and rooting. One challenge with liquid culture systems is the difficulty of keeping micro-shoots upright during rooting. This can cause abnormal foliar growth. In vitro rooting in liquid medium is possible using test-tubes or small flasks or artificial support such as glass beads or coir 7. There are many reports. These methods are difficult to commercialize because they require large quantities of vessels, space and labor to clean support materials and glass.

    Micropropagation’s rooting stage is crucial as it directly impacts greenhouse survival and acclimatization. To maximize the faster rooting process in liquid culture systems, it would be beneficial to create a system that supports explants upright during the rooting stage. This study was designed to test and develop a scaffold system that provides support for liquid-based rooting.

    Iterative design and 3D printing were used to create a system compatible with commercially-available culture vessels. Figure 1: The scaffold is made up of two pieces. It forms a grid that can be used to hold the shoots upright. After growth, the two pieces of the grid are easily separated and can be removed gently without causing damage to the roots. The system was tested with either static liquid culture or a combination of a temporary immersion rocker and was compared to semi-solid culture for three species of banana (Musa) spp. 2 MATERIALS AND METHODS

    2.1 3D printed Rootscaffold

    A two-piece rooting structure was created using AXIOM 3D printers (Airwolf3D CA, USA) as well as 2.85mm polycarbonate (PC), filament (Fly Thinking Material China). All units were created using Fusion 360 (Autodesk USA) software. They were exported as Stereo Lithography file. Simplify3D (Simplify3D in the United States) printing software was used to create Stereo Lithography files. These files were then exported as gcodes. There were several iterations of this design (not shown), until the final one was chosen based on its fit into the vessel and ease-of-use. Figure 1 shows the final root-stand design dimensions. They were 235 x 85x 80 x 80 mm (lxbxh) and 2.75 mm thick.

    2 In vitro culture of banana, apple rootstock, and cherry birch

    Cherry birch (Betula lenda) and apple (Malus domestica.cv.) are in vitro-grown. Geneva 41), and bananas (Musa.sp. Plantlets were taken from the University of Guelph’s germplasm collection (GRIPP). They were cultured in previously optimized media for multiplication, and then for rooting as described below.

    PRACTICAL APPLICATION

    Although there are many advantages to using liquid culture systems for different plant species, commercial micropropagation is not possible. The inability to hold individual micro-shoots upright during rooting is a major problem that prevents liquid culture systems from being used. This can cause abnormal root development. Two-piece scaffold design allows shoots to be placed in a grid and kept upright during denver pop culture con rooting. This makes micropropagation easier and more cost-effective. The root stand makes it possible to transfer plants easily without causing damage to the plantlets. This increases the survival rate. This new design can solve a major problem in plant tissue culture. It also allows for easy transfer of plants without damage to the plantlets. In vitro-grown shoots were transferred to DKW basal salt mixture containing vitamins, 20 uM indole-3,butyric acid and 3% sucrose 10. The in vitro apple shoots were then multiplied using DKW basal sodium mixture with vitamins, 2 uM BA, 3% sucrose. In vitro-grown shoots were transferred to rooting media containing DKW basal sodium mixture with vitamins, 2.5uM indole-3,butyric acid and 3% sucrose. In vitro banana shoots were grown on MS (Murashige, Skoog medium), 11 basal salt mixture containing vitamins (PhytoTechnology Shawnee Mission. KS, USA), 30 uMBA, 5 uM Kinetin and 3% sucrose. The shoots that had been grown in vitro were transferred to rooting media containing MS basal sodium mixture with vitamins, 10uM Naphthaleneacetic Acid (NAA), and 33% sucrose.

    After the pH had been adjusted to 5.75, all media were solidified with 2.2 g/L phytagel. After 20 minutes at 121°C and 118 kPa, the culture vessels were heat-stripped. Autoclaved media media (50mL) for plant multiplication were then dispensed in Magenta(r), GA7 boxes. After transferring two plants onto the culture media, the magenta boxes were sealed using Micropore tape (Fisher Scientific Canada). The shoots were jill whelan kept at 25 +-2degC for 16 hours under light intensity of 40 umol*m-2*s-1 by cool white fluorescent lamps (Osram, Mississauga)

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