Seed-sterilization of <em>Rhododendron wardii </em>for micropropagation

Abstract

Success in tissue culture of all plant species depends on the removal of exogenous and endogenous contaminating microorganisms. Rhododendron wardii is a parent of many of the best yellow hybrids and it has been identified as a Least Concern spp. An efficient protocol was developed for sterilization of seeds of R. wardii for in vitro cultivation for mass propagation and conservation. Three sterilizing agents, namely, HgCl2 (w/v), NaOCl (v/v) + Tween20 and H2O2 (v/v) at varying their concentration and time of exposure were tested. Treatment with 25% H2O2 for 20 min was the most effective with no contaminated cultures, where the germination percentage was 92.6% (ex vitro germination percentage was 58.61%) and produced 100% healthy plants. Even though the exposure to 15% NaOCl + Tween 20 for 20 min produced 100% contamination-free cultures, the percentage seed germination was 51%. Seeds could be sterilized with 5% NaOCl, for 20 min and 15% for 10 min for maximum germination (64%), but contamination rate was 1:2. Sterilization with HgCl2 did not produce any satisfactory result (maximum germination percentage = 18%).

High resolution microscopy to evaluate the efficiency of surface sterilization of Zea Mays seeds

“…This project was carried out in the framework of the priority program 2089 "Rhizosphere spatiotemporal organization-a key to rhizosphere functions" funded by DFG. Rhododendron wardii seeds due to tissue damage by increasing the sterilization time from 10 to 20 min and H 2 O 2 concentration from 20 to 30% was reported [27]. Hence, the damaging effect of H 2 O 2 on plant tissues and growth after seed sterilization can be different from species to species and should be empirically determined [28].…”

“…H 2 O 2 is a reactive component with a dual role of signaling (beneficial) and damaging (deleterious) for physiological properties and development of plants [ 26 ]. At low concentration H 2 O 2 possesses fungicidal and germicidal activities without affecting seed growth and germination [ 27 ]. Enzymes available in many plant cells such as catalases and peroxidases decompose H 2 O 2 into oxygen and water and protect cells from damaging effects of peroxides [ 27 ].…”

“…At low concentration H 2 O 2 possesses fungicidal and germicidal activities without affecting seed growth and germination [ 27 ]. Enzymes available in many plant cells such as catalases and peroxidases decompose H 2 O 2 into oxygen and water and protect cells from damaging effects of peroxides [ 27 ]. It was reported that after sterilization using 10% H 2 O 2 no damage on maize plants was found [ 17 ].…”

Development and Application of Low-Cost and Eco-Sustainable Bio-Stimulant Containing a New Plant Growth-Promoting Strain Kosakonia pseudosacchari TL13

“…cerasiforme ) were carefully de-husked without damaging the embryo and surface sterilized as described by Banik et al (2016) . Briefly, seeds were treated with 2% sodium hypochlorite (5 min) followed by washing with sterile distilled water, then seeds were treated with 75% ethanol (5 min), washed again with sterile water and treated with 30% hydrogen peroxide (2 min) as suggested by Amarasinghe et al (2018) ; finally, they were carefully rinsed ten times with sterile distilled water. Seeds sterility was checked by plating on Plate Count Agar (PCA; Oxoid).…”

Visible Light Active Silver Decorated Iron Titanate/Titanium Dioxide Nanohybrid for Sterilization of Explants Grown by In Vitro Technique

“…Micropropagation, the plant in vitro culture, is the commonly used technique to produce large quantities of uniform, true‐to‐type plants to meet the demand of commercial plant production. [ 1 ] The micro‐propagated plants are healthy, vigorous, and available at any season of the year. All plant species introduced to micropropagation are from the septic in vivo environment.…”