Sterilization of <i>Hibiscus rosa-sinensis</i> L. Vegetative Explants Sourced from Plants Grown in Open Environment and Influences of Organic Ingredients on <i>In Vitro</i> Direct Regeneration

Abstract

This paper reports on the effects of organic ingredients in facilitating direct shoot regeneration from nodal explants of Hibiscus rosa-sinensis L. This paper also compares the sterilization conditions for 3 types of explants (node, internode, and shoot tip) harvested from an open field. The optimized sterilization conditions for the explants were 40% Clorox-20 min exposure, 10% Clorox-15 min exposure, and 5% Clorox-40 min exposure for the node, internode and shoot tip, respectively. In the direct shoot regeneration using the nodal explants, we found MS medium containing 40 g/L sucrose, 0.3% (w/v) activated charcoal, and supplementations with myo-inositol, thiamine and nicotinic acid were suitable. The in vitro shoot survival rate was 30% with a mean leaf numbers of 2.68 produced, and a mean leaf length of 1.71 cm achieved after 5 weeks of culture on the modified medium.

Elicitation of the in vitro Cultures of Selected Varieties of Vigna radiata L. With Zinc Oxide and Copper Oxide Nanoparticles for Enhanced Phytochemicals Production

“…This can be related to the results of the present study, where the selection of in vitro explants decreased the chances of contamination, which subsequently increased the overall response of mung bean cultures. Shoots and internodes are already considered as better explants (Dar et al, 2012 ). Similarly, Himabindu et al ( 2014 ) developed an effective method to increase shoot multiplication in the green gram culture using cotyledonary nodes.…”

Preliminary study on in vitro shoot culture of Hibiscus coddii subsp. barnardii, an indigenous South African flowering plant

“…Disinfection with 70% ethanol (60 s) followed by treatment with 25% (v/v) bleach solution for 25 min and 50% (v/v) bleach solution for 15 min resulted in 85% and 80% contamination of in vitro cultures, respectively, and high mortality (80%) of the explants. In contrast, Dar et al (2012) reported 88% survival and no contamination of nodal explants of H. rosa-sinensis collected from an outside garden when surface-disinfected with 50% (v/v) Clorox ® [5.25% (w/v) NaOCl] bleach solution containing a higher concentration (2.63%) of NaOCl. In general, however, microbial contaminations of in vitro cultures are found to occur more readily when the explant source is taken directly from field grown plants (Lobodina et al, 2020).…”

“…In general, however, microbial contaminations of in vitro cultures are found to occur more readily when the explant source is taken directly from field grown plants (Lobodina et al, 2020). Disinfection of nodal explants is considered to be more difficult since the attachment of the petiole to the stem at an angle creates a V-shaped trough wherein contaminants can be trapped, making it more difficult to sterilize nodal explants with axillary buds (Dar et al, 2012).…”

Propagation of Dracaena Umbraculifera Plant by Tissue Culture Technique

“…Sakr et al (2011) reported that surface sterilization with 20% Clorox gave the best result of survival for shoot tips and axillary buds of Cerbera odollam (Apocynaceae). Dar et al (2012) stated that the optimized sterilization conditions for Hibiscus rosa-sinensis node explants were 40% Clorox for 20 min exposure. Dar et al (2012) on Hibiscus rosa-sinensis stated that the optimium sterilization conditions for explants were 40% Clorox-20 min exposure, 10% Clorox-15 min exposure, and 5% Clorox-40 min exposure for the node, internode and shoot tip, respectively.…”

“…Dar et al (2012) stated that the optimized sterilization conditions for Hibiscus rosa-sinensis node explants were 40% Clorox for 20 min exposure. Dar et al (2012) on Hibiscus rosa-sinensis stated that the optimium sterilization conditions for explants were 40% Clorox-20 min exposure, 10% Clorox-15 min exposure, and 5% Clorox-40 min exposure for the node, internode and shoot tip, respectively. Abd Alla et al (2013) treated stem nodes of cassava (Manihot esculenta), with different concentrations of Clorox (10, 20 and 30%).…”