Chemical sterilization of culture medium for in vitro multiplication of Cochlospermum regium

Abstract

Cochlospermum regium roots are used in popular medicine and its extract has diverse phytochemical molecules some with antimicrobial activity, consequently exposing this specie to genetic erosion risks. Thus, the objective of this study was to develop an in vitro multiplication protocol using chemical sterilization of culture medium. Therefore, explants obtained from apical buds of C. regium seedlings were inoculated into with 0.05mg L-1 NAA and 1mg L-1 BAP sterilized by chemical agent sodium hypochlorite (NaOCl) at 0.001%, 0.003% and 0.005% of active chlorine (Cl). Autoclaved culture medium was used as control. Result showed that the contamination by bacterial at 91 days of cultivation was significantly (P<0.05) controlled by autoclaving, 0.001% and 0.005% Cl. Moreover, the callus induction in the culture medium with 0.001% and 0.005% Cl was, respectively, 30% and 20% major than autoclaving sterilization. There was not significant (P<0.05) in the percentage of shoot induction among the sterilization preparations methods, and 65% of the explants survived in the presence of culture medium with 0.005% Cl. Histological analyses indicated that the Cl did not have any deleterious effects on morphogenic events. These results indicated that the chemical sterilization using 0.001% - 0.005% Cl controlled the fungal and bacterial multiplication in the culture medium and no affected the C. regium explants development, becoming it an alternative to autoclaving method.

WITHDRAWN: Reduction in Microbial Manifestation on In Vitro Culture of Eucalyptus Microcorys

“…At low active chlorine concentrations (up to 0.005%), sodium hypochlorite has proven viable in controlling bacterial manifestation in in vitro cultures of Eucalyptus benthamii at the establishment phase [16], Eucalyptus pellita at the multiplication phase [22], Sequoia sempervirens L. [21], the hybrid Gerbera hybrida cv. Essandre [24], Cochlospermum regium [25] and Bambusa vulgaris [26]. Our results for Eucalyptus mycrocoris agree with the literature: sodium hypochlorite at 0.003% produced the best response by eliminating microbial manifestations in 25% of individual 2 explants and 5% of individual 1 explants.…”

“…In the literature, evaluating the effect of chemical sterilization on the culture medium of in vitro Cochlospermum regium multiplication, we found the highest percentages of explant survival at 0.001% and 0.005% of active chlorine concentration: 75% and 65% of survival, respectively [25]. In Bambusa vulgaris, the highest survival percentage (68%) was found when sodium hypochlorite was added at 0.004% [26].…”

Emission of epicormic shoots and in vitro establishment of Cordia trichotoma selected adult trees

“…The use of active chlorine in micropropagation is an inexpensive and effective method for the disinfection of explants and its use is widespread in several species, such as E. cloeziana (Oliveira et al 2015), Cochlospermum regium (Gavilan et al 2018) and E. grandis × E. urophylla (Miranda et al 2020). In the present study, the lowest percentages of fungal and bacterial contamination occurred with the use of active chlorine at concentrations of 1.12 % and 2.12 %, showing the effectiveness of using this disinfection agent for the successful initial in vitro establishment of C. trichotoma from epicormic shoots.…”

Selected Laboratory Equipment and Kitchen Utensils as an Alternative Sterilization Method for Sterilizing Plant Tissue Culture Media

“…This includes the use of microwave oven, chemical reagents such as chlorine dioxide (ClO2), Diethylpyrocarbonate, sodium hypochlorite (NaOCl) (Youssef and Amin, 2001;Tisserat et al, 1992;Duan, et al, 2019;Cardoso, 2009;Cardoso and Inthurn. 2018;Macek et al, 1995;Gavilan, 2018). Other methods for sterilizing PTC media are not yet explored, and have not been conducted in the Philippines.…”