Clonal micro-garden formation of Bambusa vulgaris: effect of seasonality, culture environment, antibiotic and plant growth regulator on in vitro culture

Abstract

Background:We have developed a micropropagation technique methodology to clonal microgarden formation of Bambusa vulgaris selected adult plant. Collection site (i.e., stock plant cultivated in different environments) and seasonality of shoot collection (i.e., months of the year) effects on in vitro culture were evaluated. Explants that showed normal development were ex vitro rooted in miniincubator system, and three culture media (WPM, MS and deionized water + agar) supplemented with plant growth regulators (IBA, NAA and BAP) and antibiotic (streptomycin sulphate and culture medium free -control) were evaluated. Micropropagated plants were acclimatized in a shadow house and transferred to a semi-hydroponic system for establishment of a clonal micro-garden. The efficacy of the cloning protocol was determined with genetic fidelity analysis by ISSR molecular markers.Results: Considering all inoculations, 21.9% of nodal segments were in vitro established in nine shoot collections. The rooting percentage was 36.6%, and no interactions were observed between the use of culture medium and antibiotic. Culture medium free antibiotic resulted in 80.0% of survival and 50.0% of adventitious rooting. Micropropagated plants presented adequate growth and adaptation to ex vitro conditions in the clonal micro-garden. Molecular analyses by ISSR markers indicated the absence of genetic variations, and histological analyses revealed normal adventitious root formation originating from meristematic cells. Conclusion:The formation of a clonal micro-garden was demonstrated, proving the feasibility of the tested technique. Our results contribute to the development of a clonal propagation protocol for B. vulgaris selected adult plants.

CLONAL MICROPLANT PRODUCTION, MORPHOLOGICAL EVALUATION AND GENETIC STABILITY OF Dendrocalamus asper (Schult. & Schult.) Backer ex. K. Heyneke

“…Shoots from selected plant cultivated by one-year-old were collected and transported to the laboratory. Subsequently, the leaf sheaths were removed, and their remains were carefully scraped with a stylus for yolk exposition, facilitating asepsis (RIBEIRO et al, 2020;TEIXEIRA et al, 2021). The tissues were then washed with deionized and autoclaved water and neutral detergent.…”

“…However, contamination by phytopathogenic organisms is one significant obstacle to achieving success in the in vitro establishment (RIBEIRO et al, 2016;TORRES et al, 2016;FURLAN et al, 2018;RIBEIRO et al, 2020). Microorganism in vitro contamination has been reported for several bamboo species, such as Bambusa ventricosa (WEI et al, 2015), Bambusa vulgaris (FURLAN et al, 2018;RIBEIRO et al, 2020;TEIXEIRA et al, 2021), Dendrocalamus asper (SINGH et al, 2012) and Dendrocalamus strictus (PANDEY; SINGH, 2012), being an important event to be controlled during the tissue in vitro introduction. Therefore, defining asepsis protocols with antimicrobial agents is essential to minimize the contamination (BRONDANI et al, 2017;ORNELLAS et al, 2017).…”

“…There was root formation only in the treatment with 4.0 mg L -1 IBA (60.0 % of adventitious rooting, Table 2). Rooting stage is recommended for radial system obtention with standard and functional structure, which may favor plants' survival and ex vitro growth, avoiding possible losses during acclimatization (NOGUEIRA et al, 2017;KONZEN et al, 2021a;TEIXEIRA et al, 2021). However, this stage of micropropagation is one of the major obstacles during in vitro cultivation in bamboo species (SINGH et al, 2012).…”

“…Bamboo species occur naturally in tropical, subtropical, and temperate regions all over the world (SINGH et al, 2013a;CANAVAN et al, 2017;RAMAKRISHNAN et al, 2020;MUSTAFA et al, 2021). More than 4,000 traditional uses and 1,500 commercial applications with economic viability for bamboos have been described, besides its ecological and social usages -mainly, due to its morphological and silvicultural features (LIESE;KOHL, 2015;ZHAO et al, 2017;SAWARKAR et al, 2021;TEIXEIRA et al, 2021).…”

“…Separating clumps and planting stalk pieces are laborious techniques and low-efficiency (COSTA et al, 2017;KONZEN et al, 2021a). Despite the difficulties and considering some alternatives for the propagation of bamboo species, in vitro cultivation can be a viable tool for the production of a large number of clonal plants (RIBEIRO et al, 2016;FURLAN et al, 2018;HOSSAIN et al, 2018;RIBEIRO et al, 2020;TEIXEIRA et al, 2021).…”