Micropropagation, plantlets production estimation and ISSR marker-based genetic fidelity analysis of Guadua magna and G. angustifolia

Abstract

The bamboo species Guadua magna and G. angustifolia have been propagated nearly exclusively by conventional techniques of vegetative propagation. Micropropagation is a promising technique and an alternative to conventional ones. This study aimed to micropropagate plants, estimate the plantlets production and analyze the genetic fidelity of G. magna and G. angustifolia by ISSR molecular markers. Mother plants of both species were cultivated in a greenhouse, and either sprayed or not with fungicide. In the laboratory, microcuttings were disinfested and established in MS culture medium with 3.0 mL L-1 of Plant Preservative Mixture (PPM®) and 1 mL L-1 of Carbendazin®. The contamination-free shoots were multiplied in liquid or semi-solid MS medium with 3.0 mg L-1 of BAP for five subcultures. Rooting was performed in liquid or semi-solid MS1/2 medium, plus 3.0 mg L-1 of IBA. Acclimatization was performed on a commercial substrate, in a growth chamber, and the genetic fidelity of the clones produced was analyzed via ISSR markers. The addition of fungicide and PPM® to the medium reduced the contamination in G. magna, but not in G. angustifolia. The liquid medium was more efficient than the semi-solid one for the multiplication of both species, which showed production potentials between 760 and 920 plants per initial microcutting, after five subcultures. Rooted plants exhibited a survival rate of up to 100 % in acclimatization. No polymorphic regions were found in the clones analyzed by ISSR at the end of the fifth subculture, suggesting that micropropagation is a safe technique for the large-scale multiplication of bamboos.

KEYWORDS: Bambusoideae, clonal fidelity, molecular markers, vegetative propagation.