THERMOTHERAPY AS A MICROBIAL CONTAMINANT-REDUCING AGENT IN MICROPROPAGATION OF BAMBOO

ABSTRACT The aim of the research was to evaluate the effectiveness of thermotherapy by humid heat of nodal segments to reduce the incidence of microbial contaminants in the introduction phase of the in vitro propagation of Bambusa vulgaris. Two independent experiments were carried out in a completely randomized design using a 4 × 2 factorial arrangement, four levels of temperature (25, 30, 40 and 50 °C) versus two treatment time intervals (5 or 10 min). Each treatment was composed of nine replications, corresponding to one single-nodal segment obtained from primary or secondary branches, in sucrose-free Murashige and Skoog medium with half the concentration of salts, supplemented with 6.5 g L-1 agar, 50 mg L-1 citric acid, 50 mg L-1 ascorbic acid, 200 mg L-1 chloramphenicol and 2 mg L-1 N6-benzylaminopurine. Isolation of prevalent mold fungi per repetition was conducted, and identification at genus level obtained from microcultures. The treatments corresponding to 50 °C for 5 or 10 min reduced significantly the fungal contamination that occurred at 11% and 0%, respectively, in nodal segments from primary branches, and 0% and 11%, respectively, in those from secondary ones. Fungi of genera Alternaria, Bipolaris and Curvularia were isolated from both types of segment, and Cladosporium from secondary branches only. The lowest percentages of mycelial growth detected in nodal segments submitted to both binomials demonstrated the effectiveness of the technique to reduce fungal contamination despite negative interference on sprouting. RESUMO O objetivo da pesquisa foi avaliar a eficácia da termoterapia por calor úmido em segmentos nodais para reduzir a incidência de contaminantes microbianos na fase de introdução da propagação in vitro de Bambusa vulgaris. Dois experimentos independentes foram conduzidos em delineamento inteiramente casualizado, em arranjo fatorial 4 × 2, quatro níveis de temperatura (25, 30, 40 e 50 °C) × dois intervalos de tempo de exposição ao tratamento (5 ou 10 minutos). Cada tratamento foi composto por nove repetições correspondentes a um segmento nodal de uma gema obtido de ramo primário ou secundário, em meio de Murashige e Skoog sem sacarose, com metade da concentração de sais, suplementado com 6,5 mg L-1 de ágar, 50 mg L-1 de ácido cítrico, 50 mg L-1 de ácido ascórbico, 200 mg L-1 de cloranfenicol e 2 mg L-1 de N6-benzilaminopurina. Isolamento de fungos filamentosos prevalentes nas repetições foi realizado, e, a partir de microcultivos,