The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots.

RESUMO Objetivou-se avaliar o potencial alelopático de extratos aquosos de braquiária, girassol e sorgo na germinabilidade e crescimento inicial de alface (Lactuca sativa L.) visando o uso destes extratos como herbicida natural. O estudo foi realizado no Laboratório de Fisiologia Vegetal da Universidade Federal do Recôncavo da Bahia. Foi instalado um experimento no delineamento inteiramente casualizado, com 4 tratamentos e 6 repetições. Os tratamentos foram extratos de braquiária, girassol, sorgo e como controle, água destilada. Os extratos foram obtidos, utilizando 200 g de material vegetal para 1000 mL de água destilada. Os aquênios de alface foram distribuídos em caixa gerbox contendo duas folhas de papel germitest umedecidas com os tratamentos e levadas para câmara de germinação. A germinação foi acompanhada diariamente. Foram avaliados o tempo médio de germinação e, após dez dias, o crescimento inicial, medindo-se o comprimento da parte aérea (CPA) e o comprimento do sistema radicular (CSR). Os dados gerados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey. Com base nos resultados, os extratos de braquiária, girassol e sorgo não afetaram a germinação da alface, mas os de braquiária e sorgo aumentaram o tempo médio de germinação. Os extratos causaram aumento na parte aérea, diminuíram o crescimento do sistema radicular e a formação de plântulas normais. Por causarem inibição no crescimento do sistema radicular da alface, apresentam potencial para serem utilizados como herbicidas naturais.

ABSTRACT This study aimed to evaluate the allelopathic potential of aqueous extracts of Brachiaria, sunflower, sorghum germination and early growth of lettuce (Lactuca sativa L.), in the attempt of employing these extracts as a natural herbicide. The study was conducted at the Laboratory of Plant Physiology of the Universidade Federal do Recôncavo da Bahia. A trial was arranged in a completely randomized design with 4 treatments and 6 replications. The treatments were extracts of Brachiaria, sunflower, sorghum, and, as a control, distilled water. The extracts were obtained by using 200 g of plant material in 1000 mL of distilled water. First, the lettuce seeds were distributed in a seedling box containing two sheets of germitest moistened paper with treatments, and then were taken to a germination chamber. The germination was daily monitored. The average germination time and the initial growth were evaluated after ten days, through the measuring of the aerial part length (APL) and the root system length (RSL). The data generated were subjected to analysis of variance and the means were compared by the Tukey test. Based on the results, the extracts of Brachiaria, sunflower and sorghum did not affect the germination of lettuce, but, on the other hand, the Brachiaria and sorghum increased the germination time. The extracts caused an increase in the aerial part and reduced the root system growth and the formation of normal seedlings. Since they caused inhibition on the growth of the lettuce root system, have also potential to be used as natural herbicides.