RESUMO

Despite the benefits of bacterial endophytes, recent studies on the mostly Gram-negative bacteria lack of regard for formulation strategies. The encapsulation into biopolymeric materials such as amidated pectins hydrogels is a suitable alternative. Here, this research aimed at supporting the capability of the plant growth-promoting bacteria Kosakonia radicincitans DSM16656T to endophytically colonize plant seedlings. In this approach, the pre-conditioned cells through osmoadaptation and hydroxyectoine accumulation were used. In general, pre-osmoadapted and hydroxyectoine-supplemented bacteria cells formulated in amidated pectin dried beads increased the endophytic activity by 10-fold. Moreover, plant promotion in radish plants enhanced by 18.9% and 20.7% for a dry matter of tuber and leaves. Confocal microscopy studies with GFP-tagged bacteria revealed that bacterial aggregates formed during the activation of beads play an essential role in early colonization stages. This research encourages the integration of fermentation and formulation strategies in a bioprocess engineering approach for exploiting endophytic bacteria.

RESUMO

This study presents an anhydrobiotic engineering approach aiming at conferring a high degree of desiccation tolerance to the Gram-negative endophyte Kosakonia radicincitans. In particular, pre-conditioning of bacteria under high salinities provides a remarkable positive influence on drying survival. The endophytic bacteria accumulate exogenous hydroxyectoine > 500 µmol g-1 dry weight cells exerted by osmotic stress at 4% NaCl. Microfermentation research demonstrated that hydroxyectoine provides positive effects on reducing the lag phase duration and alleviates the dissolved oxygen consumption under high salinity conditions. Beyond the amassing of hydroxyectoine, this work provides evidence supporting the notion that hydroxyectoine can produce significant changes in the endogenous bacterial metabolome during the exponential growth phase at high-osmolarity. Metabolome changes include alterations on tricarboxylic acid cycle, novo-synthesis of specific intracellular metabolites such as mannitol, myo-inositol and trehalose, and fold changes on amino acids such as L-leucine, L-asparagine, L-serine, L-methionine and L-proline. The significant fold change of L-aspartate suggests a potential acidic proteome at high-osmolarity environments, extending the knowledge of salt-stressed bacterial endophytes. Thus, these findings place the metabolic salt stress response and the hydroxyectoine accumulation by K. radicincitans into a physiological context, paving the way into the interaction between cellular phenotype associated with salt stress tolerance and drying survival capacity of Gram-negative endophytes.

Assuntos

Diamino Aminoácidos/farmacologia , Endófitos/fisiologia , Enterobacteriaceae/fisiologia , Tolerância ao Sal/fisiologia , Dessecação , Fermentação , Metaboloma , Concentração Osmolar , Pressão Osmótica , Salinidade , Estresse Fisiológico/fisiologia , Trealose/metabolismo

RESUMO

Gram-negative bacterial endophytes have attracted research interest caused by their advantageous over epiphytic bacteria in plant nutrition and protection. However, research on these typically Gram-negative endophytes has deficiencies concerning the role of cultivation and pre-formulation strategies on further plant colonisation capabilities. Besides, the influence of cultivation conditions and osmotic stress within bacterial endophytes on their phosphate solubilising ability has not yet been addressed. By pre-conditioning cells with an osmoadaptation and a hydroxyectoine accumulation approach, this research aimed at enhancing the capability of the plant growth promoting bacterium Kosakonia radicincitans strain DSM 16656T to both solubilise phosphate and colonise plant seedlings. The results showed that halotolerant bacterial phenotypes increased the root-colonising capability by approximately 3-fold and presented growth-promoting effects in radish plants. Interestingly, findings also demonstrated that salt stress in the culture media along with the accumulation of hydroxyectoine led to an increase in the in vitro phosphate-solubilising ability by affecting the production of acid phosphatases, from 1.24 to 3.34 U mg-1 for non-salt stressed cells and hydroxyectoine-added cells respectively. Thus, this approach provides a useful knowledge upon which the salt stress and compatible solutes in bacteria endophytes can confer phenotypic adaptations to support the eco-physiological performance concerning phosphate-solubilising abilities and endosphere establishment.