Novel chitosan/alginate microcapsules simultaneously loaded with copper cations and Trichoderma viride have been prepared and characterized. Information about the intermolecular interactions between biopolymers and bioactive agents was obtained by Fourier transform infrared spectroscopy. Encapsulation of T. viride spores and the presence of copper cations in the same compartment does not inhibit their activity. Microcapsule loading capacity and efficiency as well as swelling behavior and release depend on both the size of the microcapsule and bioactive agents. The in vitro copper cation release profile was fitted to a Korsmeyer-Peppas empirical model. Fickian diffusion was found to be a rate-controlling mechanism of release from smaller microcapsules, whereas anomalous transport kinetics controlled release from larger microcapsules. The T. viride spore release profile exhibited exponential release over the initial lag time. The results obtained opened perspectives for the future use of chitosan/alginate microcapsules simultaneously loaded with biological and chemical agents in plant nutrition and protection.

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encapsulated bioactive agents applied to the Lactuca sativa L. present an innovative approach to stimulate the production of plant secondary metabolites increasing its nutritive value. calcium and copper ions were encapsulated in biopolymeric microparticles (microspheres and microcapsules) either as single agents or in combination with biocontrol agents, Trichoderma viride spores, a fungal plant growth mediator. Both, calcium and copper ions are directly involved in the synthesis of plant secondary metabolites and alongside, Trichoderma viride can provide indirect stimulation and higher uptake of nutrients. All treatments with microparticles had a positive effect on the enhancement of plant secondary metabolites content in Lactuca sativa L. the highest increase of chlorophylls, antioxidant activity and phenolic was obtained by calcium-based microparticles in both, conventionally and hydroponically grown lettuces. non-encapsulated fungus Trichoderma viride enhanced the synthesis of plant secondary metabolites only in hydroponics cultivation signifying the importance of its encapsulation. encapsulation proved to be simple, sustainable and environmentally favorable for the production of lettuce with increased nutritional quality, which is lettuce fortified with important bioactive compounds.Plant secondary metabolites (PSM) are natural sources of biologically active compounds used for a healthy diet, in traditional medicine and in a wide range of industrial applications 1 . The interest in enhancing PSM production is focused to obtain high yields suitable for commercial exploitation. Plant content of secondary plant metabolites is affected by genetic, environmental, and agronomic factors 2 . A variety of strategies (screening and selection of high-yielding cell lines, the culture of cells from various plant parts, suspension culture, induction by elicitors, metabolic engineering, optimizing media, plant growth regulators, etc.) 3 as well as treatments with microspheres loaded with chemical and biological agents 4 were used for enhancing PSM production in plant cell culture.PSM such as polyphenols encompasses several classes of structurally diverse natural products biogenetically arising from the shikimate-phenylpropanoids-flavonoids pathways. Plants require these compounds for pigmentation, growth, reproduction, resistance to pathogens and for many other functions and they represent the adaptive characteristics that were subjected to the natural selection during evolution. In comparison to the animals, plants synthesize a broader spectrum of PSM because of the immobility and impossibility to escape predators, thus they evolved such a chemically based defense against predators 5 . The number of plant secondary metabolites in fresh lettuce can be improved with the addition of desirable compounds during the growth which is readily available for the plant root uptake. Higher PSM share would also have an important impact on human health by improving the antioxidant and nutrient intake through the human diet 6,7 .With the...

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