The cultivation of medicinal plants and the production of bioactive compounds derived from them are of fundamental importance and interest, not only at the pharmacological level but also in nutraceutical and cosmetic industries and in functional foods, as well as plant protection in agriculture. In order to respond adequately to the increased demands of the global market from a quantitative and qualitative point of view and to guarantee environmental sustainability of the productions, it is necessary to resort to innovation tools, such as tissue culture in vitro technology. Nowadays, it is well known that the cultivation through the Temporary Immersion System (TIS) in a bioreactor has considerable advantages both for the in vitro mass production of the plants and for the production of secondary metabolites. The present review focuses on the application of TIS during the last two decades to produce biomass and bioactive compounds from medicinal plants. Indeed, almost one hundred papers are discussed, and they particularly focus on the effects of the culture system, vessel design and equipment, immersion time and frequency, and substrate composition for 88 medicinal species in TIS bioreactor culture.

Plant genetic resources conservation may be a potential option for the improvement of agricultural crops through modern biotechnologies, and in vitro conservation is a tool available to safeguard plant biodiversity. Ex situ conservation of plant genetic resources using the in vitro procedures is in progress in many countries. The slow growth storage (SGS) technique is a valid in vitro approach to preserve several vegetatively propagated species by controlling the growth and development of plantlets, economizing storage space and labor and reducing costs. Moreover, SGS prolongs the timing between subcultures, lowers the risk of losing germplasm through handling errors, such as contamination problems, and decreases the risk of genetic instability due to the reduction in the number of subcultures. SGS is applied by considering different factors: temperature, light or darkness conditions, medium composition, including mineral or sucrose concentrations, and the presence/absence of plant growth regulators, osmotic agents and growth inhibitors. SGS protocols for some fruit species have been well defined, others require additional research. The present review focuses on the effect of several factors that influence the SGS of in vitro shoots derived from temperate and tropical fruit species during the last ten years.

Origanum vulgare L. (oregano) was chosen as suitable model to investigate the ability of the endophyte-microbiome, especially that of lactic acid bacteria, to develop specific interactions with the plant, mediated by the essential oils (EOs). Combined culture-dependent and -independent approaches analyzed the bacterial dynamic and assembly of Origanum vulgare L. throughout the life cycle. Epiphyte bacteria were more abundant than the endophyte ones. The number of presumptive lactic acid bacteria increased throughout oregano life cycle, according to the plant organ. Diverse species of lactic acid bacteria populated the plant, but Lactobacillus plantarum stably dominated both epiphyte and endophyte populations. High-throughput DNA sequencing showed highest epiphyte bacterial diversity at early vegetative and full-flowering stages, with blooming signing the main microbial differentiation among plant organs. Proteobacteria, Actinobacteria and Bacteroidetes, and Firmicutes and Cyanobacteria at lower abundance were the main phyla. Various genera were detectable, but oregano harbored mainly Methylobacterium, Sphingomonas, Rhizobium and Aurantimonas throughout phenological stages. Firmicutes epiphyte and endophyte microbiotas were different, with a core microbiota consisting of Bacillus, Exiguobacterium, Streptococcus, Staphylococcus and Lactobacillus genera. Bacillus dominated throughout phenological stages. High-throughput DNA sequencing confirmed the dominance of L. plantarum within the epiphyte and endophyte populations of lactic acid bacteria. Yields of EOs varied among plant organs and throughout plant life cycle. L. plantarum strains were the most resistant to the total EOs (mainly thymol and carvacrol) as extracted from the plant. The positive correlation among endophyte lactic acid bacteria and the EOs content seems confirm the hypothesis that the colonization within plant niches may be regulated by mechanisms linked to the synthesis of the secondary metabolites.