Axenic cell culture of the seagrass Cymodocea nodosa from cotyledonary tissue

[EN] Plant Tissue Culture, also called “micropropagation”, is the propagation of plants from different tissues (or explants) in a shorter time than conventional propagation, making use of the ability that many plant cells have to regenerate a whole plant (totipotency).There are two alternative mechanisms by which an explant can regenerate an entire plant, namely organogenesis and somatic embryogenesis. Since the last decades, the number of higher terrestrial plants species from which these techniques have been successfully applied has continually increased. However, few attempts have been carried out in marine plants. Previous seagrasses authors have focused their studies on i) vegetative propagation of rhizome fragments as explants in Ruppia maritima, Halophila engelmannii, Cymodocea nodosa and Posidonia oceanica; ii) culture of meristems in Heterozostera tasmanica, C. nodosa or P. oceanica; and iii) culture of germinated seeds on aseptic conditions, in Thalassia testudinum, H. ovalis, P. coriacea, P. oceanica, and H. decipiens. All these studies determine the most adequate culture medium for each species (seawater, nutrients, vitamins, carbon sources, etc...), often supplemented with different plant growth regulators and the necessary conditions for the culture maintenance, such as light and temperature. On the other hand, several studies have previously established protocols for cell or protoplast isolation in the species Zostera marina, Z. muelleri, P. oceanica, and C. nodosa, using shoots collected from natural meadows as original vegetal source, but further cell growth was never accomplished. Due to the absence of somatic embryogenesis or organogenetic studies in seagrasses we wonder: IS THE SUCCESSFUL APPLICATION OF TISSUE CULTURE TECHNIQUES POSSIBLE IN SEAGRASSES?

Optimización del cultivo de diatomeas bentónicas para el cultivo de post-larvas de abalón ("Haliotis tuberculata coccinea" Reeve, 1846)

[EN] Diatom cell quantity and their biochemical composition vary among species and are greatly affected by harvest stage or culture conditions. Biometric parameters, growth, attachment capacity and variations in biochemical composition of four species of benthic diatoms (Amphora sp., Navicula incerta, Nitzschia sp. and Proschkinia sp.) were studied. For biochemical analysis the diatoms were harvested at different stages, in log and stationary phase of growth. The culture conditions were identical for all the experiments, benthic diatoms were cultured during 7 days in F/2 medium at 28.5 ± 1.4 ºC, at different original inoculating densities (50000, 100000, and 250000 cell mL-1), under continuous light of 5403 ± 649 Lux provided by cool white fluorescent lighting. The cultures were neither aerated nor agitated. These results show that the specific density of 10000 cell mL-1 was the best for weekly production: Proschkinia sp. reached the highest cell density of 5.81 x 106 cells mL-1 and Amphora sp. had the highest cell attachment capacity with 12000 cell mm-2, in stationary phase of growth. Protein and lipid content were higher in log phase than in stationary phase for the four diatoms. Amphora sp. in log phase of growth had the highest lipid content of 9.74% dry weight (DW). Polyunsaturated fatty acid (PUFA) content ranged from 23.25% to 38.62% of the total fatty acids (TFA), and the four diatoms tested were richer in n-3 PUFA than in n-6 PUFA. All the diatoms had significant quantities of 20:5n-3 (EPA) ranging between 12.69% and 17.68% of TFA. Benthic diatoms play an important and critical role in abalone culture as they are the principal food source of abalone post-larvae. Therefore, it is necessary to improve diatom quantity and quality to optimize post-larval nutrition and the consistency of production, resulting in an increase in growth and survival of abalones.