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- Field performance of seedlings and microproppagated plants of carob treePublication . Osório, Maria Leonor; Osório, Júlio; David, Maria Manuela; Romano, AnabelaMicropropagated plantlets of cv. Mulata and cv. Galhosa growing in pots, after acclimatization in the glasshouse and growth for several months under natural conditions, were transferred to a field trial. Carob seedlings of ‘Mulata’ were also transferred under the same conditions. The field trial has been established with 100% of success in micropropagated plants and 97% in seedlings. Three months after transfer to the field, plants showed good growth but micropropagated plants of ‘Mulata’ exhibited more vigour than ‘Galhosa’ and than seedlings, with the greatest number of branches and a larger length increase in the main stem. Before transference to the field, net photosynthetic rates (PN), water use efficiency (PN/gs) and quantum yield of PSII (fPSII) of the potted plants were higher in seedlings than in micropropagated plants. Three months after transplantation all gas exchange parameters were improved for all types of plants, particularly net photosynthetic rate, and no significant differences were observed between plants either micropropagated or seedlings. On the other hand, fPSII decreased significantly in seedlings. The results suggest that the pre-acclimation, in pots, to external environmental conditions might have improved field survival, plant vigour and enhanced the endurance to adverse conditions of micropropagated carob trees.
- A new medium formulation for in vitro rooting of carob tree based on leaf macronutrients concentrationsPublication . Gonçalves, Sandra; Correia, P. J.; Martins-Loução, M. A.; Romano, AnabelaExperiments were performed to optimize the macronutrients concentrations for in vitro rooting of Ceratonia siliqua micropropagated shoots. Several dilutions of Murashige and Skoog (MS) medium were tested: full-strength MS, halfstrength MS (½MS), and ½MS + full N. The frequency of in vitro rooting was enhanced when the ½MS was used (50 % rooted shoots). Mature leaves from 20 - 30 year-old carob trees and from 2 year-old micropropagated plants were collected and the concentrations of macronutrients (N, P, K, Ca, Mg) assessed. Based on the mineral composition of the leaves a new medium was formulated and compared with the previous ones showing an increment of the rooting frequency to 80 %. Moreover, shoots rooted in the new medium did not show symptoms of apical necrosis that occurred in the other tested media.
- Locust bean gum (LBG) as a gelling agent for plant tissue culture mediaPublication . Gonçalves, Sandra; Romano, AnabelaLocust bean gum (LBG) is a natural hydrocolloid extracted from the seeds of carob tree (Ceratonia siliqua L.). This work describes the successful use of LBG as a gelling agent in combination with agar for shoot multiplication and rooting of carob tree and Iberian rose shoots. Its presence did not affect the multiplication rate of both species. The rooting frequency of carob shoots was even significantly increased in the presence of 5 g of LBG plus 4 g of agar to the medium compared to medium solidified with 9 g of agar. Iberian rose shoots rooting was not influenced by the addition of this gum to the rooting medium. Results obtained show that LBG can be used in combination with agar in culture medium as a gelling agent without negative effect on plant material and with the advantage of reduced medium costs.
- Seasonal changes in CO2 assimilation in leaves of seedlings and micropropagated plants of Carob tree established in the fieldPublication . Osório, Maria Leonor; Osório, Júlio; David, Maria Manuela; Correia, Maria João; Romano, AnabelaIn this communication we evaluate the field performance of two micropropagated Portuguese carob cultivars (Galhosa and Mulata) throughout the season, particularly at extreme conditions of light and temperature. Two irrigated plots were established in the field: 1) micropropagated plants, vs 2) seedlings. During the first year following transplantation to the field, we followed net photosynthetic rate, stomatal conductance, transpiration rate, chlorophyll a fluorescence and leaf contents in chlorophyll, carotenoids and protein. No significant differences were detected between seedlings and micropropagated plants along the year. However, at the end of summer, despite irrigation, the photosynthetic rate (NP), the quantum yield of PSII (fPSII) and the intrinsic efficiency of open PSII reaction centers (F’v/F’m) declined, concomitantly with the increase of the thermal energy dissipation at the PSII (NPQ). As the maximal photochemical efficiency of PSII (Fv/Fm) was maintained high (0.82), these results indicate that regulated thermal dissipation in light harvesting complexes was promoted in order to avoid photoinhibition. After the first growth period in the field, data from micropropagated plants did not differ from seedlings, and those plants showed the characteristic behaviour of plants well adapted to Mediterranean climates. So, in vitro propagation could be use as a promising alternative to traditional propagation and establishment of carob orchards.