RESUMO
The limited development of photoprotective mechanisms, specifically heat dissipation capacity, found in micropropagated plants may be the result of low xanthophyll cycle pigment content and reduced de-epoxidation capacity making them highly susceptible to photodamage. The effects of gradual or sudden increase of light on Castanea sativa in vitro cultured and during their ex vitro transference was evaluated. The results were compared with those determined in nursery-grown plants. In vitro plants responded poorly to gradual increase in irradiance, exhibiting a low electron transport rate (ETR) agreeing with low non-photochemical quenching (NPQ) and a limited de-epoxidation capacity, not synthesizing detectable amounts of zeaxanthin (Z). Regarding a sudden increase in light (photoinhibition treatment, PhT); post-PhT as in vitro as well nursery plants showed a significant decrease in their maximal efficiency of PSII (F(v)/F(m)), but in vitro the decrease was very drastic (around 0.2) different from that observed in nursery (around 0.69). In vitro, NPQ was mainly determined by the slow relaxing component, NPQ(s) (80.8%), concomitant with a pronounced decrease of D1 protein post-PhT, and a lack of de-epoxidation capacity. During ex vitro transfer, PhT lead to death of some plants, specifically during root induction. The photoprotective mechanisms were activated over time in ex vitro conditions, indicating that micropropagated Castanea sativa display a potential for light acclimation, adjusting their photosynthetic apparatus to the ambient growth irradiance. Understanding the mechanisms that micropropagated plants deployed and how they face high light intensity events, will allow us to search for strategies to improve performance to possible light fluctuations that normally occur in ex vitro conditions during plant acclimation.
Assuntos
Aclimatação , Metabolismo Energético , Fagaceae/efeitos da radiação , Luz , Clorofila/análise , Proteínas de Cloroplastos/análise , Proteínas de Cloroplastos/química , Transporte de Elétrons , Fagaceae/química , Fagaceae/crescimento & desenvolvimento , Oxirredução , Processos Fotoquímicos/efeitos da radiação , Complexo de Proteína do Fotossistema II/química , Folhas de Planta/química , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/efeitos da radiação , Raízes de Plantas/química , Raízes de Plantas/crescimento & desenvolvimento , Tilacoides/química , Xantofilas/química , Zeaxantinas , beta Caroteno/análise , beta Caroteno/químicaRESUMO
Although tannin-rich forages are known to increase protein uptake and to reduce gastrointestinal nematode infections in grazing ruminants, most published research involves forages with condensed tannins (CT), while published literature lacks information on the anthelmintic capacity, nutritional benefits, and antioxidant capacity of alternative forages containing hydrolyzable tannins (HT). We evaluated the anthelmintic activity and the antioxidant capacity of plant extracts containing either mostly CT, mostly HT, or both CT and HT. Extracts were prepared with 70% acetone, lyophilized, redissolved to doses ranging from 1.0mg/mL to 25mg/mL, and tested against adult Caenorhabditis elegans as a test model. The extract concentrations that killed 50% (LC(50)) or 90% (LC(90)) of the nematodes in 24h were determined and compared to the veterinary anthelmintic levamisole (8 mg/mL). Extracts were quantified for CT by the acid butanol assay, for HT (based on gallic acid and ellagic acid) by high-performance liquid chromatography (HPLC) and total phenolics, and for their antioxidant activity by the oxygen radical absorbance capacity (ORAC) assay. Extracts with mostly CT were Lespedeza cuneata, Salix X sepulcralis, and Robinia pseudoacacia. Extracts rich in HT were Acer rubrum, Rosa multiflora, and Quercus alba, while Rhus typhina had both HT and CT. The extracts with the lowest LC(50) and LC(90) concentrations, respectively, in the C. elegans assay were Q. alba (0.75 and 1.06 mg/mL), R. typhina collected in 2007 (0.65 and 2.74 mg/mL), A. rubrum (1.03 and 5.54 mg/mL), and R. multiflora (2.14 and 8.70 mg/mL). At the doses of 20 and 25mg/mL, HT-rich, or both CT- and HT-rich, extracts were significantly more lethal to adult C. elegans than extracts containing only CT. All extracts were high in antioxidant capacity, with ORAC values ranging from 1800 µmoles to 4651 µmoles of trolox equivalents/g, but ORAC did not correlate with anthelmintic activity. The total phenolics test had a positive and highly significant (r=0.826, p ≤ 0.01) correlation with total hydrolyzable tannins. Plants used in this research are naturalized to the Appalachian edaphoclimatic conditions, but occur in temperate climate areas worldwide. They represent a rich, renewable, and unexplored source of tannins and antioxidants for grazing ruminants, whereas conventional CT-rich forages, such as L. cuneata, may be hard to establish and adapt to areas with temperate climate. Due to their high in vitro anthelmintic activity, antioxidant capacity, and their adaptability to non-arable lands, Q. alba, R. typhina, A. rubrum, and R. multiflora have a high potential to improve the health of grazing animals and must have their anthelmintic effects confirmed in vivo in both sheep and goats.