CO2-enriched atmosphere and supporting material impact the growth,morphophysiology and ultrastructure of in vitro Brazilian-ginseng [Pfaffia glomerata (Spreng.) Pedersen] plantlets

This study aimed to evaluate under photoautotrophic conditions the effect of CO₂-enriched atmosphere (360 or 1,000 μmol CO₂ mol^?1 air) combined with two substrate types (agar or Florialite^®) in vitro on plants of Pfaffia glomerata, an endangered medicinal species with promising applications in phytotherapy and phytomedicine. The effects of the treatments on the growth, stomatal density, Rubisco activity, carbon isotopic discrimination, metabolite accumulation, photosynthetic pigments and ultrastructural characteristics were investigated. After a 35-day cultivation period, the in vitro-growth of P. glomerata nodal segments under the different treatments resulted in plants with substantial differences in relation to their growth, photosynthetic pigments, stomatal density and leaf ultrastructural characteristics. The enrichment with CO₂ coupled with a porous substrate increased the growth of P. glomerata. The stomatal density in the abaxial epidermis more than doubled in response to the high CO₂ supply in both supporting types, whereas the Rubisco activity and activation state were both unresponsive to the treatments. Regardless of the CO₂ supply, the plants grown in agar displayed higher carbon isotope discrimination than their counterparts grown in Florialite^®. We propose that the long-term photosynthetic performance was improved using Florialite^® as a growth support in combination with a high CO₂ supply. No apparent signs of photosynthetic down-regulation could be found under elevated CO₂ conditions. The enrichment of in vitro atmospheres with CO₂ coupled with a porous substrate offers new possibilities for improving the growth and production on a commercial scale of high morphological and physiological quality Pfaffia plants.     

Survey radiography and computerized tomography imaging of the thorax in female dogs with mammary tumors

Background  

Accurate early diagnosis of lung metastases is important for establishing therapeutic measures. Therefore, the present study aimed to compare survey thoracic radiographs and computerized tomography (CT) scans to specifically identify lung metastases in female dogs with mammary tumors.     

Expression of Melocactus glaucescens SERK1 sheds new light on the mechanism of areolar activation in cacti

Plant Cell, Tissue and Organ Culture (PCTOC) - Areolar activation is the most popular in vitro propagation method for cacti. Even though it is relatively simple, few established protocols exist....     

Involvement of glutathione metabolism in Eichhornia crassipes tolerance to arsenic

• Aquatic macrophytes are potentially useful for phytoremediation programmes in environments contaminated by arsenic (As). Biochemical and physiological modification analyses in different plant parts are important to understand As tolerance mechanisms.
• The objective was to evaluate glutathione metabolism in leaves and roots of Eichhornia crassipes (Mart.) Solms treated to As. Specimens of E. crassipes were cultured for 3 days in Clark's nutrient solution containing 7 μm As. The enzymes ATP sulphurylase (ATPS), glutathione reductase (GR), glutathione peroxidase (GSH‐Px), glutathione sulphotransferase (GST) and γ‐glutamylcysteine synthetase (γ‐ECS) activity, glutathione content, total protein and non‐protein thiols were evaluated.
• The ATPS activity increased in roots. GR activity in leaves and GSH‐Px in roots were lower. GST activity was higher in roots and lower in leaves, and γ‐ECS activity was higher in leaves. Glutathione levels were lower, total thiol levels were higher and non‐protein levels did not change in E. crassipes leaves and roots. Exposure to As increased enzyme activity involved with sulphur metabolism, such as ATPS. Higher GR activity and lower GSH‐Px indicate increased glutathione conjugation to As due to increased GSH availability. The higher GST activity indicates its participation in As detoxification and accumulation through As GSH conjugation. Changes in glutathione and thiol levels suggest high phytochelatin synthesis.
• In conclusion, the increments in ATPS, GR, GST and γ‐ECS activity indicate that these enzymes are involved in GSH metabolism and are part of the E. crassipes As detoxification mechanism.

     

Development of mycorrhized vitroplants of Jatropha curcas L. at different rooting stages

In this work, we attempted to assess the effects of inoculation of arbuscular mycorrhizal fungus (AMF), Glomus clarum, on the survival and development of micropropagated Jatropha curcas plantlets at different rooting stages. Elongated shoots (average 3?cm) of J. curcas, maintained for 0, 14, or 21?days on rooting medium in the presence or absence of 1?mg?L^?1 indole-3-butyric acid (IBA), were transferred to a sand:soil:vermiculite (1:1/2:1) (v:v:v) substrate. At the time of transplantation, the plantlets were either inoculated or not inoculated with G. clarum that had been monoaxenically produced in Ri-transformed carrot roots. After a 2-week acclimatization period, 100?% of the plants kept for 0 or 14?days in rooting medium survived. However, those that remained for 21?days in rooting medium displayed post-acclimatization survival rates of 93 and 89?% for plants inoculated and non-inoculated with G. clarum, respectively. Colonization rates ranged from 70 to 93?%, and the stimulatory effects of AMF were evidenced by increased phosphorus uptake by plants and increases in all evaluated growth parameters, except plant height. Plants that were not subjected to the rooting stage showed growth similar to or higher than those subjected to the rooting stage, regardless of the addition of IBA. It can be concluded that stimulatory effects of mycorrhizal fungi were observed, and that the inoculation of J. curcas proved effective during the initial period of the acclimatization phase.     

CO2 enrichment and supporting material impact the primary metabolism and 20-hydroxyecdysone levels in Brazilian ginseng grown under photoautotrophy

Plant cell and organ cultures via the implementation of effective elicitation strategies can offer attractive biotechnological platforms for the enhanced production of phytochemicals of pharmaceutical interest. For the first time, the elicitation of exogenous signal molecules was conducted to enhance the production of pharmacologically active alkaloids and flavonoids in Isatis tinctoria L. hairy root cultures (ITHRCs). ITHRCs III and V correspondingly possessing high alkaloid and flavonoid productivity were adopted for elicitation treatments. The maximum accumulation of alkaloids in ITHRCs III elicited by 142.61 µM salicylic acid for 28.18 h and flavonoids in ITHRCs V elicited by 179.54 µM methyl jasmonate for 41.87 h increased 5.89- and 11.21-folds as compared with controls, respectively. Moreover, expressions of 11 genes involved in alkaloid and flavonoid biosynthetic pathways were significantly up-regulated following elicitation, among which YUCCA, CHI and F3′H genes might play a crucial role in the target phytochemical augmentation. Overall, two effective elicitation protocols were provided here to improve the yields of bioactive alkaloids and flavonoids in ITHRCs, which was useful for the scale-up production of these valuable compounds to meet the demands for natural bioactive ingredients by pharmaceutical industries.

     

In vitro propagation of Herreria salsaparilha Martius (Herreriaceae) as affected by different sealing materials and gaseous exchanges

The effects of sealing material on the gaseous composition inside flasks containing Herreria salsaparilha in vitro cultures, and on axillary shoot propagation of this species were evaluated. Axillary shoots (1.5 cm in size) were cultured in 200 ml flasks containing 45 ml MS medium supplemented with 30 g l⁻¹sucrose, 100 mg l⁻¹ myo-inositol, MS vitamins and 5 g l⁻¹ agar. Flasks were sealed with polypropylene rigid closure (C), polypropylene rigid closure with a filter (CF), or single- or double-layer polyvinylchloride films (PVC1 and PVC2, respectively). Cultures were maintained at 26 ± 1°C, and a 16/8 h light/dark photoperiod with 35 μmol m⁻² s⁻¹ irradiance. After 60 days, water loss (%), internal O₂ concentration (%), plant height (cm), leaf and root fresh and dry weights (g), and root number and length (cm) were evaluated. Using an open system infrared gas analyzer, the photosynthetic and respiratory rates of in vitro H. salsaparilha cultures were also estimated. The sealing material influenced water loss in flasks, with CF closure showing higher percentage of water loss. O₂ concentration was significantly higher in PVC-sealed flasks than in other treatments, leading to the formation of a small dome on top of the flask. Despite the high oxygen accumulation, shoot development was not affected.