In vitro callus-induction in globe artichoke (Cynara cardunculus L. var. scolymus) as a system for the production of caffeoylquinic acids

Summary

Globe artichoke (Cynara cardunculus L. var. scolymus) provides a rich dietary source of bio-active compounds derived from phenylpropanoid metabolism, notably caffeoylquinic acids (CQAs) and flavonoids. Micropropagation techniques have been established for this species, but in vitro cultures have not yet been extended to generate an efficient system for the induction of callus tissue. In this study, we compared more than 100 combinations of media supplements (e.g., phytohormones, absorbers of polyphenols, and inhibitors of polyphenol oxidase), along with various light regimes, and three different genotypes of globe artichoke to define the optimal conditions for callus induction from leaf explants. This led to the elaboration of an in vitro culture protocol which resulted in a high frequency of callus induction after just 1 week in culture. The procedure used leaf explants from virus-free, meristem culturederived plantlets. Quantitative HPLC analysis revealed that, as in globe artichoke leaves, the predominant phenolic esters present in callus were mono- and di-caffeoylquinic acids (diCQA). The concentration of diCQA was three- to five-fold higher in calli than in leaves. The exposure of calli to UV-C light further enhanced the levels of CQAs. In vitro callus culture combined with UV-C irradiation may thus represent a viable production system for diCQA that is suitable for the synthesis of pharmacologically-active compounds.     

能源利用效率、环境和食品安全

为了满足营养需求和后代对环境的要求,对农业实践的深刻反思势在必行。像可持续发展和能源效率标准将会在动物饲料领域发挥越来越重要的作用。     

Correlation of Rutin Accumulation with 3-O-Glucosyl Transferase and Phenylalanine Ammonia-lyase Activities During the Ripening of Tomato Fruit

In tomato, the predominant flavonoid is quercetin-3-rutinoside (rutin). In this study, we aim to investigate the phenylalanine ammonia-lyase (PAL) and the quercetin-3-O-glucosyl transferase (3-GT) reactions in the formation of rutin during tomato fruit ripening. Tomatoes of the Moneymaker variety at different development stages (green, breaker, turning, pink, red, and deep red) were divided into flesh and peel fractions. In each sample, both the content of rutin and the enzymatic activities for PAL and 3-GT were recorded. The highest activities of PAL were recorded in the peel of turning fruit (3,000 μkat/mg fresh weight). In fruit flesh, maximal activity was observed in red fruit (917.3 μkat/mg). For both tissues, PAL activity strongly decreased at the final (deep red) fruit stage. The activity of 3-GT in peel peaked in the turning fruit stage (50.7 pkat/mg), while in flesh maximal activity (33.4 pkat/mg) was observed in green fruit, which rapidly declined at the turning stage. Higher levels of rutin were detected in the tomato peel compared to the flesh part with the highest level being found at the green stage. The relation of PAL and 3-GT activities to rutin content is also evaluated.     

Testing the shade tolerance of selected crops under Parkia biglobosa (Jacq.) Benth. in an agroforestry parkland in Burkina Faso, West Africa

In Western Africa, interactions between trees and agricultural crops are a key element in determining parkland management in an agricultural environment that is rapidly changing. Eggplant (Solanum melongena), chilli pepper (Capsicum annuum), taro (Colocasia esculenta) and pearl millet (Pennisetum glaucum) were tested for their shade tolerance under Parkia biglobosa trees in south-central Burkina Faso using a split-plot design. Soil characteristics, chlorophyll fluorescence and crop growth and yield were measured to quantify the effect of P. biglobosa on the crops and their environment. The experiment ran during 2?years. P. biglobosa suppressed the vegetative growth and yield of pearl millet in both years. Eggplant and chilli pepper were severely injured by the rains and produced fruits only during the first year. Eggplant yields were suppressed by trees to between one third and one tenth of the yield in the control plots. However, chilli pepper yields increased by up to 150% when grown under the tree canopy compared to the control. In both years, the vegetative growth and yield of taro was higher when grown in the shade than outside the tree canopy.     

Independent lines of evidence of a genetic relationship between acoustic wave velocity and kraft pulp yield in <Emphasis Type="Italic">Eucalyptus globulus</Emphasis>

Key message

Multiple lines of evidence suggest acoustic wave velocity (AWV) would provide a rapid and efficient method to indirectly select for superior pulp yield in Eucalyptus globulus breeding programs.

Context

Eucalyptus globulus is one of the most widely planted hardwood species in temperate regions of the world and is primarily grown for pulpwood.

Aims

To determine if acoustic wave velocity (AWV) can be used to indirectly select for kraft pulp yield in E. globulus.

Methods

Genetic group effects, additive and non-additive variance components, and genetic correlations were estimated for AWV and pulpwood traits, including Kraft pulp yield. In a separate trial, the relative position of quantitative trait loci (QTL) for these traits was compared.

Results

Estimated narrow-sense heritabilities for AWV and pulp yield were both 0.26, and these traits were strongly genetically correlated (0.84). Furthermore, co-located QTL for these traits were identified. Further evidence that AWV could be used to indirectly select for pulp yield was provided by the ranking of genetic groups—Otways and King Island had the highest AWV and pulp yield and Strzelecki and Tasmania the lowest. There was no evidence of dominance variation in wood property traits.

Conclusion

Together, these findings suggest that AWV could be used as a selection criterion for kraft pulp yield in E. globulus breeding programs.

     

Stress-induced biosynthesis of dicaffeoylquinic acids in globe artichoke

Leaf extracts from globe artichoke ( Cynara cardunculus L. var. scolymus) have been widely used in medicine as hepatoprotectant and choleretic agents. Globe artichoke leaves represent a natural source of phenolic acids with dicaffeoylquinic acids, such as cynarin (1,3-dicaffeoylquinic acid), along with its biosynthetic precursor chlorogenic acid (5-caffeoylquinic acid) as the most abundant molecules. This paper reports the development of an experimental system to induce caffeoylquinic acids. This system may serve to study the regulation of the biosynthesis of (poly)phenolic compounds in globe artichoke and the genetic basis of this metabolic regulation. By means of HPLC-PDA and accurate mass LC-QTOF MS and MS/MS analyses, the major phenolic compounds in globe artichoke leaves were identified: four isomers of dicaffeoylquinic acid, three isomers of caffeoylquinic acid, and the flavone luteolin 7-glucoside. Next, plant material was identified in which the concentration of phenolic compounds was comparable in the absence of particular treatments, with the aim to use this material to test the effect of stress application on the regulation of biosynthesis of caffeoylquinic acids. Using this material, the effect of UV-C, methyl jasmonate, and salicylic acid treatments on (poly)phenolic compounds was tested in different globe artichoke genotypes. UV-C exposure consistently increased the levels of dicaffeoylquinic acids in all genotypes, whereas the effect on compounds from the same biosynthetic pathway, for example, chlorogenic acid and luteolin-7-glucoside, was much less pronounced and was not statistically significant. No effect of methyl jasmonate or salicylic acid was found. Time-response experiments indicated that the level of dicaffeoylquinic acids reached a maximum at 24 h after UV radiation. On the basis of these results a role of dicaffeoylquinic acids in UV protection in globe artichoke is hypothesized.     

Assessing the recovery of a long-lived herb following logging: Trillium ovatum across a 424-year chronosequence

We investigated the long-term recovery of Trillium ovatum (Liliaceae) following clear-cut logging by assessing demographic trends across a forest chronosequence that ranged in age from 2 to 424 years. In each of 20 sites across the chronosequence, we used 50 × 50 m plots to assess the population size, age and stage structure, seed production, and the spatial distribution of seedlings relative to reproductive plants. We found that Trillium populations were significantly reduced in younger sites and a significant positive relationship of population size and stand age, suggesting that population recovery following stand-replacement disturbances may take centuries. Slow recovery rates of Trillium can partly be explained by short seed dispersal distances. We found 67% of new Trillium recruits within only one meter of potential parent plants. Despite the general trend of slow recovery, a few recently disturbed populations showed fast population growth. In these cases, high rates of post-disturbance recruitment were related to the number of individuals surviving the disturbance event. We found that local, within-site seed production can most likely mitigate limited dispersal abilities of Trillium. The rate of Trillium recovery following disturbance, therefore, depends on the number of plants that have persisted through the disturbance event and serve as within-site seed sources. Our study suggests that the nature of the stand-replacement disturbance (i.e., magnitude and intensity) may be critical in determining the recovery of understory plants in managed forests.     

The relative importance of different regeneration mechanisms in a selectively cut savanna-woodland in Burkina Faso,West Africa

Savanna plant species are capable of regenerating both sexually and asexually. The importance of each regeneration mechanism depends on the species, the type and the intensity of disturbance. This paper describes the relative importance of sexual and asexual regeneration for the restoration of a selectively cut savanna woodland in Burkina Faso. Plantlets regenerated sexually or asexually were determined by excavating the below ground system and assessing basal and aerial sprouts within 144 quadrats along transects in 2, 5, 10 year-cut and control plots. The result shows that seedling sprouts were predominant (83%) compared to root sucker (4%), coppice (5%), water sprout (2%) and layering (less than 1%). True seedlings constituted a minor component (5%) of the plantlet population. A total of 54 woody species was found in the plantlet population with 52 presenting seedling sprouts. Twelve species were found regenerating from both seeds and root buds; of which Dichrostachys cinerea, Pteleopsis suberosa and Detarium microcarpum had the highest number of plantlets. Feretia apodanthera was the only layering species encountered in the sampled area. Coppices, root suckers and water sprouts showed higher height and bigger collar diameter compared to seedling sprouts and true seedlings. It can be concluded that sexual reproduction is the dominant mechanism of seedling recruitment in this disturbed savanna-woodland, and that its success relies on the ability of plantlets of seed origin to resprout abundantly.     

Efficacy of a canarypox-vectored recombinant vaccine expressing the hemagglutinin gene of equine influenza H3N8 virus in the protection of ponies from viral challenge

OBJECTIVE: To determine onset and duration of immunity provided by a 2- or 3-dose series of a new canarypox-vectored recombinant vaccine for equine influenza virus (rCP-EIV vaccine) expressing the hemagglutinin genes of influenza H3N8 virus strains A/eq/Kentucky/94 and A/eq/Newmarket/2/93 in ponies. ANIMALS: Forty-nine 1- to 3-year-old male Welsh Mountain Ponies that were seronegative for equine influenza virus. PROCEDURES: Vaccinated and control ponies were challenged with aerosolized influenza virus A/eq/Sussex/89 (H3N8), representative of the Eurasian lineage of circulating influenza viruses. In trial 1, control ponies and ponies that received rCP-EIV vaccine were challenged 2 weeks after completion of the 2-dose primary vaccination program. In trial 2, ponies were challenged 5 months after 2 doses of rCP-EIV vaccine or 1 year after the first boosting dose of rCP-EIV vaccine, administered 5 months after completion of the primary vaccination program. After challenge, ponies were observed daily for clinical signs of influenza and nasal swab specimens were taken to monitor virus excretion. RESULTS: The challenge reliably produced severe clinical signs consistent with influenza infection in the control ponies, and virus was shed for up to 7 days. The vaccination protocol provided clinical and virologic protection to vaccinates at 2 weeks and 5 months after completion of the primary vaccination program and at 12 months after the first booster. CONCLUSION AND CLINICAL RELEVANCE: The rCP-EIV vaccine provided protection of ponies to viral challenge. Of particular importance was the protection at 5 months after the second dose, indicating that this vaccine closes an immunity gap between the second and third vaccination.     

Inter- and intra-provenances variations in seed size and seedling characteristics of Khaya senegalensis A. Juss in Burkina Faso

Within- and between-provenance variations in seed and seedling traits of Khaya senegalensis A. Juss were studied at INERA in Burkina Faso. Nursery grown seedlings from four provenances in Burkina Faso were used for the study. The studies revealed significant variability in all traits evaluated. Seed length and weight significantly varied among provenances and families within provenances, where Bopiel and Koyenga had the highest mean values. Height and root collar diameter of 1 year old seedlings significantly varied among families within provenances. Except leaf biomass ratio and carbon isotope ratio which varied significantly among provenances but not among families within provenances, all other seedling biomass traits—total plant biomass, stem biomass ratio, leaf biomass ratio, root biomass ratio, root shoot ratio, specific leaf area, and leaf area ratio were significantly affected by provenances and families within provenances. The magnitude of variation due to family effect ranged from 65 to 93 % for seed size traits, and from 4.5 to 17.8 % for seedling characters. Estimates of family heritability were moderate to high (0.67–0.95) for seed traits, but low to moderate (0.19–0.59) for seedling characters, indicating that much of the total variation in seed traits is due to the genetic effect. The two most prominent provenances, Koyenga and Bopiel, with higher seed size and greater seedling growth could be considered for an eventual K. senegalensis improvement program in Burkina Faso.