Nonlinear plant-plant interactions modulate impact of extreme drought and recovery on a Mediterranean ecosystem.

Interaction effects of different stressors, such as extreme drought and plant invasion, can have detrimental effects on ecosystem functioning and recovery after drought. With ongoing climate change and increasing plant invasion, there is an urgent need to predict the short- and long-term interaction impacts of these stressors on ecosystems. We established a combined precipitation exclusion and shrub invasion (Cistus ladanifer) experiment in a Mediterranean cork oak (Quercus suber) ecosystem with four treatments: (1) Q. suber control; (2) Q. suber with rain exclusion; (3) Q. suber invaded by shrubs; and (4) Q. suber with rain exclusion and shrub invasion. As key parameter, we continuously measured ecosystem water fluxes. In an average precipitation year, the interaction effects of both stressors were neutral. However, the combination of imposed drought and shrub invasion led to amplifying interaction effects during an extreme drought by strongly reducing tree transpiration. Contrarily, the imposed drought reduced the competitiveness of the shrubs in the following recovery period, which buffered the negative effects of shrub invasion on Q. suber. Our results demonstrate the highly dynamic and nonlinear effects of interacting stressors on ecosystems and urges for further investigations on biotic interactions in a context of climate change pressures.

Influence of the Storage of Cistus ladanifer L. Bales from Mechanised Harvesting on the Essential Oil Yield and Qualitative Composition.

Cistus ladanifer is a Mediterranean native plant from which valuable products, such as essential oil, are obtained. Manual harvesting of the plants in wild shrublands is usual during short periods of time. Their mechanised harvesting could increase the volume of harvested plants and prevent fires, further storage of the plants collected being necessary. The objective of this work is to study the influence of the storage period of mechanically harvested bales on the essential oil yield and qualitative composition. The harvesting trials were carried out with an adapted commercial harvester baler and the storage of the bales was performed indoors during 1-7 days, 15-30 days and 100-120 days. Afterwards, the bales were crushed (30 mm) and distilled in a 30 litre stainless steel still with saturated steam (0.5 bar). The essential oil components were identified by GC-MS and quantified by GC-FID. The storage of mechanically harvested Cistus ladanifer does not decrease the oil yield of steam distillation on a pilot scale. However, it leads to differences in the quantitative composition of the essential oils, decreasing the total monoterpene compounds content and increasing that of oxygenated sesquiterpenes, especially when the biomass is stored for 100-120 days, without affecting its qualitative composition.

Permittivity Measurements for Cypress and Rockrose Biomass Versus Temperature, Density, and Moisture Content.

Permittivity of materials is of utmost importance for microwave applicators' design and to predict high-frequency dielectric heating of materials. In the case of aromatic plant biomass, however, there are few data that help researchers design microwave applicators for microwave-assisted extraction. In this work, the permittivity of cypress and rockrose biomass samples were measured versus temperature, density, and moisture content. A resonant technique based on a coaxial bi-reentrant microwave cavity was employed to obtain the complex permittivity of biomass samples as a function of those magnitudes around the 2.45 GHz ISM frequency. The obtained measurements show that large variations for permittivity values can be found with moisture content and density changes for both cypress and rockrose biomass. Temperature also has effects in a lesser degree, although it has an important influence on the cypress biomass loss factor. Polynomial expressions fitting the experimental data were provided in order to facilitate the estimation of intermediate values, which were not explicitly arranged in this work. As a general trend, the permittivity of cypress and rockrose biomass increases with increasing values of moisture content and density, whereas the biomass loss factor increases when temperature rises.

Neglected Mediterranean plant species are valuable resources: the example of Cistus ladanifer.

MAIN CONCLUSION: The combination of genotypic selection, targeted and improved cultivation, and processing techniques for specific applications gives C. ladanifer the potential to be used as a valuable resource in Mediterranean areas with poor agronomic advantages. Cistus ladanifer (rockrose) is a perennial shrub, well adapted to the Mediterranean climate and possibly to upcoming environmental changes. As a sequence to a thorough review on taxonomic, morphological, chemical and competitive aspects of C. ladanifer, the research team focuses here on the economic potential of C. ladanifer: from production to applications, highlighting also known biological activities of extracts and their compounds. The use of this natural resource may be a viable solution for poor and contaminated soils with no need for large agricultural techniques, because this species is highly resistant to pests, diseases and extreme environmental factors. In addition, this species reveals interesting aptitudes that can be applied to food, pharmaceutical, phytochemical and biofuel industries. The final synthesis highlights research lines toward the exploitation of this neglected resource, such as selection of plant lines for specific applications and development of agronomic and processing techniques.

Cistus ladanifer (Cistaceae): a natural resource in Mediterranean-type ecosystems.

MAIN CONCLUSION: Cistus ladanifer has a well-defined taxonomic identity. 2,2,6-trimethylcyclohexanone may be an authenticity and taxonomic marker. Its traits and applications make it a possible economic resource fitted for Mediterranean areas. Cistus ladanifer is a dominant shrub species endemic to the western Mediterranean region. Due to its dominant nature and its potential ecological, aromatic or pharmacological applications, C. ladanifer has been the object of numerous studies. In this review current knowledge on different aspects of this species is summarized, from its taxonomy to its chemical characterisation or its competitive traits. There are no doubts about the taxonomic entity of C. ladanifer, although the recognition of infraspecific taxa deserves more attention. Given that the fragrant exudate of C. ladanifer holds a very specific composition, one species specific carotenoid, 2,2,6-trimethylcyclohexanone, derivative is proposed as an authenticity marker for uses of C. ladanifer in pharmacological or aromatic industries. Evidence is also gathered on the extreme adaptation of C. ladanifer to stressful conditions in the Mediterranean region, such as the ability to survive in low hydric and high solar exposition conditions, presistence in poor and contaminated soils, and growth inhibition of several other plants through the release of allelochemicals. Thus, the finding of potential applications for this plant may contribute to enhance the economic dimension of derelict lands, such as mine tailings or poor agricultural Mediterranean areas.

The Influence of the Long-Term Outdoor Storage of Rockrose (Cistus laurifolius L.) Shrub Biomass on Biofuel's Quality, Pre-Treatment and Combustion Processes.

Biomass storage is an essential requirement in the supply chain of bio-refineries and power plants. This research aims to evaluate the influence of long-term outdoor storage (1 year) of baled rockrose (Cistus laurifolius L.) shrub biomass on biofuel's quality, pre-treatment processes and on combustion emissions in an industrial boiler. The raw material was obtained from different rockrose shrublands in north central Spain. A total of 233 tWM (tones of wet matter) of biomass were used to produce biofuels (30 mm of milled biomass and Ø 8 mm pellets) in the pre-treatment pilot plants at CEDER-CIEMAT. The combustion tests were conducted in an industrial moving grate boiler with a thermal power of 50 MWth, in a 17 MWe power plant. Outdoor storage improved some biofuel quality parameters, mainly the reduction in ash content, which allowed 30 mm of milled material to be classified as class I1 (ISO 17225-9:2022) and pellets as class I3 (ISO 17225-2-2021). No significant differences were observed in the total specific mass flow and energy consumption in the pre-treatment processes. The combustion tests had similar results, with the emissions being below the limits established in the directive (EU) 2015/2193. The results obtained indicated that the 1-year outdoor storage of rockrose-baled biomass, under Mediterranean conditions, was feasible for its subsequent use as biofuel.

Labdanum Resin from Cistus ladanifer L.: A Natural and Sustainable Ingredient for Skin Care Cosmetics with Relevant Cosmeceutical Bioactivities.

Labdanum resin from Cistus ladanifer L. (Cistaceae) is an abundant natural resource in the Iberian Peninsula worth being explored in a sustainable manner. It is already used in the cosmetic industry; mainly by the fragrances/perfumery sector. However, given the highest market share and traditional uses, labdanum resin also has the potential to be used and valued as a cosmetic ingredient for skincare. Aiming to evaluate this potential, labdanum methanolic absolute and fractions purified by column chromatography were characterized by UPLC-DAD-ESI-MS and then evaluated for UV-protection, antioxidant, anti-elastase, anti-inflammatory, and antimicrobial activities. Labdanum absolute represented ~70% of the resin; diterpenoid and flavonoid fractions represented ~75% and 15% of the absolute, respectively. Labdane-type diterpenoids and methylated flavonoids were the main compounds in labdanum absolute and in diterpenoid and flavonoid fractions, respectively. Labdanum absolute showed a spectrophotometric sun protection factor (SPF) near 5, which is mainly due to flavonoids, as the flavonoids' SPF was 13. Low antioxidant activity was observed, with ABTS radical scavenging being the most significant (0.142 ± 0.017, 0.379 ± 0.039 and 0.010 ± 0.003 mgTE/mgExt, for the absolute and flavonoid and terpene fractions, respectively). Anti-aging and anti-inflammatory activity are reported here for the first time, by the inhibition of elastase activity (22% and 13%, by absolute and flavonoid extract at 1 mg/mL), and by the inhibition of nitric oxide production in LPS-induced RAW 264.7 cells (84% to 98%, at 15 µg/mL extracts, flavonoid fraction the most active), respectively. Antimicrobial activity, against relevant skin and cosmetic product microorganisms, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Escherichia coli, revealed that only S. aureus was susceptible to labdanum absolute (MIC: 1.2 mg/mL) and its fractions (MIC: <0.3 mg/mL). In conclusion, labdanum resin showed potential to be used in sunscreen cosmetics, anti-inflammatory skincare cosmeceuticals or medicines but has low potential as a cosmetic product preservative given the low antioxidant and low-spectrum antimicrobial activities.

Polyphenol Diversity and Antioxidant Activity of European Cistus creticus L. (Cistaceae) Compared to Six Further, Partly Sympatric Cistus Species.

This investigation focused on the qualitative and quantitative composition of polyphenolic compounds of Mediterranean northern shore Cistus creticus and six further, partly sympatric Cistus species (C. albidus, C. crispus, C. ladanifer, C. monspeliensis, C. parviflorus, C. salviifolius). Aqueous extracts of 1153 individual plants from 13 countries were analyzed via high performance liquid chromatography (HPLC). The extracts of C. creticus were primarily composed of two ellagitannins (punicalagin and punicalagin gallate) and nine flavonol glycosides (myricetin and quercetin glycosides, with m-3-O-rhamnoside as the dominant main compound). Differences in the proportions of punicalagin derivatives and flavonol glycosides allowed the classification into two chemovariants. Plants containing punicalagin derivatives and flavonol glycosides were especially abundant in the western and central Mediterranean areas and in Cyprus. From Albania eastwards, punicalagin and punicalagin gallate were of much lesser importance and the predominant chemovariant there was a nearly pure flavonol type. With its two chemovariants, C. creticus takes a central position between the flavonol-rich, purple-flowered clade (besides C. creticus, here represented by C. albidus and C. crispus) and the more ellagitannin-rich, white- or whitish-pink-flowered clade (here represented by C. ladanifer, C. monspeliensis, C. parviflorus and C. salviifolius). The median antioxidative capacity of C. creticus plant material was, with 166 mg Trolox equivalents/g dry wt, about half of the antioxidative capacity of C. ladanifer (301 mg te/g dry wt), the species with the highest antioxidative potential.

Pullulan Films Containing Rockrose Essential Oil for Potential Food Packaging Applications.

Active packaging is designed to control the development of decay- and disease-causing microorganisms and is emerging as a promising technology for extending shelf-life, maintaining food safety, reducing waste, and minimizing the risks for foodborne diseases. The goal of this work was to develop and characterize bioactive pullulan-based films, containing rockrose (Cistus ladanifer) essential oil. Among other abundant compounds (camphene, bornyl acetate and trans-pinocarveol), α-pinene was identified as the major compound of rockrose essential oil (39.25%). The essential oil presented stronger antibacterial activity against Gram-positive than against Gram-negative bacteria. The antioxidant results indicate the potential of the developed films to be used to package foods susceptible to oxidation and rancification, thus improving their shelf-life. Also, this study reflects the potential of rockrose essential oil, free or incorporated in pullulan, as a promising quorum sensing inhibitor, since it was able to interrupt intercellular communication, inhibiting violacein production. Electronic microscopy images showed the antibiofilm activity of the films with rockrose essential oil that were able to influence bacterial adhesion, which may be explained by the differences in the surface free energy of the films, as also determined.

Antibacterial potential of the Cistus incanus L. phenolics as studied with use of thin-layer chromatography combined with direct bioautography and in situ hydrolysis.

The main aim of this study was to detect and identify antibacterial components of fraction I derived from eleven commercial C. incanus herbal teas. Fraction I obtained by a well-established phytochemical protocol of a multi-step extraction was expected to contain flavonoid aglycons alone. Antibacterial profile of fraction I was demonstrated by means of thin-layer chromatography - direct bioautography (TLC-DB) using a Gram positive B. subtilis and a Gram negative A. fischeri strain. Six chromatographic zones of fraction I exhibited a well pronounced antibacterial potential. In qualitative terms, a good agreement was observed among chromatographic fingerprints and the corresponding bioautograms of the eleven samples. The compounds isolated from the six zones were analyzed by HPLC- diode array detector (DAD)-electrospray ionization (ESI)-MS. High numerical m/z values valid for certain constituents of these isolates suggested that some selected antibacterial components are, unexpectedly, flavonoid glycosides. In order to confirm this suggestion, three independent HPTLC methods (multi-development on amino phase and two two-dimensional developments on silica gel phase) were devised to in situ hydrolyze flavonoid glycosides and then separate and visualize the liberated glucose and some other building blocks of the zones' components. Additionally, the sensitivity of glucose detection with p-aminobenzoic acid reagent was enhanced by paraffin. In that way, the presence of the kaempferol glycosides (and not only the aglycones alone) in fraction I was confirmed. Beside kaempferol, p-coumaric acid as a building block unit was shown by HPLC-DAD-MS analysis of the hydrolyzed isolates. Results proved apigenin, kaempferide and acylated kaempferol glycosides (cis- and trans-tiliroside and their conjugates with p-coumaric acid) to be antibacterial components of fraction I. Because isomers of the coumaric acid conjugated tiliroside were detected only in fraction I and not in the crude C. incanus extract, they are regarded as artifacts produced through fractionation.

Inter- and intraspecific diversity in Cistus L. (Cistaceae) seeds, analysed with computer vision techniques.

This work aims to discriminate among different species of the genus Cistus, using seed parameters and following the scientific plant names included as accepted in The Plant List. Also, the intraspecific phenotypic differentiation of C. creticus, through comparison with three subspecies (C. creticus subsp. creticus, C. c. subsp. eriocephalus and C. c. subsp. corsicus), as well as the interpopulation variability among five C. creticus subsp. eriocephalus populations was evaluated. Seed mean weight and 137 morphocolorimetric quantitative variables, describing shape, size, colour and textural seed traits, were measured using image analysis techniques. Measured data were analysed applying step-wise linear discriminant analysis. An overall cross-validated classification performance of 80.6% was recorded at species level. With regard to C. creticus, as case study, percentages of correct discrimination of 96.7% and 99.6% were achieved at intraspecific and interpopulation levels, respectively. In this classification model, the relevance of the colorimetric and textural descriptive features was highlighted, as well as the seed mean weight, which was the most discriminant feature at specific and intraspecific level. These achievements proved the ability of the image analysis system as highly diagnostic for systematic purposes and confirm that seeds in the genus Cistus have important diagnostic value.