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The efficacy of ultrasound-assisted extraction (UAE) of fucoidan from four frozen thalli of Arctic brown algae (Fucales, Phaeophyceae) Fucus vesiculosus, Fucus distichus, Fucus serratus , and Ascophyllum nodosum was studied for the first time and compared with the heat-assisted extraction (HAE). The chemical composition and molecular weight distribution of fucoidans were characterized using HPLC-RID, HPSEC, ferric reducing antioxidant power (FRAP) assay, total antioxidant capacity (TAC). The composition of fucoidan was heterogeneous and depended on the extraction method and the species: F. vesiculosus , F. distichus , F. serratus and A. nodosum . UAE was more efficient for fucoidan extraction when compared with HAE. Two-level quarter fractional factorial design (FFD) was applied to optimize the extraction UAE process of fucoidan from frozen F. vesiculosus . Five parameters of extraction were studied: extraction time (20–60 min), ultrasonic amplitude (60–100 %), pH (2–6), liquid-to-raw-material ratio (10–30 mL g -1 ), and particle size (1–3 mm). The highest yield was achieved at the extraction time of 55 min, the ultrasonic amplitude of 80%, pH 3.4, liquid-to-raw-material ratio of 25 mL g -1 and particle size of 1.4 mm. It was found that the power law and the Peleg’s model describe the kinetics of fucoidan extraction with the highest accuracy. The environmental friendliness of the proposed extraction technique is shown. Overall, a novel biorefinery process for fresh frozen algal biomass was established to produce a lipid fraction, mannitol, and two distinctive polysaccharide products such as fucoidan and alginic acid from a single seaweed feedstock and optimized fucoidan UAE extraction. Ultrasound was effective to enhance the extraction of fucoidan at a relatively low temperature from frozen algae thalli.

Bacteria and marine macroalgae form close associations, while various bacteria affect the morphogenesis and growth of macroalgae. Hyphomonas strains exhibit normal morphogenetic activity in protoplasts of the red alga Pyropia yezoensis (nori). However, the effects of the bacteria on the growth of Pyropia from protoplast cells to regenerated thalli remain unknown. Here, we assessed the growth of P. yezoensis and Pyropia tenera using combined cultures of three Hyphomonas strains (LNM10-16, SCM-2, and LNM-9) and three algal media (artificial seawater with vitamins, artificial seawater, and natural seawater) over 7 weeks. Third week after culture, the three Hyphomonas strains showed almost similar levels of normal growth activity for both Pyropia species. However, at 7 weeks, significant differences were observed among the three Hyphomonas strains in terms of length, length-to-width ratio, and normal morphology of Pyropia thalli. LNM10-16 significantly promoted the thalli length and length-to-width ratios of both Pyropia species in artificial seawater without vitamins and natural seawater, compared with the other two Hyphomonas strains. P. yezoensis cultured in artificial seawater with vitamins showed a much higher demand for LNM10-16 in development of the thalli length than P. tenera . These results may be explained by differences in the growth activities of Hyphomonas strains and the nutrient requirements of Pyropia species. Furthermore, the bacteria were more specifically attached to the rhizoid surfaces of both species. This study is the first to reveal that Hyphomonas strains affect the growth of Pyropia species by attaching to their rhizoids.

Thalli sheep is a significant breed reared under tropical region of Punjab province of Pakistan. The present study was conducted to predict live body weight (LBW) by means of from some body measurements, i.e., chest girth (CG), belly girth (BG), rump height (RH), withers height (WH), neck girth (NG), and body length (BL) taken from 155 Thalli indigenous sheep of Pakistan. Age factor is determined to be a significant source of variation for BL, BG, CG, BG, WH, and NG (p < 0.05). LBW is correlated significantly with BL (0.850), CG (0.825), BG (0.849), RH (0.579), WH (0.547), and NG (0.7760), respectively (p < 0.01). For LBW prediction, CART and MARS data mining algorithms were comparatively used based on ten cross-validation method. Among 185 candidate MARS models with 1–5 degrees of interaction and 2–38 terms, the MARS model with 7 terms and no interaction effect in R software was the best model for LBW prediction on the basis of the smallest cross-validated RMSE value. Also, the optimal CART tree structure was obtained with 9 terminal nodes for the smallest cross-validated RMSE value. MARS algorithm outperformed CART in LBW prediction and explained 90.3 (%) of variability in LBW of Thalli sheep. Results of the optimal CART structure reflected that Thalli sheep with BL > 75 cm, RH > 83 cm, and NG > 55 cm has the heaviest LBW of 72 kg. The optimal MARS model displays that the heaviest LBW can be produced by Thalli sheep with BL > 71.12 cm, BG > 106.68 cm, WH > 76.2 cm, NG > 50.8 cm in 5th age group. In conclusion, it coud be recommended that MARS predictive modeling may enable animal breeders to obtain elite Thalli sheep population and to detect body measurement positively influencing LBW as indirect selection criteria for not only describing breed characterization and developing flock management standards, but also ensuring sustainable meat production and rural development in Pakistan.

Copper ion (Cu 2+ ) and copper oxide (CuO) nanoparticle (NP) ecotoxicity are of increasing concern as they are considered to be a potential risk to marine systems. This study represents the first attempt to evaluate CuO NP impacts on the seaweeds and Cu 2+ on the chlorophyte Ulva rigida . Effects on oxidative stress, antioxidant defence markers, photosystem II function, thalli growth, and cell viability in U. rigida exposed for 4 up 72 h to1 and 5 mg L −1 Cu 2+ and CuO NPs were examined. Hydrogen peroxide (H 2 O 2 ) generation, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, and growth inhibition seemed to be reliable and early warning markers of toxicity. The most important variables of the principal component analysis (PCA): H 2 O 2 generation, antioxidant stress markers, and growth-based toxicity index, were higher at 1 mg L −1 CuO NPs compared to CuSO 4 and at 5 mg L −1 CuSO 4 compared to CuO NPs. Intracellular uptake kinetics fit well to the Michaelis–Menten equation. The higher toxicity at 5 mg L −1 CuSO 4 compared to 1 mg L −1 was due to the higher Cu uptake with increasing concentration, suggesting and higher accumulation ability. On the contrary, 1 mg L −1 CuO NPs induced more strongly toxicity effects than 5 mg L −1 . The relatively stronger effect of CuO NPs at 1 mg L −1 than the respective CuSO 4 concentration could be attributed to the higher rate of initial uptake ( V c ) and the mean rate of Cu uptake [ C max /(2 ×  K m )] at CuO NP treatment. The intracellular seaweed experimental threshold of Cu, which coincided with the onset of oxidative stress, was within the Cu concentration range recorded in Mediterranean Ulva spp., indicating that it may pose a substantial risk to marine environments.

Endolichenic fungi are diverse groups of predominantly filamentous fungi that reside asymptomatically in the interior of lichen thalli. Natural products from endolichenic fungi, isolated from a variety of different lichen species, have been attracting increased attention for their potential to produce bioactive metabolites possessing new structures and representing different structural classes. This is evident from the steady increase of publications devoted to endolichenic fungal metabolites over the past decade, since the first report of endolichenic secondary metabolites. The bioactive metabolites produced by endolichenic fungi originate from multiple biosynthetic pathways and occupy different chemical structure classes, including steroids, quinones, terpenoids, peptides, xanthones, sulfur-containing chromenones, etc. Endolichenic fungal metabolites possess a diverse array of bioactivities, such as anticancer, antiviral, antibacterial, antifungal, and anti-Alzheimer’s disease. This review provides the first thorough assessment of endolichenic fungi, their biodiversity, secondary metabolites, and associated bioactivity. This review will highlight the bioactive metabolites reported in recent years from endolichenic fungi, as well as discussing the potential of these symbiotic fungi as sources of new, diverse natural products with varying bioactivities.

An understanding of how biotic interactions shape species’ distributions is central to predicting host–symbiont responses under climate change. Switches to locally adapted algae have been proposed to be an adaptive strategy of lichen-forming fungi to cope with environmental change. However, it is unclear how lichen photobionts respond to environmental gradients, and whether they play a role in determining the fungal host’s upper and lower elevational limits. Deep-coverage Illumina DNA metabarcoding was used to track changes in the community composition of Trebouxia algae associated with two phylogenetically closely related, but ecologically divergent fungal hosts along a steep altitudinal gradient in the Mediterranean region. We detected the presence of multiple Trebouxia species in the majority of thalli. Both altitude and host genetic identity were strong predictors of photobiont community assembly in these two species. The predominantly clonally dispersing fungus showed stronger altitudinal structuring of photobiont communities than the sexually reproducing host. Elevation ranges of the host were not limited by the lack of compatible photobionts. Our study sheds light on the processes guiding the formation and distribution of specific fungal–algal combinations in the lichen symbiosis. The effect of environmental filtering acting on both symbiotic partners appears to shape the distribution of lichens.

Phenylpropanoids play important roles in plant physiology and the enzyme 4-coumarate: coenzyme A ligase (4CL) catalyzes the formation of thioesters. Despite extensive characterization in various plants, the functions of 4CLs in the liverwort Marchantia paleacea remain unknown. Here, four 4CLs from M . paleacea were isolated and functionally analyzed. Heterologous expression in Escherichia coli indicated the presence of different enzymatic activities in the four enzymes. Mp4CL1 and Mp4CL2 were able to convert caffeic, p-coumaric, cinnamic, ferulic, dihydro-p-coumaric, and 5-hydroxyferulic acids to their corresponding CoA esters, while Mp4CL3 and Mp4CL4 catalyzed none. Mp4CL1 transcription was induced when M . paleacea thalli were treated with methyl jasmonate (MeJA). The overexpression of Mp4CL1 increased the levels of lignin in transgenic Arabidopsis. In addition, we reconstructed the flavanone biosynthetic pathway in E . coli . The pathway comprised Mp4CL1, co-expressed with chalcone synthase (CHS) from different plant species, and the efficiency of biosynthesis was optimal when both the 4CL and CHS were obtained from the same species M . paleacea .

Lichens are of great ecological importance but mechanisms regulating lichen symbiosis are not clear. Umbilicaria muhlenbergii is a lichen-forming fungus amenable to molecular manipulations and dimorphic. Here, we established conditions conducive to symbiotic interactions and lichen differentiation and showed the importance of UMP1 MAP kinase in lichen development. In the initial biofilm-like symbiotic complexes, algal cells were interwoven with pseudohyphae covered with extracellular matrix. After longer incubation, fungal-algal complexes further differentiated into primitive lichen thalli with a melanized cortex-like and pseudoparenchyma-like tissues containing photoactive algal cells. Mutants deleted of UMP1 were blocked in pseudohyphal growth and development of biofilm-like complexes and primitive lichens. Invasion of dividing mother cells that contributes to algal layer organization in lichens was not observed in the ump1 mutant. Overall, these results showed regulatory roles of UMP1 in symbiotic interactions and lichen development and suitability of U. muhlenbergii as a model for studying lichen symbiosis. The mechanisms regulating fungal-algal interactions during the formation of lichen symbioses are not clear. Here, Wang et al. establish conditions conducive to symbiotic interactions and lichen differentiation using a fungus amenable to genetic manipulation, showing the importance of a MAP kinase in lichen development.

The development of golden tides is potentially influenced by global change factors, such as ocean acidification and eutrophication, but related studies are very scarce. In this study, we cultured a golden tide alga, Sargasssum muticum, at two levels of pCO2 (400 and 1000 µatm) and phosphate (0.5 and 40 µM) to investigate the interactive effects of elevated pCO2 and phosphate on the physiological properties of the thalli. Higher pCO2 and phosphate (P) levels alone increased the relative growth rate by 41 and 48 %, the net photosynthetic rate by 46 and 55 %, and the soluble carbohydrates by 33 and 62 %, respectively, while the combination of these two levels did not promote growth or soluble carbohydrates further. The higher levels of pCO2 and P alone also enhanced the nitrate uptake rate by 68 and 36 %, the nitrate reductase activity (NRA) by 89 and 39 %, and the soluble protein by 19 and 15 %, respectively. The nitrate uptake rate and soluble protein was further enhanced, although the nitrate reductase activity was reduced when the higher levels of pCO2 and P worked together. The higher pCO2 and higher P levels alone did not affect the dark respiration rate of the thalli, but together they increased it by 32 % compared to the condition of lower pCO2 and lower P. The neutral effect of the higher levels of pCO2 and higher P on growth and soluble carbohydrates, combined with the promoting effect on soluble protein and dark respiration, suggests that more energy was drawn from carbon assimilation to nitrogen assimilation under conditions of higher pCO2 and higher P; this is most likely to act against the higher pCO2 that caused acid–base perturbation via synthesizing H+ transport-related protein. Our results indicate that ocean acidification and eutrophication may not boost golden tide events synergistically, although each one has a promoting effect.

The aim of this study was to evaluate the multivariate traits in Thalli sheep for phenotypic characterization and their association with the live bodyweight of the animals for the selection of superior individuals by using the principal component analysis (PCA) technique and regression equation. Twenty-one (21) multivariate traits were taken of young Thalli sheep aged 0-12 months in Thall area of Punjab, Pakistan. Morphometric traits measured were withers height (WH), head width (HW), head length (HL), body length (BL), ear width (EW), ear length (EL), neck width (NW), neck length (NL), heart girth (HG), rump length (RL), tail length (TL), rump width (RW), barrel depth (BD), birth weight (BiW), live body weight (BW), sacral pelvic width (SPW), teat diameter (TED), teat length (TEL), testes length (TsL), scrotal circumference (SC) and testes width (TsW). Positive and highly significant (P≤0.01) correlations among different morphometric traits were present in overall animals of 0-12-month age groups of Thalli sheep such as WH, BL, HL, HW, EL, EW, NL, NW, HG, RL, BD, SPW, BiW and BW. The results of principal component analysis of data for all age groups of young Thalli sheep were showed that three principal components (PCs) were extracted as PC1, PC2, and PC3. For the 0-3-month age group, PC1 showed high variance before and after rotation (44.33% and 37.68%), for 4-6-month age group, PC1 had a high value of variance as 58.45% and PC2 had 15.17%, for 7-9-month age group, three principal components were extracted with high eigenvalues (7.56) for PC1 and high variance 50.45% and for 10-12-month age group, PC1 had a high value of variance as 50.05% and PC2 and PC3 had values as 10.67% and 9.05% respectively and their cumulative variance was 69.79%. Regression equations for all age groups of young Thalli sheep presented a very close relationship between predicted bodyweight and actual live bodyweight. The present study showed that hearth girth, body length, and withers height are very important parameters for the estimation of body weight and phenotypical characterization.