Preharvest methyl jasmonate application delays cell wall degradation and upregulates phenolic metabolism and antioxidant activities in cold stored raspberries.

The effects of preharvest methyl jasmonate (MeJA) spray application on the physicochemical quality, metabolism of phenolics, and cell wall components in raspberries were investigated during a 10-day cold storage period. MeJA spray reduced firmness loss, decay incidence, and weight loss, while maintained higher levels of soluble solids content, ascorbic acid, anthocyanins and flavonoids in raspberries. Furthermore, MeJA application resulted in increased total pectin and protopectin levels, as well as lowered water-soluble pectin, and activities of pectin methyl esterase, polygalacturonase and cellulase enzymes. Additionally, MeJA treatment upregulated the phenylpropanoid pathway, leading to higher endogenous phenolics and activities of phenylalanine-ammonia lyase and shikimate dehydrogenase. In conclusion, preharvest MeJA spray application could be adopted to enhance the storage potential of cold-stored raspberries for 10 days by maintaining higher firmness, assuring better physicochemical quality, and increasing phenolic metabolism, while reducing cell wall hydrolysis.

A comparison of raspberry freezing-damage during preservation in isochoric and isobaric conditions.

Introduction: Fruits are perishable, thus it's crucial to have an efficient preservation technique that can increase storage time while keeping physical quality and nutritional attributes in order to avoid wastage. The majority of methods for long-term storage require refrigeration. Methods: In this investigation, we assess the viability of isochoric freezing as a different technique of raspberry (Rubus idaeus L.) preservation. Raspberries were subjected to different storage conditions: isochoric freezing at -4°C, conventional isobaric settings at +3°C (refrigerator), -21°C (freezer), and -4°C with a trehalose solution in a plastic bag. The study assessed changes in weight loss, visual appearance, color, hardness, °Brix values, and pH over a seven-day period. Results: Key findings reveal that raspberries subjected to isochoric freezing below the freezing point of water experienced minimal weight loss after seven days. Visual appearance, color, hardness, °Brix values, and pH were comparable to fresh raspberries, indicating minimal alteration. Discussion: These results suggest that isochoric freezing shows potential as a preservation method that maintains the physical and chemical properties of raspberries similar to fresh fruit. Implementing diverse preservation techniques tailored to raspberries may contribute to environmental sustainability by reducing food wastage and the associated environmental impact.

High-throughput color determination of red raspberry puree and correlation of color parameters with total anthocyanins.

BACKGROUND: Red raspberry fruit color is a key driver of consumer preference and a major target of breeding programs worldwide. Screening for fruit color typically involves the determination of anthocyanin content and/or the assessment of color through a colorimeter. However, both procedures are time-consuming when the analyses involve hundreds or thousands of samples. The objectives of this study were to develop a high-throughput method for red raspberry puree color measurement and to test the correlations between color parameters and total anthocyanin content. Color coordinates were collected with a colorimeter on 126 puree samples contained in Petri dishes and with the Tomato Analyzer Color Test (TACT) module to assess the same samples prepared in Petri dishes and in 96-well plates. An additional 425 samples were analyzed using only 96-well plates. Total anthocyanins were extracted from all 551 samples. RESULTS: Regression models for L*, a*, b* measured with the colorimeter and TACT using Petri dishes were all significant (p < 0.001), but very consistent only for L* (R2 = 0.94). Significant (p < 0.001) and very consistent regressions (R2 = 0.94 for L* and b*, R2 = 0.93 for a*) were obtained for color parameters measured with TACT using Petri dishes and TACT using plates. Of the color parameters measured with the colorimeter, only L*, a*/b*, and hue significantly correlated with total anthocyanins (p < 0.05), but, except for L* (R = - 0.79), the correlations were weak (R = - 0.23 for a*/b* and R = 0.22 for hue). Conversely, all correlations with total anthocyanins and color parameters measured with TACT were significant (p < 0.001) and moderately strong (e.g., R = - 0.69 for L* and R = 0.55 for a*/b*). These values were indicative of darker colors as total anthocyanin content increased. CONCLUSIONS: While the colorimeter and TACT-based methods were not fully interchangeable, TACT better captured color differences among raspberry genotypes than the colorimeter. The correlations between color parameters measured with TACT and total anthocyanins were not strong enough to develop prediction models, yet the use of TACT with 96-well plates instead of Petri dishes would enable the high-throughput measurement of red raspberry puree color.

Bacteria Associated with Spores of Arbuscular Mycorrhizal Fungi Improve the Effectiveness of Fungal Inocula for Red Raspberry Biotization.

Intensive crop production leads to the disruption of the symbiosis between plants and their associated microorganisms, resulting in suboptimal plant productivity and lower yield quality. Therefore, it is necessary to improve existing methods and explore modern, environmentally friendly approaches to crop production. One of these methods is biotization, which involves the inoculation of plants with appropriately selected symbiotic microorganisms which play a beneficial role in plant adaptation to the environment. In this study, we tested the possibility of using a multi-microorganismal inoculum composed of arbuscular mycorrhizal fungi (AMF) and AMF spore-associated bacteria for biotization of the red raspberry. Bacteria were isolated from the spores of AMF, and their plant growth-promoting properties were tested. AMF inocula were supplemented with selected bacterial strains to investigate their effect on the growth and vitality of the raspberry. The investigations were carried out in the laboratory and on a semi-industrial scale in a polytunnel where commercial production of seedlings is carried out. In the semi-industrial experiment, we tested the growth parameters of plants and physiological response of the plant to temporary water shortage. We isolated over fifty strains of bacteria associated with spores of AMF. Only part of them showed plant growth-promoting properties, and six of these (belonging to the Paenibacillus genus) were used for the inoculum. AMF inoculation and co-inoculation of AMF and bacteria isolated from AMF spores improved plant growth and vitality in both experimental setups. Plant dry weight was improved by 70%, and selected chlorophyll fluorescence parameters (the contribution of light to primary photochemistry and fraction of reaction centre chlorophyll per chlorophyll of the antennae) were increased. The inoculum improved carbon assimilation, photosynthetic rate, stomatal conductance and transpiration after temporary water shortage. Raspberry biotization with AMF and bacteria associated with spores has potential applications in horticulture where ecological methods based on plant microorganism interaction are in demand.

The Effect of Rubus idaeus Polyphenols Extract in Induced Endometriosis in Rats.

Endometriosis is a common gynecological condition with a complex physio-pathological background. This study aimed to assess the role of Rubus idaeus leaf extract (RiDE) as a potential therapeutic agent in reducing the size of the endometriotic lesions and modulate the plasma expression of MMP-2, MMP-9, and TGF-ß1. The endometriotic lesions were induced in a rat model by the autologous transplant of endometrium. Thirty-six female rats, Wistar breed, with induced endometriosis, were divided into four groups and underwent treatment for 28 days. The CTRL group received 0.5 mL/day of the vehicle; the DG group received 1 mg/kg b.w./day dienogest; the RiDG group received 0.25 mL/kg b.w./day RiDE and the D+RiDG group received 1 mg/kg b.w./day dienogest and 0.25 mL/kg b.w./day RiDE, respectively. Rats' weight, endometriotic lesion diameter and grade, and plasma levels of MMP-2, MMP-9, and TGF-ß1 were assessed before and after treatment. The administration of RiDE in association with dienogest vs. dienogest determined a lower weight gain and a reduction in diameter of the endometriotic lesions. RiDE administration restored MMP2 and MMP9 plasma levels to initial conditions. Rubus idaeus extract may help in reducing dienogest-associated weight gain, lower the size of endometriotic lesions, and have anti-inflammatory effects through MMP2 and MMP9 reduction.

Quality Characteristics of Raspberry Fruits from Dormancy Plants and Their Feasibility as Food Ingredients.

The raspberry (Rubus idaeus L.) is a soft red fruit consumed worldwide due to its bitter-sweet taste and phenolics-associated health benefits. During plant dormancy, raspberry fruits are discarded. However, this work hypothesised that these fruits have the chemical quality to be valorised, which would mitigate their waste if adequately stabilised. This can be achieved by drying. The Pacific Deluxe and Versailles varieties were dried by freeze- and convective-drying (30 °C and 40 °C). The freeze-dried fruits preserved their colour, drupelets structure, and phenolic content. Convective-drying promoted a significant fruit darkening, which was more evident at 30 °C due to the longer drying process, and a loss of drupelets structure. Both temperatures promoted a similar decrease in phenolic content, as determined by HPLC, although the ABTS●+ antioxidant activity at 40 °C was lower (IC50 = 9 compared to 13 µg AAE/mg dry weight). To incorporate dried raspberries into muffin formulations, while keeping their red colour, it was necessary to change the raising agent from sodium bicarbonate to baker's yeast. Sensory analysis by a non-trained panel revealed good acceptance, showing that fresh or dried raspberry fruits from dormancy had suitable characteristics for use as food ingredients.

Therapeutic Effects of Oral Application of Menthol and Extracts from Tormentil (Potentilla erecta), Raspberry Leaves (Rubus idaeus), and Loosestrife (Lythrum salicaria) during Acute Murine Campylobacteriosis.

Human food-borne infections with the enteropathogen Campylobacter jejuni are becoming increasingly prevalent worldwide. Since antibiotics are usually not indicated in campylobacteriosis, alternative treatment regimens are important. We here investigated potential disease-alleviating effects of menthol and of extracts from tormentil, raspberry leaves, and loosestrife in acute murine campylobacteriosis. Therefore, C. jejuni-infected microbiota-depleted IL-10-/- mice were orally treated with the compounds alone or all in combination from day 2 until day 6 post-infection. Whereas neither treatment regimen affected gastrointestinal pathogen loads, the combination of compounds alleviated C. jejuni-induced diarrheal symptoms in diseased mice on day 6 post-infection. Furthermore, the therapeutic application of tormentil and menthol alone and the combination of the four compounds resulted in lower colonic T cell numbers in infected mice when compared to placebo counterparts. Notably, pro-inflammatory cytokines measured in mesenteric lymph nodes taken from C. jejuni-infected mice following tormentil, menthol, and combination treatment did not differ from basal concentrations. However, neither treatment regimen could dampen extra-intestinal immune responses, including systemic pro-inflammatory cytokine secretion on day 6 post-infection. In conclusion, the combination of menthol and of extracts from tormentil, raspberry leaves, and loosestrife constitutes an antibiotic-independent approach to alleviate campylobacteriosis symptoms.

Fine-mesh exclusion netting reduces Drosophila suzukii (Diptera: Drosophilidae) infestation and improves organic fall-bearing raspberry yields.

The economical production of small fruits has been significantly complicated by the spotted-wing drosophila, Drosophila suzukii Matsumura, throughout its invaded rage. Fall-bearing red raspberries are especially susceptible to D. suzukii, and significant efforts to mitigate their damage are undertaken by growers. Exclusion barriers made from fine-mesh netting has emerged as an organic compliant strategy to mitigate damage from D. suzukii. Identifying less susceptible raspberry cultivars may also alleviate D. suzukii damage. Key pollination complications arise from exclusion-based tactics, but properly timing exclusion establishment may provide a remedy. In a 2-yr-old fall-bearing raspberry planting containing 3 cultivars, exclusion barriers were erected at 2 phenological timings. Spinosad- and pyrethrin-based pesticides were also applied to separate rows throughout harvest and a fourth treatment group matured without management. A subsection of raspberries was harvested and examined for marketability or damage, and D. suzukii adult populations were sampled with baited traps. An exclusion barrier applied early in fruit development was the most effective pest management strategy of those we tested. Its use decreased D. suzukii captures by 75%, decreased D. suzukii-infested fruit weights by 48%, and increased marketable yield by 63% compared to the no management control; however, it also increased pollination deficient fruit weights. Exclusion netting applied later in fruit maturation also decreased D. suzukii captures, infested fruit weights, and the weight of fruits damaged by other arthropod pests but did not significantly increase marketable yield. Our results indicate that netting exclusions may hold promise for the economical production of fall-bearing raspberries.

Arbuscular mycorrhizae influence raspberry growth and soil fertility under conventional and organic fertilization.

Introduction: Introducing beneficial soil biota such as arbuscular mycorrhizal fungi (AMF) to agricultural systems may improve plant performance and soil fertility. However, whether bioinocula species composition affects plant growth and soil fertility, and whether fertilizer source influences AMF colonization have not been well characterized. The objectives of this research were to: (1) assess if AMF bioinocula of different species compositions improve raspberry (Rubus idaeus L.) performance and characteristics of soil fertility and (2) evaluate the impact of fertilizer source on AMF colonization. Methods: Five bioinocula with different AMF species compositions and three fertilizer sources were applied to tissue culture raspberry transplants in a randomized complete block design with eight replicates. Plants were grown in a greenhouse for 14 weeks and plant growth, tissue nutrient concentrations, soil fertility, and AMF root colonization were measured. Results: Shoot K and Zn concentrations as well as soil pH and K concentration increased in the Commercial Mix 1 treatment (Glomus, Gigaspora, and Paraglomus AMF species) compared to the non-inoculated control. RFI (raspberry field bioinoculum; uncharacterized AMF and other microbiota) increased soil organic matter (SOM), estimated nitrogen release (ENR), and soil copper (Cu) concentration compared to the non-inoculated control. Furthermore, plants receiving the Mix 1 or RFI treatments, which include more AMF species, had greater AMF root colonization than the remaining treatments. Plants receiving organic fertilizer had significantly greater AMF colonization than conventionally fertilized plants. Conclusion: Taken together, our data indicate that coupling organic fertilizers and bioinocula that include diverse AMF species may enhance raspberry growth and soil fertility.

Volatilomics of raspberry fruit germplasm by combining chromatographic and direct-injection mass spectrometric techniques.

The application of direct-injection mass spectrometric (DI-MS) techniques, like Proton Transfer Reaction Time of Flight Mass Spectrometry (PTR-ToF-MS) has been suggested as a reliable phenotyping tool for fruit volatilome assessment in both genetic and quality-related studies. In this study the complexity of raspberry aroma was investigated by a comprehensive untargeted VOC analysis, done by combining SPME-GC-MS and PTR-ToF-MS assessments with multi-block discriminant analysis using the DIABLO mixOmics framework. The aim was to acquire an exhaustive characterization of the raspberry volatilome according to different fruit ripening stages (pink, ripe, and overripe) and genetic variances (50 accessions), as well as to investigate the potential of PTR-ToF-MS as a rapid and high throughput VOC phenotyping tool to address issues related to raspberry fruit quality. Results of this study demonstrated the complementarity between SPME-GC-MS and PTR-ToF-MS techniques to evaluate the raspberry aroma composition. PTR-ToF-MS generates reliable raspberry VOC fingerprints mainly due to a reduced compound fragmentation and precise content estimation. In addition, the high collinearity between isomers of monoterpenes and norisoprenoids, discovered by GC analysis, reduces the main analytic limitation of PTR-ToF-MS of not being able to separate isomeric molecules. The high similarity between the VOC matrices obtained by applying PTR-ToF-MS and SPME-GC-MS confirmed the possibility of using PTR-ToF-MS as a reliable high throughput phenotyping tool for raspberry volatiolome assessment. In addition, results provided by the germplasm collection investigation enabled to distinguish the best performing accessions, based on VOCs composition, to be used as superior parental lines for future breeding programs.

Abscisic Acid Synthesis and Signaling during the Ripening of Raspberry (Rubus idaeus 'Heritage') Fruit.

The raspberry (Rubus idaeus L.) fruit is characterized by its richness in functional molecules and high nutritional value, but the high rate of fruit softening limits its quality during postharvest. Raspberry drupelets have a particular ripening regulation, depending partially on the effect of ethylene produced from the receptacle. However, the possible role of abscisic acid (ABA) in the modulation of quality parameters during the ripening of raspberry is unclear. This study characterized the fruit quality-associated parameters and hormonal contents during fruit development in two seasons. The quality parameters showed typical changes during ripening: a drastic loss of firmness, increase in soluble solids content, loss of acidity, and turning to a red color from the large green stage to fully ripe fruit in both seasons. A significant increase in the ABA content was observed during the ripening of drupelets and receptacles, with the higher content in the receptacle of ripe and overripe stages compared to the large green stage. Moreover, identification of ABA biosynthesis-(9-cis-epoxycarotenoid dioxygenase/NCED) and ABA receptor-related genes (PYRs-like receptors) showed three genes encoding RiNCEDs and nine genes for RiPYLs. The expression level of these genes increased from the large green stage to the full-ripe stage, specifically characterized by a higher expression of RiNCED1 in the receptacle tissue. This study reports a consistent concomitant increase in the ABA content and the expression of RiNCED1, RiPYL1, and RiPYL8 during the ripening of the raspberry fruit, thus supporting the role for ABA signaling in drupelets.

Investigating Biochemical and Histopathological Responses between Raspberries and Aculeastrum americanum.

Late leaf rust is a fungal disease in raspberries caused by Aculeastrum americanum (Farl.) M. Scholler U. Braun (syn. Thekopsora americana (Farl.) Aime McTaggart) leading to early defoliation and yield losses. Red raspberries (Rubus idaeus L.) are susceptible to this pathogen, although this susceptibility varies among cultivars. In contrast, black raspberries were previously reported to be more resistant (Rubus occidentalis L.) and immune (Rubus niveus Thunb.) to this pathogen, raising their importance in plant breeding programs. However, what features make them respond differently to the same pathogen? In this study, we characterize for the first time the pre- and post-formed structural and biochemical defense mechanisms of R. idaeus cv. Autumn Bliss, R. occidentalis and R. niveus. Ultrastructural and histopathological analyses were used to uncover the interactions between these raspberries and A. americanum. The ultrastructural results indicate that the pathogen germinates on both leaf surfaces but can only form appressoria on the stomata. Although the three raspberry species were infected and colonized by A. americanum, a clear difference in susceptibility was observed between them. A compact mesophyll, pre- and post-formed phenolic compounds, and post-formed pectic compounds were the main plant defense mechanisms against fungal colonization. These findings provide new information about raspberries' defense mechanisms in response to A. americanum and elucidate the interactions occurring in these pathosystems.

Ethnopharmacology of Rubus idaeus Linnaeus: A critical review on ethnobotany, processing methods, phytochemicals, pharmacology and quality control.

ETHNOPHARMACOLOGICAL RELEVANCE: Rubus idaeus Linnaeus (RI) is a Chinese herbal medicine that has been widely used in China for a long time to reinforce the kidney, nourish the liver, improve vision, and arrest polyuria. AIM OF THE STUDY: This work aims to evaluate the recent progress of the chemical composition, pharmacological activity, pharmacokinetics, metabolism, and quality control and of Rubus idaeus, which focuses on the insufficiency of existing research and will shed light on future studies of Rubus idaeus. METHODS: Literatures about "Rubus idaeus","Red raspberry" and "Fupenzi"are retrieved by browsing the database, such as Web of Science (http://www.webofknowledge.com/wos), Pubmed (https://pubmed.ncbi.nlm.nih.gov/), CNKI (http://www.cnki.net/), and Wanfang Data (http://www.wanfangdata.com.cn). In addition, related textbooks and digital documents are interrogated to provide a holistic and critical review of the topic. The period of the literature covered from 1981 to 2022. RESULTS: Approximately 194 compounds have been isolated from Rubus idaeus, which is rich in phenols, terpenoids, alkaloids, steroids, and fatty acids. Numerous investigations have demonstrated that Rubus idaeus exhibits many pharmacological activities, including hypoglycemic and hypolipidemic, anti-Alzheimer effect, anti-osteoporosis, hepatoprotective, anti-cancer, neuroprotective, anti-bacteria and skin care, etc. However, it is worth noting that most of the research is not associated with the conventional effect, such as reducing urination and treating opacity of the cornea. CONCLUSION: The effectiveness of Rubus idaeus has been proved by its long-term clinical application. The research on the pharmacological activity of Rubus idaeus has flourished. In many pharmacological experiments, only the high-dose group can achieve the corresponding efficacy, so the efficacy of Rubus idaeus needs to be further interrogated. Meanwhile, the relationship between pharmacological activity and specific compounds of Rubus idaeus has not been clarified yet. Last but not least, studies involving toxicology and pharmacokinetics are very limited. Knowledge of bioavailability and toxicological behavior of Rubus idaeus can help understand the herb's pharmacodynamic and safety profile.

Integrated 16s RNA sequencing and network pharmacology to explore the effects of polyphenol-rich raspberry leaf extract on weight control.

Introduction: Obesity is recognized as a chronic low-grade inflammation associated with intestinal flora imbalance, leading to dyslipidemia and inflammation. Modern research has found that polyphenols have anti-obesity effects. However, the mechanism of action of raspberry leaf extract (RLE) with high polyphenols in regulating obesity is still unknown. This study investigated the improvement effect of supplementing RLE on high-fat diet (HFD) induced obesity in mice. Methods: RLE was used to intervene in HFD induced C57BL/6J male mice during prevention stage (1-16 weeks) and treatment stage (17-20 weeks). Their weight changes and obesity-related biochemical indicators were measured. The changes in intestinal flora were analyzed using 16S rRNA sequencing, and finally the targets and pathways of the 7 typical polyphenols (quercetin-3-O-glucuronide, ellagic acid, kaempferol-3-O-rutinoside, chlorogenic acid, brevifolin carboxylic acid, quercetin-3-O-rutinoside, and quercetin) of RLE in the regulation of obesity were predicted by network pharmacology approach. Results and discussion: The results showed that RLE effectively prevented and treated weight gain in obese mice induced by HFD, alleviated adipocyte hypertrophy, reduced Interleukin-6 and Tumor Necrosis Factor Alpha levels, and improved intestinal flora, especially Muriaculaceae, Alistipes and Alloprevotella, and decreased the Firmicutes/Bacteroidota ratio. Network pharmacology analysis selected 60 common targets for 7 RLE polyphenols and obesity. Combined with protein-protein interaction network, enrichment analysis and experimental results, TNF, IL-6, AKT1, and PPAR were predicted as potential key targets for RLE polyphenols. Conclusion: The potential mechanism by which polyphenol-rich RLE regulates obesity may be attributed to the specific polyphenols of RLE and their synergistic effects, therefore RLE has a great anti-obesity potential and may be used as a means to alleviate obesity and related diseases.

Mineral composition of repair raspberry (Rubus idaeus L.) fruits.

In recent years, raspberry breeding has shifted its emphasis from agronomic performance to characteristics related to the sensory qualities of the fruit and its potential health benef its. The therapeutic and preventive properties of raspberries are related to their biochemical composition. In this regard, the purpose of the work was to determine the content of macro- and micronutrients in fruits of different cultivars of repair raspberry using modern high-tech analytical methods and the selection of genetic sources of the analyzed elements for further breeding. The objects of the research were 17 cultivars of repair raspberry of different ecological and geographical origin from the genetic plant bioresource collection of FSBSO ARHCBAN. It was found that the ash residue of berries contains 12 major elements, which form the following descending series: K > P > Mg ≥ Mo > Ca > S ≥ Ni > Zn > Mn > Se > Fe ≥ Co. The largest proportion of ash residue in raspberry fruits is K. Depending on the cultivar, its quantity averaged from 12.81 wt % (Samorodok and Karamelka) to 22.37 wt % (Atlant). The minimum K content was observed in the ash of the Carolina cultivar (5.62 wt %), while in berries of this cultivar Mg (2.91), Ca (2.62) and Zn (0.14 wt %) accumulated above average. Among the group of early maturing cultivars, the cultivar Yubileinaya Kulikova stands out with a high content of Mo (4.63), Ca (2.19), Fe (0.25) and Co (0.21 wt %). The cultivar Pingvin is characterized by a high content of K (22.65) and Se (0.31 wt %). The medium maturity cultivar Samorodok is characterized by a higher content of P (4.08), S (0.47), Ni (0.51) and Zn (0.26 wt %). Among the late maturing cultivars, the cultivar Poranna Rosa stands out with the preferential accumulation of nine elements: Mg (2.98), P (4.42), S (0.36), K (20.34), Ca (1.71), Mn (0.14), Co (0.13), Se (0.21) and Mo (3.08 wt %). Correlation relationships between the elements have been established. Samples with the highest accumulation of macro- and microelements in berries represent genetic sources for further selection of raspberry for improvement of the mineral composition of fruits.

Relationship between Phenolic Compounds and Antioxidant Activity in Berries and Leaves of Raspberry Genotypes and Their Genotyping by SSR Markers.

The red raspberry is one of the world's most popular berries. The main direction of its breeding has switched to nutritional quality, and the evaluation of raspberry germplasm for antioxidant content and activity is very important. As berries, raspberry leaves contain valuable bioactive compounds, but the optimal time for their collection is unknown. We evaluated 25 new breeding lines and standard raspberry cultivars for their polyphenolic content and antioxidant capacity. The antioxidant activity of berries correlated better with the content of total phenolics (0.88 and 0.92) and flavonoids (0.76 and 0.88) than with anthocyanins (0.37 and 0.66). Two breeding lines were significantly superior to the standard cultivars and can be used in further breeding. Leaves collected in three phenological phases of the raspberry contained more phenolics (5.4-fold) and flavonoids (4.1-fold) and showed higher antioxidant activities (2.4-fold in FRAP assay, 2.2-fold in ABTS) than berries. The optimal time for harvesting raspberry leaves is the fruit ripening stage, with exceptions for some cultivars. Genetic diversity analysis using microsatellite (SSR) markers from flavonoid biosynthesis genes divided the genotypes into five clusters, generally in agreement with their kinships. The relationship between genetic data based on metabolism-specific SSR markers and the chemical diversity of cultivars was first assessed. The biochemical and genetic results show a strong correlation (0.78). This study is useful for further the improvement of raspberry and other berry crops.

Biologically Active Preparations from the Leaves of Wild Plant Species of the Genus Rubus.

The plants of the genus Rubus (R.) are applied as antiseptic agents in the treatment of skin diseases. Despite the great interest in plants of this genus, there are few reports on the antioxidant and biological activities of preparations obtained from the leaves of these plants. Therefore, we decided to evaluate the antioxidant activity of preparations from leaves of wild plant species of the genus Rubus using the frequently applied DPPH, ABTS, and FRAP methods, as well as to determine the total polyphenol content using the Folin−Ciocalteau method and perform qualitative evaluation by gas chromatography−mass spectrometry (GC-MS). The bactericidal and fungicidal activities of the obtained preparations were evaluated by applying laboratory tests: using the disc and the well methods based on the standards EN 13697:2019, EN 13697:2015, and EN 1500:2013. Microbiological tests of the plant preparations against bacteria, fungi, and yeasts isolated from the environment and against reference strains were performed. Moreover, antimicrobial testing of antibiotics against the tested strains was performed for comparison. The n-octanol/water partition coefficient of the obtained preparations was determined by the shake-flask method to determine their lipophilicity. According to the results, a high content of polyphenols and other antioxidant and biologically active compounds can be thought of as the parameter responsible for the effective activity of plant preparations obtained from wild plant species of the genus Rubus. The methods for determining bactericidal and fungicidal activity clearly demonstrates that preparations with reduced ethanol content exhibit bactericidal and fungicidal activity on surfaces. Testing of hand disinfection by means of rubbing with the preparations confirmed their antimicrobial activity against Escherichia coli K12 NCTC 10538. The obtained results show that the tested preparations exhibit on average two times lower activity against the reference bacterial strains than comparable antibiotics. The preparations obtained from the leaves of R. idaeus L. and R. fruticosus L. could complement classical antibiotics. While environmental bacteria showed a similar response to the preparations and antibiotics, their sensitivity was about one-third less than that of the reference strains. Our studies have shown that the obtained preparations are highly hydrophilic (logP < 0). Thus, these preparations can only be used in lipid bilayers in the aqueous core of liposomes, not in the lipid envelope.

Protection of Erythrocytes and Microvascular Endothelial Cells against Oxidative Damage by Fragaria vesca L. and Rubus idaeus L. Leaves Extracts-The Mechanism of Action.

The aim of this work is to determine the biological activity of ellagitannins rich extracts from leaves of raspberry (Rubus idaeus L.) and wild strawberry (Fragaria vesca L.) in relation to cells and cell membranes. Detailed qualitative and quantitative analysis of phenolic compounds of the extract was made using chromatographic methods. Cytotoxic and antioxidant activities of tested extracts in relation to erythrocytes and human vascular endothelial cells (HMEC-1) were determined by using fluorimetric and spectrophotometric methods. In order to establish the influence of the extracts on the physical properties of the membrane, such as osmotic resistance and erythrocytes shapes, mobility and/or hydration of polar heads and fluidity of hydrocarbon chains of membrane lipids, microscopic and spectroscopic methods were used. The results showed that the extracts are non-toxic for erythrocytes and HMEC-1 cells (up to concentration of 50 µg/mL), but they effectively protect cells and their membranes against oxidative damage. The increase in osmotic resistance of erythrocytes, formation of echinocytes and changes only in the polar part of the membrane caused by the extracts demonstrate their location mainly in the hydrophilic part of the membrane. The results indicate that tested extracts have high biological activities and may be potentially used in delaying the ageing process of organisms and prevention of many diseases, especially those associated with oxidative stress.

Evaluation of real-time nutrient analysis of fertilized raspberry using petiole sap.

The time delay in receiving conventional tissue nutrient analysis results caused red raspberry (Rubus idaeus L.) growers to be interested in rapid sap tests to provide real-time results to guide immediate nutrient management practices. However, sap analysis has never been conducted in raspberry. The present work aimed to evaluate the relationship of petiole sap nitrate (NO3 -), potassium (K+), and calcium (Ca2+) concentrations measured using compact ion meters and leaf tissue total nitrogen (TN), potassium (K), and calcium (Ca) concentrations measured using conventional tissue nutrient analysis. The relationship of petiole sap NO3 - and leaf tissue TN concentrations with plant growth and production variables was also explored. Fertilizer treatments of urea were surface applied to raised beds of established "Meeker" floricane red raspberry plots at control, low, medium, and high rates (0, 34, 67, and 101 kg N ha-1, respectively) in 2019 and 2020. The experiment was arranged in a randomized complete block design with three replications. Whole leaves were collected from representative primocanes in mid- and late- July and August 2019 and 2020 (i.e., four sampling time points per year). At each sampling time point, a subsample of leaves was used for petiole sap analyses of NO3 -, K+, and Ca2+ concentrations using compact ion meters, and conventional tissue testing of leaf tissue TN, K, and Ca concentrations, respectively. There were no interactions between N fertilizer rate and year nor between N fertilizer rate and sampling time. No significant differences were found due to N fertilizer rate for petiole sap NO3 -, K+, Ca2+ nor leaf tissue TN, K, Ca concentrations. However, significant year and sampling time effects occurred in measured petiole sap and leaf tissue nutrient concentrations. Overall, the correlations between petiole sap NO3 - and leaf tissue TN, petiole sap Ca2+ and leaf tissue Ca, petiole sap K+ and leaf tissue K concentrations were non-strong and inconsistent. Future research is warranted as the interpretation of correlations between raspberry petiole sap and leaf tissue nutrient concentrations were inconclusive.

Rubus idaeus extract improves symptoms in knee osteoarthritis patients: results from a phase II double-blind randomized controlled trial.

BACKGROUND: Osteoarthritis (OA) is the most frequent cause of disability in elderly people. In daily practice, the main objective of the physician is to reduce patient symptoms using treatments without adverse effects. However, the most prescribed treatment to manage OA symptoms remains nonsteroidal anti-inflammatory drugs which are associated with severe adverse effects. Therefore, we need a safe alternative to managing OA. One candidate is Rubus idaeus leaf extracts known to inhibit inflammatory responses. OBJECTIVE: This study aimed to evaluate the effects of a 12-weeks intervention with an ethanolic extract from Rubus idaeus leaf on symptoms of knee osteoarthritis. METHOD: The study was a randomized, double-blind, placebo-controlled, monocentric trial of 198 participants with femorotibial osteoarthritis. Participants were randomized equally to receive one daily during 3 months either 1 capsule of Rubus idaeus leaf extract 400 mg, 1 capsule of Rubus idaeus leaf extract 200 mg, or 1 capsule of placebo. The participants were assessed at baseline and after one and three months of treatment. The primary endpoint was an absolute change of the Western Ontario McMaster osteoarthritis index (WOMAC) pain subscale. The secondary endpoints were WOMAC global score, stiffness and function sub-scales, knee pain VAS score at walking, the Short Form (SF)-36, the Short Physical Performance Battery (SPPB), the 20-m walk test, and the International Physical Activity Questionnaire (IPAQ) and Outcome Measures in Rheumatology Clinical Trials and Osteoarthritis Research Society International (OMERACT-OARSI) responders rate. Statistical analyses were conducted on the intent-to-treat (ITT) population. RESULTS: In the Intention-to-treat population, WOMAC pain was not significantly modified by Rubus idaeus leaf extract compared to placebo. In contrast, Rubus idaeus leaf extract 400 mg after 12 weeks of treatment significantly reduced pain measured by the VAS. The mean pain decrease induced by Rubus ideaus leaf extract was over -7 mm which is clinically relevant and reached a clinically statistical difference compared to placebo with the highest dose. Rubus Ideaus was not significantly more efficient than the placebo on WOMAC global score, stiffness, and physical function subscores, IPAQ, SF-36, walking distance in treadmill test, SPPB, and evaluation of associated treatments needed to manage OA. CONCLUSION: Rubus idaeus leaf extract was well tolerated and effective to relieve pain in a patient with knee osteoarthritis. TRIAL REGISTRATION: NCT03703024  (11/10/2018).