Плоды малины Rubus idaeus L. как источник функциональных ингредиентов

Lack of variation among black raspberry cultivars is thought to be a limiting factor in fruit production and in breeding improved cultivars. An assessment of the available diversity in black raspberry is needed to effectively develop improved cultivars. Such an assessment was done to estimate the genetic similarities for RAPD markers in 16 black raspberry genotypes and to determine the genetic diversity among these genotypes based on these markers. In addition, the ability to distinguish between the black raspberry genotypes, two red raspberry cultivars ( Rubus idaeus L.), and a blackberry cultivar ( Rubus hybrid) was determined. A similarity matrix from 379 RAPD markers was calculated, and a phylogenetic tree was constructed using the PHYLIP suite of phylogeny software, which revealed the relationship among the genotypes. An average of 81% similarity was calculated among 16 black raspberry genotypes with a maximum similarity of 98% and a minimum of 70%. The average similarity between black raspberry and red raspberry was 41% and was 26% between black raspberry and blackberry. Combined marker profiles from six RAPD primers could be used to distinguish between the 16 black raspberry genotypes. Red raspberry and blackberry could be distinguished from black raspberry by 27 and 29 of 30 RAPD primers tested, respectively. Genetic diversity was most prominent in genotypes from the extremes of the black raspberry indigenous range. Diversifying the germplasm pool for black raspberry cultivar improvement can be achieved through utilizing genotypes from the extremes of the black raspberry range and through interspecific hybridization.

Researchers developing new cultivars of red raspberry (Rubus idaeus subsp. idaeus L.) and black raspberry (R. occidentalis L.) observe progeny of breeding populations for several seasons to identify those that perform reliably. If a portion of any breeding population could be eliminated based on a qualitative character or molecular marker, resources used for that portion could be used for other progeny. Our objective is to identify such molecular markers for red raspberry and black raspberry. A black raspberry × red raspberry cross was made to develop a map of each parent, and an F2 population was generated to join the maps. Simple sequence repeat (SSR) markers derived from red raspberry and strawberry were used. The level of homozygosity for the red raspberry was 40%, and the level for the black raspberry was 80%. Severe segregation skewing was observed in the F2 generation and indicates problems with transmission. Our findings help quantify the relative levels of homozygosity previously reported for red raspberry and black raspberry. In addition, the severe skewing observed in the F2 generation provides a molecular perspective to the fertility problems previously reported for the black raspberry × red raspberry hybrids (purple raspberry). Since black raspberry is highly homozygous, purple raspberry has transmission and fertility problems, and black raspberry breeders have reported a frustratingly low level of diversity in this subgroup, development of a black raspberry map is expected to require twice the markers as for a red raspberry map, emphasizing the need for a black raspberry sequence from which to develop molecular markers.

WHAT'S GOING ON IN THE WORLD OF RUBUS BREEDING?

A comparative study of raspberry structural and biochemical responses to late leaf rustRosaceae is a family composed of important crops such as apple, peach, strawberry, and raspberry.Among these species, red and black raspberries belong to the genus Rubus and are high-value crops with increasing worldwide demand.However, these plants are affected by several diseases that impact their quality and productivity.Late leaf rust is a fungal disease in raspberries caused by Aculeastrum americanum (Farl.)M. Scholler & U. Braun (syn.Thekopsora americana (Farl.)Aime & McTaggart).While red raspberries (Rubus idaeus L.) are susceptible, black raspberries were previously reported as more resistant (R. occidentalis L.) and immune (R. niveus Thunb.) to this pathogen.Since genetic resistance is a promising way to manage this disease, hybridizing red and black raspberries can provide new cultivars with interesting traits.However, it is important to understand how plants respond to pathogens before developing expensive and long breeding programs.The aim of this thesis was to investigate the histopathological and biochemical responses of red and black raspberries and a hybrid between them to A. americanum colonization.It was observed that the pathogen germinated and colonized all studied raspberries, however, black raspberries and the hybrid had distinctive responses compared to R. idaeus.The compact mesophyll, the pre-and post-formed phenolic compounds, post-formed pectic compounds, and cell collapse in the lesioned area were the main defense mechanisms against A. americanum.Additionally, to structural and biochemical studies of raspberries defense responses, it was performed a comparative genomics analysis to identify the calcium-dependent protein kinases (CDPKs), focusing on the CPK28 orthologs in black raspberry (R. occidentalis) and strawberry (Fragaria vesca L.) as representative of Rosaceae.Both have 26 CDPKs together.The orthologs FvCPK28 and RoCPK28 were cloned and expressed in Nicotiana benthamiana and Arabidopsis cpk28 mutants.It was found that both Rosaceae CPK28 orthologs are plasmamembrane localized, and their overexpression dampened the oxidative burst upon elicitation.These results provided a proof-of-concept to investigate the functional similarities between these proteins and Arabidopsis CPK28.

Glycation is associated with several neurodegenerative disorders, including Alzheimer’s disease (AD), where it potentiates the aggregation and toxicity of proteins such as β-amyloid (Aβ). Published studies support the anti-glycation and neuroprotective effects of several polyphenol-rich fruits, including berries, which are rich in anthocyanins. Herein, blackberry, black raspberry, blueberry, cranberry, red raspberry, and strawberry extracts were evaluated for: (1) total phenolic and anthocyanins contents, (2) free radical (DPPH) scavenging and reactive carbonyl species (methylglyoxal; MGO) trapping, (3) anti-glycation (using BSA-fructose and BSA-MGO models), (4) anti-Aβ aggregation (using thermal- and MGO-induced fibrillation models), and, (5) murine microglia (BV-2) neuroprotective properties. Berry crude extracts (CE) were fractionated to yield anthocyanins-free (ACF) and anthocyanins-enriched (ACE) extracts. The berry ACEs (at 100 μg/mL) showed superior free radical scavenging, reactive carbonyl species trapping, and anti-glycation effects compared to their respective ACFs. The berry ACEs (at 100 μg/mL) inhibited both thermal- and MGO-induced Aβ fibrillation. In addition, the berry ACEs (at 20 μg/mL) reduced H2O2-induced reactive oxygen species production, and lipopolysaccharide-induced nitric oxide species in BV-2 microglia as well as decreased H2O2-induced cytotoxicity and caspase-3/7 activity in BV-2 microglia. The free radical scavenging, reactive carbonyl trapping, anti-glycation, anti-Aβ fibrillation, and microglial neuroprotective effects of these berry extracts warrant further in vivo studies to evaluate their potential neuroprotective effects against AD.

Influence of Variety and Location of Growth on Resulting Bramble Dried Leaf Volatiles

The triacylglycerol (TAG) compositions of blackberry, red raspberry, black raspberry, blueberry and cranberry seed oils were examined using ultra-performance convergence chromatography-quadrupole time-of-flight mass spectrometry (UPC2-QTOF MS). A total of 52, 53, 52, 59 and 58 TAGs were detected and tentatively identified from the blackberry, red raspberry, black raspberry, blueberry and cranberry seed oils, respectively, according to their accurate molecular weight in MS1 and fragment ion profiles in MS2. OLL was the most abundant TAG in the blackberry, red raspberry and black raspberry seed oils. Furthermore, the fatty acid compositions of the five berry seed oils were directly determined by gas chromatography coupled with mass spectrometry (GC-MS). In addition, the seed oils had total phenolic contents ranging 13.68–177.06 µmol GAE (gallic acid equivalent)/L oil, and significant scavenging capacities against DPPH, peroxyl, and ABTS+ radicals. These results indicated that the combination of UPC2 and QTOF MS could effectively identify and semi-quantify the TAGs compositions of the berry seed oils with sn-position information for the fatty acids. Understanding the TAGs compositions of these berry seed oils could improve the utilization of these potentially high nutritional value oils for human health.

Amphidiploid production of hybrid between black raspberry and Rubus parvifolius L., a wild Asian species, using colchicine treatment

Vegetable are widespread throughout the world and is a major part of the human diet. From the perspective of agricultural crops that belong to the group of Beta vulgaris (beetroot, mangold, sugar beets, fodder beet) are first-rate vegetables. Especially popular is used in the food industry for the production of sugar, various vegetable juices, coloring agents, and many other products. Beetroot (Beta vulgaris L.) is considered one of the ten most important vegetable in the world, thanks to the content of rare natural pigments (betalains), polyphenols, flavonoids, antioxidants, vitamins, minerals and fiber. In this work we evaluated content of bioactive substances, especially the content of total polyphenols, anthocyanins and antioxidant activity in several varieties (Cylindra, Kahira, Chioggia, Crosby Egyptian) of beetroot (Beta vulgaris L.). Samples of plant material were collected at full maturity stages from areas of Zeleneč (Czech Republic). Zeleneč is area without negative influences and emission sources. Samples of fresh beetroot (Beta vulgaris L.) were homogenized and were prepared as an extract: 50 g cut beetroot (Beta vulgaris L.) extracted by 100 mL 80% ethanol for sixteen hours. These extracts were used for analyses. The content of the total polyphenols was determined by using the Folin-Ciocalteu reagent (FCR). The absorbance was measured at 765 nm of wavelength against blank. The content of total anthocyanins was measured at 520 nm wavelength of the blank. Antioxidant activity was measured using a compound DPPH˙ (2.2-diphenyl-1-picrylhydrazyl) at 515.6 nm in the spectrophotometer. In the present experiment it was detected, that total polyphenols content in samples ranges from 218.00 mg.kg-1 to 887.75 mg.kg-1, total anthocyanins content in samples ranges from 14.48 ±0.40 mg.kg-1 to 84.50 ±4.71 mg.kg-1 and values of antioxidant activity were in interval from 8.37 ±0.29% to 21.83 ±0.35%.

Chapter 29 - Chemical Composition of Raspberry (Rubus spp.) Cultivars

The influence of growth regulator prohexadione-Ca (ProCa) concurrently with young canes removal on the modification of photosynthetic pigments content and antioxidant enzymes (peroxidase, POD; catalase, CAT; polyphenol oxidase, PPO; superoxide dismutase, SOD) activities in leaves and fruits of raspberry (Rubus idaeus L.) cultivar 'Willamette' was studied. ProCa increased while canes removal decreased chlorophylls and carotenoids content compared to control. POD, CAT, and PPO activities in leaves after removal of young canes were higher compared to control (2-4 times) which was visually confirmed for POD by isoelectrofocusing. Removal of young canes slithly increased, while ProCa significantly enhanced SOD activity in leaves compared to control (475.10 and 218.38 nkat mg-1 prot, respectively). Pattern of SOD activity in fruit was similar as in leaf with substantial increase compared to control (about 15 times). Combination of implemented measures increased activity of all enzymes in the leaves and fruits. Our study could provide a better knowledge of the ProCa and canes removal influences on the action of enzymes in order to regulate their activities in fruit products.

Anthocyanins in black raspberry extracts may play a key role in the regulation of oncogene expression in cancer cell cultures. Variations in anthocyanin levels of `Jewel', `Mac Black', and `Bristol' black raspberries grown at seven commercial farms in Ohio were investigated using HPLC and uv-vis spectrometry. Cyanidin-3-rutinoside (cy-3-rut) and cyanidin-3-(2G-xylorutinoside) (cy-3-2-xyl), the two major compounds present in all cultivars (≈2:1), were highly correlated with total anthocyanin contents. Sample variation in total anthocyanin, cy-3-rut, and cy-3-2-xyl levels was greater among commercial farms than among cultivars grown at the same location. The antioxidant activities of cy-3-rut, cyanidin-3-glucoside, cyanidin-3,5-diglucoside (cy-3,5-diglc), and pelargonidin-3-glucoside from purified extracts were determined using the free radical scavenging assays DPPH and ABTS, and the ferric reducing power assay FRAP. All pure anthocyanins showed strong antioxidant potentials except for cy-3,5-diglc. Cy-3-rut was identified and quantified as the dominant anthocyanin in black raspberries and was also the most potent antioxidant. Results suggest that anthocyanins, cy-3-rut in particular, may function as the primary antioxidants in black raspberries. Genetic and environmental variation in the anthocyanin contents necessitate characterization of the antioxidant and anthocyanin levels in fruits from any given source prior to measuring biological and medicinal activities.

Optimization of experimental conditions for bioactive compounds recovery from raspberry fruits (Rubus idaeus L.) by using combinations of ultrasound-assisted extraction and deep eutectic solvents

Due to the lack of effective and stable reference genes, studies on functional genes in Rubus, a genus of economically important small berry crops, have been greatly limited. To select the best internal reference genes of different types, we selected four representative cultivars of blackberry and raspberry (red raspberry, yellow raspberry, and black raspberry) as the research material and used RT-qPCR technology combined with three internal stability analysis software programs (geNorm, NormFinder, and BestKeeper) to analyze 12 candidate reference genes for the stability of their expression. The number of most suitable internal reference genes for different cultivars, tissues, and fruit developmental stages of Rubus was calculated by geNorm software to be two. Based on the results obtained with the three software programs, the most stable genes in the different cultivars were RuEEF1A and Ru18S. Finally, to validate the reliability of selected reference genes, the expression pattern of the RuCYP73A gene was analyzed, and the results highlighted the importance of appropriate reference gene selection. RuEEF1A and Ru18S were screened as reference genes for their relatively stable expression, providing a reference for the further study of key functional genes in blackberry and raspberry and an effective tool for the analysis of differential gene expression.

Total monomeric anthocyanins from six commercial plum varieties were extracted using two protocols, viz methanol/water extraction through end-over-end shaking and ethanol/water extraction in a shaking water bath; adapted from the Australian Wine Research Institute (AWRI) standard method. Anthocyanins were determined using the Association of Official Analytical Chemists (AOAC) standard protocol and a modification of the AWRI method. The mean relative standard deviation was found to be similar between methods, at 13.5% and 9.9%, respectively. On average, the anthocyanin concentrations given by the AWRI method were 27% higher than those obtained using the AOAC standard method. This was attributed to the AWRI method not correcting for haze or matrix interference, but estimating the anthocyanin concentration from the absorbance at a single wavelength. Anthocyanin measurements on the two extractions using the same measurement protocol supported this, indicating that the methanolic extracts gave a higher anthocyanin yield than the ethanolic extracts. Furthermore, HPLC profiling of the anthocyanin content demonstrated significantly more anthocyanins were extracted through the methanol extraction protocol. The methanol/water extraction protocol and AOAC standard method are much simpler to perform than the AWRI method and appear to be at least as precise. Thus this extraction and spectrophotometric protocol is well suited to future work on plum matrices. The extraction method is also more suitable for the subsequent photometric and/or HPLC determination of phenolic and total antioxidant contents.