Towards smart and sustainable development of modern berry cultivars in Europe.

Fresh berries are a popular and important component of the human diet. The demand for high-quality berries and sustainable production methods is increasing globally, challenging breeders to develop modern berry cultivars that fulfill all desired characteristics. Since 1994, research projects have characterized genetic resources, developed modern tools for high-throughput screening, and published data in publicly available repositories. However, the key findings of different disciplines are rarely linked together, and only a limited range of traits and genotypes has been investigated. The Horizon2020 project BreedingValue will address these challenges by studying a broader panel of strawberry, raspberry and blueberry genotypes in detail, in order to recover the lost genetic diversity that has limited the aroma and flavor intensity of recent cultivars. We will combine metabolic analysis with sensory panel tests and surveys to identify the key components of taste, flavor and aroma in berries across Europe, leading to a high-resolution map of quality requirements for future berry cultivars. Traits linked to berry yields and the effect of environmental stress will be investigated using modern image analysis methods and modeling. We will also use genetic analysis to determine the genetic basis of complex traits for the development and optimization of modern breeding technologies, such as molecular marker arrays, genomic selection and genome-wide association studies. Finally, the results, raw data and metadata will be made publicly available on the open platform Germinate in order to meet FAIR data principles and provide the basis for sustainable research in the future.

Evaluation of Some Phenological and Biochemical Characteristics of Selected New Late Flowering Dried Apricot Cultivars.

In this study, promising candidates of six apricot genotypes were compared with our local and major cultivar (Hacihaliloglu) based on their phenological, pomological, and biochemical characteristics. Fruit weight, stone weight, flesh firmness, flesh fruit/stone ratio, TSS (Total soluble solids), glucose, fructose, sucrose, total phenols, total antioxidant activity (FRAP and ABTS assays) and bud break, first bloom, full bloom, and harvesting date of apricot cultivar candidates were compared with cultivar Hacihaliloglu 'HH.' According to the obtained results, the higher content of total phenolic compounds and total antioxidant activity was determined in the N95 genotype (96.87 µmol TE/g) fruit, whereas the 'HH' cultivar was characterized by the lower value (94.6 µmol TE/g) especially determined by FRAP method. Individual sugars and the soluble solids content of fruit differed between selected genotypes and 'HH' cultivar. Glucose, fructose, and sucrose contents were higher in all selected genotypes than in Hacihaliloglu cultivar.

Identification of sex-linked SNP markers using RAD sequencing suggests ZW/ZZ sex determination in Pistacia vera L.

BACKGROUND: Pistachio (Pistacia vera L.) is a dioecious species that has a long juvenility period. Therefore, development of marker-assisted selection (MAS) techniques would greatly facilitate pistachio cultivar-breeding programs. The sex determination mechanism is presently unknown in pistachio. The generation of sex-linked markers is likely to reduce time, labor, and costs associated with breeding programs, and will help to clarify the sex determination system in pistachio. RESULTS: Restriction site-associated DNA (RAD) markers were used to identify sex-linked markers and to elucidate the sex determination system in pistachio. Eight male and eight female F1 progenies from a Pistacia vera L. Siirt × Bagyolu cross, along with the parents, were subjected to RAD sequencing in two lanes of a Hi-Seq 2000 sequencing platform. This generated 449 million reads, comprising approximately 37.7 Gb of sequences. There were 33,757 polymorphic single nucleotide polymorphism (SNP) loci between the parents. Thirty-eight of these, from 28 RAD reads, were detected as putative sex-associated loci in pistachio. Validation was performed by SNaPshot analysis in 42 mature F1 progenies and in 124 cultivars and genotypes in a germplasm collection. Eight loci could distinguish sex with 100% accuracy in pistachio. To ascertain cost-effective application of markers in a breeding program, high-resolution melting (HRM) analysis was performed; four markers were found to perfectly separate sexes in pistachio. Because of the female heterogamety in all candidate SNP loci, we report for the first time that pistachio has a ZZ/ZW sex determination system. As the reported female-to-male segregation ratio is 1:1 in all known segregating populations and there is no previous report of super-female genotypes or female heteromorphic chromosomes in pistachio, it appears that the WW genotype is not viable. CONCLUSION: Sex-linked SNP markers were identified and validated in a large germplasm and proved their suitability for MAS in pistachio. HRM analysis successfully validated the sex-linked markers for MAS. For the first time in dioecious pistachio, a female heterogamety ZW/ZZ sex determination system is suggested.

Comparison of lipids, fatty acids and volatile compounds of various kumquat species using HS/GC/MS/FID techniques.

BACKGROUND: Kumquat (Fortunella spp.) is one of the important fruit species that has been introduced into Turkey recently. It is well adapted to the Mediterranean region in Turkey. Early research of kumquat was started on adaptation and pomological traits, and only a few studies have been concerned with chemical content. After understanding the health benefits of kumquat fruits farmers started their plantations in Turkey. Thus in this study the fruits of five kumquat species - Fortunella margarita (Lour.) Swingle, F. crassifolia Swingle, F. obovata Hort. ex Tanaka, F. hindsii (Champ. ex Benth.) Swingle, and limequat [Citrus aurantifolia × F. japonica (Thumb.)] - were compared based on their lipids, fatty acids and volatile profiles. The lipids, fatty acids and volatile profiles were determined by using gas chromatography with flame ionisation detection (GC/FID) and headspace gas chromatography mass spectrometry (HS-GC/MS) techniques. Volatile constituents of kumquat fruits were isolated by a headspace method. RESULTS: The total lipid content (%) of kumquat species ranged from 0.26% to 0.37%. The polyunsaturated fatty acids were detected in a higher amount compared to the monounsaturated fatty acids and saturated fatty acids, respectively. The headspace technique led to the identification of 39 compounds and among a total of 39 constituents identified, d-limonene was the most abundant compound, comprising 67.78-88.72%. The results indicated that kumquats are rich in dietary and health benefits. CONCLUSION: Thirty-nine compounds were identified and terpenes were the major chemical group in all kumquat genotypes. Esters, alcohols, aldehydes and ketones were also detected. The present study revealed that the kumquat fruits have a valuable levels of health beneficial chemo-preventive effects and genotypes varied based on their lipid, fatty acids and volatile constituents.

Phenolic compounds of apple cultivars resistant or susceptible to Venturia inaequalis.

CONTENT: Phenolic compounds play an important role in the plant defense mechanism and are responsible for antioxidant capacity in fruits and vegetables. It is known that the phenolics can determine in the leaves of plants which are resistant/susceptible to fungal infections. OBJECTIVE: This study investigated the total phenolic compounds, content of shikimic acid from 33 different apple cultivars leaves infected with Venturia inaequalis [(Cke). Wint.] cultured in Fruit Research Station, in Egirdir, Isparta, Turkey. MATERIALS AND METHODS: Leaves of apple cultivars were collected three times in an interval of 30 d from July to September in 2010, and analyzed using HPLC methods to detect changes in the amount of the phenolic compounds and shikimic acid. RESULTS: Total phenolic compounds and shikimic acid in resistant/moderate susceptible apple cultivars were higher than susceptible apple cultuvars, although not statistically different between resistant and susceptible apples. The content of shikimic acid was statistically higher only in the leaves of the domestic cultivar Ankara güzeli on all three dates. DISCUSSION AND CONCLUSION: Recently, there have been increased studies trying to explain the resistance mechanism in plants. Natural resistance genes are investigated in some apple cultivars and new resistance varieties which have resistant genes are identified daily. Our study hold to determine the relationship between the phenolic compounds and the expression of resistance seems to be promising.

Sugar, Invertase Enzyme Activities and Invertase Gene Expression in Different Developmental Stages of Strawberry Fruits.

The cultivated strawberry (Fragaria × ananassa) is octoploid (2n = 8x = 56) and has been the focused fruit species of which an increasing number of molecular and genetic research has been conducted in recent years. The aim of this study is to identify the relationships between sucrose metabolism, invertase enzyme activity and gene expression in four different fruit development periods (red, pink, green and white) of two commercially important strawberry varieties 'Rubygem' and 'Fortuna'. The metabolite profiles (glucose, fructose, sucrose and total sugar content) of two varieties were discovered to be extremely similar. The highest amount of total sugar was found in red fruits, while the lowest was obtained from green fruits. Invertase represents one of the key enzymes in sucrose metabolism. The lowest invertase activity was obtained from the green fruits in 'Rubygem' and 'Fortuna' during four developmental periods. In these varieties, the amount of sucrose was found to be close to glucose and fructose and the lowest amount was detected in green period, while invertase activity was relatively high during red and pink periods and invertase gene expression was determined at high levels in both primers (St-4 and St-6) in the green period. The results of the study indicated that sugar content and invertase activity were positively correlated while enzyme activity and gene expression were negatively correlated.

Genetic characterization of pomegranate (Punica granatum L.) genotypes by AFLP markers.

The Coruh Valley, located in Northeastern Turkey, is one of the most important centers of diversity in pomegranate in Turkey. In this study, we attempted to characterize 19 promising pomegranate genotypes originating from the Coruh Valley in using fluorescent dye AFLP markers and capillary electrophoresis. Four AFLP primer combinations were used, generating a total of 297 fragments, 213 of which were polymorphic (73.0%). Resolving powers of the AFLP primers ranged from 0.700 to 1.018, with a total of 3.440, while polymorphism information contents ranged from 0.707 to 0.837 with an average of 0.764. UPGMA clustering of the genotypes showed two major groups. Most of the fruit characteristics of the genotypes within the same group were variable. Therefore, the results showed that molecular characterization is necessary to get reliable relationships among pomegranate genotypes and AFLP markers can be used effectively in pomegranate.

Evaluation of Polyphenolic Profile and Antibacterial Activity of Pomegranate Juice in Combination with Rifampin (R) against MDR-TB Clinical Isolates.

BACKGROUND: The global rise of multi-drug resistant M. tuberculosis demands unconventional treatment to enhance the efficiency of current drugs. Punica granatum, which is known as pomegranate, is considered as a member of the Punicaceae family. Pomegranate, which is broadly documented for its activity against a wide spectrum of bacterial pathogens, deserves further scrutiny in this respect. METHODS: Within this scope, this study investigated the effect of fresh pomegranate juice (FPJ) on the antibacterial activity of anti-tuberculosis drugs (Rifampin (R) and Isoniazid (INH)) against MDR-TB clinical isolates. The drug resistance profiles in M. tuberculosis clinical isolates were determined by susceptibility test using BACTEC MGIT 960 system. Four concentrations of fresh pomegranate juice (FPJ) (5%, 10%, 15%, and 20%) were evaluated in combination with R and INH at a dose range of (1.0 µg/ml) and (0.1 µg/ml), respectively against the MDR-TB isolates by the BACTEC MGIT 960 system. Moreover, this study scrutinized individual phenolic compounds of FPJ by using highperformance liquid chromatography (HPLC). The total polyphenols (TP), total flavonoid (TF), total anthocyanins content (TAC), and the antioxidant capacity were also assessed in FPJ. RESULTS: Synergistic effects were observed between R and INH with FPJ against all tested strains. However, combination therapy of rifampin was more effective than isoniazid one. Therefore, the combination of R and FPJ has been used against (27) MDR-TB clinical isolates. 5% of FPJ plus R (1.0 µg/ml) were found to suppress the growth of one isolates for first group (INH and R resistant). However, 5% of FPJ demonstrated no synergistic impact with R for second (SM, R and INH resistant) and third group (INH, EMB, R and SM resistant). Moreover, 10% of FPJ and R (1.0 µg/ml) inhibited the bacterial growth of three isolates of first group and two isolates and one isolate for second and third group, respectively. Remarkably, 15% of FPJ plus R (1.0 µg/ml) appeared to inhibit the growth of MDR-TB isolates for all tested groups indicating a strong synergistic effect. Regarding H37RV, the complete inhibition of the bacterial growth was found to occur at 15% and 20% concentrations of FPJ only. Minimum inhibitory concentration (MIC) of FPJ ranged from (4% to13%) for first group and from (10% to15%) for second and third group. Thus, FPJ at 15% inhibited 100% of bacteria for all tested isolates (MIC100% =15%). Phenolic compounds identified in FPJ were gallic acid, benzoic acid, syringic, folic acid, pelargonidin, naringin+ellagic acid, naringenin, chlorogenic acid, caffeic acid, catechin, myricetin, kaempferol, quercetin, cyanidin-3-glycoside, p-cummaric acid, ferulic acid, and rutin. Total phenolic (TP), total flavonoid (TF), and total anthocyanin (TA) content were 841.5 mg/L, 638.73 mg RE/L, and 47.43 mg/L, accordingly. CONCLUSION: Overall, FPJ displayed synergistic effect with R against MDR-TB clinical isolates due to its high content of polyphenol and antioxidant capability.

Health and taste related compounds in strawberries under various irrigation regimes and bio-stimulant application.

Strawberry has a unique status within the fruit species in terms of health and taste related compounds. This experimental study concerned the application of a bio-stimulant at various drip irrigation levels (IR125, IR100, IR75 and IR50). The effects of the bio-stimulant (seaweed extract) on the eating quality, i.e., the taste-related (TSS, fructose, glucose, sucrose and citric, malic, l-ascorbic acid), and health-related (antioxidant activity, total phenol, myricetin and quercetin) compounds were studied in two strawberry cultivars. The 'Rubygem' with its higher sugar and lower acid content has been more preferable than the 'Kabarla' cultivar. The bio-stimulant contributes to taste by improving the TSS, fructose, sucrose and also to health by increasing the quercetin content of the fruit which is associated to the cardiovascular properties and cancer reducing agents. The experiment conducted revealed significant increases only in the TSS contents and antioxidant activity under the IR50 and IR75 deficit irrigation treatments.

Phenolic composition of strawberry genotypes at different maturation stages.

The effects of maturation (green, pink, and ripe) on phenolic composition of strawberry cultivars Camarosa, Dorit, Chandler, and Osmanli and their hybrids were investigated using a high-pressure liquid chromatography (HPLC) method. p-Hydroxybenzoic acid, p-coumaric acid, ellagic acid, cyanidin-3-glucoside, pelargonidin-3-glucoside, kaempferol, quercetin, and myricetin were individually quantified for each stage. The highest amounts of anthocyanins were obtained from ripe fruits whereas ellagic acid was found as the main phenolic in the green fruits. Phenolic concentrations were found statistically different in green and ripe fruits. One hybrid was found to have higher phenolic contents than the other genotypes. The p-hydroxybenzoic and p-coumaric acid levels changed during maturation, but no differences in contents of flavonoids in green and ripe fruit were detected.

Aroma biosynthesis in strawberry: s-adenosylmethionine:furaneol o-methyltransferase activity in ripening fruits.

Among the most important volatile compounds in the aroma of strawberries are 2,5-dimethyl-4-hydroxy-3(2H)-furanone (Furaneol) and its methoxy derivative (methoxyfuraneol, mesifuran). Three strawberry varieties, Malach, Tamar, and Yael, were assessed for total volatiles, Furaneol, and methoxyfuraneol. The content of these compounds sharply increased during fruit ripening, with maximum values at the ripe stage. An enzymatic activity that transfers a methyl group from S-adenosylmethionine (SAM) to Furaneol sharply increases during ripening of strawberry fruits. The in vitro generated methoxyfuraneol was identified by radio-TLC and GC-MS. The partially purified enzyme had a native molecular mass of approximately 80 kDa, with optimum activity at pH 8.5 and 37 degrees C. A high apparent K(m) of 5 mM was calculated for Furaneol, whereas this enzyme preparation apparently accepted as substrates other o-dihydroxyphenol derivatives (such as catechol, caffeic acid, and protocatechuic aldehyde) with much higher affinities (K(m) approximately 105, 130, and 20 microM, respectively). A K(m) for SAM was found to be approximately 5 microM, regardless of the acceptor used. Substrates that contained a phenolic group with only one OH group, such as p-coumaric and trans-ferulic acid, as well as trans-anol and coniferyl alcohol, were apparently not accepted by this activity. It is suggested that Furaneol methylation is mediated by an O-methyltransferase activity and that this activity increases during fruit ripening.

Comparitive study on volatile aroma compounds of two different garlic types (Kastamonu and Chinese) using gas chromatography mass spectrometry (HS-GC/MS) technique.

BACKROUND: The medicinal use of garlic is much older than its usage as a food. The medical importance of garlic comes forward for its sulfur-containing components. In this study, it was aimed to compare Kastamonu garlic type with Chinese garlic type based on their aroma profiles. MATERIALS AND METHODS: Fresh Kastamonu garlic samples harvested from Kastamonu region of Turkey and Chinese garlic samples obtained from Turkish market were used as plant material. Volatile aroma compounds were determined using Headspace Gas Chromatography Mass Spectrometry (HS-GC/MS). RESULTS: Sixteen and twenty aroma components were identified in Kastamonu and Chinese garlic types, respectively. Kastamonu garlic type was found to be richer than Chinese garlic types in terms of sulfur-containing compounds. Diallyl disulphide, which is one of these components, was detected at level of 41.87% and 34.95% in the Kastamonu and Chinese garlic types, respectively. Also di-2-propenyl trisulfide was found only in Kastamonu garlic types. Disulfide, methyl 2-propenyl was determined at similar levels in both garlic types. CONCLUSION: The majority of garlic grown in Kastamonu region of Turkey is assessed by medical companies. CONCLUSION: The results of the current study showed that Kastamonu garlic type has important medical properties. Therefore, this garlic can also be used in the medical field, as well as the consumption as food.

Determination of volatile aroma compounds of Ganoderma lucidum by gas chromatography mass spectrometry (HS-GC/MS).

This study was conducted at Horticulture Department of Cukurova University, Adana, Turkey during 2010-2011. Fresh sample of Ganoderma lucidum collected from Mersin province of Turkey was used as material. Volatile aroma compounds were performed by Headspace Gas Chromatography (HS-GC/MS). Alcohols, aldehydes, acids, phenol, L-Alanine, d-Alanine, 3Methyl, 2-Butanamine, 2-Propanamine were determined. 1-Octen-3-ol (Alcohol) and 3-methyl butanal (Aldehyde) were identified as major aroma compounds.

Genetic characterization of pomegranate (Punica granatum L.) genotypes by AFLP markers

The Coruh Valley, located in Northeastern Turkey, is one of the most important centers of diversity in pomegranate in Turkey. In this study, we attempted to characterize 19 promising pomegranate genotypes originating from the Coruh Valley in using fluorescent dye AFLP markers and capillary electrophoresis. Four AFLP primer combinations were used, generating a total of 297 fragments, 213 of which were polymorphic (73.0%). Resolving powers of the AFLP primers ranged from 0.700 to 1.018, with a total of 3.440, while polymorphism information contents ranged from 0.707 to 0.837 with an average of 0.764. UPGMA clustering of the genotypes showed two major groups. Most of the fruit characteristics of the genotypes within the same group were variable. Therefore, the results showed that molecular characterization is necessary to get reliable relationships among pomegranate genotypes and AFLP markers can be used effectively in pomegranate.