Modeling the Impact of Agricultural Mitigation Measures on the Spread of Sharka Disease in Sweet Cherry Orchards.

Sharka is a disease affecting stone fruit trees. It is caused by the Plum pox virus (PPV), with Myzus persicae being one of the most efficient aphid species in transmitting it within and among Prunus orchards. Other agricultural management strategies are also responsible for the spread of disease among trees, such as grafting and pruning. We present a mathematical model of impulsive differential equations to represent the dynamics of Sharka disease in the tree and vector population. We consider three transmission routes: grafting, pruning, and through aphid vectors. Grafting, pruning, and vector control occur as pulses at specific instants. Within the model, human risk perception towards disease influences these agricultural management strategies. Model results show that grafting with infected biological material has a significant impact on the spread of the disease. In addition, detecting infectious symptomatic and asymptomatic trees in the short term is critical to reduce disease spread. Furthermore, vector control to prevent aphid movement between trees is crucial for disease mitigation, as well as implementing awareness campaigns for Sharka disease in agricultural communities that provide a long-term impact on responsible pruning, grafting, and vector control.

Self-Emulsifying Micellization of Crude Extracts from Apple (Malus domestica cv. Anna), Plum (Prunus domestica cv. Satsuma), and Guava (Psidium guajava L.) Fruits.

Micellar microemulsions are thermodynamically stable self-emulsifying systems that have been used to successfully improve the low oral bioavailability of several bioactive phytochemicals, such as antioxidant polyphenols. However, most studies have reported the micellization of single-compounds or purified chemical fractions; thus, the stability, phytochemical-loading efficiency, and bioactivity of complex crude extracts remain largely unexplored. In this study, we evaluated the effects of micellar emulsification of tropical apple (Malus domestica cv. Anna), plum (Prunus domestica cv. Satsuma), and guava (Psidium guajava L.) extracts regarding particle size and stability, polyphenol-loading efficiency, antioxidant capacity, and cytotoxic activity in human and murine cells. Simple food-grade extraction protocols were implemented to obtain apple, plum, and guava extracts. Total polyphenols, flavonoids, and antioxidant activity (DPPH) were determined in the fruit extracts, and their polyphenol profile was further characterized by liquid chromatography (HPLC-DAD). The dried extracts were mixed into a food-grade, self-emulsifying system, and their cytotoxicity in human and murine cell lines was compared. Our research showed that complex fruit matrixes were successfully emulsified into thermodynamically stable polysorbate-based nanometric micelles with uniform size distribution and consistent pH stability, with potential applications in food and biomedical industries.

An Upgraded, Highly Saturated Linkage Map of Japanese Plum (Prunus salicina Lindl.), and Identification of a New Major Locus Controlling the Flavan-3-ol Composition in Fruits.

Japanese plum fruits are rich in phenolic compounds, such as anthocyanins and flavan-3-ols, whose contents vary significantly among cultivars. Catechin (C) and epicatechin (EC) are flavan-3-ol monomers described in the fruits of this species and are associated with bitterness, astringency, antioxidant capacity, and susceptibility to enzymatic mesocarp browning. In this study, we aimed to identify quantitative trait loci (QTL) associated with the content of flavan-3-ol in Japanese plum fruits. We evaluated the content of C and EC in the mesocarp and exocarp of samples from 79 and 64 seedlings of an F1 progeny (<'98-99' × 'Angeleno'>) in the first and second seasons, respectively. We also constructed improved versions of linkage maps from '98-99' and 'Angeleno,' presently called single-nucleotide polymorphisms (SNPs) after mapping the already available GBS reads to Prunus salicina Lindl. cv. 'Sanyueli' v2.0 reference genome. These data allowed for describing a cluster of QTLs in the cultivar, 'Angeleno,' associated with the flavan-3-ol composition of mesocarp and exocarp, which explain up to 100% of the C/EC ratio. Additionally, we developed a C/EC metabolic marker, which was mapped between the markers with the highest log of odds (LOD) scores detected by the QTL analysis. The C/EC locus was located in the LG1, at an interval spanning 0.70 cM at 108.30-108.90 cM. Our results suggest the presence of a novel major gene controlling the preferential synthesis of C or EC in the Japanese plum fruits. This study is a significant advance in understanding the regulation of synthesizing compounds associated with fruit quality, postharvest, and human health promotion.

Silverleaf (Chondrostereum purpureum) Effects on Japanese Plum (Prunus salicina).

Silverleaf is an important fungal trunk disease of fruit crops, such as Japanese plum (Prunus salicina). It is known that infection by Chondrostereum purpureum results in discolored wood, "silvered" foliage, and tree decline. However, effects on fruit yield and quality have not been assessed. Therefore, the objectives of this study were to determine C. purpureum pathogenicity on P. salicina and the effects on physiology, fruit yield, and quality, in Chile, in 2019 and 2020. Wood samples from affected plum trees were collected in the Chilean plum productive area. Fungi were isolated by plating wood sections from the necrosis margin on culture media. Morphological and molecular characteristics of the isolates corresponded to C. purpureum (98%). Representative isolates were inoculated from healthy plum plants and after 65-d incubation, wood necrotic lesions and silver leaves were visible. Fungi were reisolated, fulfilling Koch's postulates. To determine Silverleaf effects, xylem water potential and fruit yield and quality were measured in healthy and Silverleaf-diseased plum trees 'Angeleno'. Water potential was altered in diseased trees, and fruit yield was reduced by 51% (2019) and by 41% (2020) compared to fruit from healthy trees. Moreover, cover-colour, equatorial-diameter, and weight were reduced, and fruit were softer, failing to meet the criteria to be properly commercialized and exported to demanding markets.

Polyphenolic QTOF-ESI MS Characterization and the Antioxidant and Cytotoxic Activities of Prunus domestica Commercial Cultivars from Costa Rica.

There is an increased interest in plum research because of their metabolites' potential bioactivities. In this study, the phenolic profiles of Prunus domestica commercial cultivars (Methley, Pisardii and Satsuma) in Costa Rica were determined by Ultra Performance Liquid Chromatography coupled with High Resolution Mass Spectrometry using a quadrupole-time-of-flight analyzer (UPLC-ESI-QTOF MS) on enriched phenolic extracts obtained through Pressurized Liquid Extraction (PLE) under acidic and neutral extraction conditions. In total, 41 different phenolic compounds were identified in the skin and flesh extracts, comprising 11 flavan-3-ols, 14 flavonoids and 16 hydroxycinnamic acids and derivatives. Neutral extractions for the skins and flesh from all of the cultivars yielded a larger number of compounds, and were particularly rich in the number of procyanidin trimers and tetramers when compared to the acid extractions. The total phenolic content (TPC) and antioxidant potential using the DPPH and ORAC methods exhibited better results for neutral extracts with Satsuma skins and Methley flesh, which showed the best values (685.0 and 801.6 mg GAE/g extract; IC50 = 4.85 and 4.39 µg/mL; and 12.55 and 12.22 mmol TE/g extract, respectively). A Two-Way ANOVA for cytotoxicity towards AGS gastric adenocarcinoma and SW620 colon adenocarcinoma indicated a significant difference (p < 0.05) for PLE conditions, with better results for neutral extractions, with Satsuma skin delivering the best results (IC50 = 60.7 and 46.7 µg/mL respectively) along with Methley flesh (IC50 = 76.3 and 60.9 µg/mL, respectively). In addition, a significant positive correlation was found between TPC and ORAC (r = 0.929, p < 0.05), as well as a significant negative correlation (p < 0.05) between TPC and cytotoxicity towards AGS and SW620 cell lines (r = -0.776, and -0.751, respectively). A particularly high, significant, negative correlation (p < 0.05) was found between the number of procyanidins and cytotoxicity against the AGS (r = -0.868) and SW620 (r = -0.855) cell lines. Finally, the PCA clearly corroborated that neutral extracts are a more homogenous group exhibiting higher antioxidant and cytotoxic results regardless of the part or cultivar; therefore, our findings suggest that PLE extracts under neutral conditions would be of interest for further studies on their potential health benefits.

First Report of Fruit Rot of Sweet Cultivars of Japanese Plum caused by Alternaria alternata, A. arborescens, and A. tenuissima in Chile.

During the last two seasons, an unusual fruit rot was observed in four orchards of sweet Japanese plum (Prunus salicina) cultivars located in the Chilean Central Valley (30°00'S, 70°42'W). The incidence was 5% in Black Majesty, 4% in Red Lyon, and 6% in Sweet Mary cultivars in 2020. Fruits in the field showed a firm, dehydrated, and slightly sunken rot on the blossom end, along with rough and irregular epidermis in the affected area. Internally, the fruit flesh appeared light to dark-brown or olive-green. Symptomatic fruits (n=119) were superficially disinfected (75% ethanol) and, pieces of the pericarp (3 x 3 mm) were removed and placed on potato dextrose agar (PDA). Isolates of Alternaria spp. were obtained and 9 of these were selected for identification. Colonies were dark olive to gray-brown with white margins, small, catenulate and muriform conidia, produced in single or branched conidiophores. Isolates produced brown to golden-brown, ovoid, ellipsoidal to obclavate conidia with dimensions of 19.7 to 26.7 × 10.0 to 11.9 µm with two to four transverse and zero to three longitudinal septa on 0.05× PDA (Pryor and Michailides 2002) after 7 d at 20°C under 10/14 h light/dark cycles. A molecular analysis was performed by sequencing the nuclear genes RNA polymerase II subunit (RPB2), plasma membrane ATPase (ATP), and the calmodulin (Cal) gene using primers RPB2-5F2/fRPB2-7cR, ATPDF1/ATPDR1, and CALDF1/CALDR1, respectively (Lawrence et al. 2013; Woudenberg et al. 2013). A BLAST search revealed the presence of Alternaria spp. with a 99% to 100% identity with the reference sequences of A. alternata (JQ905182, JQ671874, JQ646208), A. arborescens (JQ646487, JQ671880, JQ646214), and A. tenuissima (JQ811961, JQ811989, JQ646209). Maximum parsimony phylogenetic analysis confirmed the identifications. Sequences were deposited in GenBank as numbers MW514249 to MW514257, MT872324 to MT872332, and MT872314 to MT872322 for RPB2, ATP, and Cal sequences, respectively. All these Alternaria isolates were deposited in the Colección Chilena de Recursos Genéticos Microbianos - INIA, Chillán Chile (RGM3069 to RGM3077). Pathogenicity of A. alternata (n=4), A. arborescens (n=3) and, A. tenuissima (n=2) was tested in Red Lyon plum fruits. Plums were disinfected in 1% sodium hypochlorite for 2 min, rinsed in sterile distilled water for 1 min and dried on absorbent towels in a laminar flow hood. Then, the plums were wounded on the blossom end with a sterile needle (1 x 0.5 mm), inoculated with 10 µl of a conidial suspension (106 conidia/ml), wrapped with Parafilm and maintained in a humid chamber (>95% relative humidity). An equal number of fruits wounded and inoculated with sterile water were used as a control. After 7 days at 20°C, all inoculated fruits developed a dark-brown firm rot with lesion lengths of 24.4 (±3.0) mm, 19.6 (±0.7) mm, and 16.8 (±2.4) mm for A. alternata, A. arborescens and A. tenuissima, respectively. A. alternata was the most aggressive species (P < 0.001). Control fruits remained asymptomatic. Koch's postulates were fulfilled after the re-isolating the causal agent from the border of the lesions. Leaf spots and fruit rots caused by Alternaria isolates have been reported in stone fruits, including plums (Kim et al. 2005; Long et al. 2021; Moosa et al., 2019; Yang et al. 2014). To our knowledge, this is the first report of A. alternata, A. arborescens, and A. tenuissima associated with fruit rot in sweet Japanese plum cultivars in the field, in Chile.

Effect of treatment with sunflower oil and in combination with pomegranate and plum sauce on the chemical, microbiological and sensory properties of marinated Carp fillets (Cyprinus carpio) / Efeito do tratamento com óleo de girassol e em combinação com molho de romã e ameixa nas propiedades químicas, microbiológicas e sensoriais de filetes de carpa marinados (Cyprinus carpio)

The effect of sunflower oil (SO), sunflower oil-pomegranate sauce (PS1) and sunflower oil-plum sauce (PS2) on shelf life of marinated carp fillets was investigated in terms of sensory evaluation, chemical (peroxide value (POV), thiobarbituric acid (TBA), free fatty acids (FFA), total volatile basic nitrogen (TVB-N) and pH), and microbiological properties (Total viable count (TVC), Escherichia coli, Total coliform, Staphylococci/Micrococci, Salmonella spp.) during 30 days of storage. Proximate composition, which includes total crude protein%, fat%, moisture% and ash% of fresh carp fillets before marination process was analyzed and found 18.69±0.86, 4.08±0.19, 74.33±0.63 and 2.17±0.45, respectively. Sensory evaluation analysis showed that total appearance, odor-taste and texture scores decreased during storage. TVB-N significantly increased in all group after 1 month of storage (P 0.05). At the end of storage, the lowest TBA value (1.38 mg MA/kg carp fillet) was determined in the PS1 group. During storage, the highest POV value was observed in the control group (8.49 mEq O2/kg) compared to other groups. Initial TVC of 1.5 log CFU/g, 1.7 log CFU/g and 1.8 log CFU/g increased to 3.7 log CFU/g, 2.9 log CFU/g and 3.2 log CFU/g in SO, PS1 and PS2 groups, respectively. Results showed that the shelf life of marinated carp fillets treated with sunflower oil and in combination with pomegranate and plum sauce was more than 1 month.(AU)

O efeito do óleo de girassol (SO), molho de óleo de girassol-romã (PS1) e molho de óleo de girassol-ameixa (PS2) na vida útil de filetes de carpa marinados foi investigado em termos sensorial, químico (valor de peróxido (PV), ácido tiobarbitúrico) (TBA), ácidos graxos livres (AGL), nitrogênio básico volátil total (TVB-N) e pH) e propriedades microbiológicas (contagem viável total (TVC), Escherichia coli, coliforme total, estafilococos/Micrococos, Salmonella spp.). Durante 30 dias de armazenamento. A composição aproximada, que inclui % de proteína bruta total, % de gordura, % de umidade e % de cinzas de filés de carpa frescos antes do processo de marinação foi analisada e encontrada 18,69 ± 0,86, 4,08 ± 0,19, 74,33 ± 0,63 e 2,17 ± 0,45, respectivamente. A análise sensorial mostrou que os escores totais de aparência, odor e sabor diminuíram durante o armazenamento. TVB-N aumentou significativamente (P 0,05) em todos os grupos após 1 mês de armazenamento. No final do armazenamento, o menor valor de TBA (1,38 mg MA / kg de filé de carpa) foi determinado no grupo PS1. Durante o armazenamento, o maior valor de POV foi observado no grupo controle (8,49 mEq O2/kg) em comparação com outros grupos. O TVC inicial de 1,5 log CFU/g, 1,7 log UFC/g, 1,8 log CFU/g aumentou para 3,7 log CFU/g, 2,9 log CFU/g, 3,2 log CFU/g nos grupos SO, PS1 e PS2, respectivamente. Os resultados mostraram que a vida útil dos filés de carpa marinados tratados com óleo de girassol e em combinação com molho de romã e ameixa era superior a um mês.(AU)

Qualidade físico-química de ameixas ‘Letícia’ produzidas sobre porta-enxertos clonais e em plantas autoenraizadas, no meio-oeste de Santa Catarina / Physicochemical quality of ‘Letícia’ plumsproduced on clonal rootstocks and own-rooted trees, in the Midwest of Santa Catarina state, Brazil

Objetivou-se com esse trabalho avaliar os efeitos de porta-enxertos clonais e de plantasautoenraizadas na qualidade físico-química de ameixas ‘Letícia’, no meio-oeste de Santa Catarina-SC. Foram avaliados frutos da cv. Letícia produzidos nos ciclos produtivos de 2018, 2019 e 2020, provenientes de pomar experimental daEstação Experimental daEpagri em Videira-SC. As plantas foram enxertadas sobre 19 diferentes acessos (cultivares, híbridos interespecíficos e seleções) utilizados como porta-enxertos clonais, além da cv. Letícia autoenraizada (sem porta-enxerto). Avaliou-se acidez total, teor de sólidos solúveis, ratio, pH, firmeza de polpa e cor da epiderme dos frutos. Os valores de pH oscilaram de 2,98 para o porta-enxerto ‘Barrier’ (2018) a 3,68 para plantas sobre ‘Santa Rosa’ (2020). ‘Santa Rosa’ como porta-enxerto destacou-se pelo maior ratio(19,46) na safra 2020, seguido pelo ‘Flordaguard em 2018 (14,92). Os maiores valores de firmeza foram obtidos para GxN9 em 2018 e Nemared em 2020, e os menores por ‘Santa Rosa’ em 2019 e 2020. A resposta dos porta-enxertos não serepetiu ao longo das safras avaliadas, demonstrando que o fator safra teve maior influência sobre a qualidade dos frutos, com exceção de ‘Santa Rosa’. Os porta-enxertos Rigitano, Clone 15 (Prunus mume), Nemared e I-67-52-4 (P. persica) apresentaram incompatibilidade com a cultivar-copa Letícia. Plantas autoenraizadas de ‘Letícia’ produzem frutos com características físico-químicas semelhantes aos produzidos em plantas enxertadas nos porta-enxertos testados, tornando-se uma opção tecnicamente interessante de cultivo sem o uso de porta-enxertos.

The objective of this work was to evaluate the effects of clonal rootstocks and self-rooted plants on the physicochemical quality of'Letícia' plumsin themid-west of Santa Catarina. The evaluated fruits were from the 2018 to 2020 crops, from an experimental orchard of cv. Letícia, whose plants were grafted onto 19 different accessions (cultivars, interspecific hybrids,and selections) used as clonal rootstocks, in addition to own-rooted Letícia, at the Epagri Experimental Station in Videira-SC. Here, we evaluated total acidity, soluble solids content, ratio, pH, flesh firmness, and epidermis color of the fruits.The pH values ranged from 2.98 for the 'Barrier'rootstock (2018) to 3.68 for plants on 'Santa Rosa' (2020). ‘Santa Rosa’ as rootstock stood out for the highest ratio (19.46) in the 2020 harvest, followed by ‘Flordaguard in 2018 (14.92). The highest firmness values were obtained forGxN9 in 2018 and Nemared in 2020, and the lowest for 'Santa Rosa' in 2019 and 2020. The performance of the rootstocks was not repeated throughout the evaluated crops, demonstrating that the yield factor had a greater influence on fruit quality, except for'Santa Rosa'. The fruits of plants grafted onto 'Santa Rosa' showed more advanced maturation at harvest. 'Letícia' plums produced on 'Santa Rosa' presented precocity of maturation in relation to the other tested clonal rootstocks. The rootstocks Rigitano,Clone 15 (Prunus mume), Nemared and I-67-52-4 (P. persica) showed incompatibility with the 'Letícia' canopy cultivar. Own-rooted 'Leticia' plants produce fruits with physicochemical characteristics similar to those produced on plants grafted on the testedrootstocks, indicating that the own-root cultivation is a technically interesting option for this cultivar.

Qualidade físico-química de ameixas ‘Letícia produzidas sobre porta-enxertos clonais e em plantas autoenraizadas, no meio-oeste de Santa Catarina / Physicochemical quality of ‘Letícia plumsproduced on clonal rootstocks and own-rooted trees, in the Midwest of Santa Catarina state, Brazil

Objetivou-se com esse trabalho avaliar os efeitos de porta-enxertos clonais e de plantasautoenraizadas na qualidade físico-química de ameixas ‘Letícia, no meio-oeste de Santa Catarina-SC. Foram avaliados frutos da cv. Letícia produzidos nos ciclos produtivos de 2018, 2019 e 2020, provenientes de pomar experimental daEstação Experimental daEpagri em Videira-SC. As plantas foram enxertadas sobre 19 diferentes acessos (cultivares, híbridos interespecíficos e seleções) utilizados como porta-enxertos clonais, além da cv. Letícia autoenraizada (sem porta-enxerto). Avaliou-se acidez total, teor de sólidos solúveis, ratio, pH, firmeza de polpa e cor da epiderme dos frutos. Os valores de pH oscilaram de 2,98 para o porta-enxerto ‘Barrier (2018) a 3,68 para plantas sobre ‘Santa Rosa (2020). ‘Santa Rosa como porta-enxerto destacou-se pelo maior ratio(19,46) na safra 2020, seguido pelo ‘Flordaguard em 2018 (14,92). Os maiores valores de firmeza foram obtidos para GxN9 em 2018 e Nemared em 2020, e os menores por ‘Santa Rosa em 2019 e 2020. A resposta dos porta-enxertos não serepetiu ao longo das safras avaliadas, demonstrando que o fator safra teve maior influência sobre a qualidade dos frutos, com exceção de ‘Santa Rosa. Os porta-enxertos Rigitano, Clone 15 (Prunus mume), Nemared e I-67-52-4 (P. persica) apresentaram incompatibilidade com a cultivar-copa Letícia. Plantas autoenraizadas de ‘Letícia produzem frutos com características físico-químicas semelhantes aos produzidos em plantas enxertadas nos porta-enxertos testados, tornando-se uma opção tecnicamente interessante de cultivo sem o uso de porta-enxertos.(AU)

The objective of this work was to evaluate the effects of clonal rootstocks and self-rooted plants on the physicochemical quality of'Letícia' plumsin themid-west of Santa Catarina. The evaluated fruits were from the 2018 to 2020 crops, from an experimental orchard of cv. Letícia, whose plants were grafted onto 19 different accessions (cultivars, interspecific hybrids,and selections) used as clonal rootstocks, in addition to own-rooted Letícia, at the Epagri Experimental Station in Videira-SC. Here, we evaluated total acidity, soluble solids content, ratio, pH, flesh firmness, and epidermis color of the fruits.The pH values ranged from 2.98 for the 'Barrier'rootstock (2018) to 3.68 for plants on 'Santa Rosa' (2020). ‘Santa Rosa as rootstock stood out for the highest ratio (19.46) in the 2020 harvest, followed by ‘Flordaguard in 2018 (14.92). The highest firmness values were obtained forGxN9 in 2018 and Nemared in 2020, and the lowest for 'Santa Rosa' in 2019 and 2020. The performance of the rootstocks was not repeated throughout the evaluated crops, demonstrating that the yield factor had a greater influence on fruit quality, except for'Santa Rosa'. The fruits of plants grafted onto 'Santa Rosa' showed more advanced maturation at harvest. 'Letícia' plums produced on 'Santa Rosa' presented precocity of maturation in relation to the other tested clonal rootstocks. The rootstocks Rigitano,Clone 15 (Prunus mume), Nemared and I-67-52-4 (P. persica) showed incompatibility with the 'Letícia' canopy cultivar. Own-rooted 'Leticia' plants produce fruits with physicochemical characteristics similar to those produced on plants grafted on the testedrootstocks, indicating that the own-root cultivation is a technically interesting option for this cultivar.(AU)

Effect of treatment with sunflower oil and in combination with pomegranate and plum sauce on the chemical, microbiological and sensory properties of marinated Carp fillets (Cyprinus carpio) / Efeito do tratamento com óleo de girassol e em combinação com molho de romã e ameixa nas propiedades químicas, microbiológicas e sensoriais de filetes de carpa marinados (Cyprinus carpio)

ABSTRACT: The effect of sunflower oil (SO), sunflower oil-pomegranate sauce (PS1) and sunflower oil-plum sauce (PS2) on shelf life of marinated carp fillets was investigated in terms of sensory evaluation, chemical (peroxide value (POV), thiobarbituric acid (TBA), free fatty acids (FFA), total volatile basic nitrogen (TVB-N) and pH), and microbiological properties (Total viable count (TVC), Escherichia coli, Total coliform, Staphylococci/Micrococci, Salmonella spp.) during 30 days of storage. Proximate composition, which includes total crude protein%, fat%, moisture% and ash% of fresh carp fillets before marination process was analyzed and found 18.69±0.86, 4.08±0.19, 74.33±0.63 and 2.17±0.45, respectively. Sensory evaluation analysis showed that total appearance, odor-taste and texture scores decreased during storage. TVB-N significantly increased in all group after 1 month of storage (P<0.05). At the end of storage, the lowest TBA value (1.38 mg MA/kg carp fillet) was determined in the PS1 group. During storage, the highest POV value was observed in the control group (8.49 mEq O2/kg) compared to other groups. Initial TVC of 1.5 log CFU/g, 1.7 log CFU/g and 1.8 log CFU/g increased to 3.7 log CFU/g, 2.9 log CFU/g and 3.2 log CFU/g in SO, PS1 and PS2 groups, respectively. Results showed that the shelf life of marinated carp fillets treated with sunflower oil and in combination with pomegranate and plum sauce was more than 1 month.

RESUMO: O efeito do óleo de girassol (SO), molho de óleo de girassol-romã (PS1) e molho de óleo de girassol-ameixa (PS2) na vida útil de filetes de carpa marinados foi investigado em termos sensorial, químico (valor de peróxido (PV), ácido tiobarbitúrico) (TBA), ácidos graxos livres (AGL), nitrogênio básico volátil total (TVB-N) e pH) e propriedades microbiológicas (contagem viável total (TVC), Escherichia coli, coliforme total, estafilococos/Micrococos, Salmonella spp.). Durante 30 dias de armazenamento. A composição aproximada, que inclui % de proteína bruta total, % de gordura, % de umidade e % de cinzas de filés de carpa frescos antes do processo de marinação foi analisada e encontrada 18,69 ± 0,86, 4,08 ± 0,19, 74,33 ± 0,63 e 2,17 ± 0,45, respectivamente. A análise sensorial mostrou que os escores totais de aparência, odor e sabor diminuíram durante o armazenamento. TVB-N aumentou significativamente (P <0,05) em todos os grupos após 1 mês de armazenamento. No final do armazenamento, o menor valor de TBA (1,38 mg MA / kg de filé de carpa) foi determinado no grupo PS1. Durante o armazenamento, o maior valor de POV foi observado no grupo controle (8,49 mEq O2/kg) em comparação com outros grupos. O TVC inicial de 1,5 log CFU/g, 1,7 log UFC/g, 1,8 log CFU/g aumentou para 3,7 log CFU/g, 2,9 log CFU/g, 3,2 log CFU/g nos grupos SO, PS1 e PS2, respectivamente. Os resultados mostraram que a vida útil dos filés de carpa marinados tratados com óleo de girassol e em combinação com molho de romã e ameixa era superior a um mês.

The purification of pectin from commercial fruit flours results in a jaboticaba fraction that inhibits galectin-3 and colon cancer cell growth.

Fruits are a prime source of nutrients, bioactive compounds, and dietary fibers. Some products available on the Brazilian market use fruit by-products and claim to have useful effects on human health due to their dietary fiber content. The study aimed to extract and purify the total (28-47 w/w yield) and soluble dietary fiber (4-7 w/w yield) from jaboticaba, papaya, and plum commercial flours sold in Brazil and to study the in vitro biological effects of the fractions. The purified water-soluble fractions consisted mainly of pectin-derived oligosaccharides (5-15 KDa molecular weight) with a negligible content of polyphenols, protein, ashes, and starch. Jaboticaba sample was 95% galacturonic acid while plum and papaya samples were 40% galacturonic acid and 40% galactose (mol%), approximately. The samples were tested for recombinant human galectin-3 inhibition and changes in the cell viability of human colorectal cancer cells. Only the jaboticaba sample inhibited galectin-3 and decreased HCT116 cell viability after 48 h of treatment (p ≤ 0.01) while the plum sample decreased the cell viability after 24 h treatment (p ≤ 0.05). The results obtained in this study demonstrate the relationship between the structure of the soluble fibers extracted from jaboticaba flour and the possible beneficial effects of their consumption.

Silencing of one copy of the translation initiation factor eIFiso4G in Japanese plum (Prunus salicina) impacts susceptibility to Plum pox virus (PPV) and small RNA production.

BACKGROUND: In plants, host factors encoded by susceptibility (S) genes are indispensable for viral infection. Resistance is achieved through the impairment or the absence of those susceptibility factors. Many S genes have been cloned from model and crop species and a majority of them are coding for members of the eukaryotic translation initiation complex, mainly eIF4E, eIF4G and their isoforms. The aim of this study was to investigate the role of those translation initiation factors in susceptibility of stone fruit species to sharka, a viral disease due to Plum pox virus (PPV). RESULTS: For this purpose, hairpin-inducing silencing constructs based on Prunus persica orthologs were used to generate Prunus salicina (Japanese plum) 4E and 4G silenced plants by Agrobacterium tumefaciens-mediated transformation and challenged with PPV. While down-regulated eIFiso4E transgenic Japanese plums were not regenerated in our conditions, eIFiso4G11-, but not the eIFiso4G10-, silenced plants displayed durable and stable resistance to PPV. We also investigated the alteration of the si- and mi-RNA profiles in transgenic and wild-type Japanese plums upon PPV infection and confirmed that the newly generated small interfering (si) RNAs, which are derived from the engineered inverted repeat construct, are the major contributor of resistance to sharka. CONCLUSIONS: Our results indicate that S gene function of the translation initiation complex isoform is conserved in Prunus species. We discuss the possibilities of using RNAi silencing or loss-of-function mutations of the different isoforms of proteins involved in this complex to breed for resistance to sharka in fruit trees.

Plum (Prunus salicina) peel and pulp microparticles as natural antioxidant additives in breast chicken patties.

Fiber microparticles (MPCs) separately obtained from peel and pulp of Japanese plum residues contained co-extracted ß-carotene, lutein, and α- and γ-tocopherols, as well as polyphenols (cyanidins, quercetin derivatives, pentameric proanthocyanidins). Peel and pulp MPCs were then separately evaluated as natural antioxidant additives (2.0% w/w level) in raw breast chicken patty, susceptible to oxidation. Their effect on technological properties was also analyzed. MPCs reduced in 50% the formation of thiobarbituric acid reactive substances (TBARS) in raw patties during 10-days storage at 4.0°C. Ferric reducing power (FRAP) was 77-157% higher in MPCs-added patties, especially with peel MPCs, being then attributed to the antioxidants supplied by these MPCs. It can be also associated to the highest α- and γ-tocopherol levels found in the peel MPCs-added patties, which remained high after cooking as well. Also, higher pectin and low lignin contents of pulp MPCs determined greater hydration, stabilized the cyanidins and, hence, the red color transferred to raw patties, and increased springiness of cooked patties. Plum peel and pulp MPCs are efficient additives for chicken meat products.

Polyphenolic Characterization and Antioxidant Activity of Malus domestica and Prunus domestica Cultivars from Costa Rica.

The phenolic composition of skin and flesh from Malus domestica apples (Anna cultivar) and Prunus domestica plums (satsuma cultivar) commercial cultivars in Costa Rica, was studied using Ultra Performance Liquid Chromatography coupled with High Resolution Mass Spectrometry (UPLC-DAD-ESI-MS) on enriched-phenolic extracts, with particular emphasis in proanthocyanidin and flavonoids characterization. A total of 52 compounds were identified, including 21 proanthocyanidins ([(+)-catechin and (-)-epicatechin]) flavan-3-ols monomers, five procyanidin B-type dimers and two procyanidin A-type dimers, five procyanidin B-type trimers and two procyanidin A-type trimers, as well as one procyanidin B-type tetramer, two procyanidin B-type pentamers, and two flavan-3-ol gallates); 15 flavonoids (kaempferol, quercetin and naringenin derivatives); nine phenolic acids (protochatechuic, caffeoylquinic, and hydroxycinnamic acid derivatives); five hydroxychalcones (phloretin and 3-hydroxyphloretin derivatives); and two isoprenoid glycosides (vomifoliol derivatives). These findings constitute the first report of such a high number and diversity of compounds in skins of one single plum cultivar and of the presence of proanthocyanidin pentamers in apple skins. Also, it is the first time that such a large number of glycosylated flavonoids and proanthocyanidins are reported in skins and flesh of a single plum cultivar. In addition, total phenolic content (TPC) was measured with high values observed for all samples, especially for fruits skins with a TPC of 619.6 and 640.3 mg gallic acid equivalents/g extract respectively for apple and plum. Antioxidant potential using 2,2-diphenyl-1-picrylhidrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) methods were evaluated, with results showing also high values for all samples, especially again for fruit skins with IC50 of 4.54 and 5.19 µg/mL (DPPH) and 16.8 and 14.6 mmol TE/g (ORAC) respectively for apple and plum, indicating the potential value of these extracts. Significant negative correlation was found for both apple and plum samples between TPC and DPPH antioxidant values, especially for plum fruits (R = -0.981, p < 0.05) as well as significant positive correlation between TPC and ORAC, also especially for plum fruits (R = 0.993, p < 0.05) and between both, DPPH and ORAC antioxidant methods (R = 0.994, p < 0.05).

A fast and efficient protocol for small RNA extraction in Japanese plum and other Prunus species

Background: Small ribonucleic acids represent an important repertoire of mobile molecules that exert key roles in several cell processes including antiviral defense. Small RNA based repertoire includes both small interfering RNA (siRNA) and microRNA (miRNA) molecules. In the Prunus genus, sharka disease, caused by the Plum pox virus (PPV), first occurred on European plum (Prunus domestica) and then spread over among all species in this genus and thus classified as quarantine pathogen. Next-generation sequencing (NGS) was used for the study of siRNA/miRNA molecules; however, NGS relies on adequate extraction protocols. Currently, knowledge of PPV-Prunus interactions in terms of siRNA populations and miRNA species is still scarce, and siRNA/miRNA extraction protocols are limited to species such as peach, almond, and sweet cherry. Results: We describe a reliable procedure for siRNA/miRNA purification from Prunus salicina trees, in which previously used protocols did not allow adequate purification. The procedure was based on a combination of commercially available RNA purification kits and specific steps that yielded high quality purifications. The resulting molecules were adequate for library construction and NGS, leading to the development of a pipeline for analysis of both siRNAs and miRNAs in the PPV­P. salicina interactions. Results showed that PPV infection led to altered siRNA profiles in Japanese plum as characterized by decreased 24-nt and increased 21- and 22-nt siRNAs. Infections showed miR164 and miR160 generation and increased miR166, miR171, miR168, miR319, miR157, and miR159. Conclusion: We propose this protocol as a reliable and reproducible small RNA isolation procedure for P. salicina and other Prunus species.

Genotyping by Sequencing for SNP-Based Linkage Analysis and Identification of QTLs Linked to Fruit Quality Traits in Japanese Plum (Prunus salicina Lindl.).

Marker-assisted selection (MAS) in stone fruit (Prunus species) breeding is currently difficult to achieve due to the polygenic nature of the most relevant agronomic traits linked to fruit quality. Genotyping by sequencing (GBS), however, provides a large quantity of useful data suitable for fine mapping using Single Nucleotide Polymorphisms (SNPs) from a reference genome. In this study, GBS was used to genotype 272 seedlings of three F1 Japanese plum (Prunus salicina Lindl) progenies derived from crossing "98-99" (as a common female parent) with "Angeleno," "September King," and "September Queen" as male parents. Raw sequences were aligned to the Peach genome v1, and 42,909 filtered SNPs were obtained after sequence alignment. In addition, 153 seedlings from the "98-99" × "Angeleno" cross were used to develop a genetic map for each parent. A total of 981 SNPs were mapped (479 for "98-99" and 502 for "Angeleno"), covering a genetic distance of 688.8 and 647.03 cM, respectively. Fifty five seedlings from this progeny were phenotyped for different fruit quality traits including ripening time, fruit weight, fruit shape, chlorophyll index, skin color, flesh color, over color, firmness, and soluble solids content in the years 2015 and 2016. Linkage-based QTL analysis allowed the identification of genomic regions significantly associated with ripening time (LG4 of both parents and both phenotyping years), fruit skin color (LG3 and LG4 of both parents and both years), chlorophyll degradation index (LG3 of both parents in 2015) and fruit weight (LG7 of both parents in 2016). These results represent a promising situation for GBS in the identification of SNP variants associated to fruit quality traits, potentially applicable in breeding programs through MAS, in a highly heterozygous crop species such as Japanese plum.

De novo transcriptome assembly of 'Angeleno' and 'Lamoon' Japanese plum cultivars (Prunus salicina).

Japanese plum (Prunus salicina L.) is a fruit tree of the Rosaceae family, which is an economically important stone fruit around the world. Currently, Japanese plum breeding programs combine traditional breeding and plant physiology strategies with genetic and genomic analysis. In order to understand the flavonoid pathway regulation and to develop molecular markers associated to the fuit skin color (EST-SSRs), we performed a next generation sequencing based on Illumina Hiseq2000 platform. A total of 22.4 GB and 21 GB raw data were obtained from 'Lamoon' and 'Angeleno' respectively, corresponding to 85,404,726 raw reads to 'Lamoon' and 79,781,666 to 'Angeleno'. A total of 139,775,975 reads were filtered after removing low-quality reads and trimming the adapter sequences. De novo transcriptome assembly was performed using CLC Genome Workbench software and a total of 54,584 unique contigs were generated, with an N50 of 1343 base pair (bp) and a mean length of 829 bp. This work contributed with a specific Japanese plum skin transcriptome, providing two libraries of contrasting fruit skin color phenotype (yellow and red) and increasing substantially the GB of raw data available until now for this specie.

Stabilization by multipoint covalent attachment of a biocatalyst with polygalacturonase activity used for juice clarification.

Derivatized-agarose supports are suitable for enzyme immobilization by different methods, taking advantage of different physical, chemical and biological conditions of the protein and the support. In this study, agarose particles were modified with MANAE, PEI and glyoxyl groups and evaluated to stabilize polygalacturonase from Streptomyces halstedii ATCC 10897. A new immobilized biocatalyst was developed using glyoxyl-agarose as support; it exhibited high performance in degrading polygalacturonic acid and releasing oligogalacturonides. Maximal enzyme activity was detected at 5h of reaction using 0.05g/mL of immobilized biocatalyst, which released 3mg/mL of reducing sugars and allowed the highest product yield conversion and increased stability. These results are very favorable for pectin degradation with reusability up to 18 successive reactions (90h) and application in juice clarification. Plum (4.7°Bx) and grape (10.6°Bx) juices were successfully clarified, increasing reducing sugars content and markedly decreasing turbidity and viscosity.

Correlación del contenido de fenoles y antocianinas con la capacidad antioxidante Syzygium cumini (L. ) Skeels, (jambolan) / Phenol and anthocyanin content and correlation with the antioxidant capacity of Syzygium cumini (L.) Skeels, (jambolan)

Introducción: la especie Syzygium cumini (L.) Skeels, (jambolam), sin. Eugenia jambolana L, es una planta rica en metabolitos secundarios con un elevado potencial biológico, medicinal, entre otros y que pueden estar asociados con las propiedades terapéuticas reportadas para S. cumini. Objetivo: comparar capacidad antioxidante de extractos del fruto, pulpa y semilla de S. cumini, con respecto al contenido de polifenoles y antocianinas. Métodos: fruto, pulpa y semillas S. cumini deshidratados y macerados se sometieron a extracción sólido-líquido; el contenido de fenoles se evaluó por el método de Folin-Ciocalteu, el de antocianinas por el método de diferencial de pH y el potencial antioxidante in vitro se evaluó con base en los métodos del radical estable 1,1-difenil­2-picrilhidracil, el radical catión 2,2´-azino-bis(3-etilbenzotiazolin-6-sulphonic ácido) y poder antioxidante de reducción del ion férrico. Resultados: el contenido de fenoles obtenidos tanto en los extractos metanólicos como etanólicos fue muy similar en fruto y pulpa, mientras que en las semillas se observó un incremento hasta 5 veces con respecto a la pulpa, presentó correlación con el efecto de protección antioxidante entre los extractos evaluados. Conclusiones: la elevada presencia de fenoles determinada en todos los extractos afectó de manera positiva la capacidad protectora contra radicales libres, y los frutos de S. cumini, podrían ser considerados como una fuente potencial de compuestos biaoctivos con aplicaciones terapéuticas(AU)

Introduction: Syzygium cumini (L.) Skeels, (jambolam), sin. Eugenia jambolana L, is considered as a source of secondary metabolites with a biological and medicinal potential and these can be associated with its therapeutics properties. Objetive: to correlated the content of polyphenols and anthocyanins with antioxidant capacity in ethanolic and methanolic extracts of S. cumini. Methods: dehydrated and powered fruits, pulp and seeds of S. cumini was subjected to solid-liquid extraction. Total phenolis content was determined by Folin-Cicoulteau method, anthocyanin content was determined by the pH differential method and the potential antioxidant was evaluated using 1,1-difenil­2-picrilhidracil radical stable, 2,2´-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical cation and ferric reducing/antioxidant power tests. Results: all extracts showed an important correlation between total phenolis content and antioxidant capacity values, however, in the seed total phenolis content was almost 5 times higher than that obtained in fruit and pulp. Conclusions: all extracts exhibited a considerable antioxidant potential against free radical, and S. cumini could be used as an alternative source of natural bioactive compounds with application in therapeutic research area(AU)

Syzygium cumini (L.) skeels: a prominent source of bioactive molecules against cardiometabolic diseases.

Syzygium cumini (Myrtaceae) is a worldwide medicinal plant traditionally used in herbal medicines due to its vaunted properties against cardiometabolic disorders, which include: antihyperglycemic, hypolipemiant, antiinflammatory, cardioprotective, and antioxidant activities. These properties have been attributed to the presence of bioactive compounds such as phenols, flavonoids, and tannins in different parts of the plant, albeit the knowledge on their mechanisms of action is scarce. This mini-review highlights the cardiometabolic properties of S. cumini by correlating its already identified phytochemicals with their described mechanisms of action. Data herein compiled show that some compounds target multiple metabolic pathways; thereby, becoming potential pharmacological tools. Moreover, the lack of clinical trials on S. cumini usage makes it a fruitful field of interest for both scientific community and pharmaceutical industry.