Metabolomics approach for phenolic compounds profiling of soursop (Annona muricata L.) fruit during postharvest storage.

INTRODUCTION: Soursop (Annona muricata L.) is a crop with medicinal properties and numerous bioactive compounds. Ripening is a complex process that regulates fruit quality and changes in metabolite content, such as flavonoids, polyphenols, and organic acids. OBJECTIVES: This study aimed to analyze the phenolic profiling of soursop fruit ripening. METHODS: The metabolic changes in different days of storage of soursop fruits were investigated using a semi-metabolomic approach based on ultra-performance liquid chromatography coupled to electrospray ionization quadrupole-time of flight mass spectrometry (UPLC-ESI-QTOF-MS). Further, multivariate analysis such as supervised partial least squares discriminant analysis (PLS-DA) was conducted to identify differential metabolites. RESULTS: A total of 68 metabolites were identified in soursop fruit during postharvest storage. A higher number of metabolites were identified in the Day zero (D0) compared to the Day one (D1), Day three (D3), and Day five (D5), belonging to flavonoids, other polyphenols, phenolic acids, and organic acids. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the pathways of flavone and flavonol biosynthesis, flavonoid biosynthesis, and biosynthesis of secondary metabolites were mostly enriched. Additionally, we included all the compounds and their postharvest storage in the public Phenolics profile database. CONCLUSIONS: Here, we show that the stage of ripening has a significant effect on the phenolic content, highlighting the point of cut (D0) and the onset of senescence (D5). The findings of this study provide new insights into the soursop fruit quality and may contribute to the identification of metabolic markers for its storage.

Bacillus mojavensis enhances the antioxidant defense mechanism of soursop (Annona muricata L.) fruits during postharvest storage.

Rapid softening of soursop (Annona muricata L.) fruit results in postharvest losses. Bacillus genus is one of the most studied antagonistic biological control agents against postharvest diseases. Nevertheless, information about how this bacterium acts on the fruits is still not understood. The objective of this study aims to gain an insight into the effect of Bacillus mojavensis on the activity and gene expression of antioxidant defense enzymes in soursop fruits during postharvest storage. Our findings indicate different responses in the fruits inoculated with B. mojavensis at biochemical and molecular levels. On day one, fruits inoculated with B. mojavensis presented a mean value of 79.09 GAE/100 gFW in total phenols, and higher superoxide dismutase (SOD) and catalase (CAT) activities (1.35 and 1.78-fold higher, respectively). On the other hand, on the third day of storage, the ferric reducing/antioxidant power (FRAP) reached its highest level, including an increase in the expression of SOD, and PPO genes by 18.7-fold and 4.5-fold in fruits inoculated with B. mojavensis. Finally, on the fifth day of storage, soursop fruits inoculated with B. mojavensis had the highest mean values for 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH·), 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonate (ABTS· +), with values of 194.68 EAA/100 gFW, and 172.33 EAA/100 gFW, respectively. Indeed, higher polyphenol oxidase (PPO), and peroxidase (POD) activities (2.17-fold and 1.27-fold higher, respectively) were recorded compared to the control fruits. We show that depending on the stage of ripening, the antagonist bacteria B. mojavensis enhanced the antioxidant capacity, enzymatic activity, and gene expression of soursop fruits.

Salpianthus macrodontus Extracts, a Novel Source of Phenolic Compounds with Antibacterial Activity against Potentially Pathogenic Bacteria Isolated from White Shrimp.

This study aimed to evaluate the antibacterial activity in vitro of Salpianthus macrodontus and Azadirachta indica extracts against potentially pathogenic bacteria for Pacific white shrimp. Furthermore, the extracts with higher inhibitory activity were analyzed to identify compounds responsible for bacterial inhibition and evaluate their effect on motility and biofilm formation. S. macrodontus and A. indica extracts were prepared using methanol, acetone, and hexane by ultrasound. The minimum inhibitory concentration (MIC) of the extracts was determined against Vibrio parahaemolyticus, V. harveyi, Photobacterium damselae and P. leiognathi. The polyphenol profile of those extracts showing the highest bacterial inhibition were determined. Besides, the bacterial swimming and swarming motility and biofilm formation were determined. The highest inhibitory activity against the four pathogens was found with the acetonic extract of S. macrodontus leaf (MIC of 50 mg/mL for Vibrio spp. and 25 mg/mL for Photobacterium spp.) and the methanol extract of S. macrodontus flower (MIC of 50 mg/mL for all pathogens tested). Both extracts affected the swarming and swimming motility and the biofilm formation of the tested bacteria. The main phenolic compounds related to Vibrio bacteria inhibition were naringin, vanillic acid, and rosmarinic acid, whilst hesperidin, kaempferol pentosyl-rutinoside, and rhamnetin were related to Photobacterium bacteria inhibition.

Transcriptome Analysis of Soursop (Annona muricata L.) Fruit under Postharvest Storage Identifies Genes Families Involved in Ripening.

Soursop (Annona muricata L.) is climacteric fruit with a short ripening period and postharvest shelf life, leading to a rapid softening. In this study, transcriptome analysis of soursop fruits was performed to identify key gene families involved in ripening under postharvest storage conditions (Day 0, Day 3 stored at 28 ± 2 °C, Day 6 at 28 ± 2 °C, Day 3 at 15 ± 2 °C, Day 6 at 15 ± 2 °C, Day 9 at 15 ± 2 °C). The transcriptome analysis showed 224,074 transcripts assembled clustering into 95, 832 unigenes, of which 21, 494 had ORF. RNA-seq analysis showed the highest number of differentially expressed genes on Day 9 at 15 ± 2 °C with 9291 genes (4772 up-regulated and 4519 down-regulated), recording the highest logarithmic fold change in pectin-related genes. Enrichment analysis presented significantly represented GO terms and KEGG pathways associated with molecular function, metabolic process, catalytic activity, biological process terms, as well as biosynthesis of secondary metabolites, plant hormone signal, starch, and sucrose metabolism, plant-pathogen interaction, plant-hormone signal transduction, and MAPK-signaling pathways, among others. Network analysis revealed that pectinesterase genes directly regulate the loss of firmness in fruits stored at 15 ± 2 °C.

DIVERSIDAD GENÉTICA DE POBLACIONES DE GUANÁBANA (Annona muricata L.) EN NAYARIT, MÉXICO MEDIANTE MARCADORES SSR Y SRAP / Genetic diversity of soursop populations (Annona muricata L.) in Nayarit, Mexico using SSR and SRAP markers

RESUMEN La guanábana (Annona muricata L.) es un cultivo de importancia económica para Nayarit, México. Los frutos han tenido una excelente aceptación en el mercado regional, dificultando su comercialización a lugares lejanos porque la producción es altamente perecedera, aunado a que los árboles de los huertos de guanábana son en su mayoría ecotipos o fenotipos sin ningún plan de mejoramiento genético. Debido a la falta de variedades comerciales y de un banco de germoplasma, es importante conocer la diversidad genética para identificar y seleccionar genotipos; una de las herramientas para este propósito es el uso de marcadores moleculares. El objetivo de esta investigación fue analizar la diversidad genética de guanábana de las principales zonas productoras de Nayarit. Se extrajo ADN genómico de hojas de guanábana, las cuales fueron recolectadas de 11 huertos (poblaciones) de las siguientes zonas: Compostela (cinco poblaciones), Tepic (tres poblaciones) y San Blas (tres poblaciones). Posteriormente, se realizó un análisis mediante marcadores moleculares SSR y SRAP. Los resultados indicaron que los SSR no mostraron polimorfismo entre las poblaciones. Por otro lado, en los marcadores SRAP se obtuvieron 116 loci polimórficos con un promedio de porcentaje de loci polimórfico (P) entre las zonas productoras de 29,55 %. Asimismo, se realizó un AMOVA, el cual mostró que el mayor porcentaje de varianza se encuentra dentro de las poblaciones. Además, los análisis de agrupamiento demostraron la formación de tres grupos independientes. Por tanto, se obtuvo una alta homocigocidad y baja diversidad genética de guanábana entre las zonas y poblaciones estudiadas.

ABSTRACT Soursop (Annona muricata L.) is a crop of economic importance for Nayarit, Mexico. Soursop fruits have had an excellent acceptance in the regional market, making it difficult its commercialization to distant places because the production is highly perishable, in addition to the fact that the trees in the soursop orchards are mostly ecotypes or phenotypes without any genetic improvement plan. Due to the lack of commercial varieties and a germplasm bank, it is important to know the genetic diversity to identify and select genotypes; one of the tools for this purpose is the use of molecular markers. The objective of this research was to analyze the genetic diversity of soursop in the main producing areas of Nayarit. Genomic DNA was extracted from soursop leaves from 11 orchards (populations) in the following areas: Compostela (five populations), Tepic (three populations) and San Blas (three populations). Subsequently, we performed molecular analysis using SSR and SRAP molecular markers. The results indicated that the SSRs showed no polymorphism between the populations. On the other hand, we found 116 polymorphic loci in the SRAP markers with an average percentage of polymorphic loci (P) among the producing areas of 29.55 %. Likewise, an AMOVA was performed, showing that the highest percentage of variance is found within the populations. Furthermore, cluster analyzes demonstrated the formation of three independent groups. Therefore, a high homozygosity and low genetic diversity of soursop were obtained between the areas and populations studied.

Growth Promoting Activity of Annona muricata L. Leaf Extracts on Lactobacillus casei.

Soursop leaves are a source of phytochemical compounds, such as phenolic acids, flavonoids, hydrolyzable tannins, and acetogenins. These compounds can have several types of biological activities. Lactic acid bacteria can uptake phenolic compounds present in plants or fruits. The aim of the present work was to investigate the in vitro effect of hexane, acetone, methanolic, and aqueous extracts of soursop leaves (Annona muricata L.) on the growth, motility, and biofilm formation of Lactobacillus casei, and to determine compounds related to growth. The minimum concentration promoting growth, motility (swimming, swarming, and twitching), and biofilm-forming capacity (crystal violet) were evaluated. The results showed the growth-promoting capacity of acetone and aqueous extracts at low doses 25-50 mg/L, and an inhibition in the four extracts at higher doses of 100 mg/L. The L. casei growth is related to ellagic acid, quercetin rhamnoside, kaempferol dihexoside, quercetin hexoside, secoisolariciresinol, and kaempferol hexoside-rhamnoside. Hexane extract increased the three types of motility, while aqueous maintained swimming and twitching motility similar to control. The four extracts inhibited the biofilm formation capacity.

Differential Responses of Antioxidative System during the Interaction of Soursop Fruits (Annona muricata L.) and Nectria haematococca at Postharvest Storage.

Soursop fruit (Annona muricata L.) production is diminished by the attack of pathogens such as Nectria haematococca. However, the fruit-pathogen interaction at the biochemical and molecular levels is still unknown. The objective of this study was to analyze the response of the soursop fruit to the presence of N. haematococca during postharvest storage. Soursop fruits were inoculated with the pathogen and total phenolic compounds, antioxidant capacity by Ferric reducing/antioxidant power (FRAP), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS•+), and 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH•), as well as enzymatic activity and transcript levels of polyphenol oxidase (PPO) and superoxide dismutase (SOD), were evaluated at 1, 3, and 5 days of storage. The noninoculated fruits were the controls of the experiment. The highest total phenol content was recorded on day one in the inoculated fruits. FRAP, ABTS, and DPPH activity presented the highest values on day three in the control fruits. Inoculated fruits recorded the highest PPO activity on day five and a five-fold induction in the PPO transcript on day three. SOD activity showed a decrease during the days of storage and 10-fold induction of SOD transcript on day three in the inoculated fruits. Principal component analysis showed that total phenols were the variable that contributed the most to the observed variations. Furthermore, a positive correlation between total phenols and SOD activity, PPO expression, and SOD expression, as well as between DPPH and FRAP, was recorded. The results showed a differential response in antioxidant capacity, enzymatic activity, and gene expression during the interaction of soursop fruits-N. haematococca at postharvest storage.

Quality and shelf life of the gonad of lion's paw scallop transported and stored whole in refrigeration / Qualidade e vida de prateleira da gônada do bivalve pata de leão, transportado e armazenado inteiramente em refrigeração

The effect of refrigerated 48h transport and 4 days storage on the quality and shelf life of the whole lion's paw scallop Nodipecten subnodosus gonad was evaluated. Proximal composition, adenosine 5´triphosphate (ATP) and related products, K-value, total volatile bases (TVB-N), trimethylamine (TMA-N), pH, fatty acid profile and microbiological analyses were quantified. Gonad holds a significant composition of essential fatty acids while levels of gonadal ATP were initially low; moreover, K-value of the gonad remained constant. With respect to TVB-N and TMA-N, only the former exceeded allowed limits. The pH level showed no significant variation during storage and, despite the high level of TVB-N, according to the TMA-N as well as microbiological analyses it was demonstrated innocuity after 4 days under the transportation and storage conditions utilized.

Avaliou-se o efeito do transporte em refrigeração por 48 horas e quatro dias de armazenamento sobre a qualidade e vida de prateleira da gônada do bivalve pata de leão, Nodipecten subnodosus. Determinou-se a composição centesimal, a adenosina 5'trifosfato (ATP) e afins, o índice K, bases voláteis totais (TVB-N), trimetilamina (TMA-N), pH, perfil de ácidos graxos e análise microbiológica. A Gônada apresentou uma importante composição de ácidos graxos essenciais e baixos níveis iniciais de ATP, enquanto o índice K manteve-se constante. Quanto a TVB -N e TMA- N, apenas as primeiras ultrapassaram os limites admissíveis. Os valores de pH não mostraram nenhuma mudança significativa durante o armazenamento e, apesar dos altos níveis de TVB -N, de acordo com a análise quantitativa e microbiológica TMA- N, a segurança do produto foi demonstrada após quatro dias sob as condições de transporte e armazenamento utilizado.

Diversity of unavailable polysaccharides and dietary fiber in domesticated nopalito and cactus pear fruit (Opuntia spp.).

The aim of this study was to quantify mucilages, pectins, hemicelluloses, and cellulose of nopalitos (edible, as vegetable, young cladodes of flat-stemmed spiny cacti) of most consumed Mexican cultivars, and sweet and acid cactus pear fruits of Opuntia spp. The hypothesis is that, regardless of their unavailable polysaccharides diversity, nopalitos and cactus pear fruits are rich sources of soluble and insoluble dietary fiber. Twelve cultivars of Opuntia spp. were used. Nopalitos had a significant variation in structural polysaccharides among the cultivars: mucilages (from 3.8 to 8.6% dry matter (DM)) averaged near a half of pectins content (from 6.1 to 14.2% DM) and tightly bound hemicelluloses (from 2.2 to 4.7% DM), which were the less abundant polysaccharides, amounted 50% of the loosely bound hemicelluloses (from 4.3 to 10.7% DM). Acid fruits (or 'xoconostle') had significantly higher unavailable polysaccharides content than sweet fruit, and contain similar proportions than nopalitos. Unavailable polysaccharides represent a high proportion of dry tissues of nopalitos and cactus pear fruits, composition of both of these soluble and insoluble polysaccharides (total dietary fiber) widely vary among cultivars without an evident pattern. Nopalitos and cactus pear fruit can be considered an excellent source of dietary fiber.