ABSTRACT
Banana lectin (BanLec) is a dimeric protein occurring in fruit pulp that modulates immune cell functioning in vitro. In order to assess the immune response in vivo, BanLec from ripe banana (Musa acuminata) fruit was purified and orally given to mice for seven days. The analysis of cytokines in the mice peripheral blood revealed increased IL-10, IL-17 and TNFα, and a reduction of IFNγ and IL-6. In the thymus, an increase of CD4+ and a decrease of CD8+ T-cells were observed after oral administration of BanLec. The modulation of pro- and anti-inflammatory cytokines and T-cells in the peripheral blood and thymus of mice demonstrated the immunomodulatory properties of natural BanLec in vivo. This research brings new data on a protein from a fresh fruit consumed worldwide that may act as an immunomodulator, potentially affecting the host response to infections, immune diseases and cancer.
Subject(s)
Cytokines/metabolism , Lectins/administration & dosage , Lectins/pharmacology , Musa/chemistry , T-Lymphocytes/metabolism , Administration, Oral , Amino Acid Sequence , Animals , Body Weight/drug effects , Cytokines/blood , Feeding Behavior/drug effects , Lectins/chemistry , Lymphocyte Count , Male , Mice, Inbred BALB C , T-Lymphocytes/drug effects , Thymus Gland/cytologyABSTRACT
BACKGROUND: Papaya (Carica papaya L.) is a commercially important crop that produces climacteric fruits with a soft and sweet pulp that contain a wide range of health promoting phytochemicals. Despite its importance, little is known about transcriptional modifications during papaya fruit ripening and their control. In this study we report the analysis of ripe papaya transcriptome by using a cross-species (XSpecies) microarray technique based on the phylogenetic proximity between papaya and Arabidopsis thaliana. RESULTS: Papaya transcriptome analyses resulted in the identification of 414 ripening-related genes with some having their expression validated by qPCR. The transcription profile was compared with that from ripening tomato and grape. There were many similarities between papaya and tomato especially with respect to the expression of genes encoding proteins involved in primary metabolism, regulation of transcription, biotic and abiotic stress and cell wall metabolism. XSpecies microarray data indicated that transcription factors (TFs) of the MADS-box, NAC and AP2/ERF gene families were involved in the control of papaya ripening and revealed that cell wall-related gene expression in papaya had similarities to the expression profiles seen in Arabidopsis during hypocotyl development. CONCLUSION: The cross-species array experiment identified a ripening-related set of genes in papaya allowing the comparison of transcription control between papaya and other fruit bearing taxa during the ripening process.
Subject(s)
Carica/genetics , Fruit/physiology , Oligonucleotide Array Sequence Analysis/methods , Transcriptome , Arabidopsis/genetics , Carica/physiology , Gene Expression Regulation, Plant , Genes, Plant , Solanum lycopersicum/genetics , RNA, Plant/genetics , Transcription Factors/genetics , Transcription Factors/physiology , Vitis/geneticsABSTRACT
Papayas have a very short green life as a result of their rapid pulp softening as well as their susceptibility to physical injury and mold growth. The ripening-related changes take place very quickly, and there is a continued interest in the reduction of postharvest losses. Proteins have a central role in biological processes, and differential proteomics enables the discrimination of proteins affected during papaya ripening. A comparative analysis of the proteomes of climacteric and pre-climacteric papayas was performed using 2DE-DIGE. Third seven proteins corresponding to spots with significant differences in abundance during ripening were submitted to MS analysis, and 27 proteins were identified and classified into six main categories related to the metabolic changes occurring during ripening. Proteins from the cell wall (alpha-galactosidase and invertase), ethylene biosynthesis (methionine synthase), climacteric respiratory burst, stress response, synthesis of carotenoid precursors (hydroxymethylbutenyl 4-diphosphate synthase, GcpE), and chromoplast differentiation (fibrillin) were identified. There was some correspondence between the identified proteins and the data from previous transcript profiling of papaya fruit, but new, accumulated proteins were identified, which reinforces the importance of differential proteomics as a tool to investigate ripening and provides potentially useful information for maintaining fruit quality and minimizing postharvest losses.
Subject(s)
Carica/metabolism , Electrophoresis, Gel, Two-Dimensional/methods , Fruit/metabolism , Proteomics/methods , Cell Wall/metabolism , Chromatography, Liquid/methods , Ethylenes/chemistry , Gene Expression Profiling/methods , Gene Expression Regulation, Plant , Image Processing, Computer-Assisted , Mass Spectrometry/methods , Plastids/metabolism , Time FactorsABSTRACT
Papaya is a climacteric fruit that has high amounts of benzylglucosinolates (BG) and benzylisothiocyanates (BITC), but information regarding levels of BG or BITC during fruit development and ripening is limited. Because BG and BITC are compounds of importance from both a nutritional and a crop yield standpoint, the aim of this work was to access data on the distribution and changes of BG and BITC levels during fruit development and ripening. BG and BITC levels were quantified in peel, pulp, and seeds of papaya fruit. Volatile BITC was also verified in the internal cavity of the fruit during ripening. The influence of the ethylene in BG and BITC levels and mirosinase activity was tested by exposing mature green fruits to ethylene and 1-methylcyclopropene (1-MCP). The highest BG levels were detected in seeds, followed by the peel and pulp being decreased in all tissues during fruit development. Similarly, the levels of BITC were much higher in the seeds than the peel and pulp. The levels of BG for control and ethylene-treated fruit were very similar, increasing in the pulp and peel during late ripening but not changing significantly in seeds. On the other hand, fruit exposed to 1-MCP showed a decrease in BG amount in the pulp and accumulation in seed. The treatments did not result in clear differences regarding the amount of BITC in the pulp and peel of the fruit. According to the results, ethylene does not have a clear effect on BITC accumulation in ripening papaya fruit. The fact that BG levels in the pulp did not decrease during ripening, regardless of the treatment employed, and that papaya is consumed mainly as fresh fruit, speaks in favor of this fruit as a good dietary source for glucosinolate and isothiocyanates.
Subject(s)
Carica/chemistry , Carica/growth & development , Fruit/growth & development , Glycoside Hydrolases/genetics , Isothiocyanates/analysis , Plant Proteins/genetics , Thiocyanates/analysis , Thioglucosides/analysis , Amino Acid Sequence , Carica/enzymology , Carica/genetics , Cyclopropanes/metabolism , Ethylenes/metabolism , Fruit/chemistry , Fruit/enzymology , Fruit/physiology , Gene Expression , Glycoside Hydrolases/chemistry , Glycoside Hydrolases/metabolism , Molecular Sequence Data , Phylogeny , Plant Proteins/chemistry , Plant Proteins/metabolism , Sequence AlignmentABSTRACT
Ripening affects the quality and nutritional contents of fleshy fruits, and papayas are climacteric fruits very susceptible to postharvest losses due to the fast softening caused by ethylene. This paper reports the changes in respiration, ethylene production, and pulp color and firmness, along with the contents of soluble sugars and major carotenoids, during ripening of 'Golden' papaya, an important Brazilian cultivar that has been exported to North American and European markets. The results obtained for nontreated and ethylene- or 1-MCP-treated papaya suggest that 1-MCP can decrease the quality of treated fruit and that even the use of ethylene for triggering or inducing homogeneous ripening can result in lower quality when compared to that of fruit allowed to ripe naturally.
Subject(s)
Carica , Cyclopropanes/pharmacology , Ethylenes/pharmacology , Fruit/drug effects , Fruit/growth & development , Plant Growth Regulators/pharmacology , Carbohydrates/analysis , Carotenoids/analysis , Fruit/chemistryABSTRACT
Starch phosphorylases are enzymes that can use starch as substrate, and they are supposed to act in both in starch synthesis and degradation. This paper reports the effects of ethylene and 1-methylcyclopropene (1-MCP) on the degradation of starch and phosphorylase activity and gene expression. The results indicate that phosphorylase activity is induced during ripening and that it is associated with the onset of starch degradation. The regulation of banana phosphorylase activity is mainly dependent on gene expression, and the absence of ethylene perception by 1-MCP had a positive effect. However, this effect can be precluded by increased levels of ethylene, both autocatalytic and exogenous.
Subject(s)
Cyclopropanes/pharmacology , Ethylenes/pharmacology , Fruit/enzymology , Musa/enzymology , Phosphorylases/genetics , Phosphorylases/metabolism , Fruit/growth & development , Gene Expression/drug effects , Musa/growth & development , RNA, Messenger/analysisABSTRACT
Unripe bananas have a high content of starch (almost 20%) that is metabolized during fruit ripening with a concomitant synthesis of soluble sugars. Since starch granules are composed of amylose and amylopectin, several enzymes have to be involved in its mobilization during banana ripening, with a necessary participation of one starch-debranching enzyme (DBE) to hydrolyze the alpha-1,6-branches of amylopectin. Banana DBE seems to be an isoamylase-type enzyme, as indicated by substrate specificity and the cloning of a 1575 bp cDNA, similar to the isoamylase sequences from potato, Arabdopsis, and maize. The assays for DBE indicated only minor changes in activity during ripening, and the results of the northern and western blots with antiserum against the recombinant banana isoamylase were in agreement with the steady-state level of activity, since no significant changes in gene expression were observed. The high activity on beta-limit dextrin and the similarity to the potato isoform 3 suggest that during banana ripening the hydrolysis of alpha-1,6-linkage of amylopectin results from the activity of a pre-existing isoamylase-type debranching enzyme in coordination with other amylolitic enzymes. To the best of our knowledge, this is the first evaluation of activity and expression of a DBE from a fruit.
Subject(s)
Cloning, Molecular , Fruit/enzymology , Gene Expression , Isoamylase/genetics , Isoamylase/metabolism , Musa/enzymology , Amino Acid Sequence , Base Sequence , Isoamylase/chemistry , Molecular Sequence Data , Sequence Alignment , Starch/metabolism , Substrate SpecificityABSTRACT
Six strawberry cultivars grown on the same commercial plantation in Brazil were evaluated for their chemical composition and quality attributes at the ripe stage. The profiles of the main soluble sugars, ascorbic acid, and anthocyanins were also obtained during the developmental stages. Results showed significant differences among cultivars in all of the investigated parameters. Cv. Campineiro showed an average value for texture of 0.63 N, half the value found for cv. Oso Grande. Anthocyanin content ranged from 13 (cv. Campineiro) to 55 (cv. Mazi) mg/100 g. Total ascorbic acid found for cv. Campineiro (85 mg/100 g) was twice the amount found in cv. Dover (40 mg/100 g). Fructose was the predominant soluble sugar in almost all cultivars. The proportion among the main soluble sugars (fructose, sucrose, and glucose) was similar for Oso Grande and Toyonoka cultivars. The flavonol content (quercetin plus kaempferol derivatives) ranged from 2.7 to 7.1 mg/100 g, with a mean value of 6.1 mg/100 g, whereas free ellagic acid ranged from 0.9 to 1.9 and total phenolics varied from 159 to 289 (mean 221) mg/100 g.
