Effect of Aloe vera gel coating on organoleptic and nutritional quality of minimally processed carrot.

Demand for fresh vegetables has led to development studies in postharvest area mainly focused on minimizing and look for alternatives to chemical additives for food preservation. The use of natural derived edible coatings emerges as a promising alternative for maintaining quality of vegetables. The objective of this study was to evaluate the effect of Aloe vera gel in minimally processed carrot during postharvest storage. Samples with different degrees of processing were immersed in Aloe vera gel, packaged polyolefin bags, and stored in refrigerated chambers at 5 °C for 12 days. Different organoleptic and quality parameters were evaluated. In general, the samples treated with Aloe vera gel showed less quality loss and a lower increment in the bleaching index. Moreover, sensory analysis allowed to establish that carrots processed in slices and shredded and coated with the gel had a more flavorful taste and higher moisture content. Aloe vera treatment did not influence the microbiological growth of bacteria and fungi during storage. Regarding nutritional quality, the treated samples showed a higher accumulation or lower degradation of phenols, flavonoids, and carotenoids, probably generating in this way, a higher antioxidant capacity in these samples. Finally, Aloe vera gel treatment did not influence sugar dynamics in any of the samples. It can be concluded that the treatment with Aloe vera gel allows maintaining a better organoleptic and nutritional quality of carrots with different degrees of processing during refrigerated storage.

Postharvest shelf life extension of minimally processed kale at ambient and refrigerated storage by use of modified atmosphere.

Kale is becoming an important vegetable worldwide, mainly due to its nutritional properties. Kale leaves can be marketed whole, although minimal processing is also in demand. In this article, it was analyzed the effect of packaging in a modified atmosphere of fresh-cut kale leaves stored at 20 °C and 4 °C. Kale leaves were cut into 4 × 4 cm strips and stored in low-density polyethylene bags. Samples processed in the same way but stored in PVC were used as controls. Leaves kept in a modified atmosphere showed a delay in color change with Hue values from about 130 to 120 under PMA against 130 to 100 in control group (CTR) leaves. Chlorophyll degradation was also delayed in both storage temperatures. Samples stored under PMA showed about two times the levels of total chlorophylls with respect to CTR samples at the end of the storage. No changes in total sugar content were detected during storage and no differences were detected between control and modified atmospheres stored samples. Samples maintained in a modified atmosphere showed a lower decrement in soluble proteins and a lower rate of RUBISCO degradation at both temperatures. The relation of RUBISCO content PMA/CTR ranged from 1 to about 3 toward the end of storage No changes in phenols content were found when comparing control and treated samples. However, flavonoid and the antioxidant contents increased in samples stored in modified atmospheres with respect to their controls. We demonstrated that storage in modified atmospheres could be an adequate and simple methodology to extend postharvest life of this minimally processed product at both ambient and refrigerated storage.

Antioxidant response and quality of sunburn Beurré D'Anjou pears (Pyrus communis L.).

Sunburn is a physiological fruit disorder induced by exposure to excessive solar radiation. This disorder leads to significant losses in the yield of marketable fruits by negatively affecting quality parameters such as maturity and external color of the fruits. The purpose of this work was to characterize the physiological and biochemical aspects related to oxidative metabolism in Beurré D'Anjou pear fruit with different sunburn levels. Fruits were collected and classified into three sunburn levels at harvest: no sunburn (S0), mild sunburn (S1), and moderate sunburn (S2). On sunburned area, the maturity indices were measured on the fruit flesh, while external color, photosynthetic and photoprotective pigments, total phenols, electrolyte leakage, lipid peroxidation, antioxidant capacity and antioxidant enzymatic activities were determined on fruit peel. The hue angle and saturation of peel color of pears with different sunburn levels showed significant reduction with increasing damage. These changes in peel color were associated with a reduction in chlorophyll content and variations in carotenoid and anthocyanin levels. Due to metabolic changes resulting from defense and adaptive responses to high solar radiation, sunburned tissues showed significantly increased firmness, soluble solids content, and starch degradation, and lower acidity compared to undamaged fruits. We observed also increased antioxidant capacity in the peel of S1 and S2 fruit, related to higher phenolic contents and increased SOD and APX activities. Consistent with previous reports in apple, our study demonstrates that sunburn affects pear fruit quality traits and maturity state by enhancing oxidative metabolism.

Evidence of glucosinolates translocation from inflorescences to stems during postharvest storage of broccoli.

Broccoli is a vegetable appreciated by consumers for its nutritional properties, particularly for its high glucosinolate (GLS) content. However, broccoli shows a high rate of senescence during postharvest and the GLS content in inflorescences decreases sharply. Usually, postharvest studies on broccoli focus on inflorescences, ignoring the other tissues harvested such as the stems and main stalk. In this work, GLS metabolism in whole heads of broccoli (including inflorescences, small stems and stalk) was analysed during postharvest senescence. The content of GLS content, expression of GLS metabolic genes, and expression of GLS transport-associated genes were measured in the three parts of harvested broccoli. A marked decrease in the content of all GLSs was detected in inflorescences, but an increase in the stems and stalk. Also, decreased expressions of GLS biosynthesis and degradation genes were detected in all tissues analysed. On the other hand, an increase in the expression of one of the genes involved in GLS transport was observed. These results suggest that GLSs would be transported from inflorescences to stems during postharvest senescence. From a commercial point of view, broccoli stems are usually discarded and not used as food. However, the accumulation of GLSs in the stems is an important factor to consider when contemplating potential commercial use of this part of the plant.

Phenylalanine ammonia lyase is more relevant than Chalcone synthase and Chalcone isomerase in the biosynthesis of flavonoids during postharvest senescence of broccoli.

Broccoli contains a high content of nutraceutical compounds, such as glucosinolates and flavonoids. In this work, the effect of different treatments that modulate postharvest senescence of broccoli was evaluated and flavonoid metabolism during postharvest storage was analyzed at 20°C. A decrease in hue angle (HUE°) and chlorophylls and an increase in flavonoid content were detected during senescence. It observed that most of the treatments that delayed senescence also decreased flavonoid content, except visible light and UV-C treatments. In all cases, a direct correlation between those treatments that increased flavonoid biosynthesis and BoPAL gene expression was detected. This response was not detected in the expression of the other two flavonoid synthesis relevant genes BoCHS and BoCHI, suggesting that BoPAL has a greater influence on the regulation of the via, during broccoli senescence. PRACTICAL APPLICATIONS: Broccoli is a vegetable with valuable nutritional properties. Because it is in full development at the time of harvest, it has a short shelf life. In this work, it is showed that visible light and UV-C treatments not only delayed the senescence of broccoli, but also increased flavonoid content. Our results suggest that the most important enzyme in the phenylpropanoid biosynthesis pathway during broccoli postharvest is phenylalanine ammonia lyase, and that this may be a key point in regulating the biosynthesis of these nutritionally valuable compounds.

Expression of BoNOL and BoHCAR genes during postharvest senescence of broccoli heads.

BACKGROUND: Chlorophyll is the most abundant pigment on Earth, essential for the capture of light energy during photosynthesis. During senescence, chlorophyll degradation is highly regulated in order to diminish toxicity of the free chlorophyll molecule due to its photoactivity. The first step in the chlorophyll degradation pathway is the conversion of chlorophyll b to chlorophyll a by means of two consecutive reactions catalyzed by enzymes coded by NYC1 (NON-YELLOW COLORING 1), NOL (NYC1-LIKE) and HCAR. RESULTS: In this work, we studied the expression of NOL and HCAR genes during postharvest senescence of broccoli. We found that the expression of BoNOL increase during the first days of storage and then decrease. In the case of BoHCAR, its expression is maintained during the first days and then it also diminishes. Additionally, the effect of different postharvest treatments on the expression of these genes was also analyzed. It was observed that the expression of BoNOL is lower in the treatments performed with 1-methylcyclopropene (1-MCP), 6-benzylaminopurine (6-BAP) and modified atmospheres, while BoHCAR expression showed an increase in these same treatments, and a decrease in the treatment with ethylene. There were no variations in the expression of both genes in heat treatment, UV-C treatment and visible light treatment. CONCLUSIONS: These results suggest that both BoHCAR and BoNOL show a lower regulation of their expression than other genes involved in chlorophyll degradation during senescence. © 2020 Society of Chemical Industry.

Expression profiling of endo-xylanases during ripening of strawberry cultivars with contrasting softening rates. Influence of postharvest and hormonal treatments.

BACKGROUND: Softening is one of the main features that determine fruit quality during strawberry (Fragaria x ananassa, Duch.) ripening and storage. Being closely related to textural changes, the molecular and biochemical bases underlying strawberry cell-wall metabolism is a matter of interest. Here we investigated the abundance of transcripts encoding putative strawberry endo-xylanases in plant tissues, during fruit ripening and under postharvest and hormonal treatments. Total xylanase activity and expression of related genes in strawberry varieties with contrasting firmness were analyzed. RESULTS: FaXynA and FaXynC mRNA abundance was significantly higher than FaXynB in each plant tissue studied. Higher total xylanase activity was detected at the end of the ripening of the softer cultivar ('Toyonoka') in comparison with the firmer one ('Camarosa'), correlating with the abundance of FaXynA and FaXynC transcripts. Postharvest 1-methylcyclopropene treatment up-regulated FaXynA and FaXynC expressions. FaXynC mRNA abundance decreased with heat treatment but the opposite was observed for FaXynA. Calcium chloride treatment down-regulated FaXynA and FaXynC expression. Both genes responded differently to plant growth regulators' exposure. FaXynC expression was down-regulated by auxins and gibberellins treatment and up-regulated by abscisic acid. FaXynA was up-regulated by auxins, while no changes in mRNA levels were evident by abscisic acid and gibberellins treatment. Ethephon exposure did not change FaXynA and FaXynC expressions. CONCLUSION: New knowledge about the presence of xylanases in ripening strawberry fruit and their response to postharvest and hormonal treatments is provided. Our findings suggest a role for endo-xylanases in hemicelluloses depolymerization and possibly in strawberry fruit softening. © 2020 Society of Chemical Industry.

Harvesting at different time-points of day affects glucosinolate metabolism during postharvest storage of broccoli.

The consumption of broccoli provides a large quantity of compounds with nutraceutical properties to the human diet. Broccoli has a high content of glucosinolates, compounds of the specialized metabolism with anticarcinogenic activity. In a previous work, we found that harvesting different time-points during the day affects the rate of senescence of broccoli heads during postharvest storage. In this work, we tested the same cultural practice to evaluate glucosinolate content and expression of genes involved in glucosinolate metabolism. Broccoli heads were harvested at 8:00, 13:00 and 18:00 h and stored for 5 d at 20 °C in darkness. We found that content and composition of the glucosinolate pool was affected by the time of harvest. Levels of indolic glucosinolates decreased with the time of harvest on the day whereas indolic glucosinolate showed only a moderate decrease. The expression of genes associated to the biosynthesis of aliphatic glucosinolates was variable during the day. In relation to indolic glucosinolates, an increase in the expression of the transcription factor BolMYB51 was detected around 13:00 h, which strongly correlated with the increase in expression of genes associated to their biosynthesis towards the end of the day. During postharvest, the storage in darkness affected differently the metabolisms of indolic and aliphatic glucosinolates. The content of aliphatics decreased during the postharvest period, as well as the expression of the genes associated with their biosynthesis. In contrast, in the case of indolics, their content remained constant or varied slightly, while the expression of the associated biosynthetic genes decreased only slightly. Finally, the genes related to the degradation of glucosinolates appeared to be strongly regulated by light conditions, since their expression increased during the course of the day and decreased markedly during postharvest storage in darkness. These results suggest that harvesting of broccolis close to noon would be convenient to maintain higher levels of glucosinolates during postharvest storage.

The time of the day to harvest affects the degreening, antioxidant compounds, and protein content during postharvest storage of broccoli.

Harvesting of broccoli at several moments of the day affects the rate of senescence during storage. In this work, broccoli heads were harvested at several moments and then kept at 20°C in order to analyze protein metabolism and antioxidant compounds. Almost no differences were detected in the contents of total and soluble proteins, and free amino acids. Only an increment in free amino acids was detected by day 3 in samples obtained at 8:00 hr. With reference to antioxidants, the contents of ascorbic acid, carotenoids and xanthophylls, phenols, and flavonoids were similar in samples harvested at different moments. However, an increment was detected in carotenoids, phenols, and flavonoids during storage of samples collected at 18:00 hr on day 3 and samples collected at 13:00 hr on day 5. The combination of delay of senescence and increment in antioxidants suggest harvesting at 12:00 or 18:00 hr. PRACTICAL APPLICATION: Broccoli is a vegetable with an important level of nutrients. However, it is also highly perishable and suffers a high rate of senescence and loss of quality during postharvest. In this work, it is demonstrated that the simple practice of harvests in different moments of the day can affect the postharvest behavior of broccoli, and it is suggested to carry out the harvest toward the end of the day.

Calcium chloride treatment modifies cell wall metabolism and activates defense responses in strawberry fruit (Fragaria × ananassa, Duch).

BACKGROUND: Fruit dips in calcium ions solutions have been shown as an effective treatment to extend strawberries (Fragaria × ananassa, Duch) quality during storage. In the present work, strawberry fruit were treated with 10 g L-1 calcium chloride solution and treatment effects on cell wall enzymes activities and the expression of encoding genes, as well as enzymes involved in fruit defense responses were investigated. RESULTS: Calcium treatment enhanced pectin methylesterase activity while inhibited those corresponding to pectin hydrolases as polygalacturonase and ß-galactosidase. The expression of key genes for strawberry pectin metabolism was up-regulated (for FaPME1) and down-regulated (for FaPG1, FaPLB, FaPLC, FaßGal1 and FaAra1) by calcium dips. In agreement, a higher firmness level and ionically-bound pectins (IBPs) amount were detected in calcium-treated fruit compared with controls. The in vitro and in vivo growth rate of fungal pathogen Botrytis cinerea was limited by calcium treatment. Moreover, the activities of polyphenol oxidases, chitinases, peroxidases and ß-1,3-glucanases were enhanced by calcium ion dips. CONCLUSION: News insights concerning the biochemical and molecular basis of cell wall preservation and resistance to fungal pathogens on calcium-treated strawberries are provided. © 2019 Society of Chemical Industry.

Effects of heat treatment on enzyme activity and expression of key genes controlling cell wall remodeling in strawberry fruit.

Modification of cell wall polymers composition and structure is one of the main factors contributing to textural changes during strawberry (Fragaria x ananassa, Duch.) fruit ripening and storage. The present study aimed to provide new data to understand the molecular basis underlying the postharvest preservation of strawberry cell wall structure by heat treatment. Ripe fruit (cv. Aroma) were heat-treated in air oven (3 h at 45 °C) and then stored 8 days at 4 °C + 2 days at 20 °C, while maintaining a set of non-treated fruit as controls. The effect of heat stress on the expression pattern of key genes controlling strawberry cell wall metabolism, as well as some enzymatic activities was investigated. The expression of genes proved to be relevant for pectin disassembly and fruit softening process (FaPG1, FaPLB, FaPLC, FaAra1, FaßGal4) were down-regulated by heat treatment, while the expression of genes being involved in the reinforcement of cell wall as pectin-methylesterase (FaPME1) and xyloglucan endo-transglycosilase (FaXTH1) was up-regulated. Total cell wall amount as well as cellulose, hemicellulose, neutral sugars and ionically and covalently bounded pectins were higher in heat-stressed fruit compared to controls, which might be related to higher firmness values. Interestingly, heat stress was able to arrest the in vitro cell wall swelling process during postharvest fruit ripening, suggesting a preservation of cell wall structure, which was in agreement with a lower growth rate of Botrytis cinerea on plates containing cell walls from heat-stressed fruit when compared to controls.

Measurement of expansin activity and plant cell wall creep by using a commercial texture analyzer

Background: Expansins play an important role in cell wall metabolism and fruit softening. Determination of expansin activity is a challenging problem since it depends on measuring cell wall properties by using ad hoc extensometers, a fact that has strongly restricted its study. Then, the objective of the work was to adapt a methodology to measure cell wall creep and expansin activity using a commercial texture meter, equipped with miniature tensile grips and an ad hoc cuvette of easy construction. Results: It was possible to measure hypocotyls acid growth and expansin activity in a reliable and reproducible way, using a commercial texture meter, common equipment found in laboratories of food science or postharvest technology. Expansin activity was detected in protein extracts from cucumber hypocotyls, tomato and strawberry fruits, and statistical differences in expansin activity were found in both fruit models at different ripening stages. Conclusions: The possibility of measuring expansin activity following this adapted protocol with a commercial texture meter could contribute to ease and increase the analysis of expansin in different systems, leading to a better understanding of the properties of these proteins under different experimental conditions.

Novel insights of ethylene role in strawberry cell wall metabolism.

Due to its organoleptic and nutraceutical qualities, strawberry fruit (Fragaria x ananassa, Duch) is a worldwide important commodity. The role of ethylene in the regulation of strawberry cell wall metabolism was studied in fruit from Toyonoka cultivar harvested at white stage, when most changes associated with fruit ripening have begun. Fruit were treated with ethephon, an ethylene-releasing reagent, or with 1-methylcyclopropene (1-MCP), a competitive inhibitor of ethylene action, maintaining a set of non-treated fruit as controls for each condition. Ethephon treated-fruit showed higher contents of hemicelluloses, cellulose and neutral sugars regarding controls, while 1-MCP-treated fruit showed a lower amount of those fractions. On the other hand, ethephon-treated fruit presented a lower quantity of galacturonic acid from ionically and covalently bound pectins regarding controls, while 1-MCP-treated fruit showed higher contents of those components. We also explored the ethylene effect over the mRNA accumulation of genes related to pectins and hemicelluloses metabolism, and a relationship between gene expression patterns and cell wall polysaccharides contents was shown. Moreover, we detected that strawberry necrotrophic pathogens growth more easily on plates containing cell walls from ethephon-treated fruit regarding controls, while a lower growth rate was observed when cell walls from 1-MCP treated fruit were used as the only carbon source, suggesting an effect of ethylene on cell wall structure. Around 60% of strawberry cell wall is made up of pectins, which in turns is 70% made by homogalacturonans. Our findings support the idea of a central role for pectins on strawberry fruit softening and a participation of ethylene in the regulation of this process.

Overexpression of the carbohydrate binding module of strawberry expansin2 in Arabidopsis thaliana modifies plant growth and cell wall metabolism.

Several cell wall enzymes are carbohydrate active enzymes that contain a putative Carbohydrate Binding Module (CBM) in their structures. The main function of these non-catalitic modules is to facilitate the interaction between the enzyme and its substrate. Expansins are non-hydrolytic proteins present in the cell wall, and their structure includes a CBM in the C-terminal that bind to cell wall polymers such as cellulose, hemicelluloses and pectins. We studied the ability of the Expansin2 CBM (CBMFaEXP2) from strawberry (Fragaria x ananassa, Duch) to modify the cell wall of Arabidopsis thaliana. Plants overexpressing CBMFaEXP2 were characterized phenotypically and biochemically. Transgenic plants were taller than wild type, possibly owing to a faster growth of the main stem. Cell walls of CBMFaEXP2-expressing plants were thicker and contained higher amount of pectins. Lower activity of a set of enzymes involved in cell wall degradation (PG, ß-Gal, ß-Xyl) was found, and the expression of the corresponding genes (AtPG, Atß-Gal, Atß-Xyl5) was reduced also. In addition, a decrease in the expression of two A. thaliana Expansin genes (AtEXP5 and AtEXP8) was observed. Transgenic plants were more resistant to Botrytis cinerea infection than wild type, possibly as a consequence of higher cell wall integrity. Our results support the hypothesis that the overexpression of a putative CBM is able to modify plant cell wall structure leading to modulation of wall loosening and plant growth. These findings might offer a tool to controlling physiological processes where cell wall disassembly is relevant, such as fruit softening.

The carbohydrate-binding module of Fragaria × ananassa expansin 2 (CBM-FaExp2) binds to cell wall polysaccharides and decreases cell wall enzyme activities "in vitro".

A putative carbohydrate binding module (CBM) from strawberry (Fragaria × ananassa Duch.) expansin 2 (CBM-FaExp2) was cloned and the encoding protein was over-expressed in Escherichia coli and purified in order to evaluate its capacity to bind different cell wall polysaccharides "in vitro". The protein CBM-FaExp2 bound to microcrystalline cellulose, xylan and pectin with different affinities (K(ad) = 33.6 ± 0.44 mL g(-1), K(ad) = 11.37 ± 0.87 mL g(-1), K(ad) = 10.4 ± 0.19 mL g(-1), respectively). According to "in vitro" enzyme assays, this CBM is able to decrease the activity of cell wall degrading enzymes such as polygalacturonase, endo-glucanase, pectinase and xylanase, probably because the binding of CBM-FaExp2 to the different substrates interferes with enzyme activity. The results suggest that expansins would bind not only cellulose but also a wide range of cell wall polymers.

Effects of ethylene, cytokinin and physical treatments on BoPaO gene expression of harvested broccoli.

BACKGROUND: Broccoli is a highly perishable vegetable that shows enhanced postharvest senescence and intense de-greening caused by chlorophyll degradation. One of the key steps of chlorophyll catabolism is the opening of chlorophyll tretrapyrrole catalysed by pheophorbide a oxygenase (PaO). In this study the expression of a gene encoding a putative PaO was characterised under several chemical and physical treatments. RESULTS: A fragment of a gene encoding a PaO from broccoli (BoPaO) was cloned. The expression of BoPaO showed an important increment during postharvest senescence, in correlation with chlorophyll degradation. Furthermore, broccoli heads were treated with the hormones cytokinin and ethylene. Cytokinin delayed the increment in BoPaO expression, while ethylene accelerated the process. Also, several postharvest treatments were applied in order to evaluate their effect on BoPaO expression. Samples treated with modified atmosphere, hot air, UV-C or white light showed a delay in chlorophyll degradation and de-greening. In most cases the treatments also delayed the increment in BoPaO expression during senescence. CONCLUSION: A close correlation between chlorophyll degradation and BoPaO expression was found during broccoli senescence. This relationship was corroborated in samples treated with different hormonal and physical applications.

Effect of visible light treatments on postharvest senescence of broccoli (Brassica oleracea L.).

BACKGROUND: Broccoli (Brassica oleracea L.) is a rapidly perishable vegetable crop. Several postharvest treatments have been applied in order to delay de-greening. Since light has been shown to have an effect on pigment accumulation during development and darkness is known to induce senescence, the effect of continuous and periodic exposure to low-intensity white light at 22 °C on postharvest senescence of broccoli heads was assayed. RESULTS: Exposure to a constant dose of 12 micromol m(-2) s(-1) was selected as the most suitable treatment and was employed for subsequent experiments. During the course of the treatments, hue and L* values as well as chlorophyll content and visual observation of florets indicated an evident delay in yellowing in treated samples compared with controls. No statistically significant differences in total protein content were found, but soluble protein content was higher in treated samples. Total and reducing sugar as well as starch levels decreased during postharvest senescence, with lower values in control samples. CONCLUSION: The results of this study indicate that storage under continuous low-intensity light is an efficient and low-cost treatment that delays postharvest senescence while maintaining the quality of harvested broccoli florets.

Chlorophyllase versus pheophytinase as candidates for chlorophyll dephytilation during senescence of broccoli.

Degradation of chlorophylls during senescence is a highly regulated process which requires the concerted action of several enzymes. Traditionally, it has been stated that the dismantling process of the chlorophyll molecule begins with a dephytilation step, followed by Mg(2+) removal and other breakdown reactions. Recently, new evidence suggests the possibility of a rearrangement in the first two steps of this process, occurring Mg(2+) removal prior to the loss of the phytol side chain. With the purpose of approximating to the real sequential order of these reactions and to assess if dephytilation occurs on intact (catalyzed by chlorophyllase) or Mg-free (catalyzed by pheophytinase) chlorophyll, expression of both genes was analyzed in broccoli tissue during senescence. Samples of broccoli florets treated with plant hormones, such as cytokinin and ethylene were utilized, as to assess the effect of such compounds on the expression of these genes. Results showed that chlorophyllase expression did not correlate to typical expression patterns for genes related to senescence, since a decrease in expression during senescence was found for one of the two chlorophyllase genes analyzed, and the hormonal-treatment effects on gene expression did not match those observed on chlorophyll content for both chlorophyllase genes. Pheophytinase expression patterns, on the other hand, displayed an increase in the first 3 days of induced senescence, followed by lower expression values towards the end of the experiment. Samples subjected to postharvest treatments mostly showed an inhibition of pheophytinase expression, especially in samples in which degradation of chlorophylls had been delayed. These results suggest that pheophytinase expression correlates to the visual manifestation of postharvest treatments, supporting the possibility that this enzyme is responsible for the dephytilation step in chlorophyll breakdown.

Effect of ethylene and 1-MCP treatments on strawberry fruit ripening.

BACKGROUND: Strawberry is a soft fruit, considered as non-climacteric, being auxins the main hormones that regulate the ripening process. The role of ethylene in strawberry ripening is currently unclear and several studies have considered a revision of the possible role of this hormone. RESULTS: Strawberry fruit were harvested at the white stage and treated with ethephon, an ethylene-releasing reagent, or 1-methylcyclopropene (1-MCP), a competitive inhibitor of ethylene action. The effects of the treatments on fruit quality parameters and on the activity of enzymes related to anthocyanin synthesis and cell wall degradation were evaluated. Some aspects of ripening were accelerated (anthocyanin accumulation, total sugar content and increment of phenylalanine ammonia-lyase (PAL; EC 4.3.1.24) and beta-galactosidase (EC 3.2.1.23) activities), while others were repressed (chlorophyll levels and increment of endo-1,4-beta-glucanase (EC 3.2.1.4) and beta-xylosidase (EC 3.2.1.37) activities) or unchanged (reducing sugar content, pH, titratable acidity and alpha-L-arabinofuranosidase (EC 3.2.1.55) activity) by ethylene. 1-MCP treatment caused the opposite effect. However, its effects were more pronounced, particularly in anthocyanin accumulation, phenolics, PAL and polygalacturonase (EC 3.2.1.15 and EC 3.2.1.67) activities. CONCLUSION: These observations probably indicate that strawberry produces low levels of ethylene that are sufficient to regulate some ripening aspects.

alpha-l-Arabinofuranosidase from strawberry fruit: cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness.

Softening of fleshy fruits during ripening is associated to catabolism of cell wall components. In strawberry, pectin degradation, as well as loss of neutral sugars (mainly arabinose), increases during ripening, and probably contributes to fruit softening. In this work, we report the activity of alpha-l-arabinofuranosidase (alpha-l-arafase) and the expression of related genes in strawberry. Activity of alpha-l-arafase was measured during ripening of cultivars with contrasting firmness. An important increment in the specific activity of alpha-l-arafase was detected during ripening in both cultivars. However, in the softest one (Toyonoka) the specific activities were higher than in the firmest (Camarosa). A combination of semi quantitative reverse transcriptase-PCR (RT-PCR) with degenerate primers and a screening of a cDNA library allowed the isolation and cloning of three cDNAs encoding putative alpha-l-arafases (FaAra1, FaAra2 and FaAra3). The deduced proteins revealed that FaAras belong to the glycoside hydrolase family 51 and not to glycoside hydrolase family 3. Expression studies, carried out by means of Northern-blot and semi quantitative RT-PCR, revealed that FaAras were predominantly expressed in fruit tissue and detected over the entire ripening process. Due to similarity of FaAras sequences, Northern-blot analysis probably grouped the expression of the three genes. The expression was high at small green stage, decreased at white stage and increased thereafter. The increment of the expression from white to 50% red stage was more evident in the softest cultivar (Toyonoka). Semi quantitative RT-PCR analysis allowed determining the expression of individual FaAras. The expression of the three genes was detected in all developmental and ripening stages. However, differences in expression levels could be detected between cultivars. In the softest cultivar, the expression of the three FaAras was higher at 50% and 75% red stages, and in the case of FaAra3 a higher expression was found also at 100% red stage. Overall, specific activity of alpha-l-arafase was higher in the softest cultivar; such activity reflects the expression of at least three putative FaAra genes.