Metabolic Profile of Strawberry Fruit Ripened on the Plant Following Treatment With an Ethylene Elicitor or Inhibitor.

Strawberry is the most studied nonclimacteric fruit for understanding the role ethylene has in ripening regulation. However, previous studies on the effects of ethylene on strawberry ripening were conducted with detached fruit. Thus, the aim of this work was to determine the effect of ethylene and the ethylene-action inhibitor 1-methylcyclopropene (1-MCP) applied at different developmental stages on important physical-chemical attributes of ripe 'Albion' strawberry. Fruit at four developmental stages that remained attached to the plant were dipped in one of three treatment solutions (Ethephon, 1-methylcyclopropene, and water), plus one absolute control that received no dip. Following treatment, when immature fruit were fully red or 24 h after treatment for red-treated fruit, strawberry fruit were assessed for physicochemical properties (mass, length, diameter, firmness, color, titratable acidity, soluble solids, pH, total phenolics, sugar, organic acid, amino acid, and volatile composition). The days following treatment required for fruit to ripen were also recorded. Treatments did not affect the rate of ripening nor fruit color, titratable acidity, pH, soluble solids, total phenolics, sugars, or organic acids of ripe fruit. Ethephon affected fruit mass, diameter, length, firmness, anthocyanins, amino acids, and volatiles, but these effects were dependent on fruit developmental stage at which the treatment was applied. When green fruit were treated with ethephon, ripe fruit had larger diameter and mass. Ethephon treatment of white fruit resulted in ripe fruit having greater anthocyanin content. Treatment of pink fruit resulted in ripe fruit having smaller diameter, length, and mass and greater firmness. Treatment of red fruit with ethephon altered fruit volatile composition, increasing concentrations of ethyl- and acetate-esters, which were reduced by the 1-MCP treatment. Ethephon treatment increased concentrations of 11 of the 19 free amino acids measured in ripe fruit with treatment of green and white fruit having the greatest effect. A total of 41 volatile compounds had significant correlations with 14 amino acids. While ethylene did not stimulate typical ripening of strawberry fruit, it does appear to alter fruit development and metabolism. The physiological effects of ethylene on strawberry fruit appear to depend on the developmental stage of the fruit.

Early histological, hormonal, and molecular changes during pineapple (Ananas comosus (L.) Merrill) artificial flowering induction.

Natural flowering can cause serious scheduling problems in the pineapple (Ananas comosus) industry and increase harvest costs. Pineapple flowering is thought to be triggered by increased ethylene levels and artificial forcing of pineapple flowering is a common practice to promote flowering synchronisation. However, little is known about the early hormonal and molecular changes of pineapple flowering induction and development. Here, we aimed to analyse the molecular, hormonal, and histological changes during artificial pineapple flowering by Ethrel®48 treatment. Histological analyses of the shoot apical meristem, leaf gibberellic acid (GA3), and ethylene quantification were carried out during the first 72h after Ethrel®48 treatment. Expression profiles from ethylene biosynthesis (AcACS2 and AcACO1), gibberellin metabolism (AcGA2-ox1 and AcDELLA1), and flower development (FT-like gene (AcFT), LFY-like gene (AcLFY), and a PISTILLATA-like gene (AcPI)) genes were analysed during the first 24h after Ethrel®48 treatment. Differentiation processes of the shoot apical meristem into flower buds were already present in the first 72h after Ethrel®48 treatment. Ethrel®48 lead to a reduction in GA3 levels, probably triggered by elevated ethylene levels and the positive regulation AcGA2-ox1. AcLFY activation upon Ethrel®48 may also have contributed to the reduction of GA3 levels and, along with the up-regulation of AcPI, are probably associated with the flower induction activation. AcFT and AcDELLA1 do not seem to be regulated by GA3 and ethylene. Decreased GA3 and increased ethylene levels suggest an accumulation of AcDELLA1, which may display an important role in pineapple flowering induction. Thus, this study shows that molecular, hormonal, and histological changes are present right after Ethrel®48 treatment, providing new insights into how pineapple flowering occurs under natural conditions.

Ethrel-stimulated prolongation of latex flow in the rubber tree (Hevea brasiliensis Muell. Arg.): an Hev b 7-like protein acts as a universal antagonist of rubber particle aggregating factors from lutoids and C-serum.

Ethrel is the most effective stimuli in prolonging the latex flow that consequently increases yield per tapping. This effect is largely ascribed to the enhanced lutoid stability, which is associated with the decreased release of initiators of rubber particle (RP) aggregation from lutoid bursting. However, the increase in both the bursting index of lutoids and the duration of latex flow after applying ethrel or ethylene gas in high concentrations suggests that a new mechanism needs to be introduced. In this study, a latex allergen Hev b 7-like protein in C-serum was identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS). In vitro analysis showed that the protein acted as a universal antagonist of RP aggregating factors from lutoids and C-serum. Ethrel treatment obviously weakened the effect of C-serum on RP aggregation, which was closely associated with the increase in the level of the Hev b 7-like protein and the decrease in the level of the 37 kDa protein, as revealed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), western blotting analysis and antibody neutralization. Thus, the increase of the Hev b 7-like protein level or the ratio of the Hev b 7-like protein to the 37 kDa protein in C-serum should be primarily ascribed to the ethrel-stimulated prolongation of latex flow duration.

Comparative proteomic analysis of latex from Hevea brasiliensis treated with Ethrel and methyl jasmonate using iTRAQ-coupled two-dimensional LC-MS/MS.

UNLABELLED: Ethrel (ET) is an effective and widely used latex yield stimulant of Hevea brasiliensis (Pará rubber tree), and jasmonate (JA) is a key inducer of laticifer differentiation in this plant. To examine variations in the latex proteome caused by these phytohormones, ET and methyl jasmonate (MeJA) were applied to Reyan 7-33-97 rubber tree clones, and comparative proteomic analyses were conducted. On the basis of a transcriptome shotgun assembly (TSA) sequence database and an iTRAQ-coupled two-dimensional LC-MS/MS approach, 1499 latex proteins belonging to 1078 clusters were identified. With a 1.5-fold cut-off value to determine up- and down-regulated proteins, a total of 101 latex proteins were determined to be regulated by ET and/or MeJA via pairwise comparisons among the three exposure durations (0 h, 6 h, and 48 h). Proteins associated with latex regeneration, including phosphoenolpyruvate carboxylase and acetyl-CoA C-acetyltransferase, and those associated with latex flow, such as chitinase and a sieve element occlusion protein, were affected by the application of ET. Chitinase and polyphenol oxidase were also found to be regulated by MeJA. The findings of this study may provide new insight into the roles of phytohormones in latex yield and the causative mechanisms of laticifer differentiation in rubber trees. SIGNIFICANCE: On the basis of a transcriptome shotgun assembly (TSA) sequence database and an iTRAQ-coupled two-dimensional LC-MS/MS approach, the most comprehensive proteome of the latex was profiled, and the ethylene-/jasmonate-responsive proteins were identified in the latex of H. brasiliensis. The findings of this study may provide new insight into the role of phytohormones in latex yield and the causative mechanisms of laticifer differentiation in rubber trees.

Cloning, heterologous expression and characterization of ascorbate peroxidase (APX) gene in laticifer cells of rubber tree (Hevea brasiliensis Muell. Arg.).

Ascorbate peroxidases (APXs) are a kind of crucial enzymes for removing reactive oxygen species (ROS) in plant cell. In the present study, a full-length cDNA encoding an APX, designated HbAPX, was isolated from Hevea brasiliensis by the rapid amplification of cDNA ends (RACE) method. HbAPX was 1174-bp in length and contained a 912-bp open reading frame (ORF) encoding a putative protein of 304 amino acids. The predicted molecular mass of HbAPX was 27.6 kDa (kDa) with an isoelectric point (pI) of 6.73. The phylogenetic analysis showed that HbAPX belonged to the cytosolic subgroup and was more relative to PtAPX and MdAPX2. By using PlantCare online analysis, such cis-acting elements as W-box and MRE were detected in the promoter region of HbAPX. Overproduction of recombinant HbAPX protein either in Escherichia coli or yeast enhanced their tolerance to such abiotic stresses as Cu(2+), Zn(2+), Na(2+) and hydrogen peroxide (H2O2). Ethrel application significantly down-regulated the expression of HbAPX and inhibited the activity of HbAPX in vivo. The ethrel-caused down-regulation of HbAPX may disturb the redox homeostasis in laticifer cells of rubber tree.

Characterization of HbWRKY1, a WRKY transcription factor from Hevea brasiliensis that negatively regulates HbSRPP.

Small rubber particle protein (SRPP) is a major component of Hevea brasiliensis (H. brasiliensis) latex, which is involved in natural rubber (NR) biosynthesis. However, little information is available on the regulation of SRPP gene (HbSRPP) expression. To study the transcriptional regulation of HbSRPP, the yeast one-hybrid experiment was performed to screen the latex cDNA library using the HbSRPP promoter as bait. One cDNA that encodes the WRKY transcription factor, designated as HbWRKY1, was isolated from H. brasiliensis. HbWRKY1 contains a 1437 bp open reading frame that encodes 478 amino acids. The deduced HbWRKY1 protein was predicted to possess two conserved WRKY domains and a C2H2 zinc-finger motif. HbWRKY1 was expressed at different levels, with the highest transcription in the flower, followed by the bark, latex, and leaf. Furthermore, the co-expression of pHbSRP::GUS with CaMV35S::HbWRKY1 significantly decreased the GUS activity in transgenic tobacco, indicating that HbWRKY1 significantly suppressed the HbSRPP promoter. These results suggested that HbWRKY1 maybe a negative transcription regulator of HbSRPP involved in NR biosynthesis in H. brasiliensis.

Anticipation of 'ubá' mango ripening with preharvest ethephon application / Antecipação do amadurecimento de manga 'ubá' com a aplicação pré-colheita de ethephon

Preharvest application of ethephon, a compound that releases ethylene when hydrolyzed, can anticipate and standardize mango harvesting. In this study, the effects of different doses of ethephon, applied at preharvest, on harvest anticipation of 'Ubá' mango were evaluated. On the 17th week after anthesis, 25 mango trees, five per treatment, were sprayed with ethephon at 0, 250, 500, 750 and 1000mg L-1, plus 0.5 percent of mineral oil. Five fruits from each plant were harvested daily until full ripening, and evaluated. Except, for dose 0mg L-1, in which six days after ethephon application (DAAE), harvest was carried out weekly up to 41DAAE The treated fruit presented a more intense flesh color than the non-treated, particularly at the doses of 500 and 750mg L-1. Fruit treated with 500, 750 and 1000mg L-1showed abrupt firmness reduction from the 1st DAAE and TSS higher than 18°Brix on the 6th DAAE. Non-treated fruit presented slower firmness reduction and reached 18°Brix on the 35th DAAE. The dose of 1000mg L-1 of ethephon resulted in premature fall and low fruit quality. Application of 500 or 750mg L-1 of ethephon allowed to anticipate and to standardize mango harvesting. These doses also allowed harvesting fruit with the best commercial quality and without the need of climatization after harvest.

A aplicação pré-colheita de ethephon, composto que libera etileno quando hidrolisado, pode antecipar e uniformizar a colheita de manga. Neste trabalho foram avaliados os efeitos de diferentes doses de ethephon, aplicadas na pré-colheita, sobre a antecipação da colheita de manga 'Ubá'. Na 17ª semana após a antese, 25 mangueiras, sendo cinco por tratamento, foram pulverizadas com ethephon nas doses 0, 250, 500, 750 e 1000L-1, acrescido de 0,5 por cento de óleo mineral. Foram colhidos 10 frutos de cada planta, diariamente, até o completo amadurecimento, com exceção da dose 0L-1, em que, a partir de 6 dias após aplicação do ethephon (DAAE), a colheita foi semanal até 42DAAE. Dos 10 frutos colhidos, cinco foram analisados no dia da colheita e cinco armazenados a 20±080°C e 90±5 por cento UR e avaliados após o completo amadurecimento. Frutos tratados apresentaram coloração de polpa mais intensa que os não tratados, especialmente na dose 500L-1. Frutos tratados com 500, 750 e 1000L-1 apresentaram brusca queda da firmeza a partir do 1ºDAAE, e SST superiores a 18°brix no 6ºDAAE, enquanto nos não tratados, a queda da firmeza foi mais lenta e atingiram 18°brix aos 35ºDAAE. A dose de 1000L-1 de ethephon resultou em queda prematura e má qualidade final dos frutos. Frutos colhidos no 3oDAAE e armazenados atingiram o completo amadurecimento com 12, 7, 6, 4 e 6 dias para as doses 0, 250, 500, 750 e 1000L-1, respectivamente. A aplicação de 500 ou 750L-1 de ethephon, seguida da colheita no 3oDAAE, permitiu a obtenção de frutos com qualidade comercial, além de dispensar a climatização após a colheita.

Anticipation of 'ubá' mango ripening with preharvest ethephon application

Preharvest application of ethephon, a compound that releases ethylene when hydrolyzed, can anticipate and standardize mango harvesting. In this study, the effects of different doses of ethephon, applied at preharvest, on harvest anticipation of 'Ubá' mango were evaluated. On the 17th week after anthesis, 25 mango trees, five per treatment, were sprayed with ethephon at 0, 250, 500, 750 and 1000mg L-1, plus 0.5% of mineral oil. Five fruits from each plant were harvested daily until full ripening, and evaluated. Except, for dose 0mg L-1, in which six days after ethephon application (DAAE), harvest was carried out weekly up to 41DAAE The treated fruit presented a more intense flesh color than the non-treated, particularly at the doses of 500 and 750mg L-1. Fruit treated with 500, 750 and 1000mg L-1showed abrupt firmness reduction from the 1st DAAE and TSS higher than 18°Brix on the 6th DAAE. Non-treated fruit presented slower firmness reduction and reached 18°Brix on the 35th DAAE. The dose of 1000mg L-1 of ethephon resulted in premature fall and low fruit quality. Application of 500 or 750mg L-1 of ethephon allowed to anticipate and to standardize mango harvesting. These doses also allowed harvesting fruit with the best commercial quality and without the need of climatization after harvest.

A aplicação pré-colheita de ethephon, composto que libera etileno quando hidrolisado, pode antecipar e uniformizar a colheita de manga. Neste trabalho foram avaliados os efeitos de diferentes doses de ethephon, aplicadas na pré-colheita, sobre a antecipação da colheita de manga 'Ubá'. Na 17ª semana após a antese, 25 mangueiras, sendo cinco por tratamento, foram pulverizadas com ethephon nas doses 0, 250, 500, 750 e 1000L-1, acrescido de 0,5% de óleo mineral. Foram colhidos 10 frutos de cada planta, diariamente, até o completo amadurecimento, com exceção da dose 0L-1, em que, a partir de 6 dias após aplicação do ethephon (DAAE), a colheita foi semanal até 42DAAE. Dos 10 frutos colhidos, cinco foram analisados no dia da colheita e cinco armazenados a 20±080°C e 90±5% UR e avaliados após o completo amadurecimento. Frutos tratados apresentaram coloração de polpa mais intensa que os não tratados, especialmente na dose 500L-1. Frutos tratados com 500, 750 e 1000L-1 apresentaram brusca queda da firmeza a partir do 1ºDAAE, e SST superiores a 18°brix no 6ºDAAE, enquanto nos não tratados, a queda da firmeza foi mais lenta e atingiram 18°brix aos 35ºDAAE. A dose de 1000L-1 de ethephon resultou em queda prematura e má qualidade final dos frutos. Frutos colhidos no 3oDAAE e armazenados atingiram o completo amadurecimento com 12, 7, 6, 4 e 6 dias para as doses 0, 250, 500, 750 e 1000L-1, respectivamente. A aplicação de 500 ou 750L-1 de ethephon, seguida da colheita no 3oDAAE, permitiu a obtenção de frutos com qualidade comercial, além de dispensar a climatização após a colheita.

Anticipation of 'ubá' mango ripening with preharvest ethephon application

Preharvest application of ethephon, a compound that releases ethylene when hydrolyzed, can anticipate and standardize mango harvesting. In this study, the effects of different doses of ethephon, applied at preharvest, on harvest anticipation of 'Ubá' mango were evaluated. On the 17th week after anthesis, 25 mango trees, five per treatment, were sprayed with ethephon at 0, 250, 500, 750 and 1000mg L-1, plus 0.5% of mineral oil. Five fruits from each plant were harvested daily until full ripening, and evaluated. Except, for dose 0mg L-1, in which six days after ethephon application (DAAE), harvest was carried out weekly up to 41DAAE The treated fruit presented a more intense flesh color than the non-treated, particularly at the doses of 500 and 750mg L-1. Fruit treated with 500, 750 and 1000mg L-1showed abrupt firmness reduction from the 1st DAAE and TSS higher than 18°Brix on the 6th DAAE. Non-treated fruit presented slower firmness reduction and reached 18°Brix on the 35th DAAE. The dose of 1000mg L-1 of ethephon resulted in premature fall and low fruit quality. Application of 500 or 750mg L-1 of ethephon allowed to anticipate and to standardize mango harvesting. These doses also allowed harvesting fruit with the best commercial quality and without the need of climatization after harvest.

A aplicação pré-colheita de ethephon, composto que libera etileno quando hidrolisado, pode antecipar e uniformizar a colheita de manga. Neste trabalho foram avaliados os efeitos de diferentes doses de ethephon, aplicadas na pré-colheita, sobre a antecipação da colheita de manga 'Ubá'. Na 17ª semana após a antese, 25 mangueiras, sendo cinco por tratamento, foram pulverizadas com ethephon nas doses 0, 250, 500, 750 e 1000L-1, acrescido de 0,5% de óleo mineral. Foram colhidos 10 frutos de cada planta, diariamente, até o completo amadurecimento, com exceção da dose 0L-1, em que, a partir de 6 dias após aplicação do ethephon (DAAE), a colheita foi semanal até 42DAAE. Dos 10 frutos colhidos, cinco foram analisados no dia da colheita e cinco armazenados a 20±080°C e 90±5% UR e avaliados após o completo amadurecimento. Frutos tratados apresentaram coloração de polpa mais intensa que os não tratados, especialmente na dose 500L-1. Frutos tratados com 500, 750 e 1000L-1 apresentaram brusca queda da firmeza a partir do 1ºDAAE, e SST superiores a 18°brix no 6ºDAAE, enquanto nos não tratados, a queda da firmeza foi mais lenta e atingiram 18°brix aos 35ºDAAE. A dose de 1000L-1 de ethephon resultou em queda prematura e má qualidade final dos frutos. Frutos colhidos no 3oDAAE e armazenados atingiram o completo amadurecimento com 12, 7, 6, 4 e 6 dias para as doses 0, 250, 500, 750 e 1000L-1, respectivamente. A aplicação de 500 ou 750L-1 de ethephon, seguida da colheita no 3oDAAE, permitiu a obtenção de frutos com qualidade comercial, além de dispensar a climatização após a colheita.

Influência do etileno sobre características químicas de frutos de manga var. Keitt, colhidos em estádio pré-climatérico

Mango fruits (Mangtfera indica L.) Keitt variety, harvested in the pre-climateric stage were treated with 500 and 1,000 ppm of Ethrel (except the control). Soluble carbohydrates, vitamin C, proteins and humidity were evaluated on the samples at 3, 6, 9 and 16 days post-harvest. The results obtained, showed that Ethrel did not have influence on the chemical characteristics of the fruits. Fruits showed modificátions during the maturation only for the parameters of soluble carbohydrate and vitamin C.

Frutos de manga (Mangifera indica L.) variedade Keitt, colhidos em estádio pré-climatérico, receberam tratamento com 500 e 1.000 ppm de Ethrel (com exceção da testemunha), sendo submetidos a determinações de carboidratos solúveis, vitamina C, proteínas e umidade, de acordo com amostragens realizadas 3, 6, 9 e 16 dias pós-colheita. Os resultados obtidos, revelaram que o Ethrel não excerceu nenhuma influência nas características químicas dos frutos e que, somente em termos de carboidratos solúveis e vitamina C, os frutos mostraram alterações durante a maturação.

Influência do etileno sobre características químicas de frutos de manga var. Keitt, colhidos em estádio pré-climatérico

Mango fruits (Mangtfera indica L.) Keitt variety, harvested in the pre-climateric stage were treated with 500 and 1,000 ppm of Ethrel (except the control). Soluble carbohydrates, vitamin C, proteins and humidity were evaluated on the samples at 3, 6, 9 and 16 days post-harvest. The results obtained, showed that Ethrel did not have influence on the chemical characteristics of the fruits. Fruits showed modificátions during the maturation only for the parameters of soluble carbohydrate and vitamin C.

Frutos de manga (Mangifera indica L.) variedade Keitt, colhidos em estádio pré-climatérico, receberam tratamento com 500 e 1.000 ppm de Ethrel (com exceção da testemunha), sendo submetidos a determinações de carboidratos solúveis, vitamina C, proteínas e umidade, de acordo com amostragens realizadas 3, 6, 9 e 16 dias pós-colheita. Os resultados obtidos, revelaram que o Ethrel não excerceu nenhuma influência nas características químicas dos frutos e que, somente em termos de carboidratos solúveis e vitamina C, os frutos mostraram alterações durante a maturação.

Efeito da aplicação de etileno no pH, acidez, índice refratométrico e açúcares totais de frutos de manga, colhidos em estágio pré-climatérico

Ethylene action on mango fruits was studied in relation to the ripening physiology of these fruits. Mango (Mangifera indica L.), Keitt variety, were harvested in the pre-climateric stage. These fruits, after receiving Ethrel treatment (500 e 1,000 ppni, except for control), were analized for pH, acidity, refratometric index and total sugars. The analysis and interpretation of the results showed no statistical differences among treatments, showing that ethrel didn't improve the fruits quality.

Este trabalho teve como objetivo o estudo da ação do etileno no amadurecimento de frutos de manga, tentando contribuir para o conhecimento da fisiologia do amadurecimento destes frutos. Foram utilizadas mangas (Mangifera indica L.), variedade Keitt, colhidas em estágio pré-climatérico. Estes frutos receberam tratamento com ethrel (500 e 1.000 ppm, com exceção da testemunha) e foram submetidos às seguintes determinações: pH, acidez, índice refratométrico e açúcares totais. A análise e interpretação dos resultados mostrou a inexistência de diferença estatística entre esses 3 tratamentos, indicando que a aplicação de ethrel não trouxe nenhuma vantagem na melhoria de qualidade dos frutos.

Efeito da aplicação de etileno no pH, acidez, índice refratométrico e açúcares totais de frutos de manga, colhidos em estágio pré-climatérico

Ethylene action on mango fruits was studied in relation to the ripening physiology of these fruits. Mango (Mangifera indica L.), Keitt variety, were harvested in the pre-climateric stage. These fruits, after receiving Ethrel treatment (500 e 1,000 ppni, except for control), were analized for pH, acidity, refratometric index and total sugars. The analysis and interpretation of the results showed no statistical differences among treatments, showing that ethrel didn't improve the fruits quality.

Este trabalho teve como objetivo o estudo da ação do etileno no amadurecimento de frutos de manga, tentando contribuir para o conhecimento da fisiologia do amadurecimento destes frutos. Foram utilizadas mangas (Mangifera indica L.), variedade Keitt, colhidas em estágio pré-climatérico. Estes frutos receberam tratamento com ethrel (500 e 1.000 ppm, com exceção da testemunha) e foram submetidos às seguintes determinações: pH, acidez, índice refratométrico e açúcares totais. A análise e interpretação dos resultados mostrou a inexistência de diferença estatística entre esses 3 tratamentos, indicando que a aplicação de ethrel não trouxe nenhuma vantagem na melhoria de qualidade dos frutos.