Green Ripe Research Articles

High-throughput extraction method for phenolic compounds in olive fruit (Olea europaea)

Phenolic compounds in olive fruit significantly influence the health benefits and sensory properties of processed olive oil and table olives. A simple and rapid extraction method using homogenization with dimethyl sulfoxide (DMSO) and solid-phase extraction (SPE) cleanup was developed and optimized for ten phenolics in olive fruit. Quantification was performed using high-performance liquid chromatography with diode-array detection (HPLC-DAD). Chromatographic separation of the relevant compounds was achieved using a C18 column (5 μm, 4.6 × 250 mm). The method was validated for accuracy and precision using replicate fortifications (n = 5) in 3 olive matrices (fresh, brined and California-style olives). All compounds were recovered between 80 and 120% and had relative standard deviations below 12.5%. Limits of detection (LOD) and limits of quantification (LOQ) ranged from 0.06 to 7.4 mg/kg and 0.20 to 23.3 mg/kg, respectively. The method was used to quantify phenolic concentrations in fresh olives at different maturities, brined olives and California-style olives, including the analysis of green ripe olives and black ripe olives processed without ferrous gluconate for the first time.

MUTU CABAI RAWIT (CAPSICUM FRUTESCENS L) PADA TINGKAT KEMATANGAN DAN SUHU YANG BERBEDA SELAMA PENYIMPANAN

The objective of this study is to know the ripeness level of small chilli and the right temperature during storage as far as the quality of small chilli can to defend until marketing. The study was conducted at the laboratory of Physiology and Postharvest and laboratory of Food Science and Technology of Research in Agriculture Faculty of Sam Ratulangi University Manado. Three level of ripeness are green ripe, green reddish ripe and red ripe. Temperature of storages are storage in room temperature 28°C – 32°C (64 – 77%), 4°C – 10°C (90 - 95%), 4°C – (-22)°C (95 – 98%), and –10°C (99%). With long storage are 30 days. The Method of analyzed study result is descriptive analysis and the dataare presented by the table. The quality of small chilli during storage looked about the height of water, weight, the height of capsaicin, and colour. The right temperature and the right ripeness can to maintain quality of small chilli until storage. The result study show is in level ripeness green ripe and green reddish ripe and than storage temperature 4°C – 10°C (90 - 95%) can maintain the qualiy of small chilli during storage. It is looked from amount of the height of water, weight, the height of capsaicin, and colour during storage always increase regulary.

Silencing of the SlNAP7 gene influences plastid development and lycopene accumulation in tomato.

Ripening is an important stage of fruit development. To screen the genes associated with pigment formation in tomato fruit, a suppression subtractive hybridization (SSH) cDNA library was constructed by using tomato fruit in the green ripe and break ripe stages, and 129 differential genes were obtained. Using redness as a screening marker, virus-induced gene silencing (VIGS) of the differential genes was performed with a sprout vacuum-infiltration system (SVI). The results showed that silencing the SlNAP7 gene affected the chloroplast development of tomato leaves, manifesting as a photo-bleaching phenotype, and silenced fruit significantly affected the accumulation of lycopene, manifested as a yellow phenotype. In our study, we found that silencing the SlNAP7 gene downregulates the expression of the POR and PORA genes and destroys the normal development of the chloroplast. The expression of related genes included in the lycopene biosynthesis pathway was not significantly changed, but lycopene accumulation was significantly reduced in tomato fruit. Perhaps it was caused by the destruction of the chromoplast, which leads to the oxidation of lycopene. The results show that the SlNAP7 gene influences chloroplast development and lycopene accumulation in tomato.

Gr and hp-1 tomato mutants unveil unprecedented interactions between arbuscular mycorrhizal symbiosis and fruit ripening.

Systemic responses to an arbuscular mycorrhizal fungus reveal opposite phenological patterns in two tomato ripening mutants depending whether ethylene or light reception is involved. The availability of tomato ripening mutants has revealed many aspects of the genetics behind fleshy fruit ripening, plant hormones and light signal reception. Since previous analyses revealed that arbuscular mycorrhizal symbiosis influences tomato berry ripening, we wanted to test the hypothesis that an interplay might occur between root symbiosis and fruit ripening. With this aim, we screened seven tomato mutants affected in the ripening process for their responsiveness to the arbuscular mycorrhizal fungus Funneliformis mosseae. Following their phenological responses we selected two mutants for a deeper analysis: Green ripe (Gr), deficient in fruit ethylene perception and high-pigment-1 (hp-1), displaying enhanced light signal perception throughout the plant. We investigated the putative interactions between ripening processes, mycorrhizal establishment and systemic effects using biochemical and gene expression tools. Our experiments showed that both mutants, notwithstanding a normal mycorrhizal phenotype at root level, exhibit altered arbuscule functionality. Furthermore, in contrast to wild type, mycorrhization did not lead to a higher phosphate concentration in berries of both mutants. These results suggest that the mutations considered interfere with arbuscular mycorrhiza inducing systemic changes in plant phenology and fruits metabolism. We hypothesize a cross talk mechanism between AM and ripening processes that involves genes related to ethylene and light signaling.

Bicolored display of Miconia albicans fruits: Evaluating visual and physiological functions of fruit colors.

Most bird-dispersed fruits are green when unripe and become colored and conspicuous when ripe, signaling that fruits are ready to be consumed and dispersed. The color pattern for fruits of Miconia albicans (Melastomataceae), however, is the opposite, with reddish unripe and green ripe fruits. We (1) verified the maintenance over time of its bicolored display, (2) tested the communicative function of unripe fruits, (3) tested the photoprotective role of anthocyanins in unripe fruits, and (4) verified whether green ripe fruits can assimilate carbon. Using a paired experiment, we tested whether detection of ripe fruits was higher on infructescences with unripe and ripe fruits compared with infructescences with only ripe fruits. We also measured and compared gas exchange, chlorophyll a fluorescence, and heat dissipation of covered (to prevent anthocyanin synthesis) and uncovered ripe and unripe fruits. Although the bicolored display was maintained over time, unripe fruits had no influence on bird detection and removal of ripe fruits. Ripe and unripe fruits did not assimilate CO2, but they respired instead. Since the communicative function of unripe fruits was not confirmed, seed dispersers are unlikely to select the display with bicolored fruits. Because of the absence of photosynthetic activity in ripe and unripe fruits and enhanced photoprotective mechanisms in ripe fruits rather than in unripe fruits, we could not confirm the photoprotective role of anthocyanins in unripe fruits. As an alternative hypothesis, we suggest that the bicolored fruit display could be an adaptation to diversify seed dispersal vectors instead of restricting dispersal to birds and that anthocyanins in unripe fruits may have a defense role against pathogens.

Physico-chemical variation in peel oils of Khasi mandarin (Citrus reticulata Blanco) during ripening

Present study was conducted to investigate the variations in yield and physico-chemical characteristics of Khasi mandarin (Citrus reticulata Blanco) peel oil at three different ripening stages, viz., mature green, turning, and full ripe stages. The highest amount of essential oil was obtained in peels at turning stage fruits (8.1 %), and the lowest in peel at full ripe fruit stage (5 %). Peel oil at the turning stage showed higher aldehyde content (2.97 %), and lower specific gravity (0.843 d25°), refractive index (1.4714 nD20°) and ester number (3.77) than full ripe fruits. A decrease in oxygenated monoterpenes and an increase in monoterpene hydrocarbon were also observed across fruit maturity. The peel oil at turning stage contained the highest amount of organoleptically important oxygenated constituents (1.20 %) which contributed to the overall quality and characteristic aroma of khasi mandarin oil rendering it more suitable for commercial applications.

Força requerida para o desprendimento de frutos de tomate industrial em diferentes estádios de maturação

A colheita do tomate destinado ao processamento, atualmente, é feita com colhedoras automotrizes; para isso, são necessários estudos que viabilizem a melhoria destas colhedoras, reduzindo assim perdas no campo. Este trabalho teve como objetivo quantificar a força necessária para o desprendimento do fruto de tomate de seu pedúnculo em diferentes estágios de maturação. O experimento foi realizado na Fazenda Madeira, no Município de Gameleira de Goiás-GO, e no Laboratório de Engenharia Agrícola da Universidade Estadual de Goiás. Nas condições de campo, os frutos foram retirados avaliando-se a força de tração no sentido axial do pedúnculo ao fruto e, em laboratório, nos sentidos axial e transversal. O trabalho foi realizado no delineamento experimental inteiramente casualizado, com fator único, com dez repetições. Os tratamentos foram os estádios de maturação, considerando tomates verdes, tomates pintados e tomates maduros. Os resultados foram submetidos à análise de variância, aplicando-se o teste de F e, quando significativo, o teste de Tukey, a 5% de probabilidade. A força axial média requerida para os desprendimentos foi de 14,69 N, com maior valor para os frutos maduros. Para o esforço transversal, os valores médios foram 0,98; 1,37 e 1,86 N para os frutos verdes, pintados e maduros, respectivamente.

Problems of breeding f1 hybrids rape and solutions

Purpose. Study the characteristics of sterile and fertile forms of winter rape for winter hardiness, duration of interphase periods, the sugar content in the nectar of flowers to address seed production hybrids F 1 . Methods, material. The studies were conducted during the 2010-2014. On experimental fields and the laboratory of the Plant Production Institute nd. V. Ya. Yuryev, Kharkov. Studied the original form of sterile and fertility restorer winter rape. Sowing was carried out at the optimum time, the end of the third decade of August. Observations and surveys carried out by conventional methods in scientific institutions. Micromethod used for determining free and invert sugar ferricyanide method. Results. Found that the sterile and fertile forms differ in terms of the passage of the interphase periods. The duration of the period of shoots - leaf rosette, resumption of the spring growing season - ramification, ramification - bud, budding - flowering, - wax ripeness was almost the same as that of sterile and fertile in forms. Differences over the years have been significant. During the flowering - green ripeness and the growing season in sterile form longer than the fertile forms an average of 17 and 23 days. Our investigations have established that forms fertile flowers compared with sterile synthesized more monosaccharides, sucrose and total sugars. The average fertile synthesized form monosaccharides 369 mkg / flower, sucrose 190 mkg / flower and total sugars 559 mkg / flower sterile form monosaccharides 250 mkg / flower, sucrose 108 mkg / flower and total sugars 360 mkg / flower. Concluded that the fertile form yavlyayutsya higher priority for pollination bee. When the studies were developed and proposed sites of hybridization schemes. Сonclusions. For pureness of hybrid seeds necessary to create a fertility restorer with simultaneous growth and development as a sterile lines. Necessary to develop schemes hybridization sites for oilseed rape.

Quality and Hygienic Conditions of White Lupin Silage, Affected by Forage Stage of Growth and Use of Silage Additives

A two-factor field experiment with white lupin cv. Butan was carried out. The first factor was the green forage harvest date (the flat pod stage – Cut 1 and the stage of green ripe seeds – Cut 2), while the second one – application of silage additives: biological (strains of lactic acid bacteria) and chemical (a mixture of organic acids), and the control treatment (without additives). In Cut 2 higher fresh matter (FM) and dry matter (DM) yields were obtained. Silage inoculated with the biological additive contained a significantly greater count of lactic acid bacteria. Both additives reduced counts the Clostridium bacteria, yeasts and mould fungi. The silage with the chemical additive had a three-fold higher content of water-soluble carbohydrates (WSC), while the biological additive increased lactic acid (LA) levels. White lupin can be used as a silage raw material, but plants before ensiling should be partially wilted and silage additives should be applied

The effect of forage harvest date and inoculation on the yield and fermentation characteristics of narrow-leaved lupin (Lupinus angustifolius) when ensiled as a whole crop

A experiment was conducted during seasons of the year 2005, 2006 and 2007 to evaluate the effects of the stage of maturity and inoculant application on the fermentation quality of silage produced from narrow leaved lupin. The trial was carried out on the experimental farm in Gorzyn, belonging to Poznan University of Life Sciences (520 33’53 N, 150 53’42 E) in central part of Europe in Poland. A two-factor field experiment was carried out on narrow-leaved lupin cv. Zeus. The first factor was the green forage harvest dates (Cut 1 - at the flat pod stage, Cut 2 – at the stage of green ripe seeds), the other factor – silage inoculants: biological (strains of lactic acid bacteria), chemical (a mixture of organic acids) and a control object without inoculants. The silage raw material was closed in mini-silos for 10 weeks. The crops had a fresh matter yield from 15.8 to 30.8 t ha-1 corresponding to a dry matter yield from 2.2 to 8.8 t ha-1. The dry matter content in unwilted forage per kilogram ranged from 178.8 to 233.8 g. 24-hour wilting caused an increase in dry matter content from 205.5 to 349.5 g kg-1. Both inoculates improved the quality of silages, because they increased the number of lactic acid bacteria and decreased the number of unfavourable groups of bacteria. The narrow-leaved lupin can be used as a silage raw material, but before ensilage plants should be wilted and silage inoculants should be applied.

Modeling Some Physico-Mechanical Properties of Tomato

The purpose of this study was measuring some physico-mechanical properties of tomato varieties (‘Kariz’, ‘Darbigo’, ‘Falkato’, ‘Newton’, and ‘Shaqayeq’) for three ripeness levels as a completely random factorial test; furthermore estimating the mass of the samples, using some new mathematical models, and volume, using image processing techniques. It was signified that the ‘Kariz’ had the most volume, total soluble solids, and sphericity among the varieties and was suitable for paste industry, furthermore due to its highest elasticity modulus (E=0.167 Mpa), had the most resilient. After comparing the relations of estimating the mass, the regression model using three dimensions (length, width, and thickness) had the highest accuracy (M=-183.298+2.272T+1.286W+1.414T with R2=0.96 and RSE=4.44). The external ‘Falcato’ stiffness was the lowest among the varieties ranged from 6.31-7.01 N.mm-1 whereas the most stiffness was related to the green ripeness.

Characterization of Two Orthologs of REVERSION-TO-ETHYLENE SENSITIVITY1 in Apple

The Arabidopsis RTE1 (REVERSION-TO-ETHYLENE SENSITIVITY1) and its tomato ortholog GREEN RIPE (GR) are important in plant ethylene signaling, response and/or fruit ripening. To investigate the potential role of RTE1-like genes in apple (M. ×domestica Borkh.) fruit ripening, two orthologs of RTE1 were identified in the apple genome, designated MdRTE1a and MdRTE1b. Using yeast two-hybrid (Y2H) approach, a cDNA prey library was constructed from a pool of maturing/ripening fruit of ‘Golden Delicious’. Screening the library with MdRTE1a identified two putative NAC (NAM/ATAF1, 2/CUC2) proteins, designated MdNAC1 and MdNAC2, that interacted with MdRTE1a in yeast cells. MdRTE1b, a closely related paralog of MdRTE1a, was also confirmed to be interactive with MdNAC1 and MdNAC2 using the Y2H system. The expression of MdRTE1a and MdNAC1 was high in young fruit but low in more developed and ripening fruit. This pattern of low expression remained largely consistent and was not affected by treatment with 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception, or ethephon, an ethylene releasing compound. However, the expression of MdRTE1b and MdNAC2 rapidly increased during fruit ripening and was highly inhibited by 1-MCP. These data suggested that MdRTE1a, MdNAC1 and their possible interactions in apple cells may play roles in growth, whereas MdRTE1b, MdNAC2 and their putative interactions are involved in ethylene signaling, response and fruit ripening. The possible interactions between an RTE1-like protein and a NAC protein in plant cells have not been reported thus far. Its implications in ethylene signaling and response are discussed.

Effect of ripening stage on physicochemical properties and antioxidant profiles of a promising table fruit ‘pear-jujube’ (Zizyphus jujuba Mill.)

The present study was to investigate the effect of ripening stage on physicochemical properties and antioxidant profiles of a table fruit ‘pear-jujube’ (Zizyphus jujuba Mill.). Six ripening stages of jujubes were established: S1, green ripe; S2, white ripe; S3, representing pear jujube with red surface area of 25–35%; S4, 45–55%; S5, 65–75%; S6, 100%. Jujubes at S1 and S2 were found with high levels of ascorbic acids, proanthocyanidins, phenolics and antioxidant activity. In addition, proper texture, total soluble solids, sugars, sugar–acid ratio and antioxidants as a table jujube emerged at S4. Results obtained clearly demonstrated diverse physicochemical properties, different amounts of phenolics and various degrees of antioxidant activity depending on fruit ripening. In conclusion, un-coloring jujubes could be used as potential natural antioxidants with abundant proanthocyanidins and half red ripening stage is a proper picking period for fresh-eaten as table fruit. In a practical production context, for orchard growers, harvest at relevant ripening stages with intended commercial purpose is possible.

Modulation of ethylene responses by OsRTH1 overexpression reveals the biological significance of ethylene in rice seedling growth and development

Overexpression of Arabidopsis Reversion-To-ethylene Sensitivity1 (RTE1) results in whole-plant ethylene insensitivity dependent on the ethylene receptor gene Ethylene Response1 (ETR1). However, overexpression of the tomato RTE1 homologue Green Ripe (GR) delays fruit ripening but does not confer whole-plant ethylene insensitivity. It was decided to investigate whether aspects of ethylene-induced growth and development of the monocotyledonous model plant rice could be modulated by rice RTE1 homologues (OsRTH genes). Results from a cross-species complementation test in Arabidopsis showed that OsRTH1 overexpression complemented the rte1-2 loss-of-function mutation and conferred whole-plant ethylene insensitivity in an ETR1-dependent manner. In contrast, OsRTH2 and OsRTH3 overexpression did not complement rte1-2 or confer ethylene insensitivity. In rice, OsRTH1 overexpression substantially prevented ethylene-induced alterations in growth and development, including leaf senescence, seedling leaf elongation and development, coleoptile elongation or curvature, and adventitious root development. Results of subcellular localizations of OsRTHs, each fused with the green fluorescent protein, in onion epidermal cells suggested that the three OsRTHs were predominantly localized to the Golgi. OsRTH1 may be an RTE1 orthologue of rice and modulate rice ethylene responses. The possible roles of auxins and gibberellins in the ethylene-induced alterations in growth were evaluated and the biological significance of ethylene in the early stage of rice seedling growth is discussed.

Preparation of Polyclonal Antibody and Expression Analysis of GR in Tomato

The fruit ripening of Green-ripe (Gr) mutant tomato was inhibited dramatically. To determine the expression patterns of Gr in tomato, we first produced the polyclonal antibody of Gr protein. RT-PCR was used to amplify the Gr gene from green ripe tomato fruit. And the PCR product was subcloned into prokaryotic protein expression vectors pET-30a to generate recombinant plasmid. The Gr protein was induced by IPTG in BL21 (DE3) and purified by Ni-NTA agarose column. The anti-Gr serum was produced by immunizing rabbits, and the titer of the anti-Gr serum was above 5000 by ELISA analysis. Purified by the DEAE-52 ion-column, the high purification level of anti-Gr polyclonal antibody was obtained. Furthermore, RT-CPR was used in the RNA level to demonstrate that the expression of Gr gene was specialized in some cultures of tomato. For example, the expressions of Gr were higher in seed, flower and green ripe fruit than others, and the expression level were reduced by exogenous ethylene treatment in the flower and green ripe fruit. Moreover, Polyclonal antibody of Gr was used to investigate the expression pattern of Gr in protein level by the Western blotting. Our results show that the expression level of Gr in protein level was complied with the expressions in RNA. So, we suggested that the regulation of Gr was transcriptional.

沖縄県産マンゴー(アーウィン)の完熟果と緑熟果の最適貯蔵温度

沖縄県産マンゴー(アーウィン)の完熟果と緑熟果の最適貯蔵温度

Dynamic QTL and correlated characters of tomato soluble solid content

Two hundred and six F2:3 families from the cross between TD22 and HT-1-1-1-1 were used for dynamic QTL research of tomato soluble solid content and correlative traits, and correlation analysis of soluble solid content (SSC) with fruit weight (FW), fruit shape index (FSI), soluble sugar, vitamin C (VC), and organic acid at three different development stages. The results showed that there were differences in QTL loci for soluble solid content during the three stages of tomato fruit development. Four and eight QTLs were detected in green ripe stage and red ripe stage, respectively. These QTLs showed dynamic changes, and two markers LEaat006 and Tomato|TC162363 were detected in two stages, which might be useful in molecular-marker assisted selection (MAS). The result also showed that there was extremely significant difference in SSC at the three different stages, and its main correlative traits were different at different stages. Soluble solid content was positively correlated with soluble sugar, but negatively correlated with FW at green ripe stage; SSC was positively correlated with soluble sugar and organic acid at yellow ripe stage; SSC was positively correlated with soluble sugar and organic acid, but negatively correlated with fruit weight at red ripe stage. Based on correlation analysis of these traits, linear regression model was constructed. Non-tested varieties were used to test the fitness, and the result showed that it is well fitted, and the fitness is above 95%.

Separation and Identification of Proteins Related to Fruits Ripening in Mulberry (<i>Morus alba</i>)

To elucidate the physiological mechanism of mulberry fruit ripening in protein level, differential proteome expression of mulberry fruits was analyzed by using 2-DE and mass spectrometry in different ripening stages, green ripe stage(G), half ripe stage(R) and pan ripe stage(P). A mulberry cultivator, “Da10” was used as experimental material. The results showed that separation of proteins with 2-DE were significantly improved by using phenol/SDS buffer for protein extraction. 441, 222, 328 protein spots were detected respectively in ripening stage G, R and P. Among them, differential expression of 31 proteins was more than 2-fold and 6 proteins were stage-specific expression. 8 differential proteins were identified by MALDI-TOF/TOF MS analysis and database search, which were photosynthesis related proteins (ribulose bisphosphate carboxylase/oxygenase activase and ribulose bisphosphate carboxylase small subunit), stress related protein (18kD winter accumulating protein), glucose metabolism related protein (cell wall invertase）and so on, suggesting that these proteins may play the specific physiological role in mulberry fruits ripening.

Genetic dissection of the role of ethylene in regulating auxin-dependent lateral and adventitious root formation in tomato

In this study we investigated the role of ethylene in the formation of lateral and adventitious roots in tomato (Solanum lycopersicum) using mutants isolated for altered ethylene signaling and fruit ripening. Mutations that block ethylene responses and delay ripening -Nr (Never ripe), gr (green ripe), nor (non ripening), and rin (ripening inhibitor) - have enhanced lateral root formation. In contrast, the epi (epinastic) mutant, which has elevated ethylene and constitutive ethylene signaling in some tissues, or treatment with the ethylene precursor 1-aminocyclopropane carboxylic acid (ACC), reduces lateral root formation. Treatment with ACC inhibits the initiation and elongation of lateral roots, except in the Nr genotype. Root basipetal and acropetal indole-3-acetic acid (IAA) transport increase with ACC treatments or in the epi mutant, while in the Nr mutant there is less auxin transport than in the wild type and transport is insensitive to ACC. In contrast, the process of adventitious root formation shows the opposite response to ethylene, with ACC treatment and the epi mutation increasing adventitious root formation and the Nr mutation reducing the number of adventitious roots. In hypocotyls, ACC treatment negatively regulated IAA transport while the Nr mutant showed increased IAA transport in hypocotyls. Ethylene significantly reduces free IAA content in roots, but only subtly changes free IAA content in tomato hypocotyls. These results indicate a negative role for ethylene in lateral root formation and a positive role in adventitious root formation with modulation of auxin transport as a central point of ethylene-auxin crosstalk.

Ethylene signaling: new levels of complexity and regulation

The gaseous plant hormone ethylene plays important roles in plant growth and development. Recent discoveries have expanded our linear view of ethylene signaling by revealing an elaborate signaling network with multiple regulatory circuits. At the membrane, the ethylene receptors form heteromeric and higher order complexes providing enhanced sensitivity and fine-tuning of signaling. Ethylene sensitivity is further enhanced by the rapid degradation of ethylene receptors upon ethylene binding and by dependence on a novel protein REVERSION-TO-ETHYLENE SENSITIVITY1 (RTE1)/GREEN-RIPE (GR). In the nucleus, EIN3-BINDING F-BOX1 and 2 (EBF1/2) coordinately control 26S proteasome degradation of the critical transcription factors EIN3 and EIL1. EBF1/2 expression is repressed by ETHYLENE-INSENSITIVE5 (EIN5), which encodes the exoribonuclease XRN4. Additionally, EIN3 possesses two mitogen-activated protein kinase (MAPK) phosphorylation sites that have opposing effects on EIN3 stability.