Exploiting tropical fruit processing coproducts as circular resources to promote the growth and maintain the culturability and functionality of probiotic lactobacilli.

This study evaluated the use of acerola (Malpighia glabra L., CACE), cashew (Anacardium occidentale L., CCAS), and guava (Psidium guayaba L., CGUA) fruit processing coproducts as substrates to promote the growth, metabolite production, and maintenance of the viability/metabolic activity of the probiotics Lactobacillus acidophilus LA-05 and Lacticaseibacillus paracasei L-10 during cultivation, freeze-drying, storage, and exposure to simulated gastrointestinal digestion. Probiotic lactobacilli presented high viable counts (≥8.8 log colony-forming units (CFU)/mL) and a short lag phase during 24 h of cultivation in CACE, CCAS, and CGUA. Cultivation of probiotic lactobacilli in fruit coproducts promoted sugar consumption, medium acidification, and production of organic acids over time, besides increasing the of several phenolic compounds and antioxidant activity. Probiotic lactobacilli cultivated in fruit coproducts had increased survival percentages after freeze-drying and during 120 days of refrigerated storage. Moreover, probiotic lactobacilli cultivated and freeze-dried in fruit coproducts had larger subpopulations of live and metabolically active cells when exposed to simulated gastrointestinal digestion. The results showed that fruit coproducts not only improved the growth and helped to maintain the viability and metabolic activity of probiotic strains but also enriched the final fermented products with bioactive compounds, being an innovative circular strategy for producing high-quality probiotic cultures.

Biogeography and early diversification of Tapinotaspidini oil-bees support presence of Paleocene savannas in South America.

Worldwide distributed tropical savannas were established only in the Miocene, with climatic cooling and rise of C4 grasses. However, there is evidence for an earlier presence of savanna-like vegetation in southern parts of South America. Here we investigated the biogeographic history of a clade of solitary bees which have endemic groups in areas covered by savannas and other types of open vegetation as well as forested areas. We hypothesized that these bees originated in savanna-like biomes and that shifts to forested areas and floral host shifts increased species diversification along their evolutionary history. We reconstructed a comprehensive phylogeny for Tapinotaspidini bees based majorly on original DNA sequences. We then used macroevolutionary tools to estimate ancestral range area and reconstructed ancestral habitat (open versus forested) and host plant association to analyze the effects of shifts in vegetation type and flower hosts on their diversification. Tapinotaspidini bees originated in the Paleocene and in a savanna-type, Cerrado-like, which is reinforced by reconstruction of open vegetation as the most probable ancestral area, thus bringing additional evidence to a much earlier origin of this vegetation type in South America. Shifts to forested areas occurred at least three times in a period of 30 Ma and were responsible for slight increases in diversification rates. Malpighiaceae is the ancestral floral host; host broadening occurred only in the Miocene and at least in three occasions. Host shifts, i.e. from Malpighiaceae to other oil families, occurred in the Eocene and Miocene. Both host broadening and host shifts did not significantly alter diversification rates, however exploitation of other oil sources were important in occupying new habitats. The link between biomes and host plant shifts and changes in diversification rate brings us additional insights into the evolution of bees and associated flora in South America.

Characterization and oxidative stability of oils and bioactive compounds of the fruits of Byrsonima cydoniifolia A. Juss. at different ripening stages.

BACKGROUND: Fruits present high concentrations of bioactive compounds that are beneficial to health due to their antioxidant properties. New alternatives to vegetable oils with such properties have been studied. We determined the chemical compounds of Byrsonima cydoniifolia A. Juss. fruits in three ripening stages as well as the optical behavior and quality level of the oils. RESULTS: The ripening stage affected the chemical composition of the fruits and oils. The fruits presented high values of bioactive compounds, as ascorbic acid (1.46-1.82 g kg-1 ) and total phenols (3.54-15.91 g gallic acid equivalents kg-1 ), as well as showed excellent antioxidant activity. The ripe fruits showed high content of carotenoids (45.90 mg kg-1 ) were lutein is the major carotenoid, representing 55.56%. The oil of the ripe fruits showed high lipid content (252.6 g kg-1 ) and contained oleic (521.83 g kg-1 ), palmitic (209.13 g kg-1 ) and linoleic (195.4 g kg-1 ) fatty acids. The oil of ripe fruits showed the best oxidative stability, with longer induction period (22.29 h) than the other oils. CONCLUSION: Fruits of B. cydoniifolia A. Juss. demonstrate to possess a promising potential for the use as functional ingredients. The oil of the ripe fruits was more stable and presents a greater potential to be used for edible purposes. © 2018 Society of Chemical Industry.

Phenolic Compounds Determined by LC-MS/MS and In Vitro Antioxidant Capacity of Brazilian Fruits in Two Edible Ripening Stages.

The aim of this study was to investigate the free individual phenolics and the in vitro antioxidant capacity of blackberry, acerola, yellow guava, guabiju, jambolan and jabuticaba fruits in two edible stages. Of the thirty-three phenolics investigated by liquid chromatography - tandem mass spectrometry (LC-MS/MS), twenty-five were quantified and the major ones were catechin, isoquercitrin, epicatechin and gallic acid. The highest values for the total phenolic content (in dry matter) were observed for acerola (83.6 to 97.7 mg gallic acid equivalents g-1 DM) and blackberry (18.9 to 28.3 mg gallic acid equivalents g-1 DM); however, acerola, jabuticaba, and blackberry showed the highest antioxidant capacities (134.6 to 1120.4 mg Trolox equivalents g-1 for 2,2-diphenyl-1-picrylhydrazyl and 43.6 to 501.8 µmol Trolox equivalents g-1 for ferric reducing antioxidant power). For most fruits, the antioxidant capacity decreased during the ripening, possibly due to a decrease in the concentration of most of the phenolics.

HPLC and in vitro evaluation of antioxidant properties of fruit from Malpighia glabra (Malpighiaceae) at different stages of maturation.

Malpighia glabra L. is indicated for the treatment of diseases due presenting several medicinal properties. The aim of the present study was quantify phenolic compounds and carotenoids, additionally, evaluate the antioxidant potential of different methanolic fractions (mature stage, intermediate stage and green maturity stage) of Malpighia glabra. Methanolic fractions of the three stages of maturation were characterized using HPLC-DAD. The antioxidant activity of fractions was evaluated using in vitro assay systems: iron-chelating activity (Fe2+), inhibition of the radical 1-1-diphenyl-2-picrylhydrazyl (DPPH) and inhibition of production of thiobarbituric acid reactive substances (TBARS). In quantifying compounds by HPLC-DAD showed that the compounds rutin and quercetin showed the highest peaks in the three samples tested. Regarding carotenoids, the ß-carotene stood in the analyze. The methanolic fraction of lyophilized pulp of green fruits showed better results for iron chelation and DPPH tests; TBARS by the methanolic fraction of the ripe fruits showed lower IC50 as the basal. When in the presence of iron as an inducer of peroxidation, the methanolic fraction of the pulp of green fruits showed the lowest IC50. Our research indicates that the o fruits of M. glabra good potential in scavenging free radicals and can be an important source of antioxidant phytochemical.

Heteropterys tomentosa A. Juss: Toxicological and adaptogenic effects in experimental models.

BACKGROUND: The constant pursuit of improved athletic performance characterizes high-performance sport and the use of medicinal plants as dietary supplements is becoming widespread among athletes to enhance long-term endurance performance. AIM: The present study evaluated the toxicity of Heteropterys tomentosa (HEHt) and its acute adaptogenic effects. METHODS: The in vitro safety profile was evaluated on CHO-k1 cells using the alamar Blue assay, at concentrations ranging from 3.125 to 200 µg/mL. In vivo acute oral toxicity was conducted in male and female mice with oral administration of graded doses of HEHt from 400 to 2000 mg/kg. A subchronic oral toxicity study was completed by oral administration of HEHt (50, 200 or 1000 mg/kg) and vehicle for 30 days in male Wistar rats. Clinical observations and toxicological related parameters were determined. Blood was collected for biochemical and hematological analyses, while histological examinations were performed on selected organs. Thereafter, an adaptogenic test consisting of progressive loads until exhaustion was conducted in rats ( n = 5/group) orally pre-treated with the vehicle and HEHt (25, 100 or 400 mg/kg). RESULTS: HEHt exhibited no cytotoxic effects on the CHO-k1 cells and, apparently, no acute toxicity in mice and no subchronic toxicity in rats. An ergogenic effect was observed only at the dose of 25 mg/kg compared with the vehicle in relation to time to exhaustion and exercise load ( p = .011 and .019, respectively). HEHt is safe at up to 400 mg/kg, contains astilbin and taxifolin as the major phytochemical compounds, and exhibited a potential adaptogenic effect. CONCLUSIONS: These results justify its anecdotal usage as a tonic, show that the hydroethanolic maceration of the root does not cause toxicity, and provide scientific evidence of its potential as a source of new adaptogenic substance(s).

First record of acerola weevil, Anthonomus tomentosus (Faust, 1894) (Coleoptera: Curculionidae), in Brazil / Primeiro registro do bicudo dos frutos da acerola, Anthonomus tomentosus (Faust, 1894) (Coleoptera: Curculionidae), no Brasil

Abstract The weevil of acerola fruits, Anthonomus tomentosus (Faust, 1894) (Coleoptera: Curculionidae), is recorded for the first time in Brazil. Samples of this insect were collected in fruits of acerola, Malpighia emarginata D.C. (Malpighiaceae), in four municipalities in the north-central region of Roraima State, in the Brazilian Amazon. Information about injuries observed in fruits infested with A. tomentosus, its distribution in Roraima, and suggestions for pest management are presented.

Resumo O bicudo dos frutos da acerola, Anthonomus tomentosus (Faust, 1894) (Coleoptera: Curculionidae), é registrado pela primeira vez no Brasil. Amostras deste inseto foram coletadas em frutos de acerola, Malpighia emarginata D.C. (Malpighiaceae), em quatro municípios do Centro-Norte do Estado de Roraima, na Amazônia brasileira. Informações sobre as injúrias observadas nos frutos infestados por A. tomentosus, sua distribuição em Roraima e sugestões para o seu manejo são apresentadas.

Intestinal microbiota are involved in the immunomodulatory activities of longan polysaccharide.

It is difficult for polysaccharides to be directly absorbed through the intestine, which implies other utilization mechanisms involved in the bioactivity performance of polysaccharide. In this study, the multi-omics approach was applied to investigate the impacts of longan polysaccharide on mouse intestinal microbiome and the interaction between the polysaccharide-derived microbiome and host immune system. According to the result, the longan polysaccharide showed a significant improvement in the typical intestinal immunity index of mice. Meanwhile, at the taxonomy level, the intestinal microbiota from the control group and polysaccharide group were highly distinct in organismal structure. At the functional level, a significant decline in the microbial metabolites of pyruvate, butanoate fructose and mannose in the control group was found. Additionally, a significant increase was observed in the succinic acid and the short-chain fatty acid, including acetic acid, propionic acid and butyric acid, in the polysaccharide group. Furthermore, the multi-omic based network analysis indicated that the intake of longan polysaccharide resulted in the changes of the intestinal microbiota as well as the gut metabolites, which led to the enhancement of host's immune function under the stress conditions. These results indicated the polysaccharide-derived changes in intestinal microbiota were involved in the immunomodulatory activities.

First record of acerola weevil, Anthonomus tomentosus (Faust, 1894) (Coleoptera: Curculionidae), in Brazil.

The weevil of acerola fruits, Anthonomus tomentosus (Faust, 1894) (Coleoptera: Curculionidae), is recorded for the first time in Brazil. Samples of this insect were collected in fruits of acerola, Malpighia emarginata D.C. (Malpighiaceae), in four municipalities in the north-central region of Roraima State, in the Brazilian Amazon. Information about injuries observed in fruits infested with A. tomentosus, its distribution in Roraima, and suggestions for pest management are presented.

Response of Malpighia emarginata active germplasm bank accessions to Meloidogyne enterolobii parasitism.

Malpighia emarginata is cultivated in almost all Brazil and is considered an important agricultural crop. The root-knot nematode Meloidogyne enterolobii has been described as a major threat to this crop, causing great production losses. Due to the scarcity of information about the severity of this parasite in M. emarginata plants in Brazil, this study investigated M. enterolobii resistance of ten M. emarginata genotypes from the active germplasm bank of Universidade Federal Rural de Pernambuco. The experiment was conducted adopting a completely randomized design in a factorial arrangement of 11 x 2 x 5, where M. emarginata cuttings were inoculated with 10,000 eggs in a greenhouse. After 150 days, plants were evaluated for the following parameters: gall index, egg mass index, number of eggs per root system, number of eggs per gram of root, and reproduction factor. The accessions showed different responses depending on host x pathogen interaction, from susceptibility to moderate tolerance. Accessions 027-CMF and 031-CMF were considered tolerant to the nematode and could be of great value in new breeding programs for resistance to M. enterolobii infection.

Structure and development of 'witches' broom' galls in reproductive organs of Byrsonima sericea (Malpighiaceae) and their effects on host plants.

Galls are anomalies in plant development of parasitic origin that affect the cellular differentiation or growth and represent a remarkable plant-parasite interaction. Byrsonima sericea DC. (Malpighiaceae) is a super host of several different types of gall in both vegetative and reproductive organs. The existence of galls in reproductive organs and their effects on the host plant are seldom described in the literature. In this paper, we present a novel study of galls in plants of the Neotropical region: the 'witches' broom' galls developed in floral structures of B. sericea. The unaffected inflorescences are characterised by a single indeterminate main axis with spirally arranged flower buds. The flower buds developed five unaffected brownish hairy sepals and five pairs of elliptical yellow elaiophores, five yellow fringed petals, 10 stamens and a pistil with superior tricarpellar and trilocular ovary. The affected inflorescences showed changes in architecture, with branches arising from the main axis and flower buds. The flower buds exhibited several morphological and anatomical changes. The sepals, petals and carpels converted into leaf-like structures after differentiation. Stamens exhibited degeneration of the sporogenous tissue and structures containing hyphae and spores. The gynoecium did not develop, forming a central meristematic region, from which emerges the new inflorescence. In this work, we discuss the several changes in development of reproductive structures caused by witches' broom galls and their effects on reproductive success of the host plants.

Photosynthetic and morphological acclimation of seedlings of tropical lianas to changes in the light environment.

UNLABELLED: • PREMISE OF THE STUDY: Few studies have analyzed the physiological performance of different life stages and the expression of ontogenetic niche shifts in lianas. Here, we analyzed the photosynthetic and morphological acclimation of seedlings of Stigmaphyllon lindenianum, Combretum fruticosum, and Bonamia trichantha to distinctive light conditions in a tropical dry forest and compared their response with the acclimation response of adult canopy lianas of the same species. We expected acclimation to occur faster through changes in leaf photochemistry relative to adaptation in morphology, consistent with the life history strategies of these lianas.• METHODS: Seedlings were assigned to the following light treatments: high light (HH), low light (LL), sun to shade (HL), and shade to sun (LH) in a common garden. After 40 d, HL and LH seedlings were exposed to opposite light treatments. Light response curves, the maximum photosynthetic rate in the field (Amax), and biomass allocation were monitored for another 40 d on leaves expanded before transfer.• KEY RESULTS: Photosynthetic responses, Amax, and biomass of Stigmaphyllon and Combretum varied with light availability. Physiological characters were affected by current light environment. The previous light environment (carryover effects) only influenced Amax. Morphological characters showed significant carryover effects. Stigmaphyllon showed high morphological and physiological plasticity. Sun-exposed seedlings of this liana increased stem biomass and switched from self-supporting to climbing forms.• CONCLUSIONS: Acclimation in seedlings of these lianas is consistent with the response of adult lianas in the canopy in direction, but not in magnitude. There was no evidence for ontogenetic niche shifts in the acclimation response.

Structure of floral galls of Byrsonima sericea (Malpighiaceae) induced by Bruggmanniella byrsonimae (Cecidomyiidae, Diptera) and their effects on host plants.

Galls are anomalies in plant development from parasitic origin, and affect cellular differentiation or growth of plants. This parasite-plant interaction occurs in many environments and typically in vegetative organs of plants. The existence of galls in reproductive organs and their effects on the host plant are seldom described in the literature. In this paper, we present a novel study of galls in plants of the neotropical region. Galls of Bruggmmaniella byrsonimae develop in the flower buds of Byrsonima sericea DC. (Malpighiaceae) and affect development of the reproductive organs and the reproductive effort of these plants. The sepals and petals show hypertrophy of parenchyma tissues after differentiation, and the stamens exhibit degeneration of the sporogenic tissue. The gynoecium is not entirely developed; ovary and ovules are often absent. Changes in vascular tissues are also frequent, which may indicate high demand for nutrient resources by the new tissues initiated by the larva. We compared the amount of inflorescences, galls and fruits to evaluate possible effects on host reproduction. The results suggest that the Cecidomyiidae galls in flower organs affect fruit set and the reproductive success of B. sericea.

Shoot-tip cryopreservation by droplet vitrification of Byrsonima intermedia A. Juss.: a woody tropical and medicinal plant species from Brazilian cerrado.

Cryopreservation of plant species is poorly investigated in Brazil. The aim of this study was to cryopreserve Byrsonima intermedia shoot apical meristems through droplet vitrification. A culture medium for shoot-tips growth was established using the Woody Plant Medium supplemented with 2.22 uM 6-benzylaminopurine. Excised shoot-tips were subjected to pre-culture and/or post-culture treatments on Murashige and Skoog medium with 0.3 M sucrose for 24 h prior dehydration on PVS2 at 0°C for 15, 30 or 45 minutes prior to plunging in liquid nitrogen. The effect of 15 days of shoot pre-growth on a high osmotic medium (0.3 M sucrose or 0.21 M sorbitol + 0.09 M sucrose) prior to meristem excision and cryopreservation was also investigated. Pre-culturing shoot-tips on 0.3 M sucrose for 24 h prior to cryopreservation increased the regrowth level after thawing to 90%. Shoot-tips excised from shoots pre-grown on MS + 0.21 M sorbitol + 0.09 M sucrose for 15 days presented a satisfactory regrowth level (67%).

Anatomy and lignification status of the Lophanthera lactescens Ducke plant grown in vitro.

This aim of this study was to evaluate the lignification in young stems of the Lophanthera lactescens Ducke plant grown in vitro L. lactescens (Malpighiaceae), a species endemic in the Brazilian Amazon that possesses both medicinal properties and could be used in the forest product industry. Plants grown in vitro condition in MS medium were analyzed using Infrared (IR) microspectroscopy in a diffuse reflectance mode, fluorescence microscopy. Moreover, histochemical tests such as the Wiesner and Maüle test were run to monitor the process of lignification in cell walls. The spectra of IR microscopy recorded using cross section tissue are representative of guaiacyl/syringyl lignin, based on the 1336 and 1246 cm-1 signal. Individuals presenting stem sprains, resulting from the marked development, produced gelatinous fibers with a clear cellulose layer. Initially, fluorescence microscopy demonstrated lignin deposition in the cell corner region having progressive deposition in the secondary wall of tracheary elements of the stem.

Anatomy and lignification status of the Lophanthera lactescens Ducke plant grown in vitro / Anatomia e status da lignificação de plantas de Lophanthera lactescens Ducke crescidas in vitro

This aim of this study was to evaluate the lignification in young stems of the Lophanthera lactescens Ducke plant grown in vitro L. lactescens (Malpighiaceae), a species endemic in the Brazilian Amazon that possesses both medicinal properties and could be used in the forest product industry. Plants grown in vitro condition in MS medium were analyzed using Infrared (IR) microspectroscopy in a diffuse reflectance mode, fluorescence microscopy. Moreover, histochemical tests such as the Wiesner and Maüle test were run to monitor the process of lignification in cell walls. The spectra of IR microscopy recorded using cross section tissue are representative of guaiacyl/syringyl lignin, based on the 1336 and 1246 cm-1 signal. Individuals presenting stem sprains, resulting from the marked development, produced gelatinous fibers with a clear cellulose layer. Initially, fluorescence microscopy demonstrated lignin deposition in the cell corner region having progressive deposition in the secondary wall of tracheary elements of the stem.

Esta pesquisa teve como objetivo avaliar a lignificação em caules jovens de plantas de Lophanthera lactescens Ducke crescidas in vitro. L. lactescens (Malpighiaceae) é uma espécie endêmica da Amazônia Brasileira que, além de propriedades medicinais, apresenta potencial para a indústria de produtos florestais. As plantas foram crescidas in vitro, usando-se meio nutritivo MS. As análises foram realizadas por microespectrometria no infravermelho, no modo reflectância difusa. Microscopia com fluorescência e testes histoquímicos de Wiesner e Maüle foram realizados para o acompanhamento do processo de lignificação. Os espectros registrados a partir da microscopia no infravermelho mostraram sinais em 1336 e 1246 cm-1, característicos de ligninas siringílica e guaiacílica. Indivíduos que apresentaram entorses no caule, em decorrência do acentuado desenvolvimento, produziram fibras gelatinosas, com a camada de celulose evidente. A microscopia com fluorescência evidenciou deposição de lignina inicialmente nos ângulos das células, com depósito progressivo ocorrendo na parede secundária dos elementos traqueais do caule.

Pollination of Byrsonima coccolobifolia: short-distance isolation and possible causes for low fruit production.

Byrsonima coccolobifolia is a tropical plant from the Malpighiaceae family, distributed in the neotropical savanna fields and pollinated by bees known as "collecting-oil bees". In this study, conducted in a Cerrado area located on a farm in the city of Silvânia, GO, the following hypothesis was tested: the greater the isolation degree of a plant, the lower its fruit production due to access difficulties for pollinators. Using a linear regression analysis, it was possible to relate the fruiting rate with the degree of isolation of each B. coccolobifolia individual and consequently it was found that the isolation had no influence on the pollination rate, an unexpected event that can be explained by the distance amongst individuals not being large enough to limit the movement of pollinators, or because cross-pollination was not the predominant form of reproduction, since the rate of cross-pollination was similar to self-pollination. It was also found that the proportion of fruits produced was lower than expected, a factor which may also have influenced the results.

Pollination of Byrsonima coccolobifolia: short-distance isolation and possible causes for low fruit production / Polinização de Byrsonima cocolobifolia: isolamento de curta distância e as possíveis causas para baixa produção de frutos

Byrsonima coccolobifolia is a tropical plant from the Malpighiaceae family, distributed in the neotropical savanna fields and pollinated by bees known as "collecting-oil bees". In this study, conducted in a Cerrado area located on a farm in the city of Silvânia, GO, the following hypothesis was tested: the greater the isolation degree of a plant, the lower its fruit production due to access difficulties for pollinators. Using a linear regression analysis, it was possible to relate the fruiting rate with the degree of isolation of each B. coccolobifolia individual and consequently it was found that the isolation had no influence on the pollination rate, an unexpected event that can be explained by the distance amongst individuals not being large enough to limit the movement of pollinators, or because cross-pollination was not the predominant form of reproduction, since the rate of cross-pollination was similar to self-pollination. It was also found that the proportion of fruits produced was lower than expected, a factor which may also have influenced the results.

Byrsonima coccolobifolia é uma planta tropical da família Malpighiaceae, distribuída no Cerrado e polinizada por abelhas conhecidas como "abelhas coletoras-de-óleo". No presente estudo, realizado em área de cerrado localizada em uma fazenda no município de Silvânia, GO, foi testada a hipótese de que quanto maior o grau de isolamento de uma planta, menor sua produção de frutos, devido à dificuldade de acesso dos polinizadores. Através de uma análise de regressão linear foi possível relacionar a taxa de frutificação ao grau de isolamento de cada indivíduo de B. coccolobifolia, e verificou-se que o isolamento não exerceu influência alguma na produção de frutos, fato inesperado que pode ser explicado pela distância entre os indivíduos não ser suficientemente grande a ponto de limitar o trânsito de polinizadores, ou pelo fato da polinização cruzada não ser a forma predominante de reprodução, uma vez que a taxa de polinização cruzada foi semelhante à de autopolinização. Verificou-se ainda que a proporção de frutos produzidos foi abaixo do esperado, fator que também pode ter influenciado os resultados.

Phenolic compounds from Byrsonima crassifolia L. bark: phytochemical investigation and quantitative analysis by LC-ESI MS/MS.

Phytochemical investigation of the methanolic extract of Byrsonima crassifolia's bark led to the isolation of 8 known phenolic compounds 5-O-galloylquinic acid, 3-O-galloylquinic acid, 3,4-di-O-galloylquinic acid, 3,5-di-O-galloylquinic acid, 3,4,5-tri-O-galloylquinic acid, (+)-epicatechin-3-gallate along with (+)-catechin and (+)-epicatechin. Due to their biological value, in the present study, a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, working in multiple reaction monitoring (MRM) mode, has been developed to quantify these compounds. B. crassifolia bark resulted in a rich source of phenolic compounds and particularly of galloyl derivates. The proposed analytical method is promising to be applied to other galloyl derivatives to quantify these bioactive compounds in raw material and final products.

Cloning and expression of GDP-D-mannose pyrophosphorylase gene and ascorbic acid content of acerola (Malpighia glabra L.) fruit at ripening stages.

Acerola (Malpighia glabra L.) is one of the richest natural sources of L-ascorbic acid (AsA; vitamin C). GDP-D-mannose pyrophosphorylase (GMP; EC 2.7.7.13) was found to play a major role in the proposed AsA biosynthetic pathway in plants, considering that Arabidopsis vtc1-1 mutant with point mutation in this gene has a highly reduced AsA content. GMP cDNA was isolated from acerola fruits, designated MgGMP, using rapid amplification of cDNA ends (RACE), and its expression was monitored during fruit ripening. The full-length cDNA was found to have an ORF of 1083bp encoding a polypeptide of 361 amino acids. In silico analysis of the predicted amino acid sequence showed a pI of 6.45 and molecular mass of 39.7kD. MgGMP showed over 80% amino acid sequence identity with other plant GMP homologues. The phylogenetic tree shows the close relation of MgGMP to the GMP of other plants as against those from parasite, yeasts and mammals. Southern analysis indicated that M. glabra contains not less than two copies of GMP genes. Northern blot analysis showed the transcript abundance of MgGMP in all the organs of acerola examined, with the fruit having the highest expression. The relative transcript abundance of MgGMP mRNA levels in the fruits changes as the ripening process progresses, with the unripe green fruits having the highest relative mRNA level, and the lowest was found in the fruits at advanced ripening stage. A strong correlation was also observed between the relative MgGMP mRNA levels and the AsA contents of acerola during fruit ripening.