Whole-plant and leaf determinants of growth rates in progenies of Genipa americana L. (Rubiaceae).

Genipa americana (Rubiaceae) is a fruit tree with broad phytogeographic domain and suitable for different silvicultural systems in the tropics. The knowledge associated with the relative growth rate of species such as G. americana, provides important guidelines for the effective establishment and survival of seedlings after planting in the field. In this study we investigated differences in growth, biomass allocation and photosynthesis of seedlings originating from different mother plants of G. americana in southern Bahia, Brazil. For this, we evaluated fifteen variables associated with carbon balance at the whole plant and leaf scales of twelve G. americana progenies. All seedlings grew over a period of 198 days under similar microclimatic conditions with approximately 65% full sun. Our results showed significant differences in the relative growth rates (RGR), with the highest and lowest mean values being 29.0 and 38.0 mg g-1 day-1, respectively. Differences in RGR between G. americana progenies were highly related to differences in biomass allocation at both whole plant and leaf scales. From a practical point of view, we demonstrate that the selection of mother plants to produce seedlings with higher growth rates, and consequently greater establishment capacity in field plantings, can be made from evaluations of growth and biomass allocation variables at the whole plant scale.

Long-term strict ant-plant mutualism identity characterises growth rate and leaf shearing resistance of an Amazonian myrmecophyte.

Over 125 million years of ant-plant interactions have culminated in one of the most intriguing evolutionary outcomes in life history. The myrmecophyte Duroia hirsuta (Rubiaceae) is known for its mutualistic association with the ant Myrmelachista schumanni and several other species, mainly Azteca, in the north-western Amazon. While both ants provide indirect defences to plants, only M. schumanni nests in plant domatia and has the unique behaviour of clearing the surroundings of its host tree from heterospecific plants, potentially increasing resource availability to its host. Using a 12-year survey, we asked how the continuous presence of either only M. schumanni or only Azteca spp. benefits the growth and defence traits of host trees. We found that the continuous presence of M. schumanni improved relative growth rates and leaf shearing resistance of Duroia better than trees with Azteca. However, leaf herbivory, dry matter content, trichome density, and secondary metabolite production were the same in all trees. Survival depended directly on ant association (> 94% of trees died when ants were absent). This study extends our understanding of the long-term effects of strict ant-plant mutualism on host plant traits in the field and reinforces the use of D. hirsuta-M. schumanni as a model system suitable for eco-co-evolutionary research on plant-animal interactions.

Anatomical, histochemical, and developmental approaches reveal the long-term functioning of the floral nectary in Tocoyena formosa (Rubiaceae).

Tocoyena formosa has a persistent floral nectary that continues producing nectar throughout flower and fruit development. This plant also presents an intriguing non-anthetic nectary derived from early-developing floral buds with premature abscised corolla. In this study, we characterize the structure, morphological changes, and functioning of T. formosa floral nectary at different developmental stages. We subdivided the nectary into four categories based on the floral and fruit development stage at which nectar production started: (i) non-anthetic nectary; (ii) anthetic nectary, which follows the regular floral development; (iii) pericarpial nectary, derived from pollinated flowers following fruit development; and (iv) post-anthetic nectary that results from non-pollinated flowers after anthesis. The nectary has a uniseriate epidermis with stomata, nectariferous parenchyma, and vascular bundles, with a predominating phloem at the periphery. The non-anthetic nectary presents immature tissues that release the exudate. The nectary progressively becomes more rigid as the flower and fruit develop. The main nectary changes during flower and fruit development comprised the thickening of the cuticle and epidermal cell walls, formation of cuticular epithelium, and an increase in the abundance of calcium oxalate crystals and phenolic cells near the vascular bundles. Projections of the outer periclinal walls toward the cuticle in the post-anthetic nectary suggest nectar reabsorption. The anatomical changes of the nectary allow it to function for an extended period throughout floral and fruit development. Hence, T. formosa nectary is a bivalent secretory structure that plays a crucial role in the reproductive and defensive interactions of this plant species.

Unlocking the Complete Chloroplast Genome of a Native Tree Species from the Amazon Basin, Capirona (Calycophyllum Spruceanum, Rubiaceae), and Its Comparative Analysis with Other Ixoroideae Species.

Capirona (Calycophyllum spruceanum Benth.) belongs to subfamily Ixoroideae, one of the major lineages in the Rubiaceae family, and is an important timber tree. It originated in the Amazon Basin and has widespread distribution in Bolivia, Peru, Colombia, and Brazil. In this study, we obtained the first complete chloroplast (cp) genome of capirona from the department of Madre de Dios located in the Peruvian Amazon. High-quality genomic DNA was used to construct libraries. Pair-end clean reads were obtained by PE 150 library and the Illumina HiSeq 2500 platform. The complete cp genome of C. spruceanum has a 154,480 bp in length with typical quadripartite structure, containing a large single copy (LSC) region (84,813 bp) and a small single-copy (SSC) region (18,101 bp), separated by two inverted repeat (IR) regions (25,783 bp). The annotation of C. spruceanum cp genome predicted 87 protein-coding genes (CDS), 8 ribosomal RNA (rRNA) genes, 37 transfer RNA (tRNA) genes, and one pseudogene. A total of 41 simple sequence repeats (SSR) of this cp genome were divided into mononucleotides (29), dinucleotides (5), trinucleotides (3), and tetranucleotides (4). Most of these repeats were distributed in the noncoding regions. Whole chloroplast genome comparison with the other six Ixoroideae species revealed that the small single copy and large single copy regions showed more divergence than inverted regions. Finally, phylogenetic analyses resolved that C. spruceanum is a sister species to Emmenopterys henryi and confirms its position within the subfamily Ixoroideae. This study reports for the first time the genome organization, gene content, and structural features of the chloroplast genome of C. spruceanum, providing valuable information for genetic and evolutionary studies in the genus Calycophyllum and beyond.

Root deformation affects mineral nutrition but not leaf gas exchange and growth of Genipa americana seedlings during the recovery phase after soil flooding / A deformação de raízes afeta a nutrição mineral mas não as trocas gasosas foliares e o crescimento de mudas de Genipa americana durante a fase de recuperação após o alagamento do solo

Root deformation (RD) caused by errors in the pricking out process are irreversible and very difficult to detect in container-grown seedlings at the time of planting in the field. The objective of this study was to evaluate the effects of RD on leaf gas exchange, growth, biomass allocation and mineral nutrition of G. americana seedlings during the recovery phase after soil flooding. Four-months-old seedlings, with and without RD, were flooded for 42 days and their recovery was evaluated 28 days after soil drainage. There were no significant interactions between RD and soil flooding for all leaf gas exchange, growth and mineral nutrition after soil drainage, with the exception of leaf P concentrations. In plants with no RD, the P concentration in leaves of non-flooded plants was significantly higher than that of plants with RD. Soil flooding and RD did not influence leaf or root N concentrations or whole-plant N content. RD increased the K concentration in the roots, but not in the leaves. Changes in the nutrient concentrations in leaves and roots indicate that RD may affect physiological performance of seedlings after planting in the field.

A deformação da raiz (RD) causada por erros no processo de repicagem é irreversível e difícil de detectar em mudas produzidas em embalagens no momento do plantio no campo. O objetivo deste estudo foi avaliar os efeitos do RD nas trocas gasosas foliares, crescimento, alocação de biomassa e nutrição mineral de mudas de G. americana na fase de recuperação após o alagamento do solo. Mudas com quatro meses de idade, com e sem RD, foram alagadas por 42 dias e a sua recuperação foi avaliada 28 dias após a drenagem do solo. Não houve interação significativa entre RD e alagamento do solo nas trocas gasosas foliares, crescimento e nutrição mineral após a drenagem, com exceção das concentrações de P foliar. Em plantas sem RD, a concentração de P nas folhas de plantas não alagadas foi significativamente maior que a das plantas com RD. O alagamento do solo e a RD não influenciaram as concentrações de N nas folhas e raízes, e no conteúdo de N na planta inteira. A RD aumentou a concentração de K nas raízes, mas não nas folhas. Alterações nas concentrações de nutrientes nas folhas e raízes indicam que a RD pode afetar o desempenho fisiológico das mudas após o plantio no campo.

Root deformation affects mineral nutrition but not leaf gas exchange and growth of Genipa americana seedlings during the recovery phase after soil flooding / A deformação de raízes afeta a nutrição mineral mas não as trocas gasosas foliares e o crescimento de mudas de Genipa americana durante a fase de recuperação após o alagamento do solo

Root deformation (RD) caused by errors in the pricking out process are irreversible and very difficult to detect in container-grown seedlings at the time of planting in the field. The objective of this study was to evaluate the effects of RD on leaf gas exchange, growth, biomass allocation and mineral nutrition of G. americana seedlings during the recovery phase after soil flooding. Four-months-old seedlings, with and without RD, were flooded for 42 days and their recovery was evaluated 28 days after soil drainage. There were no significant interactions between RD and soil flooding for all leaf gas exchange, growth and mineral nutrition after soil drainage, with the exception of leaf P concentrations. In plants with no RD, the P concentration in leaves of non-flooded plants was significantly higher than that of plants with RD. Soil flooding and RD did not influence leaf or root N concentrations or whole-plant N content. RD increased the K concentration in the roots, but not in the leaves. Changes in the nutrient concentrations in leaves and roots indicate that RD may affect physiological performance of seedlings after planting in the field.(AU)

A deformação da raiz (RD) causada por erros no processo de repicagem é irreversível e difícil de detectar em mudas produzidas em embalagens no momento do plantio no campo. O objetivo deste estudo foi avaliar os efeitos do RD nas trocas gasosas foliares, crescimento, alocação de biomassa e nutrição mineral de mudas de G. americana na fase de recuperação após o alagamento do solo. Mudas com quatro meses de idade, com e sem RD, foram alagadas por 42 dias e a sua recuperação foi avaliada 28 dias após a drenagem do solo. Não houve interação significativa entre RD e alagamento do solo nas trocas gasosas foliares, crescimento e nutrição mineral após a drenagem, com exceção das concentrações de P foliar. Em plantas sem RD, a concentração de P nas folhas de plantas não alagadas foi significativamente maior que a das plantas com RD. O alagamento do solo e a RD não influenciaram as concentrações de N nas folhas e raízes, e no conteúdo de N na planta inteira. A RD aumentou a concentração de K nas raízes, mas não nas folhas. Alterações nas concentrações de nutrientes nas folhas e raízes indicam que a RD pode afetar o desempenho fisiológico das mudas após o plantio no campo.(AU)

Antitumoral and Anticholinesterasic Activities of the Seven Species from Rubiaceae.

BACKGROUND: The genus Psychotria and Palicourea are reported as a source of alkaloids and iridoids, which exhibit biological activities. This study aimed to evaluate antiproliferative and anticholinesterase activities and quantification of the alkaloids of seven species among the genus found in Mato Grosso do Sul region in Brazil. METHODS: Concentrations of alkaloids were measured spectrophotometrically. The extracts were submitted to antiproliferative activity against ten cell lines. The anticholinesterase activity of the extracts was developed using brain structures of male Wistar rats: cerebral cortex, hippocampus, hypothalamus and striatum by the Ellman method. RESULTS: Alkaloids from Psychotria and Palicourea species were quantified which showed values of 47.6 to 21.9 µg/g. Regarding the antiproliferative potential, Palicourea crocea demonstrated selectivity against the 786-0 cell line (GI50: 22.87 µg/mL). Psychotria leiocarpa inhibited cell growth against OVCAR-3 (GI50: 3.28 µg/mL), K-562 (GI50: 5.26 µg/mL), HaCaT (GI50: 27.20 µg/mL), PC-3 (GI50: 34.92 µg/mL), MCF-7 (GI50: 35.80 µg/mL) and P. capillacea showed activity against OVCAR-3 (GI50: 2.33 µg/ml) and U251 (GI50: 16.66 µg/ml). The effect of acetylcholinesterase inhibition was more effective in the hippocampus, demonstrating inhibition for Paliourea crocea, Psychotria deflexa, P. brachybotrya and P. leiocarpa of 70%, 57%, 50% and 40%, respectively, followed by P. poeppigiana and P. capillacea, inhibiting 21%, compared to the control. CONCLUSION: Herein, the present work showed for the first time, anticholinesterasic and antiproliferative activities of extracts of Palicourea and Psychotria seem to be mainly associated with the levels of alkaloids in the leaves of these species.

Germination and initial development of Simira gardneriana M.R. Barbosa & Peixoto (Rubiaceae) seedlings under different temperatures and salinity levels / Germinação e desenvolvimento inicial de plântulas de Simira gardneriana M.R. Barbosa & Peixoto (Rubiaceae) em diferentes temperaturas e níveis de salinidade

Simira gardneriana M.R. Barbosa & Peixoto (Rubiaceae) is a species exclusive to the Caatinga biome. It is used as forage, firewood, and in the construction of fence posts. Species from this biome are constantly subjected to abiotic stresses, including salt and thermal stress. Thus, we evaluated the effect of irrigation water salinity on the germination and initial development of S. gardneriana seedlings at different temperatures. The experiment used seeds with an initial moisture content of about 12.32%, arranged in a factorial scheme with eight levels of salinity, obtained by the addition of sodium chloride (NaCl), diluted in distilled water at concentrations of 0.0 (control), 1.5, 2.5, 3.5, 4.5, 5.5, 6.5, and 7.5 dS m-1, and the four temperatures 25, 30, 35, and 20-30°C, in four replicates of 25 seeds each. Salinity and temperature effects were evaluated based on germination, germination speed index, seedling shoot length, root length, shoot dry matter, root dry matter, and total dry matter. Increasing water salinity levels interfered with all variables analyzed in the germination of S. gardneriana seeds, especially at a temperature of 35°C. At temperatures of 25 and 30°C, seed germination and initial development were less affected by salinity. Simira gardneriana seeds are sensitive to salinity caused by the increase in NaCl from 1.5 dS m-1, which indicates a low tolerance of this species to saline environments. Our results show that the negative effect on the germination and initial development of S. gardneriana seedlings can be explained by the increase in salt levels and temperature, reducing their capacity to absorb water and nutrients, which indicates sensitivity responses of this species to salinity and thermal stress.

Simira gardneriana M.R. Barbosa & Peixoto (Rubiaceae), é uma espécie exclusiva do bioma Caatinga, utilizada como forrageira, e a madeira usada para lenha e estacas na construção de cercas. Espécies desse bioma são submetidas, constantemente, a estresses abióticos, entre eles o salino e o térmico. Dessa forma, objetivou-se avaliar o efeito da salinidade da água de irrigação na germinação e no desenvolvimento inicial de plântulas de S. gardneriana em diferentes temperaturas. Nesse estudo foram utilizadas sementes com teor de água inicial em torno de 12,32% às quais foram arranjadas em esquema fatorial com oito níveis de salinidades, obtidos a partir da adição de cloreto de sódio (NaCl) e diluídos em água destilada nas concentrações de 0,0 (testemunha); 1,5; 2,5; 3,5; 4,5; 5,5; 6,5 e 7,5 dS m-1 e quatro temperaturas de 25, 30, 35 e 20-30 °C, em quatro repetições de 25 sementes cada. Os efeitos da salinidade e da temperatura foram avaliados por meio da germinação, índice de velocidade de germinação, comprimento da parte aérea e da raiz, massa seca da parte aérea e da raiz e total de plântulas. O aumento dos níveis de salinidade da água interfere em todas as variáveis analisadas na germinação de sementes de S. gardneriana, principalmente sob a temperatura de 35 °C. Nas temperaturas de 25 e 30 °C a germinação e o desenvolvimento inicial das sementes são menos afetados pela salinidade. As sementes de S. gardneriana são sensíveis à salinidade causada pelo aumento do NaCl a partir de 1,5 dS m-1, o que indica baixa tolerância desta espécie em ambientes salinos. Os resultados obtidos neste trabalho mostram que o efeito negativo na germinação e desenvolvimento inicial de plântulas de S. gardneriana pode ser explicado pelo aumento dos níveis salinos e temperatura, reduzindo o poder da planta em absorver água e nutrientes, o que indica respostas de sensibilidade dessa espécie a salinidade e ao estresse térmico.

Germination and initial development of Simira gardneriana M.R. Barbosa & Peixoto (Rubiaceae) seedlings under different temperatures and salinity levels / Germinação e desenvolvimento inicial de plântulas de Simira gardneriana M.R. Barbosa & Peixoto (Rubiaceae) em diferentes temperaturas e níveis de salinidade

Simira gardneriana M.R. Barbosa & Peixoto (Rubiaceae) is a species exclusive to the Caatinga biome. It is used as forage, firewood, and in the construction of fence posts. Species from this biome are constantly subjected to abiotic stresses, including salt and thermal stress. Thus, we evaluated the effect of irrigation water salinity on the germination and initial development of S. gardneriana seedlings at different temperatures. The experiment used seeds with an initial moisture content of about 12.32%, arranged in a factorial scheme with eight levels of salinity, obtained by the addition of sodium chloride (NaCl), diluted in distilled water at concentrations of 0.0 (control), 1.5, 2.5, 3.5, 4.5, 5.5, 6.5, and 7.5 dS m-1, and the four temperatures 25, 30, 35, and 20-30°C, in four replicates of 25 seeds each. Salinity and temperature effects were evaluated based on germination, germination speed index, seedling shoot length, root length, shoot dry matter, root dry matter, and total dry matter. Increasing water salinity levels interfered with all variables analyzed in the germination of S. gardneriana seeds, especially at a temperature of 35°C. At temperatures of 25 and 30°C, seed germination and initial development were less affected by salinity. Simira gardneriana seeds are sensitive to salinity caused by the increase in NaCl from 1.5 dS m-1, which indicates a low tolerance of this species to saline environments. Our results show that the negative effect on the germination and initial development of S. gardneriana seedlings can be explained by the increase in salt levels and temperature, reducing their capacity to absorb water and nutrients, which indicates sensitivity responses of this species to salinity and thermal stress.(AU)

Simira gardneriana M.R. Barbosa & Peixoto (Rubiaceae), é uma espécie exclusiva do bioma Caatinga, utilizada como forrageira, e a madeira usada para lenha e estacas na construção de cercas. Espécies desse bioma são submetidas, constantemente, a estresses abióticos, entre eles o salino e o térmico. Dessa forma, objetivou-se avaliar o efeito da salinidade da água de irrigação na germinação e no desenvolvimento inicial de plântulas de S. gardneriana em diferentes temperaturas. Nesse estudo foram utilizadas sementes com teor de água inicial em torno de 12,32% às quais foram arranjadas em esquema fatorial com oito níveis de salinidades, obtidos a partir da adição de cloreto de sódio (NaCl) e diluídos em água destilada nas concentrações de 0,0 (testemunha); 1,5; 2,5; 3,5; 4,5; 5,5; 6,5 e 7,5 dS m-1 e quatro temperaturas de 25, 30, 35 e 20-30 °C, em quatro repetições de 25 sementes cada. Os efeitos da salinidade e da temperatura foram avaliados por meio da germinação, índice de velocidade de germinação, comprimento da parte aérea e da raiz, massa seca da parte aérea e da raiz e total de plântulas. O aumento dos níveis de salinidade da água interfere em todas as variáveis analisadas na germinação de sementes de S. gardneriana, principalmente sob a temperatura de 35 °C. Nas temperaturas de 25 e 30 °C a germinação e o desenvolvimento inicial das sementes são menos afetados pela salinidade. As sementes de S. gardneriana são sensíveis à salinidade causada pelo aumento do NaCl a partir de 1,5 dS m-1, o que indica baixa tolerância desta espécie em ambientes salinos. Os resultados obtidos neste trabalho mostram que o efeito negativo na germinação e desenvolvimento inicial de plântulas de S. gardneriana pode ser explicado pelo aumento dos níveis salinos e temperatura, reduzindo o poder da planta em absorver água e nutrientes, o que indica respostas de sensibilidade dessa espécie a salinidade e ao estresse térmico.(AU)

Drying and reduction in sensitivity to desiccation of seeds of Alibertia edulis: the influence of fruit ripening stage.

The intense environmental degradation in tropical regions suitable for agriculture has decreased native forest populations of plants with important fruits and medicinal properties. Alibertia edulis is a native tree from the Brazilian Cerrado. Knowledge about the effects of drying and storage on the physiological behavior of its seeds may aid in its sustainable exploitation and conservation. The goal of the present study was to determine which fruit ripening stage results in A. edulis seeds with higher tolerance to desiccation, and to investigate the effectiveness of polyethylene glycol (PEG) induced osmotic stress in combination with abscisic acid (ABA) in increasing seed desiccation tolerance during storage. Seeds were dried in activated silica gel (fast) or under ambient conditions (slow). Seeds originating from mid-ripe or fully ripe fruits exhibited better physiological performance than those obtained from green fruits. Slow drying resulted in seeds with high physiological potential. Seeds may be stored for up to 180 days without losing viability when treated with -0.73 MPa PEG without ABA.

Ontogenesis secretion and senescence of Tocoyena bullata (Vell.) Mart. (Rubiacaeae) colleters.

Colleters are secretory structure present on many families including Rubiaceae. Particular characteristics have been described about colleters secretory cells, however senescence process are still under debate. Tocoyena bullata (Vell.) Mart. (Rubiaceae) shoot apex were collected at Jardim Botânico do Rio de Janeiro, RJ/Brazil. Stipules were separated and fragments were fixed in 2.5% glutaraldehyde and 4.0% formaldehyde in 0.05 m sodium cacodylate buffer, pH 7.2, post fixed in 1.0% osmium tetroxide in the same buffer, dehydrated in acetone, critical-point-drying, sputtered coated and observed. For light microscopy fragments were fixed and dehydrated, infiltrated with historesin and stained with 1% toluidine blue. For transmission electron microscopy, the samples were infiltrated with Epoxi resin. Colleters are present on stipule adaxial surface. On the beginning of development, these structures are recognized as small projections. Later on, colleters differentiated and secrete by cuticle rupture. The colleters senescence occurs in a concomitant and indissoluble way of programmed cell death. Ultrastructural analyses during the process strongly suggest the senescence is based on a non-autolitic programmed cell death. T. bullata colleters, present at stipule abaxial surface are cylindrical secretory structures. Colleters secretory cells originated as stipule projections; differentiate; secrete and senesce by programmed cell death. The secretion and the cell dead occurs in a concomitantly and indissoluble way.

Buttonweed emergence as affected by seed burial depth and straw on the soil surface

Knowledge of the effects of seed burial depth and the presence of straw on the soil surface on weed seedling emergence provides useful information for the development of weed management tactics. Buttonweed (Borreria densiflora DC.) is a troublesome weed that occurs in large infestations in soybean and sugarcane crops from north-central Brazil. This study investigated buttonweed emergence at different seed burial depths and straw amounts present on the soil surface. The experiment was conducted in greenhouse conditions, under a factorial design between four seed burial depths and four amounts of surface straw. Percent seedling emergence and fresh biomass (g) were evaluated at twenty-five days after installation (DAI). Greater buttonweed emergence occurred in seeds that were placed on the soil surface either without surface straw or with up to 1,000 kg ha1 of straw on the soil surface. With 4,000 kg ha1 of surface straw, buttonweed emergence was prevented when seeds were placed at a depth of 0.5 cm or deeper in the soil. These data indicated emergence of this weed species was greater at depths near the soil surface and in soils with the least amounts of surface straw. Information generated in this study provides a starting point for the development of knowledge for understanding the biology of buttonweed emergence and its population dynamics. Such information may be directly transmitted to growers and lays the groundwork for an integrated management approach for this weed species.

Buttonweed emergence as affected by seed burial depth and straw on the soil surface

Knowledge of the effects of seed burial depth and the presence of straw on the soil surface on weed seedling emergence provides useful information for the development of weed management tactics. Buttonweed (Borreria densiflora DC.) is a troublesome weed that occurs in large infestations in soybean and sugarcane crops from north-central Brazil. This study investigated buttonweed emergence at different seed burial depths and straw amounts present on the soil surface. The experiment was conducted in greenhouse conditions, under a factorial design between four seed burial depths and four amounts of surface straw. Percent seedling emergence and fresh biomass (g) were evaluated at twenty-five days after installation (DAI). Greater buttonweed emergence occurred in seeds that were placed on the soil surface either without surface straw or with up to 1,000 kg ha1 of straw on the soil surface. With 4,000 kg ha1 of surface straw, buttonweed emergence was prevented when seeds were placed at a depth of 0.5 cm or deeper in the soil. These data indicated emergence of this weed species was greater at depths near the soil surface and in soils with the least amounts of surface straw. Information generated in this study provides a starting point for the development of knowledge for understanding the biology of buttonweed emergence and its population dynamics. Such information may be directly transmitted to growers and lays the groundwork for an integrated management approach for this weed species.(AU)

Influence of the stage of ripeness on the composition of iridoids and phenolic compounds in genipap (Genipa americana L.).

Genipap fruits, native to the Amazon region, were classified in relation to their stage of ripeness according to firmness and peel color. The influence of the part of the genipap fruit and ripeness stage on the iridoid and phenolic compound profiles was evaluated by HPLC-DAD-MS(n), and a total of 17 compounds were identified. Geniposide was the major compound in both parts of the unripe genipap fruits, representing >70% of the total iridoids, whereas 5-caffeoylquinic acid was the major phenolic compound. In ripe fruits, genipin gentiobioside was the major compound in the endocarp (38%) and no phenolic compounds were detected. During ripening, the total iridoid content decreased by >90%, which could explain the absence of blue pigment formation in the ripe fruits after their injury. This is the first time that the phenolic compound composition and iridoid contents of genipap fruits have been reported in the literature.

Phenological and geographical influence in the concentration of selected bioactive 4-phenylcoumarins and chlorogenic acid in Hintonia latiflora leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Hintonia latiflora is a Mexican medicinal plant with well-documented ethnomedical record comprising more than 400 years; in modern Mexico is used for treating several maladies such as diabetes and gastric ulcers. Although the pharmacological actions of the stem-bark and leaves have been demonstrated, the phenological and geographical effect on the concentration of active principles remains unexplored. AIM OF THE STUDY: The main goals of this study were to analyze the amount of selected 4-phenylcoumarins and chlorogenic acid in the leaves in order to assess the best harvesting period, and consequently their pharmacological efficacy. In addition, the preclinical antidiabetic efficacy of the infusion of the leaves was corroborated using standard pharmacological tests. MATERIALS AND METHODS: The aqueous extracts from the leaves of Hintonia latiflora were prepared by infusion. For phenological and geographical comparison, leaves of Hintonia latiflora were collected in two different regions in Chihuahua and Michoacán. The material was analyzed by UPLC applying an analytical method that developed and validated for this purpose following the ICH guidelines. Investigation of the antidiabetic action was accomplished using an acute hypoglycemic test and oral glucose and sucrose tolerance tests. RESULTS: The validated analytical method was successfully applied for quantifying chlorogenic acid (1) and 4-phenylcoumarins (2-5) in the leaves of 12 different batches (1-12) during one-year period, and seven different batches for each geographical region; the concentration of the metabolites at the phenological cycle was significantly different, their concentration increased during the pre-senescence phase whereas in the leaf renovation stage the highest concentration of 2-5 was reached. The overall analysis of the active compounds concentration between the two populations investigated seems to be less important than the phenological variations. The aqueous extract of the leaves of Hintonia latiflora exerted its antidiabetic effect by different mechanisms showing comparable effect to the organic extract. CONCLUSIONS: The findings of the present investigation reveal that the best harvest season for the leaves of Hintonia latiflora is between the leaves renovation and senescence stages avoiding the flowering period. In addition, no significant differences were found among the two different geographical populations analyzed. The infusions of the leaves, rich in 4-phenylcoumarins and chlorogenic acid, showed comparable antidiabetic action than the organic extract.

Morphology, ecophysiology and germination of seeds of the neotropical tree Alibertia patinoi (Rubiaceae).

Alibertia patinoi (Rubiaceae) is of economic and cultural importance for communities in the Colombian Pacific and Amazon regions, where it is cultivated and mature fruits are highly appreciated and consumed. Since there is a lack of knowledge of the seed physiology of this species, we describe here the germination behavior and morphometry of seeds of Alibertia patinoi, and relate them to its habitat. Fruits were collected from a mixed food crop and a commercial plantation in Guaimía village, Buenaventura, Colombia, a tropical rain forest area. We measured length, width, thickness, mass (n = 1 400), and moisture content of seeds (n = 252). Primary dormancy tests were conducted (n = 200), followed by imbibition (n=252) and germination dynamics, under different conditions of light and temperature specific to understory and forest clearings (n = 300 seeds). Finally, seed storage behavior was established (n = 100 seeds). We observed that size and mass of seeds had a narrow range of values that did not differ within or among fruits and that the species did not exhibit primary dormancy. The seeds are recalcitrant, and recently harvested seeds exhibited higher seed moisture content (ca. 44%) and continuous metabolism. The seed germination percentage was observed to be higher under the specific dense canopy forest light and temperature conditions; furthermore, neither enriched far-red light nor darkness conditions inhibited germination. We concluded that rapid germination could be the establishment strategy of this species. Also, the physiological traits (i.e., rapid germination rate, low germination requirements, absence of primary dormancy, and recalcitrant behavior) and seed size and mass, suggest that A. patinoi is adapted to conditions of mature tropical rain forests.

Morphology, ecophysiology and germination of seeds of the Neotropical tree Alibertia patinoi (Rubiaceae) / Morfología, ecofisiología y germinación de semillas del árbol neotropical

Alibertia patinoi (Rubiaceae). Alibertia patinoi (Rubiaceae) is of economic and cultural importance for communities in the Colombian Pacific and Amazon regions, where it is cultivated and mature fruits are highly appreciated and consumed. Since there is a lack of knowledge of the seed physiology of this species, we describe here the germination behavior and morphometry of seeds of Alibertia patinoi, and relate them to its habitat. Fruits were collected from a mixed food crop and a commercial plantation in Guaimía village, Buenaventura, Colombia, a tropical rain forest area. We measured length, width, thickness, mass (n=1 400), and moisture content of seeds (n=252). Primary dormancy tests were conducted (n=200), followed by imbibition (n=252) and germination dynamics, under different conditions of light and temperature specific to understory and forest clearings (n=300 seeds). Finally, seed storage behavior was established (n=100 seeds). We observed that size and mass of seeds had a narrow range of values that did not differ within or among fruits and that the species did not exhibit primary dormancy. The seeds are recalcitrant, and recently harvested seeds exhibited higher seed moisture content (ca. 44%) and continuous metabolism. The seed germination percentage was observed to be higher under the specific dense canopy forest light and temperature conditions; furthermore, neither enriched far-red light nor darkness conditions inhibited germination. We concluded that rapid germination could be the establishment strategy of this species. Also, the physiological traits (i.e., rapid germination rate, low germination requirements, absence of primary dormancy, and recalcitrant behavior) and seed size and mass, suggest that A. patinoi is adapted to conditions of mature tropical rain forests.

La germinación y morfometría de las semillas de Alibertia patinoi se describen y se relacionan con el hábitat de la especie, el Bosque Pluvial Tropical Americano. Se midió la longitud, ancho, grosor, peso y contenido de humedad de las semillas. Se realizaron pruebas de latencia, imbibición y dinámica de la germinación con diferentes condiciones de luz y temperatura específicas de claros y de sotobosque cubierto por un dosel denso. Además, se estableció el comportamiento de almacenamiento de las semillas. El tamaño y peso de las semillas no difieren dentro ni entre frutos. Las semillas recién cosechadas no tienen latencia primaria, presentan alto contenido de humedad (ca. 44%), metabolismo continuo y son recalcitrantes. La germinación tuvo porcentajes altos y en condiciones de luz y temperatura específicas de bosque con follaje denso, y ni la luz enriquecida con rojo lejano ni la oscuridad inhibieron la germinación. Se concluye que la germinación rápida podría ser la estrategia de establecimiento de esta especie. Además, los rasgos fisiológicos, el tamaño y peso de las semillas, sugieren que A. patinoi está adaptada a las condiciones del bosque lluvioso tropical maduro y se comporta como una especie que no es pionera.

Mechanism and control of Genipa americana seed germination.

Genipa americana (Rubiaceae) is important for restoration of riparian forest in the Brazilian Cerrado. The objective was to characterize the mechanism and control of germination of G. americana to support uniform seedling production. Morphology and morphometrics of seeds, embryo and endosperm were assessed by light and scanning electron microscopy during germination. Imbibition and germination curves were generated and over the same time interval endosperm digestion and resistance were measured by puncture force analysis and activity assay of endo-ß-mannanase (EBM) in water and in abscisic acid (ABA). The gene encoding for EBM was partially cloned and its expression monitored by quantitative real-time-polymerase chain reaction. Embryos displayed growth prior to radicle protrusion. A two-phase increase in EBM activity coincided with the two stages of weakening of the micropylar endosperm. The second stage also coincided with growth of the embryo prior to radicle protrusion. Enzyme activity was initiated in the micropylar endosperm but spread to the lateral endosperm. ABA completely inhibited germination by inhibiting embryo growth, the second stage of weakening and expression of the EBM gene, but EBM activity was not significantly inhibited. This suggests that a specific isoform of the enzyme is involved in endosperm weakening. EBM may cause a general 'softening' of micropylar endosperm cell walls, allowing the embryo to puncture the endosperm as the driving force of the decrease in puncture force.

TNT removal from culture media by three commonly available wild plants growing in the Caribbean.

Plants growing in the Caribbean, Rubia tinctorum, Lippia dulcis and Spermacoce remota, were used in vitro to remove TNT from culture media. Plants were found to be resistant to high TNT levels. S. remota was able to remove TNT in less than 48 h. Part of the TNT was physically removed from the culture media by evaporation.

Chloroplast DNA phylogeography of a distylous shrub (Palicourea padifolia, Rubiaceae) reveals past fragmentation and demographic expansion in Mexican cloud forests.

Several phylogeographic studies in northern Mesoamerica have examined the influence of Pleistocene glaciations on the genetic structure of temperate tree species with their southern limit by the contact zone between species otherwise characteristic of North or South America, but few have featured plant species that presumably colonized northern Mesoamerica from South America. A phylogeographical study of Palicourea padifolia, a fleshy-fruited, bird dispersed distylous shrub, was conducted to investigate genetic variation at two chloroplast regions (trnS-trnG and rpl32-trnL) across cloud forest areas to determine if such patterns are consistent with the presence of Pleistocene refugia and/or with the historical fragmentation of the Mexican cloud forests. We conducted population and spatial genetic analyses as well as phylogenetic and isolation with migration analyses on 122 individuals from 22 populations comprising the distribution of P. padifolia in Mexico to gain insight of the evolutionary history of these populations. Twenty-six haplotypes were identified after sequencing 1389 bp of chloroplast DNA. These haplotypes showed phylogeographic structure (N(ST) = 0.508, G(ST) = 0.337, N(ST) > G(ST), P < 0.05), including a phylogeographic break at the Isthmus of Tehuantepec, with private haplotypes at either side of the isthmus, and a divergence time of the split in the absence of gene flow dating back c. 309,000-103,000 years ago. The patterns of geographic structure found in this study are consistent with past fragmentation and demographic range expansion, supporting the role of the Isthmus of Tehuantepec as a biogeographical barrier in the dispersal of P. padifolia. Our data suggest that P. padifolia populations were isolated throughout glacial cycles by the Isthmus of Tehuantepec, accumulating genetic differences due to the lack of migration across the isthmus in either direction, but the results of our study are not consistent with the existence of the previously proposed Pleistocene refugia for rain forest plant species in the region.