Species Limits In The Kielmeyera Coriacea Complex (calophyllaceae) - A Multidisciplinary Approach

Kielmeyera coriacea is widely known as one of the most peculiar species of the Cerrado, a savanna-like vegetation of Central Brazil, and it has been the subject of several studies, some including a pharmacological focus. Together with related taxa, it is usually treated as the 'K. coriacea complex'. The genus was revised approximately 30 years ago. However, the revision could not clarify the situation of this complex, and the problematic taxonomic circumscription raises doubts about various scientific studies on plants identified as K. coriacea. In the present paper, we use a multidisciplinary approach in an attempt to understand the species complex and the causes of the current taxonomic confusion better and to provide diagnostic characters to support correct identifications in the group. A genetic population approach was applied using microsatellite markers. Macromorphological, anatomical and micromorphological characters with diagnostic power were investigated in fresh plant material and herbarium specimens. The molecular analysis supports the circumscription of two species in the complex, but they were found to share various morphological and anatomical characters, and hybrid individuals were more similar to K. coriacea than to K. grandiflora. The most powerful diagnostic characters found were: the colour of fresh leaves, the prominence of the inter-secondary veins in fresh leaves, the presence and distribution of phenolic compounds evidenced by formalin-ferrous sulphate staining and the micromorphological pattern of epicuticular structures on the surfaces of leaves related to the colour of the fresh leaves. In spite of occasional hybridization, at least two species should be recognized in the K. coriacea complex. The surest way to identify these species is with fresh material. 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Rooting Capacity Of Maytenus Muelleri Schwacke Cuttings With Indolebutyric Acid Application Related To Anatomical Aspects [capacidade De Enraizamento De Estacas De Maytenus Muelleri Schwacke Com A Aplicação De ácido Indol Butírico Relacionada Aos Aspectos Anatômicos]

"Espinheira-santa" (Maytenus muelleri - Celastraceae) is a medicinal plant native to Southern Brazil, the leaves of which are traditionally used in popular medicine for the treatment of stomach ulcers and other gastric problems. There are few published studies about seedling production and vegetative propagation techniques for this species. The propagation of "espinheira-santa" by cuttings could be an efficient method to obtain homogeneous material, with desirable genetic characteristics, produced from selected mother plants. This paper aimed to study the effects of indolebutyric acid (IBA) application, in solution and in powder, on the rooting of "espinheira-santa" cuttings, collected in four seasons (April/2005 to January/2006), as well as to investigate, by means of cutting anatomical and histochemical analyses, the presence of possible impediments to adventitious rooting initiation. Cuttings from branches of six-year mother plants grown at "Estação Experimental do Canguiri", Pinhais, Paraná State, Brazil, were collected and treated with IBA (0, 1500, 3000 mg L-1 or mg kg-1) in alcoholic solution (50% v/v) and in powder. After 365 days, the percentages of rooted and dead cuttings, the number and mean length of roots/cutting were evaluated. Anatomical and histochemical analyses were performed with lugol and ferric chloride. The most promising season for rooting was Summer/2006, with 62.50% of rooting for the control treatment, due to the lesser lignification degree of branches in intense vegetative growth period. The mean number of roots/cutting was 6.94 (solution) and the mean length of roots/cutting was 4.82 cm in that same season. The applied IBA concentrations were not efficient in inducing root growth, regardless of the application method. An almost continuous layer of fibers and stone cells was detected, constituting an anatomical barrier for rooting induction. The histochemical tests revealed the presence of starch and phenolic compounds in cuttings, in all seasons. 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Molecular And Biochemical Characterization Of Caffeine Synthase And Purine Alkaloid Concentration In Guarana Fruit

Guarana seeds have the highest caffeine concentration among plants accumulating purine alkaloids, but in contrast with coffee and tea, practically nothing is known about caffeine metabolism in this Amazonian plant. In this study, the levels of purine alkaloids in tissues of five guarana cultivars were determined. Theobromine was the main alkaloid that accumulated in leaves, stems, inflorescences and pericarps of fruit, while caffeine accumulated in the seeds and reached levels from 3.3% to 5.8%. In all tissues analysed, the alkaloid concentration, whether theobromine or caffeine, was higher in young/immature tissues, then decreasing with plant development/maturation. Caffeine synthase activity was highest in seeds of immature fruit. A nucleotide sequence (PcCS) was assembled with sequences retrieved from the EST database REALGENE using sequences of caffeine synthase from coffee and tea, whose expression was also highest in seeds from immature fruit. The PcCS has 1083 bp and the protein sequence has greater similarity and identity with the caffeine synthase from cocoa (BTS1) and tea (TCS1). A recombinant PcCS allowed functional characterization of the enzyme as a bifunctional CS, able to catalyse the methylation of 7-methylxanthine to theobromine (3,7-dimethylxanthine), and theobromine to caffeine (1,3,7-trimethylxanthine), respectively. Among several substrates tested, PcCS showed higher affinity for theobromine, differing from all other caffeine synthases described so far, which have higher affinity for paraxanthine. When compared to previous knowledge on the protein structure of coffee caffeine synthase, the unique substrate affinity of PcCS is probably explained by the amino acid residues found in the active site of the predicted protein. © 2014 Elsevier Ltd. All rights reserved.1052536Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., Basic local alignment search tool (1990) J. Mol. 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A Model System To Study The Lignification Process In Eucalyptus Globulus

Recalcitrance of plant biomass is closely related to the presence of the phenolic heteropolymer lignin in secondary cell walls, which has a negative effect on forage digestibility, biomass-to-biofuels conversion and chemical pulping. The genus Eucalyptus is the main source of wood for pulp and paper industry. However, when compared to model plants such as Arabidopsis thaliana and poplar, relatively little is known about lignin biosynthesis in Eucalyptus and only a few genes were functionally characterized. An efficient, fast and inexpensive in vitro system was developed to study lignification in Eucalyptus globulus and to evaluate the potential role of candidate genes in this biological process. Seedlings were grown in four different conditions, in the presence or absence of light and with or without sucrose in the growth medium, and several aspects of lignin metabolism were evaluated. Our results showed that light and, to a lesser extent, sucrose induced lignin biosynthesis, which was followed by changes in S/G ratio, lignin oligomers accumulation and gene expression. In addition, higher total peroxidase activity and differential isoperoxidase profile were observed when seedlings were grown in the presence of light and sucrose. Peptide sequencing allowed the identification of differentially expressed peroxidases, which can be considered potential candidate class III peroxidases involved in lignin polymerization in E. globulus. © 2014 Scandinavian Plant Physiology Society.15211731Barcelo, A.R., Aznar-Asensio, G.J., Coniferyl alcohol oxidase activity of a cell-wall-located class III peroxidase (1999) Aust J Plant Physiol, 26, pp. 411-419Barcelo, A.R., Munoz, R., Sabater, F., Lupin peroxidases. 1. Isolation and characterization of cell wall-bound isoperoxidase activity (1987) Physiol Plant, 71, pp. 448-454Boerjan, W., Ralph, J., Baucher, M., Lignin biosynthesis (2003) Annu Rev Plant Biol, 54, pp. 519-546Bonawitz, N., Chapple, C., The genetics of lignin biosynthesis: connecting genotype to phenotype (2010) Annu Rev Genet, 44, pp. 337-363Cesarino, I., Araújo, P., Domingues, A., An overview of lignin metabolism and its effect on biomass recalcitrance (2012) Braz J Bot, 35, pp. 303-311Cesarino, I., Araújo, P., Sampaio Mayer, J., Paes Leme, A., Mazzafera, P., Enzymatic activity and proteomic profile of class III peroxidases during sugarcane stem development (2012) Plant Physiol Biochem, 55, pp. 66-76Cesarino, I., Araújo, P., Paes Leme, A., Creste, S., Mazzafera, P., Suspension cell culture as a tool for the characterization of class III peroxidases in sugarcane (2013) Plant Physiol Biochem, 62, pp. 1-10Cetinkol, O., Smith-Moritz, A., Cheng, G., Lao, J., George, A., Hong, K., Henry, R., Holmes, B., Structural and chemical characterization of hardwood from tree species with applications as bioenergy feedstocks (2012) PLoS One, 7, pp. e52820Chen, F., Dixon, R., Lignin modification improves fermentable sugar yields for biofuel production (2007) Nat Biotechnol, 25, pp. 759-761Chen, Y.-A., Shin, J.-W., Liu, Z.-H., Effect of light on peroxidase and lignin synthesis in mungbean hypocotyls (2002) Plant Physiol Biochem, 40, pp. 33-39Cheng, H., Zhang, Q., Guo, D., Genes that respond to H2O2 are also evoked under light in Arabidopsis (2012) Mol Plant, 6, pp. 226-228Cosio, C., Dunand, C., Specific functions of individual class III peroxidase genes (2009) J Exp Bot, 60, pp. 391-408Cosio, C., Dunand, C., Transcriptome analysis of various flower and silique development stages indicates a set of class III peroxidase genes potentially involved in pod shattering in Arabidopsis thaliana (2010) BMC Genomics, 11, p. 528Cosio, C., Vuillemin, L., De Meyer, M., Kevers, C., Penel, C., Dunand, C., An anionic class III peroxidase from zucchini may regulate hypocotyl elongation through its auxin oxidase activity (2009) Planta, 229, pp. 823-836Demura, T., Fukuda, H., Transcriptional regulation in wood formation (2007) Trends Plant Sci, 12, pp. 64-70Ferri, M., Righetti, L., Tassoni, A., Increasing sucrose concentrations promote phenylpropanoid biosynthesis in grapevine cell cultures (2011) J Plant Physiol, 168, pp. 189-195Foucart, C., Paux, E., Ladouce, N., San-Clemente, H., Grima-Pettenati, J., Sivadon, P., Transcript profiling of a xylem vs phloem cDNA subtractive library identifies new genes expressed during xylogenesis in Eucalyptus (2006) New Phytol, 170, pp. 739-752Friedrich, J.B., Peter, J.D., Indole-3-acetic acid levels after phytochrome-mediated changes in the stem elongation rate of dark- and light-grown Pisum seedlings (1992) Planta, 188, pp. 85-92Gion, J.-M., Carouché, A., Deweer, S., Bedon, F., Pichavant, F., Charpentier, J.-P., Baillères, H., Plomion, C., Comprehensive genetic dissection of wood properties in a widely-grown tropical tree: Eucalyptus (2011) BMC Genomics, 12, p. 301Goicoechea, M., Lacombe, E., Legay, S., Mihaljevic, S., Rech, P., Jauneau, A., Lapierre, C., Grima-Pettenati, J., EgMYB2, a new transcriptional activator from Eucalyptus xylem, regulates secondary cell wall formation and lignin biosynthesis (2005) Plant J, 43, pp. 553-567Gómez Ros, L., Gabaldón, C., Pomar, F., Merino, F., Pedreño, M., Barceló, A., Structural motifs of syringyl peroxidases predate not only the gymnosperm-angiosperm divergence but also the radiation of tracheophytes (2007) New Phytol, 173, pp. 63-78Grabber, J.H., Schatz, P.F., Kim, H., Lu, F.C., Ralph, J., Identifying new lignin bioengineering targets: 1. Monolignol-substitute impacts on lignin formation and cell wall fermentability (2010) BMC Plant Biol, 10, p. 114Guo, D., Chen, F., Wheeler, J., Winder, J., Selman, S., Peterson, M., Dixon, R., Improvement of in-rumen digestibility of alfalfa forage by genetic manipulation of lignin O-methyltransferases (2001) Transgenic Res, 10, pp. 457-464Hall, T., BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT (1999) Nucleic Acids Res, 41, pp. 95-98Harkin, J., Obst, J., Lignification in trees: indication of exclusive peroxidase participation (1973) Science, 180, pp. 296-298Hatfield, R., Fukushima, R.S., Can lignin be accurately measured? 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Tibouchina Sellowiana (cham.) Cogn,: Rooting, Anatomy And Biochemical Analyses In The Four Seasons [tibouchina Sellowiana (cham.) Cogn.: Enraizamento, Anatomia E Análises Bioquímicas Nas Quatro Estações Do Ano]

The present work tried to verify the effects of application of different concentration of indolebutyric acid (IBA) in talc in solution, in the rooting of cutting of Tibouchina sellowiana (Cham.) Cogn. collected in the four seasons, as well as its anatomical and biochemical variation. The semiharwood cuttings were treated with 0, 1500 and 3000 mg L-1 indolebutyric acid (IBA), in powder and alchoolic solutions. After 65 days in a greenhouse, rooting percentage, length and number of roots, cutting with callus percentage and survival, were evaluated. The use of the IBA was efficient in the induction radicial, being indifferent the form in which it was applied. The most promising stations in the formation of the system radicial were the spring (78,75%) with 3000 mg L-1 or mg kg-1 IBA and the summer (63,12 %) with 1500 mg L-1 or mg kg-1 IBA. (82,50%) with 3000 mg kg -1 IBA. Considering the cutting in the base of the stakes, in the moment of the cutting, no possible anatomical barrier or structural difference were detected so that it should promoted variations in the answers of the rooting uring the four seasons. Biochemical analyses done from the cuttings collected in the four seasons revealed that the largest concentrations of total sugars were obtained in the winter (83,21 mg g-1) and in the autumn (72,79 mg g-1). The same stations also presented the largest protein concentrations (4,69 and 3,95 mg g-1 respectively).182159171ALVARENGA, L.R., CARVALHO, V.D., Uso de substâncias promotoras de enraizamento de estacas frutíferas. 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