The anatomy of the seed-coat includes diagnostic characters in the subtribe Eugeniinae (Myrteae, Myrtaceae).

The subtribe Eugeniinae comprises of two genera, Eugenia (ca. 1,100 species) and Myrcianthes (ca. 40 species). Eugenia is the largest genus of neotropical Myrtaceae and its latest classification proposes 11 sections. This study describes the seed anatomy of forty-one species of Eugeniinae in order to provide possible diagnostic characteristics. Following standard anatomical techniques, flower buds, flowers, and fruits were processed and analyzed using microtome sections and light microscopy. The phylogeny used the regions ITS, rpl16, psbA-trnH, trnL-rpl32, and trnQ-rps16, following recent studies in the group. Ancestral character reconstruction uncovered that: (1) the ancestral ovule in Eugeniinae was campylotropous (98.9% probability), bitegmic (98.5% probability), and unitegmic ovules arose on more than one lineage independently within Eugenia; (2) the pachychalazal seed-coat appeared with a 92% probability of being the ancestral type; (3) non-lignified seed-coat (24,5% probability) and aerenchymatous mesotesta (45.8% probability) are diagnostic characters in Myrcianthes pungens (aerenchymatous mesotesta present in the developing seed-coat) and in the species of E. sect. Pseudeugenia until the species of E. sect. Schizocalomyrtus and it is the type of seed-coat that predominates in most basal sections on the tree; (4) the partial sclerification (only in the exotesta-exotestal seed-coat) is mainly observed in species of E. sect. Excelsae, E. sect. Jossinia (group X), and E. sect. Racemosae (22.2% probability); (5) and in the species of the recent lineages of Eugenia, with a probability of 27.2%, predominate the exomesotestal or testal construction of the seed-coat [character observed in almost all species analyzed of E. sect. Jossinia (group Y) and E. sect. Umbellatae]. A dehiscent fruit is considered as a plesiomorphic state in Myrtaceae; the ancestor of this family had seeds with a completely sclerified testa, and the other testa types described for the current species with dehiscent and indehiscent fruits are simplified versions of this ancestral type. Perhaps, this means that the sclerified layers in the seed-coat have remained in whole or in part as a plesiomorphic condition for taxa with a capsule and bacca. Maintaining the plesiomorphic condition may have represented a selective advantage at some point in the evolutionary history of the family and its groups.

Endoglucanase production by endophytic fungi isolated from vitis labrusca l. with peanut hull and sawdust as substrates / Produção de endoglucanase por fungos endofíticos isolados de Vitis labrusca L. utilizando casca de amendoim e serragem como substratos

Endoglucanases are enzymes widely employed in different industrial fields, albeit with high production costs. Studies on new microbial sources and low-cost substrates are highly relevant, including those on agro-industrial. Current analysis evaluates peanut hull (PH) and sawdust (SD) as substrates for submerged cultures of 14 endophytic fungi isolated from grapevine (Vitis labrusca L.) cultivars Bordô and Concord. Endophytes were grown on a carboxymethylcellulose (CMC) medium and the cup plate assay showed that eight strains (belonging to genera Cochliobolus, Diaporthe, Fusarium and Phoma) had positive results: enzymatic halos ranged from 10.8±0.02to 15.5±0.07 mm in diameter. Diaporthe sp. strains (GenBank accession codes KM362392, KM362368 and KM362378) and Fusariumculmorum KM362384 were highlighted as the most promising sources. Further, PH and SD as substrates for the fermentation of these fungi were evaluated by the cup plate assay and endoglucanase activity assay. Highest halo diameters were obtained for Diaporthe sp. KM362392: 16.1±0.01 mm (CMC), 14.5±0.01 mm (PH) and 14.7±0.03 mm (SD). The fungus also presented the highest levels of endoglucanase activity: analysis of variance revealed that CMC (3.52±0.98 µmol/min), PH (2.93±0.23 µmol/min) and SD (3.26±0.38 µmol/min) were similarly efficient as substrates. Results deepen knowledge on V. labrusca endophytes that may be endoglucanase sources, eventhough further optimizations in submerged cultures with PH and SD should be undertaken to increase theenzymatic production from these wastes.

Endoglucanases são enzimas amplamente empregadas em diferentes setores industriais; embora sua produção apresente custos elevados. Estudos sobre novas fontes microbianas e substratos mais baratos são de grande importância, incluindo os resíduos agroindustriais. Nesse estudo, casca de amendoim (CA) e serragem (SE) foram testadas como substratos para o cultivo submerso de 14 fungos endofíticos isolados das cultivares Bordô e Concord de videira (Vitis labrusca L.) Os endófitos foram crescidos em meio contendo carboximetilcelulose (CMC) e o ensaio cup plate mostrou resultados positivos para oito fungos (pertencentes aos gêneros Cochliobolus, Diaporthe, Fusarium and Phoma); os halos enzimáticos variaram entre 10,8±0,02 e 15,5±0,07 mm de diâmetro. Linhagens de Diaporthe sp. (códigos de acesso no GenBank KM362392, KM362368 e KM362378) e Fusariumculmorum KM362384 se destacaram como produtores mais promissores. Então, o uso de CA e SE como substratos para a fermentação desses fungos foi avaliado pelo ensaio cup plate e pela quantificação da atividade de endoglucanase. Os maiores halos enzimáticos foram obtidos para Diaporthe sp. KM362392: 16,1±0,01 mm (CMC), 14,5±0,01 mm (CA) e 14,7±0,03 mm (SE). Esse fungo também apresentou os maiores níveis de endoglucanase: a análise de variância revelou que CMC (3,52±0,98 µmol/min), CA (2,93±0,23 µmol/min) e SE (3,26±0,38 µmol/min) foram substratos similarmente eficientes. Esses resultados expandem o conhecimento sobre endófitos de V. labrusca que são fontes de endoglucanases; futuras otimizações quanto ao cultivo submerso com CA e SE podem ser utilizadas para aumentar a produção enzimática a partir do uso desses resíduos.

Agrobacterium-Mediated Transformation of Diaporthe schini Endophytes Associated with Vitis labrusca L. and Its Antagonistic Activity Against Grapevine Phytopathogens.

Fungus-caused diseases are among the greatest losses in grapevine culture. Biological control of pathogens by endophytes may be used to decrease fungicide application rates and environmental impacts. Previously, Diaporthe sp. B46-64 and C27-07 were highlighted as antagonists of grapevine phytopathogens. Herein, molecular multigene (ITS-TUB-TEF1) identification and phylogenetic analysis allowed the identification of these endophytes as belonging to Diaporthe schini species. Agrobacterium tumefaciens-mediated transformation was employed for obtaining 14 stable and traceable gfp- or DsRed-expressing transformants, with high transformation efficiency: 96% for the pFAT-GFP plasmid and 98% for pCAM-DsRed plasmid. Transformants were resistant to hygromycin B with gene hph confirmed by polymerase chain reaction and proved to be mitotically stable, expressing the fluorescent phenotype, with morphological differences in the colonies when compared with wild strains. In vitro antagonism tests revealed an increased antagonistic activity of some transformant strains. The current genetic transformation of D. schini mediated by A. tumefaciens proved to be an efficient technique within the randomized insertion of reporter genes for the monitoring of the strain in the environment.

Ontogeny of the pericarp of Serjania communis Camb. and Urvillea ulmacea Kunth (Sapindaceae) with emphasis on the dispersion apparatus / Ontogênese do pericarpo de Serjania communis Camb. e Urvillea ulmacea Kunth (Sapindaceae) com ênfase no aparelho de dispersão

The pericarp development of Serjania communis Camb. and Urvillea ulmacea Kunth was described to expand the structural knowledge of Sapindaceae fruit. Ovaries and fruits were analyzed with usual techniques in plant anatomy. Ovary is structurally similar and the occurrence of glandular and non-glandular trichomes varies between species. In S. communis, the samaroid schizocarp has proximal wings and distal cavities; in U. ulmacea, the fruit presents septicidal and septifragal passive dehiscence and each seed chamber presents a dorsal wing. The unistratified exocarp has long pluricellular, uniseriate and sparse trichomes in U. ulmacea. The mesocarp is spongy, parenchymatous and in S. communis it presents three distinguished regions. Dorsal, lateral and ventral vascular bundles are more developed in S. communis and fiber caps on the phloem in the lateral bundles have an important role on the mericarp separation. The endocarp derives from the adaxial meristem and its oblique and tangentially elongated cells become lignified. The wings originate from a more active meristem ­ at the ovarian edges ­ and by intense divisions of ovarian mesophyll in this region. Fruit of U. ulmacea can represent a transition state between the samaroid schizocarp formed by Serjania and the septifragal capsule produced by Cardiospermum.

O presente estudo descreve aspectos morfológicos e o desenvolvimento do pericarpo de Serjania communis Camb. e Urvillea ulmacea Kunth ampliando o conhecimento estrutural dos frutos de Sapindaceae. Foram coletados e analisados ovários e frutos seguindo técnicas usuais em anatomia vegetal. O ovário estruturalmente similar e a ocorrência de tricomas glandulares e não glandulares varia entre as espécies. Em S. communis, o esquizocarpo samaroide apresenta ala proximal e cavidade distal; em U. ulmacea, o fruto apresenta deiscência passiva septicida e septifraga e cada cavidade seminal apresenta uma ala dorsal. O exocarpo uniestratificado tem tricomas pluricelulares longos, unisseriados e esparsos em U. ulmacea. O mesocarpo é parenquimático e esponjoso e em S. communis apresenta três regiões distintas. Os feixes vasculares dorsais, laterais e ventrais são mais desenvolvidos em S. communis e as calotas de fibras sobre o floema nos feixes laterais participam da separação dos mericarpos. O endocarpo deriva do meristema adaxial e suas células oblíqua e tangencialmente alongadas tornam-se lignificadas. As alas originam-se de um meristema mais ativo ­ nas arestas do ovário ­ e por divisões intensas no mesofilo ovariano nessa região. O fruto de U. ulmacea pode representar a transição entre o esquizocarpo samaroide de Serjania e a cápsula septifraga de Cardiospermum.

Morfoanatomia e ontogênese dos frutos e sementes de Clusia lanceolata Cambess (Clusiaceae) / Morphology, anatomy and ontogeny of the fruits and seeds of Clusia lanceolata Cambess (Clusiaceae)

Clusia lanceolata Cambess é espécie arbustiva da Mata Atlântica. Caracteriza-se por ser dioica, com flores estaminadas e pistiladas grandes, vistosas, de coloração rosa, apresentando a parte interna vinácea. Foram descritos aspectos morfológicos e estruturais dos frutos e sementes dessa espécie, visando classificar corretamente o seu tipo de fruto e, também, confirmar a origem do apêndice carnoso que envolve as sementes. O material botânico utilizado constou de flores em antese e frutos, em quatro estádios de desenvolvimento, coletados de dois espécimes. Os estudos morfológicos e anatômicos foram realizados segundo técnicas usuais. O fruto é cápsula septífraga, esférica, de coloração verde-avermelhado. O exocarpo é unisseriado; o mesocarpo parenquimático contém numerosos ductos secretores e feixes vasculares, de disposição principalmente longitudinal. O endocarpo é derivado da epiderme interna do ovário e de três a quatro camadas subepidérmicas, cujas células alongam-se tangencialmente e a parede que delimita o lóculo sofre lignificação. A abertura se faz pela sutura carpelar, por meio de uma faixa de células parenquimáticas de tamanho reduzido, que se bifurca em direção aos lóculos no meio dos septos. Os óvulos anátropos bitegumentados, com endotélio e hipóstase, originam sementes também anátropas, exotégmicas e envolvidas por arilo de origem funicular e exostomal. O embrião é hipocotilar.

Clusia lanceolata Cambess. is a shrub species native to the Atlantic Forest. The plant is dioecious, with large, showy, pink staminate and pistillate flowers that are wine-colored internally. The morphological and structural aspects of the fruits and seeds of this species are described here in order to correctly classify the fruit type and to determine the origin of the fleshy appendage that surrounds the seeds. The botanical material utilized here contained flowers in anthesis and fruits in four stages of development, and were collected from two different plants. The morphological and anatomical studies were performed according to traditional techniques. The fruit is a septifrage, spherical capsule with a reddish-green coloration. The exocarp is uniseriate; the parenchymatous mesocarp contains numerous secretory ducts and vascular bundles arranged in predominantly longitudinal positions. The endocarp is derived from the internal epidermis of the ovary and has from three to four subepidermal layers whose component cells are tangentially elongated, and the cell walls lining the locule become lignified. The fruits open by means of the carpelar suture, along a stand of small parenchymous cells that bifurcate in the direction of the locules, in the midst of the septum. The anatropous bi-tegumented ovules, with an endothelium and hypostase, give rise to seeds that are likewise anatropous, exotegmic, and wrapped in an aril that has a funicular and exostomal origin. The embryo is hypocotylar.

Vegetative propagation in Piperaceae species

Piperaceae species of forest remnants of Maringá, Paraná, Brazil present vegetative propagation through root or stem (stolon). This reproduction type was verified in Piper amalago L., P. arboreum Aublet, P. xylosteoides (Kunth) Steud., Peperomia parnassifolia Miq. and Ottonia martiana Miq. Root and stem branches of these species were analyzed anatomically. Results indicated that the vegetative propagation in Piperaceae should be studied in other forests. New and additional studies should be made if the structural variations were induced environmentally or fixed genetically.

Espécies de Piperaceae de remanescentes florestais de Maringá, Paraná, Brasil apresentam propagação vegetativa por meio de raiz ou caule (estolão). Este tipo de reprodução foi verificado em Piper amalago L., Piper arboreum Aublet, Piper xylosteoides (Kunth) Steud., Peperomia parnassifolia Miq. and Ottonia martiana Miq. Raiz e ramos caulinares destas espécies foram analisados anatomicamente segundo técnicas usuais. É necessário enfatizar que esse tipo de propagação em Piperaceae deve ser investigado em espécies que ocorrem em outras matas. Novos e adicionais estudos devem ser feitos para se saber se essas variações estruturais são induzidas pelo ambiente ou fixadas geneticamente.