Propagação vegetativa de Campomanesia phaea pelas técnicas de alporquia e enxertia

The objective of this work was to evaluate the air-layering and grafting techniques in the production of cambuci (Campomanesia phaea) plants. Two experiments were conducted in randomized complete blocks for air-layering, using adult cambuci plants, and two completely randomized for the grafting, using seedlings. Air layering was evaluated with different rates of indolebutyric acid; the splice, side-veneer, and chip-budding techniques were used for grafting. Air-layering rooting was not favored by the application of indolebutyric acid rates, whereas side-veneer grafts ensured the fixation of 6% of the grafts. Histological analyses of the air layers revealed the action of parenchyma cells from the xylem in the formation of structures similar to calluses. In the grafted plants, these cells ensured tissue union and regeneration. The air-layering technique was unfeasible, but grafting is recommended to establish cambuci tree genotypes.O objetivo deste trabalho foi avaliar as técnicas de alporquia e enxertia na produção de plantas de cambucizeiro (Campomanesia phaea). Foram conduzidos dois experimentos em blocos ao acaso para alporquia, com plantas adultas de cambuci, e dois inteiramente casualizados para a enxertia, utilizando mudas. A alporquia foi avaliada com diferentes doses de ácido indolbutírico, e as técnicas do tipo inglês simples e fenda lateral foram utilizadas para a enxertia. O enraizamento dos alporques não foi favorecido pela aplicação de doses de ácido indolbutírico, enquanto a enxertia do tipo fenda lateral garantiu a fixação de 6% dos enxertos. Análises histológicas dos alporques revelaram atividade das células parenquimáticas do xilema na formação de estruturas similares a calos. Nas plantas enxertadas, essas células garantiram a união e a regeneração dos tecidos. A técnica da alporquia foi inviável, mas a enxertia é recomendada para fixação de genótipos de cambucizeiro

Cellular And Molecular Changes Associated With Competence Acquisition During Passion Fruit Somatic Embryogenesis: Ultrastructural Characterization And Analysis Of Serk Gene Expression.

The integration of cellular and molecular data is essential for understanding the mechanisms involved in the acquisition of competence by plant somatic cells and the cytological changes that underlie this process. In the present study, we investigated the dynamics and fate of Passiflora edulis Sims cotyledon explants that were committed to somatic embryogenesis by characterizing the associated ultrastructural events and analysing the expression of a putative P. edulis ortholog of the Somatic Embryogenesis Receptor-like Kinase (SERK) gene. Embryogenic calli were obtained from zygotic embryo explants cultured on Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzyladenine. Callus formation was initiated by the division of cells derived from the protodermal and subprotodermal cells on the abaxial side of the cotyledons. The isodiametric protodermal cells of the cotyledon explants adopted a columnar shape and became meristematic at the onset of PeSERK expression, which was not initially detected in explant cells. Therefore, we propose that these changes represent the first observable steps towards the acquisition of a competent state within this regeneration system. PeSERK expression was limited to the early stages of somatic embryogenesis; the expression of this gene was confined to proembryogenic zones and was absent in the embryos after the globular stage. Our data also demonstrated that the dynamics of the mobilization of reserve compounds correlated with the differentiation of the embryogenic callus.25

Carfentrazone-ethyl and glyphosate drift inhibits uredinial formation of Austropuccinia psidii on Eucalyptus grandis leaves

The response to infection of Austropuccinia psidii in resistant (CLR‐383) and susceptible (CLR‐384) Eucalyptus grandis clones, exposed to herbicide drift of carfentrazone‐ethyl, glyphosate and a mixture of these two herbicides, was evaluated at microscopic and physiological levels.Plants of the two clones showed symptoms of phytotoxicity caused by herbicide drift. However, net CO2 assimilation rate, height and shoot dry matter were lower in CLR‐384 than in CLR‐383. At the ultrastructure level, the leaves of both clones exposed to the herbicides showed thylakoid disorganization and accumulation of starch grains in the chloroplasts. Only plants of CLR‐384 were infected by A. psidii, but when exposed to herbicide drift, rust severity was lower than in control plants. Six days after inoculation (dai), plants of this clone exposed to the herbicides had smaller uredinia than control plants. At 12 dai, non‐herbicide treated plants showed normal uredinia, containing abundant urediniospores. By contrast, plants exposed to the herbicides were less colonized by the fungus, and the uredinia were smaller with reduced production of urediniospores, which were sometimes not even detected. Glyphosate and carfentrazone‐ethyl herbicide drift reduce infection and uredinial formation of A. psidii and to some extent induce basal resistance in a susceptible clone of E. grandis

Anatomical and Physiological Responses of Citrus Trees to Varying Boron Availability Is Dependent on Rootstock

In Citrus, water, nutrient transport and thereby fruit production, are influenced among other factors, by the interaction between rootstock and boron (B) nutrition. This study aimed to investigate how B affects the anatomical structure of roots and leaves as well as leaf gas exchange in sweet orange trees grafted on two contrasting rootstocks in response to B supply. Plants grafted on Swingle citrumelo or Sunki mandarin were grown in a nutrient solution of varying B concentration (deficient, adequate, and excessive). Those grafted on Swingle were more tolerant to both B deficiency and toxicity than those on Sunki, as revealed by higher shoot and root growth. In addition, plants grafted on Sunki exhibited more severe anatomical and physiological damages under B deficiency, showing thickening of xylem cell walls and impairments in whole-plant, leaf-specific hydraulic conductance and leaf CO2 assimilation. Our data revealed that trees grafted on Swingle sustain better growth under low B availablitlity in the root medium and still respond positively to increased B levels by combining higher B absorption and root growth as well as better organization of xylem vessels. Taken together, those traits improved water and B transport to the plant canopy. Under B toxicity, Swingle rootstock would also favor plant growth by reducing anatomical and ultrastructural damage to leaf tissue and improving water transport compared with plants grafted on Sunki. From a practical point of view, our results highlight that B management in citrus orchards shall take into account rootstock varieties, of which the Swingle rootstock was characterized by its performance on regulating anatomical and ultrastructural damages, improving water transport and limiting negative impacts of B stress conditions on plant growth

Novel insights into the early stages of infection by oval conidia of Colletotrichum sublineolum

Anthracnose, caused by Colletotrichum sublineolum Henn. ex Sacc. & Trotter, is one of the most important sorghum [Sorghum bicolor (L.) Moench] diseases in Brazil. This fungus exhibits conidial dimorphism, producing either falcate or oval conidia on solid and liquid media, respectively. We compared patterns of the initial infection events by these two types of conidia on sorghum leaves using light microscopy and scanning electron microscopy. The infection events during the first 24 h were similar for both oval and falcate conidia. Globose and melanized apressoria were formed at 24 h after inoculation (hai) regardless of the conidia type. Dense mycelium and oval conidia developed from germinated falcate conidia at 32 hai. Hyphal mass displaying acervuli filled with falcate conidia and surrounded by setae, developed from germinated oval conidia at 48 hai. Oval conidia were as capable as falcate conidia of infecting sorghum leaves. The inherent ability to grow faster and the easeness with which oval conidia can be produced in vitro as compared to falcate, make the former a preferred choice for studies on the C. sublineolum-sorghum interaction. It would be instructive to further investigate the potential role of the oval conidia in epidemics

Loss of type-IV glandular trichomes is a heterochronic trait in tomato and can be reverted by promoting juvenility

Glandular trichomes are structures with widespread distribution and deep ecological significance. In the Solanum genus, type-IV glandular trichomes provide resistance to insect pests. The occurrence of these structures is, however, poorly described and controversial in cultivated tomato (Solanum lycopersicum). Optical and scanning electron microscopy were used to screen a series of well-known commercial tomato cultivars, revealing the presence of type-IV trichomes on embryonic (cotyledons) and juvenile leaves. A tomato line overexpressing the microRNA miR156, known to promote heterochronic development, and mutants affecting KNOX and CLAVATA3 genes possessed type-IV trichomes in adult leaves. A re-analysis of the Woolly (Wo) mutant, previously described as enhancing glandular trichome density, showed that this effect only occurs at the juvenile phase of vegetative development. Our results suggest the existence of at least two levels of regulation of multicellular trichome formation in tomato: one enhancing different types of trichomes, such as that controlled by the WOOLLY gene, and another dependent on developmental stage, which is fundamental for type-IV trichome formation. Their combined manipulation could represent an avenue for biotechnological engineering of trichome development in plants

CO2 -enriched atmosphere and supporting material impact the growth, morphophysiology and ultrastructure of in vitro Brazilian-ginseng [Pfaffia glomerata (Spreng.) Pedersen] plantlets

This study aimed to evaluate under photoautotrophic conditions the effect of CO2-enriched atmosphere (360 or 1,000 μmol CO2 mol−1 air) combined with two substrate types (agar or Florialite®) in vitro on plants of Pfaffia glomerata, an endangered medicinal species with promising applications in phytotherapy and phytomedicine. The effects of the treatments on the growth, stomatal density, Rubisco activity, carbon isotopic discrimination, metabolite accumulation, photosynthetic pigments and ultrastructural characteristics were investigated. After a 35-day cultivation period, the in vitro-growth of P. glomerata nodal segments under the different treatments resulted in plants with substantial differences in relation to their growth, photosynthetic pigments, stomatal density and leaf ultrastructural characteristics. The enrichment with CO2 coupled with a porous substrate increased the growth of P. glomerata. The stomatal density in the abaxial epidermis more than doubled in response to the high CO2 supply in both supporting types, whereas the Rubisco activity and activation state were both unresponsive to the treatments. Regardless of the CO2 supply, the plants grown in agar displayed higher carbon isotope discrimination than their counterparts grown in Florialite®. We propose that the long-term photosynthetic performance was improved using Florialite® as a growth support in combination with a high CO2 supply. No apparent signs of photosynthetic down-regulation could be found under elevated CO2 conditions. The enrichment of in vitro atmospheres with CO2 coupled with a porous substrate offers new possibilities for improving the growth and production on a commercial scale of high morphological and physiological quality Pfaffia plants

Morpho-histological, histochemical, and molecular evidences related to cellular reprogramming during somatic embryogenesis of the model grass Brachypodium distachyon

The wild grass species Brachypodium distachyon (L.) has been proposed as a new model for temperate grasses. Among the biotechnological tools already developed for the species, an efficient induction protocol of somatic embryogenesis (SE) using immature zygotic embryos has provided the basis for genetic transformation studies. However, a systematic work to better understanding the basic cellular and molecular mechanisms that underlie the SE process of this grass species is still missing. Here, we present new insights at the morpho-histological, histochemical, and molecular aspects of B. distachyon SE pathway. Somatic embryos arose from embryogenic callus formed by cells derived from the protodermal-dividing cells of the scutellum. These protodermal cells showed typical meristematic features and high protein accumulation which were interpreted as the first observable steps towards the acquisition of a competent state. Starch content decreased along embryogenic callus differentiation supporting the idea that carbohydrate reserves are essential to morphogenetic processes. Interestingly, starch accumulation was also observed at late stages of SE process. Searches in databanks revealed three sequences available annotated as BdSERK, being two copies corresponding to SERK1 and one showing greater identity to SERK2. In silico analysis confirmed the presence of characteristic domains in a B. distachyon Somatic Embryogenesis Receptor Kinase genes candidates (BdSERKs), which suggests SERK functions are conserved in B. distachyon. In situ hybridization demonstrated the presence of transcripts of BdSERK1 in all development since globular until scutellar stages. The results reported in this study convey important information about the morphogenetic events in the embryogenic pathway which has been lacking in B. distachyon. This study also demonstrates that B. distachyon provides a useful model system for investigating the genetic regulation of SE in grass species