Anatomical changes induced by isolates of Trichoderma spp. in soybean plants.

In the current work we evaluated the anatomical changes induced by T. harzianum and T. asperellum in two soybean cultivars, BRSGO Caiaponia and NA 5909 RG. Soybean production represents a growing market worldwide, and new methods aimed at increasing its productivity and yield are constantly being sought. Fungi of the genus Trichoderma have been widely used in agriculture as a promising alternative for the promotion of plant growth and for biological control of various pathogens. It is known that Trichoderma spp. colonize plant roots, but the anatomical changes that this fungus can cause are still less studied. Experiment was conducted in a greenhouse to collect leaves and soybean roots to perform analysis of growth parameters, enzymatic activity of defense-related enzymes and anatomical changes. It was observed that inoculation of Trichoderma spp. caused anatomical alterations, among them, increase in stomatal index at the abaxial leaf surface, thickness of the root cortex, thickness of adaxial epidermis, mean diameter of the vascular cylinder, thickness of the mesophyll, and thickness of the spongy parenchyma of the soybean plants. These results indicate that the alterations in these factors may be related to the process of plant resistance to pathogens, and better performance against adverse conditions. This study demonstrates that the anatomical study of plants is an important tool to show the effects that are induced by biological control agents.

Gradients of metabolite accumulation and redifferentiation of nutritive cells associated with vascular tissues in galls induced by sucking insects.

Plant cells respond to abiotic and biotic stimuli, which generate adaptive phenotypes in plant organs. In the case of plant galls, cell phenotypes are adaptive for the gall inducer and assume characteristics mainly linked to its protection and nutrition. Herein, the cytological development and histochemical profile of Nothotrioza cattleiani galls, a sucking insect, on the leaves of Psidium cattleianum are compared with those of other galls, especially N. myrtoidis galls, searching for conserved and divergent alterations in cell fates and cycles. Leaf cell fates are completely changed within galls, except for epidermal cells, but the comparison between Nothotrioza spp. galls shows conserved fates. Nevertheless, cytological development of N. cattleiani galls is different from the standby-redifferentiation of N. myrtoidis galls. Starch and lignins, and reducing sugars form centrifugal and centripetal gradients of accumulation, respectively. Proteins, total phenolics, terpenoids, proanthocyanidins and reactive oxygen species are detected in bidirectional gradients, i.e. weak or undetectable reaction in the median cortical cells that is gradually more intense in the cell layers towards the inner and outer surfaces of the gall. True nutritive cells associated with vascular tissues, together with the bidirectional gradients of metabolite accumulation, are herein reported for the first time in insect galls. The globoid galls of N. cattleiani, though macro-morphologically similar to the galls of N. myrtoidis, are distinct and unique among insect galls, as far as the cellular, subcellular and histochemical traits are concerned. Thus, the traits of the galls on P. cattleianum studied herein represent the extended phenotypes of their inducers.

Could the Extended Phenotype Extend to the Cellular and Subcellular Levels in Insect-Induced Galls?

Neo-ontogenesis of plant galls involves redifferentiation of host plant tissues to express new phenotypes, when new cell properties are established via structural-functional remodeling. Herein, Psidium cattleianum leaves and Nothotrioza cattleiani galls are analyzed by developmental anatomy, cytometry and immunocytochemistry of cell walls. We address hypothesis-driven questions concerning the organogenesis of globoid galls in the association of P. cattleianum-N. cattleianum, and P. myrtoides-N. myrtoidis. These double co-generic systems represent good models for comparing final gall shapes and cell lineages functionalities under the perspective of convergent plant-dependent or divergent insect-induced characteristics. Gall induction, and growth and development are similar in both galls, but homologous cell lineages exhibit divergent degrees of cell hypertrophy and directions of elongation. Median cortical cells in P. cattleianum galls hypertrophy the most, while in P. myrtoides galls there is a centrifugal gradient of cell hypertrophy. Cortical cells in P. cattleianum galls tend to anisotropy, while P. myrtoidis galls have isotropically hypertrophied cells. Immunocytochemistry evidences the chemical identity and functional traits of cell lineages: epidermal cells walls have homogalacturonans (HGAs) and galactans, which confer rigidity to sites of enhanced cell division; oil gland cell walls have arabinogalactan proteins (AGPs) that help avoiding cell death; and parenchyma cell walls have HGAs, galactans and arabinans, which confer porosity. Variations in such chemical identities are related to specific sites of hypertrophy. Even though the double co-generic models have the same macroscopic phenotype, the globoid morphotype, current analyses indicate that the extended phenotype of N. cattleiani is substantiated by cellular and subcellular specificities.

The effects of in vitro culture on the leaf anatomy of Jatropha curcas L. (Euphorbiaceae) / Efeitos da cultura in vitro na anatomia foliar de Jatropha curcas L. (Euphorbiaceae)

Jatropha curcas (Euphorbiaceae) is an evergreen tree traditionally used in popular medicine, and also as a promising source for bio-fuel production. The in vitro propagation of this species has been studied as an alternative to conventional methods such as cutting and seedling. In vitro culture environment consists on closed flasks, with high relative humidity, reduced gas exchanges and artificial temperature and luminosity conditions. These conditions may induce physiological and anatomical alterations in cultivated plants. The occurrence of anatomical alterations on the leaves of J. curcas was examined in plants cultivated both in vitro and ex vitro, under greenhouse conditions. The stomatal index was higher on the leaves from greenhouse plants. Mesophyll thickness did not differ, but the greenhouse leaves presented an additional palisade layer, which reflects the environmental influence on cell division. The cells of the chlorophyllous parenchyma of young plants grown in greenhouse conditions have larger chloroplasts than those of the plants grown in vitro. The chloroplasts of mature leaves are similar in height, but the mitochondria are smaller. Current results indicate that the leaves of J. curcas respond distinctly to both environments. It is necessary to adjust the abiotic conditions in vitro to avoid precocious senescence, diagnosed by chloroplasts and mesophyll degradation.

Jatropha curcas (Euphorbiaceae) é uma espécie arbórea sempre-verde usada tradicionalmente na medicina popular e com potencial para produção de biodiesel. A propagação in vitro desta espécie tem sido avaliada como alternativa aos meios convencionais de estaquia e plantio. O ambiente in vitro consiste em frascos de vidro fechados, com umidade relativa elevada, trocas gasosas reduzidas, e condições artificialmente controladas de temperatura e luminosidade. Tais condições podem induzir anormalidades fisiológicas e anatômicas nas plantas cultivadas. A ocorrência de alterações anatômicas foi avaliada em folhas de J. curcas cultivadas in vitro e em casa de vegetação visando diagnosticar os efeitos do ambiente in vitro sobre o desenvolvimento das plantas. O índice estomático foi maior nas folhas de plantas crescidas em casa de vegetação. A espessura do mesofilo não apresentou alterações, mas as folhas das plantas crescidas em casa de vegetação apresentaram uma camada adicional de parênquima paliçádico. Citologicamente, as células do parênquima clorofiliano de plantas jovens crescidas em casa de vegetação possuem cloroplastos maiores do que aqueles das plantas crescidas in vitro. Nas plantas maduras, os cloroplastos apresentam tamanhos similares. As mitocôndrias possuem tamanhos reduzidos nas plantas maduras ex vitro. Nossos resultados mostram que as folhas de J. curcas respondem distintamente aos dois ambientes. Ajustes nas condições abióticas in vitro são necessários para evitar a senescência precoce, diagnosticada pela desintegração dos cloroplastos e consequentemente do mesofilo. Current results indicate that the leaves of J. curcas respond distinctly to both environments. It is necessary to adjust the abiotic conditions in vitro to avoid precocious senescence, diagnosed by chloroplasts and consequent mesophyll degradation.

Cytological and histochemical gradients on two Copaifera langsdorffii Desf. (Fabaceae)--Cecidomyiidae gall systems.

Previous ultrastructural and histochemical analysis proposed patterns in the accumulation of substances in galls of Diptera: Cecidomyiidae in some plant species of the temperate region. Similar analyses were done to verify the conservativeness of these patterns in the Neotropical region, where a great number of species of Cecidomyiidae is responsible for a wide diversity of morphotypes. Two gall morphotypes induced by Cecidomyiidae in a unique host plant, Copaifera langsdorffii, were studied. The gradients of carbohydrates and the activity of invertases and acid phosphatases were similar, but the cytological gradients and distribution of proteins evidenced that the sites of the induction as well as the amount of neoformed tissues may be peculiar to each gall system. The production of lipids just in the secretory cavities either in the non-galled or galled tissues indicated a potentiality of the host plant which could not be manipulated by the galling insects. Further, the absence of nucleus in the nutritive tissue, an exclusive feature of the horn-shaped galls, indicates cell death attributed to the feeding habit of the galling herbivore.