Diego Demarco

Laticíferos articulados anastomosados: novos registros para Apocynaceae

Laticifer presence is universal in Apocynaceae, in the classic literature the type described for this family is the non-articulated. Later researches have proved the occurrence of articulated laticifers only in four species, giving rise to controversies on their origin. The results obtained in our studies differ from those reported for most species of this family. In both Aspidosperma australe Mull. Arg. (Rauvolfioideae) and Blepharodon bicuspidatum Fourn. (Asclepiadoideae), the laticifers are of the articulated anastomosing type because they are formed by adding cells with rapidly dissolving transverse walls. Laticifers originate from ground meristem and/or procambium and form a branched system, they are in secretory phase since the early stages of formation in different organs, releasing latex only when the plant is damaged. The laticifer walls are exclusively pectic-cellulosic and their chemical characteristics probably change during their development. Vegetative organ laticifers occur in all stem and leaf tissues, except epidermis and medullary parenchyma of A. australe. In the flower, laticifers are found in all floral organs, except in the medullary parenchyma of the pedicel of A. australe and in the ovules of both species. The presence of the same type of laticifer in these two genera, which represent the most divergent subfamilies within the Apocynaceae corroborates the current circumscription of this family. The latex has protective function, allowing the species of this family to succeed in different environments.

Nitric Oxide, Ethylene, and Auxin Cross Talk Mediates Greening and Plastid Development in Deetiolating Tomato Seedlings.

Light-evoked cotyledon greening and chloroplast differentiation in deetiolating tomato seedlings are orchestrated by regulatory feedback loops involving phytochromes, nitric oxide, auxins, and ethylene. The transition from etiolated to green seedlings involves the conversion of etioplasts into mature chloroplasts via a multifaceted, light-driven process comprising multiple, tightly coordinated signaling networks. Here, we demonstrate that light-induced greening and chloroplast differentiation in tomato (Solanum lycopersicum) seedlings are mediated by an intricate cross talk among phytochromes, nitric oxide (NO), ethylene, and auxins. Genetic and pharmacological evidence indicated that either endogenously produced or exogenously applied NO promotes seedling greening by repressing ethylene biosynthesis and inducing auxin accumulation in tomato cotyledons. Analysis performed in hormonal tomato mutants also demonstrated that NO production itself is negatively and positively regulated by ethylene and auxins, respectively. Representing a major biosynthetic source of NO in tomato cotyledons, nitrate reductase was shown to be under strict control of both phytochrome and hormonal signals. A close NO-phytochrome interaction was revealed by the almost complete recovery of the etiolated phenotype of red light-grown seedlings of the tomato phytochrome-deficient aurea mutant upon NO fumigation. In this mutant, NO supplementation induced cotyledon greening, chloroplast differentiation, and hormonal and gene expression alterations similar to those detected in light-exposed wild-type seedlings. NO negatively impacted the transcript accumulation of genes encoding phytochromes, photomorphogenesis-repressor factors, and plastid division proteins, revealing that this free radical can mimic transcriptional changes typically triggered by phytochrome-dependent light perception. Therefore, our data indicate that negative and positive regulatory feedback loops orchestrate ethylene-NO and auxin-NO interactions, respectively, during the conversion of colorless etiolated seedlings into green, photosynthetically competent young plants.

Galacturonosyltransferase 4 silencing alters pectin composition and carbon partitioning in tomato

Pectin is a main component of the plant cell wall and is the most complex family of polysaccharides in nature. Its composition is essential for the normal growth and morphology pattern, as demonstrated by pectin-defective mutant phenotypes. Besides this basic role in plant physiology, in tomato, pectin structure contributes to very important quality traits such as fruit firmness. Sixty-seven different enzymatic activities have been suggested to be required for pectin biosynthesis, but only a few genes have been identified and studied so far. This study characterized the tomato galacturonosyltransferase (GAUT) family and performed a detailed functional study of the GAUT4 gene. The tomato genome harbours all genes orthologous to those described previously in Arabidopsis thaliana, and a transcriptional profile revealed that the GAUT4 gene was expressed at higher levels in developing organs. GAUT4-silenced tomato plants exhibited an increment in vegetative biomass associated with palisade parenchyma enlargement. Silenced fruits showed an altered pectin composition and accumulated less starch along with a reduced amount of pectin, which coincided with an increase in firmness. Moreover, the harvest index was dramatically reduced as a consequence of the reduction in the fruit weight and number. Altogether, these results suggest that, beyond its role in pectin biosynthesis, GAUT4 interferes with carbon metabolism, partitioning, and allocation. Hence, this cell-wall-related gene seems to be key in determining plant growth and fruit production in tomato.

Down-regulation of tomato PHYTOL KINASE strongly impairs tocopherol biosynthesis and affects prenyllipid metabolism in an organ-specific manner

Highlight Phytol kinase plays a key role in the regulation of isoprenoid metabolism in an organ-specific manner.

Silencing of the tomato Sugar Partitioning Affecting protein (SPA) modifies sink strength through a shift in leaf sugar metabolism

Limitations in our understanding about the mechanisms that underlie source-sink assimilate partitioning are increasingly becoming a major hurdle for crop yield enhancement via metabolic engineering. By means of a comprehensive approach, this work reports the functional characterization of a DnaJ chaperone related-protein (named as SPA; sugar partition-affecting) that is involved in assimilate partitioning in tomato plants. SPA protein was found to be targeted to the chloroplast thylakoid membranes. SPA-RNAi tomato plants produced more and heavier fruits compared with controls, thus resulting in a considerable increment in harvest index. The transgenic plants also displayed increased pigment levels and reduced sucrose, glucose and fructose contents in leaves. Detailed metabolic and enzymatic activities analyses showed that sugar phosphate intermediates were increased while the activity of phosphoglucomutase, sugar kinases and invertases was reduced in the photosynthetic organs of the silenced plants. These changes would be anticipated to promote carbon export from foliar tissues. The combined results suggested that the tomato SPA protein plays an important role in plastid metabolism and mediates the source-sink relationships by affecting the rate of carbon translocation to fruits.

Secretory Tissues and the Morphogenesis and Histochemistry of Pollinarium in Flowers of Asclepiadeae (Apocynaceae)

Premise of research. Flowers of some Apocynaceae present an unusual synorganization among parts and organs, which has led to the origin of new organs and a type of pollen dispersal and pollination unique to eudicots. The synorganization of androecium and gynoecium allowed the evolution of the pollinarium, a very complex and species-specific structure composed of pollen grains (pollinium) and secretions produced by the style head (translator). The aim of this study was to investigate the structure and activity of the secretory system related to the morphogenesis of pollinaria and the composition of their secretions in flowers of Asclepiadeae. Methodology. Flowers and buds of Asclepias, Gonioanthela, Matelea, and Oxypetalum were fixed, embedded in Paraplast, and sectioned in a rotary microtome for analysis under light microscopy. SEM was performed for additional structural data, and histochemical tests were carried out to reveal the main chemical composition of the secretions involved in pollinaria morphogenesis. Pivotal results. In the four species of Asclepiadeae studied, the style head possesses a palisade secretory epidermis that is responsible for the secretion of the translator, which is composed of a corpusculum and two caudicles. Mucilage, fatty acids, phenolic compounds, and proteins were detected in the corpusculum, whereas only neutral lipids and mucilage were present in the caudicles. In the anthers, tapetal cells are involved in the secretion of lipids that cover the whole pollinium. Just before anthesis, anthers dehisce, and the pollinia from adjacent anthers contact and adhere to the caudicles of the translator, forming the pollinarium. Conclusions. The translator shape is due to the differential activity of the secretory cells, which are spatially and temporarily coordinated, as well as the amount and composition of the secretion and the undulated outline of the secretory surface. The corpusculum and caudicles are composed of different chemical substances, and these features are related to the processes of removal and insertion of pollinia.

Transfer cells in trichomatous nectary in Adenocalymma magnificum (Bignoniaceae).

Transfer cells are specialized cells that play an important role where there are high energy costs due to facilitation of transmembrane flow of solutes. This study aimed to investigate the ontogenesis, histochemistry and ultrastructure of glandular trichomes focusing on stalk cells and their possible transfer function. Samples of prophylls of axillary buds, calyces of flowers in anthesis, and flower buds in different stages of development were collected, fixed and processed according to the common methods of microscopy. The glandular trichomes are composed of a secretory head with its cells in columnar format. The stalk is formed by two layers of cells, with the upper layer composed of cuboidal cells where the wall starts to thicken at the beginning of the pre-secretory phase. The secretion is heterogeneous, releasing glucose, other carbohydrates, lipids and phenolic compounds, with two types of release - eccrine and granulocrine. These trichomes are functionally termed as nectaries. The stalk cells appear as transfer cells since they have a thicker anticlinal wall with irregular ingrowths. The presence of transfer cells in the nectaries suggests a high specialization because it improves transport capacity of nectar and compensation in the high energy expenditure for its production and release.

Manipulation of a Senescence-Associated Gene Improves Fleshy Fruit Yield

Senescence-associated gene knockdown increases carbon exportation toward sink organs, increasing plant yield in Solanum lycopersicum. Senescence is the process that marks the end of a leaf’s lifespan. As it progresses, the massive macromolecular catabolism dismantles the chloroplasts and, consequently, decreases the photosynthetic capacity of these organs. Thus, senescence manipulation is a strategy to improve plant yield by extending the leaf’s photosynthetically active window of time. However, it remains to be addressed if this approach can improve fleshy fruit production and nutritional quality. One way to delay senescence initiation is by regulating key transcription factors (TFs) involved in triggering this process, such as the NAC TF ORESARA1 (ORE1). Here, three senescence-related NAC TFs from tomato (Solanum lycopersicum) were identified, namely SlORE1S02, SlORE1S03, and SlORE1S06. All three genes were shown to be responsive to senescence-inducing stimuli and posttranscriptionally regulated by the microRNA miR164. Moreover, the encoded proteins interacted physically with the chloroplast maintenance-related TF SlGLKs. This characterization led to the selection of a putative tomato ORE1 as target gene for RNA interference knockdown. Transgenic lines showed delayed senescence and enhanced carbon assimilation that, ultimately, increased the number of fruits and their total soluble solid content. Additionally, the fruit nutraceutical composition was enhanced. In conclusion, these data provide robust evidence that the manipulation of leaf senescence is an effective strategy for yield improvement in fleshy fruit-bearing species.

Laticifer development and its growth mode in Allamanda blanchetii A. DC. (Apocynaceae)

Abstract Recent studies have shown the laticifers of Apocynaceae, previously classified as nonarticulated, indeed are articulated, anastomosing laticifers whose transverse walls dissolve rapidly and entirely, although doubts about their growth mode still persist. To better understand the mode of laticifer growth and differentiation in this family, we studied its development in Allamanda blanchetii using anatomical and ultrastructural analyses. Our results showed that laticifers are formed by a row of cells that join each other through dissolution of the transverse walls from the center to the periphery. The laticifers originate from ground meristem and procambium; the laticifers in the different tissues connect through lateral fusion, generating a laticifer network. The laticifers occur in the cortex, pith, and vascular system of the shoot, mesophyll, and vascular bundles of the leaves. There is no apical growth, and all the organelles observed in the apices of the laticifers play a role in the production of latex or in the dissolution of the terminal walls between the cells that compose the laticifer. The latex is composed of many metabolites produced mainly in the endoplasmic reticulum and plastids. Mitochondria are abundant, and dictyosomes are scarce. The vacuome is prominent from the start of laticifer differentiation, and many small vacuoles and vesicles transport the secretion from cytosol into the central vacuole, where an emulsion of substances is stored. The articulated, anastomosing laticifers of A. blanchetii have no subcellular mechanism for production of a cell wall in a polarized manner or dissolution of middle lamella of the cells which surround the laticifer tip, proving that there is no intrusive growth in this secretory structure.

Ontogênese do pericarpo e histoquímica da exotesta e pseudocarúncula de Euphorbia milii (Euphorbiaceae)

Several types of fruit occur in Euphorbiaceae, notably the explosively dehiscent dry fruit, and different seed-coat anatomies with taxonomic importance. This paper aims to describe the pericarp ontogeny and structure in Euphorbia milii Desmoul., and evaluate the presence of the secretory exotesta and caruncle. The fruit is a schizocarp, whose the pericarp development begins with a periclinal division of the inner epidermal cells. The derived cells divide, forming about four layers of obliquely elongated cells. Then, the adjacent parenchyma cells elongate, giving rise to a palisade layer and finally, the cells between this layer and the vascular strands undergo mitosis, originating about four layers of elongated cells perpendicularly to the inner oblique cells. These three zones lignify, while the region between the vascular strands and the exocarp, where idioblasts, hypodermis and laticifers are present do not show significant changes. Before the dehiscence, a lysis of cells of the septa and the desiccation of the fruit occur, which causes contraction of the non-lignified tissues and tension between the lignified zones, promoting rupture of each mericarp from central columella and on the dorsal strand, ejecting the seeds. The seeds have pseudocaruncle and the exotesta secretes mucilage, facilitating their imbibition.