Epidermal and subepidermal changes during the formation of hairy galls induced by Eriophyidae on Avicennia schaueriana leaves.

Leaf-galling Eriophyidae (Acarina) may promote simple or complex alterations in the organs of their host plants, such as an increase in indumentum density or the reorganization of epidermis and ground system tissue patterns. To test if hairy galls of Eriophyidae on Avicennia schaueriana (Acanthaceae) are related to complex changes, leaf galls in distinct developmental phases were compared to non-galled leaves using anatomical, histochemical, and histometric analyses. Quantitative comparisons of preferential gall induction sites and gall area according to distinct leaf portions were made to evaluate if the impacts of gall formation can be related to the distinct potentialities of leaf microsites. The apical portion of the leaves and leaf margins were the sites with the highest occurrence of galls, but no relationship was detected between gall area and induction site. The gall anatomy revealed that epidermal features are influenced the most with the development of abnormal stomata and projected or sunken salt glands. The most striking change is the neoformation of elongated filiform trichomes on the abaxial surface (where the mites occur) that accumulate reducing sugars and proteins. The filiform trichomes may protect the inducers against abiotic stressors and enemies, and the primary metabolites that accumulate are important foods for mites. The mesophyll has simple alterations, only in the spongy parenchyma. Complex alterations occur only in abaxial epidermal cells close to feeding sites of the inducer. The number of inducers per gall seems to be the most important influence on gall size, since gall area is not related to the position in the leaves.

Megagametophyte development and female sterility in Maytenus obtusifolia Mart. (Celastraceae).

Reproduction in flowering plants is closely related to the megagametophyte, since the megagametophyte is involved in pollen tube reception and contains the two female gametes-egg cell and central cell. Previous conventional light microscopy methods have shown that female sterility in perfect flowers of Maytenus obtusifolia is associated with the occurrence of sterile ovules whose megagametophytes have hypertrophied synergids. Here, using transmission electron microscopy and cytochemical methods, we compare the megagametophytes in fertile and sterile ovules from perfect and pistillate flowers, and investigate the cellular events that result in the degradation of the megagametophyte cells from sterile ovules. In fertile ovules of perfect and pistillate flowers, mature megagametophytes have two synergids, egg cell and central cell. In fertile ovules, the synergids present an extensive rough endoplasmic reticulum (RER) profile, large populations of mitochondria, when compared to egg cells, vesicles, Golgi bodies, plastids and a nucleus with heterochromatin. Besides that, the egg cell has a small population of organelles and the central cell exhibits cytoplasm with free ribosomes, RER, vesicles originating from the RER, Golgi bodies and oil inclusions. In mature megagametophytes from sterile ovules of perfect and pistillate flowers, massive autophagy occurs by tonoplast rupture promoting hydrolase release, leading to protoplast and cell wall degradation-typical evidence of programmed cell death (PCD). Therefore, female sterility in the majority of M. obtusifolia sterile ovules is the result of PCD by massive autophagy in the megagametophyte cells. In a few other sterile ovules, sterility is due to the delayed or the absence of megagametophyte development.

Pollen grain development and male sterility in the perfect flowers of Maytenus obtusifolia Mart. (Celastraceae).

Perfect flowers of Maytenus obtusifolia have partial sterility of pollen grains, resulting in collapsed and developed free microspores. However, the cellular events resulting in partial male sterility have not been determined. In pistillate flowers of this species, male sterility is related to the premature programmed cell death (PCD) in tapetum and sporogenic cells. The process occurs through autophagy via macroautophagy and massive autophagy and is associated with sporophytic cytoplasmic male sterility (CMS). Here, we characterised the development of pollen grains and investigated the cellular events that result in tapetum cells and free microspores PCD in perfect flowers, using light and transmission electron microscopy combined with the TUNEL (Terminal deoxynucleotidyl transferase mediated dUDP end-Labeling) assay and the ZIO (Zinc iodide-osmium tetroxide) method. Pollen grain development in perfect flowers was divided into eight developmental stages based on the characteristics of the pollen grains. Tapetum cells undergo PCD at the free microspore stage, through a macroautophagic process, by formation of autophagosomes and by autophagosomes giving rise to lytic vacuoles at maturity. In the final stage of PCD, massive autophagy occurs by rupture of the tonoplast. The development of viable and inviable microspores diverges at the vacuolated microspore stage, when PCD occurs in some free microspores, causing interruption of pollen development through necrosis. These events result in the observed partial male sterility. Viable microspores undergo mitosis and develop into tricellular pollen grains. Male sterility in hermaphrodite individuals is here interpreted as gametophytic CMS.