Stefano Mosti

Programmed-cell-death events during tapetum development of angiosperms

SummaryProgrammed-cell-death events in the tapetum of two angiosperms (Lobivia rauschii Zecher andTillandsia albida Mez et Purpus) are described by ultrastructural methods. Tapetum degradation appears to be a type of programmed cell death, with the cellular remnants necessary for pollen development, acting as products of holocrine secretion. Diagnostic features of apoptosis during tapetum development are: general shrinkage of the whole cell and the nuclei; condensation of the chromatin at the periphery of the internal nuclear membrane; the enlargement of the endoplasmicreticulum cisternae to circumscribe portions of the cytoplasm; the persistence of mitochondria together with microfilament bundles until the last stages of tapetal degeneration.

Megasporogenesis and programmed cell death in Tillandsia (Bromeliaceae)

The degeneration of three of four meiotic products is a very common process in the female gender of oogamous eukaryotes. In Tillandsia (and many other angiosperms), the surviving megaspore has a callose-free wall in chalazal position while the other three megaspores are completely embedded in callose. Therefore, nutrients and signals can reach more easily the functional megaspore from the nucellus through the chalazal pole with respect to the other megaspores. The abortion of three of four megaspores was already recognized as the result of a programmed cell death (PCD) process. We investigated the process to understand the modality of this specific type of PCD and its relationship to the asymmetric callose deposition around the tetrad. The decision on which of the four megaspores will be the supernumerary megaspores in angiosperms, and hence destined to undergo programmed cell death, appears to be linked to the callose layer deposition around the tetrad. During supernumerary megaspores degeneration, events leading to the deletion of the cells do not appear to belong to a single type of cell death. The first morphological signs are typical of autophagy, including the formation of autophagosomes. The TUNEL positivity and a change in morphology of mitochondria and chloroplasts indicate the passage to an apoptotic-like PCD phase, while the cellular remnants undergo a final process resembling at least partially (ER swelling) necrotic morphological syndromes, eventually leading to a mainly lipidic cell corpse still separated from the functional megaspore by a callose layer.

Classical macroautophagy in Lobivia rauschii (Cactaceae) and possible plastidial autophagy in Tillandsia albida (Bromeliaceae) tapetum cells

The tapetum in anthers is a tissue that undergoes programmed cell death (PCD) during the production of pollen. We observed two types of autophagy prior to cell death. In Lobivia rauschii (Cactaceae), tapetum cells showed plant-type autophagosomes–autolysosomes, which have been found previously exclusively in root meristem cells. The autophagic structures were formed by a network of tubules which apparently merged laterally, thereby sequestering a portion of the cytoplasm. The organelles observed in the sequestered material included multilamellar bodies, which have not been reported earlier in these organelles. By contrast, Tillandsia albida (Bromeliaceae) tapetum cells contained no such organelles but showed plastids that might possibly carry out autophagy, as they contained portions of the cytoplasm similar to the phenomenon reported earlier in Phaseolus and Dendrobium. However, the ultrastructure of the T. albida plastids was different from that in the previous reports. It is concluded that in L. rauschii classical plant macroautophagy was involved in degradation of the cytoplasm, while in T. albida such classical macroautophagy was not observed. Instead, the data in T. albida suggested the hypothesis that plastids are able to carry out degradation of the cytoplasm.

Tracking the origin of the invading Caulerpa (Caulerpales, Chlorophyta) with Geographic Profiling, a criminological technique for a killer alga

The application of the Geographic Profiling technique (with the “Rossmo formula”) proved to be effective in assessing the spreading origin of invading species of Caulerpa in the Mediterranean. Geoprofiling is a technique more frequently used in criminology. We applied this method to an algal invasion for the first time. The method was calibrated with the distribution data of Caulerpa taxifolia, whose spreading in the Mediterranean Sea started from the aquarium of Monaco. This is the first time that Geographic Profiling is calibrated on a data set of sites of presence of a biological invader, of which the spreading origin is known. The application on Caulerpa racemosa var. cylindracea showed that the probable spreading center of the alga should be located in Western Sicily for the Mediterranean and in the southern part of Tenerife for the Canary Islands (Las Galletas, a fishing village). The spreading centers correspond to areas with ports, indicating that the spreading of this alga should be related to (fishing) boats. This result confirmed the opinion of other authors. The parameters used in the Rossmo Formula, obtained through calibration with the known origin of C. taxifolia invasion for identifying the origin of the spread of C. racemosa var. cylindracea may be extended to other algae with similar propagation mode and similar habitat requirements.

Chromium accumulation and changes in plant growth, selected phenolics and sugars of wild type and genetically modified Nicotiana langsdorffii.

Chromium accumulation, dry weight (DW) biomass yield, water content and concentrations of some selected phenolic compounds and carbohydrates were determined in root and shoot of Nicotiana langsdorffii, either wild type (WT) or genetically modified by the insertion of GR and rolC genes, in response to the presence of Cr(VI) in the growth medium. A biomass decrease was observed for WT plants, but not for GR and rolC transformations, in response to Cr(VI) in the growth medium, highlighting a stress situation only in WT line. Shoot chromium concentrations were in all cases about 300 mg kg(-1) DW. In root higher concentrations were found in rolC than in GR and WT (3843, 2600 and 2751 mg kg(-1) DW, respectively). Based on the DW biomass, GR and WT accumulated higher chromium quantities than rolC, both in root (330 and 424 versus 85 μg Cr per plant) and shoot (282 and 275 versus 121 μg Cr per plant). Therefore, GR should be preferred to WT as a promising candidate for chromium phytoremediation. Metabolic shifts of sugars and phenolics were generally observed in response to either gene insertions or exposure to Cr(VI), being the latter more related to the resistance to Cr(VI) than the former.

Ultrastructural aspects of the embryo and different endosperm compartments, in Eruca sativa Hill cv. Nemat (Brassicaceae) during Heart and Torpedo stages

Abstract These observations are the first on the ultrastructure of the embryo and the endosperm of Eruca sativa Hill. We investigated the cv. Nemat, which is characterized by a particularly high amount of lipids and glucosinolates. Our observations suggested that the thick and abundant micropylar endosperm, completely surrounding the suspensor, may be the main active source of nutrients for the embryo. This endosperm, like the central chamber endosperm, is particularly rich in functional chloroplasts and cellularizes later with respect to the other previ- ously investigated Brassicaceae. The last (distal with respect to the embryo) suspensor cell exhibits important features related to the passage of nutrients, such as wall ingrowths. In fact these ingrowths appear as highly convoluted labyrinthine wall projections. Such ultrastructural features are typical of transfer cells. The accumu- lation stage in E. sativa cv. Nemat appears to occur early (Heart stage of embryo development, as Brassica na pus). The endosperm compartment called Chalazal Endosperm Cyst (CEC), contributes actively to the embryo trophism during the Heart and Torpedo stages. This function is evident because of the high number of chloro- plasts in the cyst and for the observed continuity between the CEC and the other endosperm compartments (CC endosperm and micropylar endosperm) in cv. Nemat. The morphology of the CEC appeared to be more similar to the pyriform shape sensu Brown et al., but with a more flattened base with respect to the proposed examples, and without labyrinthine wall. The Chalazal Chamber appeared to be more similar to the Brown’s type B in E. sativa. The presence of chloroplasts with a well developed thylakoid system indicates an active photosynthetic activity by the majority of the seed tissues. E. sativa leaves are normally harvested for food, while the seeds of cv. Nemat appear to be particularly rich in oil. The premature independence of seeds and fruits from the necessity of absorbing nutrients from the rest of the plant, could indicate the possibility of harvesting both leaves (earlier) and seeds (later) in this plant without compromising a full seed maturation.

SURFACE INTERACTIONS OF THE EPIPHYTIC MACROALGA HINCKSIA MITCHELLIAE (PHAEOPHYCEAE) WITH THE SHOALGRASS HALODULE WRIGHTII (CYMODOCEACEAE)(1).

Meadows of Halodule wrightii (Cymodoceaceae) underwent a decline in a tidal flat located at Paranaguá Bay (Parana, SE Brazil). This decline appeared to be related to an overgrowth of the epiphytic macroalga Hincksia mitchelliae (Harv.) P. C. Silva (Phaeophyceae). In order to characterize the type of epiphytism between the alga and its plant host, we compared two samples from the beginning and end of the algal overgrowth via electron and optical microscopes. The investigation revealed that at both sampling periods, there was an epiphytism of type II, which is due to an infection of epiphytes strongly attached to the surface of the host but not associated to any apparent direct host‐tissue damage. The presence of plasmodesmata between the cells of Hincksia only in the late stage of the host–epiphyte interaction indicated a change in the vegetative organization of Hincksia in relation to its host to improve nutrient absorption and distribution through the epiphyte cells. This is the first report on plasmodesmata in H. mitchelliae. The proposed mechanisms with which the algal epiphytes lead seagrasses to death are shadowing by adhesion on Halodule surface and disruption of its osmoregulatory system. Our findings have implications for the conservation and management strategies of seagrass ecosystems.

Notes on Trochiscanthes Koch (Apiaceae) on the basis of ITS rDNA sequence

Abstract Trochiscanthes Koch (Apiaceae) is an european endemic monospecific genus. This paper deals with the phylogenetic analysis on molecular basis to assess the relationships of the genus. Parsimony, maximum likelihood and Bayesian support analyses on the basis of the ribosomal internal transcribed spacers were adopted. Moreover data were added to mericarp section data, one of the most important morphological markers in Apiaceae. Trochiscanthes nodiflora, Ligusticum porteri, Ligusticum canadense and Conioselinum scopulorum clustered together with 100% Bayesian and Bootstrap (parsimony) support. These clade was sister to Conioselinum chinense (93% Bayesian and 59% Bootstrap support). This clade corresponds to the “Conioselinum chinense” clade, by other authors. All these species clustered together with Meum athamanticum (97% Bayesian and 62% Bootstrap support). This result clarifies the position of genus Trochiscanthes. Moreover, since Ligusticum and Conioselinum (in traditional sense) resulted as probably not monophyletic in previous studies and the type species of these genera clustered far away from the “Conioselinum chinense” clade, the necessary taxonomical rearrangements (after a further revision of the group) could find a solution transferring the species currently inserted in Ligusticum and Conioselinum but within the “Conioselinum chinense” clade, to genus Trochiscanthes.

Stigma morphology in Aylostera subg. Mediolobivia (Cactaceae) and its importance for systematic investigation

The stigma of 31 taxa of Aylostera (Cactaceae) was studied using environmental scanning electron microscopy. Seven stigma types could be defined, based on the degree of fusion of the stigmatic lobes and, when these are completely separated (types I and II), their orientation. Four types (types IV, V, VI and VII) are present only in Aylostera subg. Mediolobivia, two of which (types IV and VII) are species-specific. Stigma morphology was found to be a good diagnostic character in Aylostera subg. Mediolobivia, allowing in some cases to distinguish species that had been confused previously. The stigmatic types here defined seem apt to characterize natural groups within the subgenus and are useful as an aid for species identification and the characterization of new taxa.