Alessio Papini

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.

Tracking the invasion of the alien fruit pest Drosophila suzukii in Europe

Biological invasions are a leading threat to native wildlife, human health and food production worldwide. Understanding the invasion history helps identifying introduction pathways and organizing integrated management strategies especially aimed at avoiding multiple reintroductions. We coupled a recently developed spatial analysis (Geographic profiling) with trade flows quantification to identify the most likely spreading centre of a recent invader of Europe, the spotted wing drosophila, Drosophila suzukii. This polyphagous vinegar fly recently colonized western countries, where it is heavily threatening fruit production causing severe economic losses. Characterized by a rapid spread and a huge impact, the invasion of this pest has a few precedents and it is becoming a model in invasion biology and pest management. Thanks to our spatial approach based on data presence of D. suzukii in European countries in the very first years of it spread, we update the current knowledge of a first spread in Spain and Italy, suggesting on the contrary that the South of France may be the most likely spreading centre of D. suzukii in Europe. Estimates of propagule pressure (fresh host fruits importation) support this finding as imports from contaminated South East Asian countries are higher in France than in Spain or Italy. Our study provides a first step in the comprehension of invasion history of this pest species and emphasizes geographic profiling as an efficient technique to track down invaders colonization patterns.

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.

Ultrastructure of autophagy in plant cells: a review.

Just as with yeasts and animal cells, plant cells show several types of autophagy. Microautophagy is the uptake of cellular constituents by the vacuolar membrane. Although microautophagy seems frequent in plants it is not yet fully proven to occur. Macroautophagy occurs farther away from the vacuole. In plants it is performed by autolysosomes, which are considerably different from the autophagosomes found in yeasts and animal cells, as in plants these organelles contain hydrolases from the onset of their formation. Another type of autophagy in plant cells (called mega-autophagy or mega-autolysis) is the massive degradation of the cell at the end of one type of programmed cell death (PCD). Furthermore, evidence has been found for autophagy during degradation of specific proteins, and during the internal degeneration of chloroplasts. This paper gives a brief overview of the present knowledge on the ultrastructure of autophagic processes in plants.

Species concepts and speciation factors in cyanobacteria, with connection to the problems of diversity and classification

The cyanobacteria are the most important prokaryotic primary producers on Earth, inhabiting a great diversity of aquatic and terrestrial environments exposed to light. However, the evolutionary forces leading to their divergence and speciation remain largely enigmatic compared to macroorganisms due to their prokaryotic nature, including vast population sizes, and largely asexual reproduction. The advent of modern molecular techniques has facilitated an understanding of the important factors shaping cyanobacterial evolution, including horizontal gene transfer and homologous recombination. We review the forces shaping the evolution of cyanobacteria and discuss the role of cohesive forces on speciation. Further, while myriad species concepts and definitions are currently used, only a limited subset might be applied to cyanobacteria due to their asexual reproduction. Additionally, concepts based solely on phenotypes provide insufficient resolution. A monophyletic species concept which is universal may be ideal for cyanobacteria. Actual identification of the cyanobacteria is difficult due to cryptic diversity, lack of morphological variability, and frequent convergent evolutionary events. Thus, applied molecular techniques such as DNA barcoding will be useful for identifications of environmental samples. Lastly, we show that the real biodiversity of the cyanobacteria is widely underestimated, due in part to low sampling efforts, sensitivity to the molecular markers employed, and the species definitions employed by researchers. In conclusion, we anticipate a rapid increase in cyanobacterial taxa described and large revisions of the system in the future as scientists adopt a common approach to cyanobacterial systematics.

Multiple genome relationships and a complex biogeographic history in the eastern range of Quercus suber L. (Fagaceae) implied by nuclear and chloroplast DNA variation.

Abstract The complex evolutionary history of Quercus suber has been throughly investigated in many recent works, but the details of its differentiation processes are still largely unknown. In addition, the geographical and evolutionary roles of the eastern parts of the species range have gained much less attention compared to other southern European areas. In order to fill this gap, new insights to infer the species diversification and range establishment of the cork oak in the east-central Mediterranean are here provided by means of inter- and intra-specific plastid DNA and nuclear ribosomal ITS phylogeographic studies. We analyzed 95 natural cork oak populations; 6 closely related, sympatric oaks were included in the study and used for comparisons. Evidence for a clear phylogeographical structure was detected with PCR-RFLP at 5 chloroplast loci, while ITS sequence variation is apparently unrelated with the geographical distribution. Five chloroplast haplotypes and three ITS main lineages were identified. Three haplotypes and all ITS lineages occur in the Italian Peninsula, stressing the importance of these territories for the evolutionary history of the species. Two divergent “Italian” haplotypes are highly shared, and one ITS variant is basal to the ingroup, revealing sister relationships within Cerris taxonomic group. Hypotheses of hybridization, lineage sorting of ancient DNA polymorphisms and of reticulate evolution of the whole species group are presented and discussed.

Plastome data reveal multiple geographic origins of Quercus Group Ilex

Nucleotide sequences from the plastome are currently the main source for assessing taxonomic and phylogenetic relationships in flowering plants and their historical biogeography at all hierarchical levels. One major exception is the large and economically important genus Quercus (oaks). Whereas differentiation patterns of the nuclear genome are in agreement with morphology and the fossil record, diversity patterns in the plastome are at odds with established taxonomic and phylogenetic relationships. However, the extent and evolutionary implications of this incongruence has yet to be fully uncovered. The DNA sequence divergence of four Euro-Mediterranean Group Ilex oak species (Quercus ilex L., Q. coccifera L., Q. aucheri Jaub. & Spach., Q. alnifolia Poech.) was explored at three chloroplast markers (rbcL, trnK/matK, trnH-psbA). Phylogenetic relationships were reconstructed including worldwide members of additional 55 species representing all Quercus subgeneric groups. Family and order sequence data were harvested from gene banks to better frame the observed divergence in larger taxonomic contexts. We found a strong geographic sorting in the focal group and the genus in general that is entirely decoupled from species boundaries. High plastid divergence in members of Quercus Group Ilex, including haplotypes shared with related, but long isolated oak lineages, point towards multiple geographic origins of this group of oaks. The results suggest that incomplete lineage sorting and repeated phases of asymmetrical introgression among ancestral lineages of Group Ilex and two other main Groups of Eurasian oaks (Cyclobalanopsis and Cerris) caused this complex pattern. Comparison with the current phylogenetic synthesis also suggests an initial high- versus mid-latitude biogeographic split within Quercus. High plastome plasticity of Group Ilex reflects geographic area disruptions, possibly linked with high tectonic activity of past and modern distribution ranges, that did not leave imprints in the nuclear genome of modern species and infrageneric lineages.

Programmed cell death in the nucellus of Tillandsia (Bromeliaceae)

Abstract We report the ultrastructural aspects and histochemical assay regarding the degenerating nucellar cells of Tillandsia during early female gametophyte development. Nucellar cells degeneration was observed to start during the somatogenesis of the gametophyte. Ultrastructural features of degenerating process and TUNEL assay indicated that the nucellar cells death mechanism is genetically controlled. Mitochondria and cytoplasmic vesicles persisted until cell degeneration was completed. This feature represens a peculiar aspect of the PCD mechanism in this specific case.

Molecular phylogeny, morphology and taxonomic re-circumscription of the generic complex Nonea/Elizaldia/Pulmonaria/Paraskevia (Boraginaceae-Boragineae)

We performed a molecular phylogenetic analysis of the generic complex Nonea/Elizaldia/Paraskevia/ Pulmonaria (Boraginaceae-Boragineae), using trn L UAA and ITS1 sequences from non-coding plastid and nuclear DNA, respectively. In the strict consensus of most parsimonious trees from combined ITS1/trnL sequences the ingroup forms a monophyletic group with two sister clades of Nonea/Elizaldia and Paraskevia/ Pulmonaria. The eastern Mediterranean Nonea ohtusifolia is sister to the rest of the ingroup and more distant to Nonea taxa than to those of Pulmonaria. Elizaldia is firmly nested in Nonea and forms a clade that includes the southern Mediterranean species N. vesicaria. Nonea as currently circumscribed is therefore paraphyletic. The molecular results are congruent with morphological, karyological and chorological features and suggest a rede-finition of Nonea s.l. based on monophyletic groups. We propose: (i) the institution of the new monotypic genus Melanortocarya gen. nov. for N. ohtusifolia, (ii) the inclusion of Elizaldia within Nonea, and (iii) the transfer of Paraskevia (Nonea) cesatiana to Pulmonaria. Nonea embergeri, previously treated at the infraspecific level, is recognized at species rank and E. calycina subsp. multicolor is placed in synonymy of N. calycina.

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.