The orchid flora of Portugal - Addendum n.2 - Chorological and systematic remarks on Dactylorhiza maculata (L.) Soó s.l. and Epipactis helleborine (L.) Crantz - First report of E. phyllanthes G.E.Sm.

Close examination of Dactylorhiza maculata s.l. populations in the Serra da Estrela lead to the proposal that all of them are referable to D. caramulensis and that D. ericetorum is probably absent from this area, although it is well represented in the north of the country. New hypotheses are formulated towards a possible hybrid origin of D. caramulensis between D. ericetorum and D. elata. Three localities with truly typical representatives of Epipactis helleborine are reported, two from the Bragança district and one from the Serra da Estrela. A single individual of E. phyllanthes was discovered close to Bragança; this is the first mention of an autogamous Epipactis in Portugal. Trend towards autogamy was also detected on one plant from the Serra da Estrela. A discussion is initiated on the need to further develop understanding on chromosome structure and mating systems in orchid species, especially in complex genera such as Dactylorhiza and Epipactis, to adequately fulfil conservation purposes

Effects of high temperature on mitotic index, microtubule and chromatin organization in rye (Secale cereale L.) root-tip cells

Stressful high temperatures on plants can limit whole-plant function and decrease crop productivity. However, little is known regarding heat stress effects on microtubule cytoskeleton and chromatin in roots from intact plants. Here we studied high temperature effects on cell division, microtubule and chromatin organization patterns in rye root tips from intact plants subjected to 40ºC for 4 h and after different recovery periods (0RT, 7RT, 24 RT). We showed that heat stress induced changes in nuclear morphology as detected by the unusual presence of interphase cells with irregularly shaped nuclei, probably associated with changes in chromosome segregation at anaphase, leading to micronuclei formation as well as changes in the mitotic index. These alterations were associated to differential effects in microtubules organization in both heat-stressed interphase and mitotic cells at 0RT and 7RT. Although no changes in the distribution of H3 phosphorylation of Ser 10 residues on chromatin were found in cells from heat-stressed plants, marked alterations in chromatin DNA methylation patterns were detected. These effects included higher agglutination of 5-methylcytosine domains in both interphase and metaphase cells compared to controls. Taken together these results seem to suggest that alterations in microtubule conformation upon heat stress influences nuclear chromatin organization and cell cycle progression. However, when seedlings recovered from stress (24RT), root tip cells presented microtubule configurations and chromatin organization patterns similar to controls. We conclude that in spite of heat stress markedly altered cell cycle progression and distribution of epigenetic marks, these responses are transient to cope with such stress conditions in the rootsinfo:eu-repo/semantics/publishedVersio

Conservation of a critically endangered endemic halophyte of West Portugal: a microcosm assay to assess the potential of soil technology for species reintroduction

Original ResearchThe soil system has been frequently overlooked during plant reintroduction planning and practice since working with soils and plant roots can be difficult, particularly in saline environments. Coastal saline environments are major contributors to regional and global biodiversity and an important source of endemic species. However, various species are in decline or considered threatened, particularly halophytes (salt tolerant) due to negative anthropic impacts. The Lusitanian endemic halophyte Limonium daveaui formerly had a large distribution range along the west coast of Portugal but currently it shows a restricted distribution in the Tagus estuary. Field surveys revealed that this critically endangered species forms few local populations with small size invaded by exotic species. In this study, we investigated the potential utilization of Technosols, an innovative sustainable, ecological engineering method combined with brackish water irrigation for potential L. daveaui reintroduction in native habitats. Seed germination percentages were evaluated in different environmental conditions. Through a microcosm assay, a Technosol was constructed using a saline Fluvisol with a mixture of low value inorganic and organic wastes, which were chemically characterized. Plants were cultivated in the Fluvisol and Technosol and irrigated with brackish water collected in the nearby area. To assess plant growth, morphometric parameters and the plants’ physiological status were assessed and the fresh and dry biomass determined. Results showed that seed germination was higher on moist filter paper with distilled water than in Fluvisol or Technosol. Plants grown in Technosol had a greater development, with higher values of photosynthetic indexes and biomass production than in Fluvisol. Our findings provide a basis for future in situ conservation studies and support the idea that eco-friendly soil technology approaches are beneficial to conserve rare halophyte speciesinfo:eu-repo/semantics/publishedVersio

Relationships between transcription, silver staining and chromatin organization of nucleolar organizers in Secale cereale

Summary. The nucleolar organizer regions (NORs) are composed of hundreds of rRNA genes, typically spanning several megabases. Cytologically, NORs include regions that are highly condensed and regions that are decondensed, the latter corresponding to regions at which associated proteins stain intensively with silver (Ag-NORs) and where active rRNA gene transcription is thought to occur. To test the relationship between rRNA gene activity, NOR silver staining, and rDNA (genes coding for rRNA) chromatin condensation, we used the DNA methyltransferase inhibitor 5-azacytidine to evaluate the correlation between the epigenetic regulation of rRNA genes and NOR silver staining in the plant Secale cereale. Following 5-azacytidine treatment, we observed an increase in rRNA gene transcription as well as a reduction in the number of cells showing a significant difference in the size of the silver-stained domains in the two NORs. These transcriptional changes occurred concomitantly with an increase in nuclear and nucleolar size and were associated with the reallocation of most of the rDNA from perinucleolar heterochromatin into the nucleolus. Collectively, these results suggest that rRNA gene transcription, silver staining, and NOR decondensation are interrelated in S. cereale

Le statut et la protection des orchidées du Portugal

Fifty-six species and subspecies are presently recognised as indigeneous in Portugal. About half of them are rare or very rare, or even probably extinct for two of them. The causes of rarity and the factors responsible for the regression that still affects many species are analysed. Besides the fact that a few species have only little Mediterranean or Atlantic affinities, there are causes linked with habitat disappearance, mainly in humid and coastal zones. Regulations applicable in Portugal are reviewed, but few of them are specifically concerned with orchids. Furthermore orchids are never considered among priority options towards the establishment of nature reserves and parcs. We propose a list of sites and habitats especially rich or favourable to the conservation of orchids, which should receive a better attention on behalf of Portuguese officers responsible for nature protection. Moreover, knowledge of demographic and genetic characteristics of orchid populations should absolutely be developed if we wish to guarantee the effectiveness of conservation strategies

Effects of individual and population parameters on reproductive success in three sexually deceptive orchid species

Reproductive success (RS) in orchids in general, and in non-rewarding species specifically, is extremely low. RS is pollinator and pollination limited in food deceptive orchids, but this has rarely been studied in sexually deceptive orchid species. Here, we tested the effects of several individual (plant height, inflorescence size, nearest neighbour distance and flower position) and population (patch geometry, population density and size) parameters on RS in three sexually deceptive Ophrys (Orchidaceae) species. Inter-specific differences were observed in RS of flowers situated in the upper versus the lower part of the inflorescence, likely due to species-specific pollinator behaviour. For all three species examined, RS increased with increasing plant height, inflorescence size and nearest neighbour distance. RS generally increased with decreasing population density and increasing patch elongation. Given these results, we postulate that pollinator availability, rather than pollinator learning, is the most limiting factor in successful reproduction for sexually deceptive orchids. Our results also suggest that olfactory ‘display’ (i.e. versus optical display), in terms of inflorescence size (and covarying plant height), plays a key role in individual RS of sexually deceptive orchids. In this regard, several hypotheses are suggested and discusse

Genome restructuring in rye affects the expression, organization and disposition of homologous rDNA loci

Research ArticleThe standard rye cultivar ‘Imperial’ and a structural variant carrying an intact 1R chromosome and two telocentric 1R chromosomes (short and long arms) were used to investigate expression patterns of homologous rDNA loci, and the influence of chromosome structural change on their interphase organisation and relative disposition. Sequential silver staining and in situ hybridization with the rDNA probe pTa71, established a correspondence between the expression and organization patterns of rDNA domains in metaphase and interphase cells. In most cells of the cultivar Imperial, nucleolar organizer region (NOR) silver staining on metaphase chromosomes with equivalent numbers of rDNA genes revealed a size heteromorphism between homologousrDNA loci, resulting from their differential expression. NOR heteromorphism in the structural variant line was significantly reduced. The preferential activity of one NOR over its homologue was found to be random within cells and independent of parental origin. Nucleotypic modifications mediated by changes in the 1R chromosome structure include increased proximity between homologous rDNA loci in interphase, and an increase in the frequency of cells with intra-nucleolar ribosomal condensed chromatin. These results seem to indicate a ‘sequence recognition’ process for the regulation of homologous loci

Ribosomal DNA heterochromatin in plants

The aim of this review is to integrate earlier results and recent findings to present the current state-of-the art vision concerning the dynamic behavior of the ribosomal DNA (rDNA) fraction in plants. The global organization and behavioral features of rDNA make it a most useful system to analyse the relationship between chromatin topology and gene expression patterns. Correlations between several heterochromatin fractions and rDNA arrays demonstrate the heterochromatic nature of the rDNA and reveal the importance of the genomic environment and of developmental controls in modulating its dynamicsFCT - Fundação para a Ciência e Tecnologi

Phosphorilation of histone H3 in plants - a dynamic affair

Histones are the main protein components of chromatin: they undergo extensive post-translational modifications, particularly acetylation, methylation, phosphorylation, ubiquitination and ADP-ribosylation which modify the structural/functional properties of chromatin. Posttranslational modifications of the N-terminal tails of the core histones within the nucleosome particle are thought to act as signals from the chromatin to the cell, for various processes. Thus, in many ways histone tails can be viewed as complex protein–protein interaction surfaces that are regulated by numerous post-translational modifications. Histone phosphorylation has been linked to chromosome condensation/segregation, activation of transcription, apoptosis and DNA damage repair. In plants, the cell cycle dependent phosphorylation of histone H3 has been described; it is hyperphosphorylated at serines 10/28 and at threonines 3/11 during both mitosis and meiosis in patterns that are specifically coordinated in both space and time. Although this post-translational modification is highly conserved, data show that the chromosomal distribution of individual modifications can differ between groups of eukaryotes. Initial results indicate that members of the plant Aurora kinase family have the capacity to control cell cycle regulated histone H3 phosphorylation, and in addition we describe other potential H3 kinases and discuss their functions

Rye Bs disclose ancestral sequences in cereal genomes with a potential role in gametophyte chromatid segregation

Research ArticleTwo sequence families, E3900 and D1100, are amplified on the subtelomeric domain of the long arm of rye B chromosomes, the region that controls its drive mechanism. In this work, polymerase chain reaction (PCR) with a number of primers spanning E3900 shows that the organization and nucleotide sequence of E3900-related portions are present and highly conserved on rye A chromosomes as well as in other cereals. Quantitative Real-Time PCR estimates two E3900 sequences to be represented in 100–150 copies on Bs and at least as single copies on As. A novel E3900- related sequence, with a deletion that results in a frameshift and subsequently an open reading frame with putative DNA binding motifs, is identified. Expression analysis of E3900 indicates identical transcription levels in leaves from plants with and without Bs, showing that the expression of these sequences must be silenced on Bs and tightly regulated on As in leaves. In contrast, E3900 transcription is upregulated during meiosis exclusively in plants with Bs, maintaining a high level of transcription in the gametophyte. Interestingly, Bs not only influence their own chromatid segregation but also that of the regular chromosome complement of both rye and wheat. There is a drastic increase in frequency of disrupted metaphase and anaphase cells in the first mitosis of pollen grains carrying Bs, which appears to be due to anomalous adherences between sister chromatids. Taken together, this work provides insight into how E3900 sequences are potentially associated with important evolutionary mechanisms involved in basic cellular processes