Combination of diffusion tensor and functional magnetic resonance imaging during recovery from the vegetative state

Background: The rate of recovery from the vegetative state (VS) is low. Currently, little is known of the mechanisms and cerebral changes that accompany those relatively rare cases of good recovery. Here, we combined functional magnetic resonance imaging (fMRI) and diffusion tensor imaging (DTI) to study the evolution of one VS patient at one month post-ictus and again twelve months later when he had recovered consciousness. Methods fMRI was used to investigate cortical responses to passive language stimulation as well as task-induced deactivations related to the default-mode network. DTI was used to assess the integrity of the global white matter and the arcuate fasciculus. We also performed a neuropsychological assessment at the time of the second MRI examination in order to characterize the profile of cognitive deficits. Results: fMRI analysis revealed anatomically appropriate activation to speech in both the first and the second scans but a reduced pattern of task-induced deactivations in the first scan. In the second scan, following the recovery of consciousness, this pattern became more similar to that classically described for the default-mode network. DTI analysis revealed relative preservation of the arcuate fasciculus and of the global normal-appearing white matter at both time points. The neuropsychological assessment revealed recovery of receptive linguistic functioning by 12-months post-ictus. Conclusions: These results suggest that the combination of different structural and functional imaging modalities may provide a powerful means for assessing the mechanisms involved in the recovery from the VS.

Combination of diffusion tensor and functional magnetic resonance imaging during recovery from the vegetative state

Background: The rate of recovery from the vegetative state (VS) is low. Currently, little is known of the mechanisms and cerebral changes that accompany those relatively rare cases of good recovery. Here, we combined functional magnetic resonance imaging (fMRI) and diffusion tensor imaging (DTI) to study the evolution of one VS patient at one month post-ictus and again twelve months later when he had recovered consciousness. Methods fMRI was used to investigate cortical responses to passive language stimulation as well as task-induced deactivations related to the default-mode network. DTI was used to assess the integrity of the global white matter and the arcuate fasciculus. We also performed a neuropsychological assessment at the time of the second MRI examination in order to characterize the profile of cognitive deficits. Results: fMRI analysis revealed anatomically appropriate activation to speech in both the first and the second scans but a reduced pattern of task-induced deactivations in the first scan. In the second scan, following the recovery of consciousness, this pattern became more similar to that classically described for the default-mode network. DTI analysis revealed relative preservation of the arcuate fasciculus and of the global normal-appearing white matter at both time points. The neuropsychological assessment revealed recovery of receptive linguistic functioning by 12-months post-ictus. Conclusions: These results suggest that the combination of different structural and functional imaging modalities may provide a powerful means for assessing the mechanisms involved in the recovery from the VS.

Vegetative and reproductive morphology of Kallymenia patens (Kallymeniaceae, Rhodophyta) in the Mediterranean Sea

Reproductive morphology of the Mediterranean red alga Kallymenia patens is described for the first time, confirming its position in the genus. K. patens is characterized by a non-procarpic female reproductive apparatus, carpogonial branch systems consisting of supporting cells bearing both three-celled carpogonial branches and subsidiary cells that lack a hypogynous cell and carpogonium; fusion cells develop numerous connecting filaments, and tetrasporangia are scattered over the thallus and are probably cruciately divided. Old fertile spathulate specimens of K. patens are morphologically similar to K. spathulata, but they can be distinguished by the length of spathulated proliferations (up to 0.6 cm and 6 cm, respectively), the length of inner cortical cells (up to 70 and 30 μm, respectively), and the gonimoblast location (in proliferations from the perennial part of the blade and over all the thallus surface, respectively)

Analysis of a Plant Complex Resistance Gene Locus Underlying Immune-Related Hybrid Incompatibility and Its Occurrence in Nature

Mechanisms underlying speciation in plants include detrimental (incompatible) genetic interactions between parental alleles that incur a fitness cost in hybrids. We reported on recessive hybrid incompatibility between an Arabidopsis thaliana strain from Poland, Landsberg erecta (Ler), and many Central Asian A. thaliana strains. The incompatible interaction is determined by a polymorphic cluster of Toll/interleukin-1 receptor-nucleotide binding-leucine rich repeat (TNL) RPP1 (Recognition of Peronospora parasitica1)-like genes in Ler and alleles of the receptor-like kinase Strubbelig Receptor Family 3 (SRF3) in Central Asian strains Kas-2 or Kond, causing temperature-dependent autoimmunity and loss of growth and reproductive fitness. Here, we genetically dissected the RPP1-like Ler locus to determine contributions of individual RPP1-like Ler (R1R8) genes to the incompatibility. In a neutral background, expression of most RPP1-like Ler genes, except R3, has no effect on growth or pathogen resistance. Incompatibility involves increased R3 expression and engineered R3 overexpression in a neutral background induces dwarfism and sterility. However, no individual RPP1-like Ler gene is sufficient for incompatibility between Ler and Kas-2 or Kond, suggesting that co-action of at least two RPP1-like members underlies this epistatic interaction. We find that the RPP1-like Ler haplotype is frequent and occurs with other Ler RPP1-like alleles in a local population in Gorzów Wielkopolski (Poland). Only Gorzów individuals carrying the RPP1-like Ler haplotype are incompatible with Kas-2 and Kond, whereas other RPP1-like alleles in the population are compatible. Therefore, the RPP1-like Ler haplotype has been maintained in genetically different individuals at a single site, allowing exploration of forces shaping the evolution of RPP1-like genes at local and regional population scales.

Vegetative propagation of bauhinia x blakeana, an ornamental sterile tree

Bauhinia x blakeana (B. purpurea x B. variegata) is a natural hybrid that has been cultivated in gardens, streets and parks. Due to its sterility, it must be vegetatively propagated. The objective of this work was to evaluate the viability of cuttings and grafting on its propagation. Semi-woody cuttings were collected during four seasons and treated with 0; 1,000; 2,000; and 3,000 mg L-1 of IBA. The experimental design was entirely randomized and the treatments were arranged in a 4x4 factorial scheme (four collecting times x four IBA concentrations) and five replications with 10 cuttings each, per collecting time and per IBA concentration. Characteristics of roots and shoots were evaluated after 90 days. The data means were compared by the Tukey test and submitted to the polynomial regression analysis. For the grafting experiment, B. variegata and B. variegata var. candida plants of six and 12 months were used as rootstocks and the splice graft and T-budding methods were tested. The experimental design was entirely randomized and the treatments were arranged in a 2x2x2 factorial scheme (two rootstock species x two grafting methods x two rootstock ages) and four replications with five plants each, per rootstock species, per grafting method and per rootstock age. Characteristics of shoots were evaluated after 90 days and the data means were compared by the Tukey test. B. x blakeana can be propagated by semi-woody cuttings collected in spring, without IBA application, or in summer, with the application of 3,000 mg L-1 of IBA. The tested grafting methods were not effective.

VEGETATIVE RESCUE AND CUTTINGS PROPAGATION OF Araucaria angustifolia (Bertol.) Kuntze

Brazilian pine or araucaria (Araucaria angustifolia) is a coniferous tree with great economic, social and environmental importance in southern Brazil, being exploited for both wood production and for its edible pine nuts. However, no efficient cloning techniques are available and, therefore, the purpose of this study was to evaluate the effectiveness of vegetative rescue methods for cuttings propagation of the species. Shoots/cuttings were generated in two ways: 26 years old trees underwent coppicing and 20 years old trees had the primary branches on the upper third of crown pruned at 2, 20 and 50 cm from the main trunk. Orthotropic shoots were rooted after application of indole-3-butyric acid (IBA) at 0, 2, 4 and 6 g.L-1. Coppicing produced 47 cuttings per plant with 90% orthotropic shoots, while pruning resulted in 182 cuttings per plant with 44% orthotropic shoots. Rooting success indexes were low with no influence of IBA, although they are slightly superior to the ones available in the literature for the species, ranging from 12 to 30% for the coppice shoots and from 0 to 28% for the branches shoots. We conclude that both vegetative rescue techniques are viable and have potentially important applications. Coppicing is recommended for the propagation aiming the production of wood, while shoots derived from the side branches of the crown are more appropriate for seeds orchards formation.

Vegetative and reproductive development of Costa Rican weedy rice compared with commercial rice (Oryza sativa)

The variability in the chronology of the vegetative and reproductive development of weedy rice complex has been little studied. However, a field trial was established to study the timing of growth stages of sixteen weedy rice morphotypes and five rice varieties of Costa Rica. Weedy rice presented a wide range of variation for all descriptors among and within morphotypes. Weedy rice was taller than the rice varieties during vegetative phase and showed a growth increase of 14-23 cm every two weeks. Six morphotypes emerged earlier than commercial rice varieties, but no differences where found between samples for the time required for starting tillering. Early emergence of weedy rice morphotypes was not associated with early flowering, thus no correlation was detected between the vegetative and reproductive phases. All weedy rice morphotypes reached anthesis and maturity earlier than the rice varieties. Nevertheless, varieties Setesa-9 and CR-5272 overlapped anthesis with eleven morphotypes and variety CR-4338 overlapped flowering with eight weedy rice morphotypes. In contrast, none of the morphotypes overlapped anthesis with varieties CR-1821 and CR-1113. The results obtained showed the competitive capacity of weedy rice and provided valuable information about flowering overlap between weedy rice morphotypes and rice varieties which will be useful in the design of gene flow studies among them.

Late-acting self-incompatibility in Capparis retusa (Capparaceae), a species of Chaco woodland in NE Argentina

The reproductive biology of Capparis retusa was studied by means of controlled pollination experiments and fluorescence microscopy observations of post-pollination events. Self-pollinated flowers mostly failed to form fruits despite the fact that self-pollen tubes grew to the ovary and penetrated ovules within 24 h. Since embryo development to globular stage was observed in some self-fertilized ovules it was concluded that control of self-fertilization in this species occurs by some kind of post-zygotic mechanism.

Vegetative organ anatomy of Ianthopappus corymbosus Roque & Hind (Asteraceae-Mutisieae)

A study on the vegetative organ anatomy of Ianthopappus corymbosus was conducted in order to provide a basis for comparison with the genus Richterago, since this species had been previously included in that genus. The anatomical characters of I. corymbosus that support its exclusion from the genus Richteragon are: epithelial cell organization of adventitious root secretory canals, non-glandular trichomes, and presence of cortical vascular bundles in the stem. In Ianthopappus corymbosus, the underground system consists of rhizophore from which adventitious roots branch off. The subapical meristem of the adventitious root revealed that the ground meristem forms the inner layer which in a meristematic phase, forms 2/3 of the cortex. This layer will differentiate in the endodermis, with Casparian strips, and is referred to as meristematic endodermis. Endodermic secretory canals, limited by four epithelial cells, appear in the region adjacent to the primary phloem.

The site of self-incompatibility action in cupuassu (Theobroma grandiflorum)

Cupuassu (Theobroma grandiflorum (Willd. ex Spreng.) Schumann) is a fruit tree that is attracting attention in Brazil and also in other tropical countries. Its pulp is used to prepare ice-cream, juices, jellies and many other home-made sweets. This species has a very low fecundity, attributed to problems with pollination and self-incompatibility, noted as being restrictive to the agronomic productivity of the species. Controlled pollinations between compatible and incompatible parents were made and flowers were collected at three times: 24, 48 and 72 h after pollinations, during the floral seasons of 1995 and 1998. In flowers collected 24 h after pollination, in both compatible and incompatible crosses, around 70% of ovules showed egg apparatus without evidences of sperm cell delivery; nevertheless some pollen tubes had been observed. Ovaries collected at 48 h showed different behaviors according to the type of cross. In the compatibles, 35% of ovules did not show evidences of gamete fusion. In incompatible crosses this number rose to 50%. Collected ovaries with 72 h, from compatible and incompatible crosses, showed respectively 1.2% and 14.2% of the ovules with the egg apparatus and polar nuclei only. In these ovaries, for the two types of crosses, the presence of sperm nuclei was common and the zygote was still undivided. The incompatibility action is occurring after fertilization, in a late self-incompatibility action.

Diurnal variations of non-structural carbohydrates in vegetative tissues of Melinis minutiflora, Echinolaena inflexa and Lolium multiflorum (Poaceae)

Echinolaena inflexa (Poir.) Chase is a native C3 grass species with high biomass production and abundance in the cerrado comparable to Melinis minutiflora Beauv., a C4 forage grass of African origin, widespread in the cerrado, displacing some native herbaceous species. These species accumulate mainly starch, differently of Lolium multiflorum Lam. ssp. italicum Beck cv. Lema (ryegrass), a C3 temperate forage grass that accumulates fructans as the major storage carbohydrate in the vegetative tissues. In the present study, we analyzed diurnal variations in the content and composition of non-structural carbohydrates (NSC) in the aerial vegetative parts of the three species. Results showed similarity in water-soluble carbohydrates (WSC) and starch contents between Melinis minutiflora and Echinolaena inflexa, with the exotic grass accumulating higher amounts of NSC. However, the levels of sucrose, glucose and fructose were 10 fold higher in Lolium multiflorum, that accumulates fructan as the main storage carbohydrate. Although the contents of WSC and starch were similar in the tropical species E. inflexa and M. minutiflora, their diurnal variations were different and could be related to starch metabolism, especially in M. minutiflora.

Vegetative propagation strategies of four rupestrian species of Leiothrix (Eriocaulaceae)

Leiothrix is endemic of South America and includes 37 species, 25 of which occur in the state of Minas Gerais. Nineteen of those occur in the "Serra do Cipó", a mountain chain, located in the southern portion of the Espinhaço mountain range. This study examines vegetative propagation strategies of four species of Leiothrix, endemic to the Minas Gerais portion of the Espinhaço mountain range. For each species we established permanent plots, where we marked 30 to 51 rosettes or clones, and then took morphological and phenological measurements. Leiothrix crassifolia (Bong.) Ruhland and L. curvifolia var. lanuginosa (Bong.) Ruhland are rhizomatous, forming compact clones. Leiothrix vivipara (Bong.) Ruhland does not produce rhizomes, but is pseudoviviparous, i.e., produces numerous ramets originating from inflorescences. These ramets are formed precociously, and the flower heads do not touch the ground. In Leiothrix spiralis (Bong.) Ruhland both of these strategies are seen: it is both rhizomatous and pseudoviviparous. In this species, the ramets are formed late, only after the flower head has touched the ground. One of the typical conditions of the rupestrian grasslands is soil water shortage in some periods of the year and nutrient scarcity all year round. These conditions might have created an ideal ecological scenario for the evolution of both pseudovivipary and rhizomatous clonal growth in Leiothrix.

A tobacco cDNA reveals two different transcription patterns in vegetative and reproductive organs

In order to identify genes expressed in the pistil that may have a role in the reproduction process, we have established an expressed sequence tags project to randomly sequence clones from a Nicotiana tabacum stigma/style cDNA library. A cDNA clone (MTL-8) showing high sequence similarity to genes encoding glycine-rich RNA-binding proteins was chosen for further characterization. Based on the extensive identity of MTL-8 to the RGP-1a sequence of N. sylvestris, a primer was defined to extend the 5' sequence of MTL-8 by RT-PCR from stigma/style RNAs. The amplification product was sequenced and it was confirmed that MTL-8 corresponds to an mRNA encoding a glycine-rich RNA-binding protein. Two transcripts of different sizes and expression patterns were identified when the MTL-8 cDNA insert was used as a probe in RNA blots. The largest is 1,100 nucleotides (nt) long and markedly predominant in ovaries. The smaller transcript, with 600 nt, is ubiquitous to the vegetative and reproductive organs analyzed (roots, stems, leaves, sepals, petals, stamens, stigmas/styles and ovaries). Plants submitted to stress (wounding, virus infection and ethylene treatment) presented an increased level of the 600-nt transcript in leaves, especially after tobacco necrosis virus infection. In contrast, the level of the 1,100-nt transcript seems to be unaffected by the stress conditions tested. Results of Southern blot experiments have suggested that MTL-8 is present in one or two copies in the tobacco genome. Our results suggest that the shorter transcript is related to stress while the larger one is a flower predominant and nonstress-inducible messenger.

Vegetative and reproductive morphology of Kallymenia patens (Kallymeniaceae, Rhodophyta) in the Mediterranean Sea

Reproductive morphology of the Mediterranean red alga Kallymenia patens is described for the first time, confirming its position in the genus. K. patens is characterized by a non-procarpic female reproductive apparatus, carpogonial branch systems consisting of supporting cells bearing both three-celled carpogonial branches and subsidiary cells that lack a hypogynous cell and carpogonium; fusion cells develop numerous connecting filaments, and tetrasporangia are scattered over the thallus and are probably cruciately divided. Old fertile spathulate specimens of K. patens are morphologically similar to K. spathulata, but they can be distinguished by the length of spathulated proliferations (up to 0.6 cm and 6 cm, respectively), the length of inner cortical cells (up to 70 and 30 μm, respectively), and the gonimoblast location (in proliferations from the perennial part of the blade and over all the thallus surface, respectively)

Pollen ultrastructure in anther cultures of Datura innoxia. I. Division of the presumptive vegetative cell.

Ultrastructural features of embryogenic pollen in Datura innoxia are described, just prior to, during, and after completion of the first division of the presumptive vegetative cell. In anther cultures initiated towards the end of the microspore phase and incubated at 28 degrees C in darkness, the spores divide within 24 h and show features consistent with those of dividing spores in vivo. Cytokinesis is also normal in most of the spores and the gametophytic cell-plate curves round the presumptive generative nucleus in the usual highly ordered way. Further differentiation of the 2 gametophytic cells does not take place and the pollen either switches to embryogenesis or degenerates. After 48-72 h, the remaining viable pollen shows the vegetative cell in division. The cell, which has a large vacuole and thin layer of parietal cytoplasm carried over from the microspore, divides consistently in a plane parallel to the microspore division. The dividing wall follows a less-ordered course than the gametophytic wall and usually traverses the vacuole, small portions of which are incorporated into the daughter cell adjacent to the generative cell. The only structural changes in the vegetative cell associated with the change in programme appear to be an increase in electron density of both plastids and mitochondria and deposition of an electron-dense material (possibly lipid) on the tonoplast. The generative cell is attached to the intine when the vegetative cell divides. Ribosomal density increases in the generative cell and exceeds that in the vegetative cell. A thin electron-dense layer also appears in the generative-cell wall. It is concluded that embryogenesis commences as soon as the 2 gametophytic cells are laid down. Gene activity associated with postmitotic synthesis of RNA and protein in the vegetative cell is switched off. The data are discussed in relation to the first division of the embryogenic vegetative cells in Nicotiana tabacum.