植物中的细胞程序性死亡

细胞程序性死亡（ＰＣＤ）对于维持植物的正常生长发育非常重要,目前已成为植物学研究的一个热点。本文综合评述了近年来植物ＰＣＤ研究的某些进展,包括植物ＰＣＤ的特征,植物的营养生长、生殖生长以及与环境互作过程中存在的各种ＰＣＤ及其证据,植物ＰＣＤ发生的分子机制及其调控等等。对植物ＰＣＤ研究中有待进一步解决的问题和可能意义提出了自己的见解。     

Early Features of Apoptosis Detected by Four Different Flow Cytometry Assays

The objective of this study was to investigate the sensitivity, specificity and reproducibility of some frequently used apoptosis assays. The degree of apoptosis was tested in two T-lymphoblastoid cell lines, HSB and Jurkat, in which apoptosis was induced by ionizing radiation. HSB and Jurkat samples were taken before, and 0, 2, 4, 6, 8 and 24 h after irradiation with 6 and 10 Gray, or with 10 and 14 Gray, respectively. Four frequently used flow cytometric techniques were evaluated: (i) Annexin V/Propidium Iodide assay, detecting the translocation of phosphatidylserine to the outer leaflet of the plasma membrane, simultaneously with preservation of the membrane integrity; (ii) Terminal deoxynucleotidyl Transferase (TdT) Uridine triphosphate (UTP) nick end labelling (TUNEL), revealing the presence of DNA strand breaks; (iii) DNA-flow cytometry, measuring DNA-stainability (DNA-fragmentation assay) and (iv) Phycoerythrin-labelled (PE) Apo2.7-assay, a monoclonal antibody against 7A6 antigen, a protein, which becomes exposed upon the mitochondrial membrane during apoptosis. As a general standard for identifying that apoptosis had occurred, the cells were assessed for the presence of DNA-laddering on agar gel electrophoresis and by demonstration of characteristic cell morphology. Results were as follows: Fluorescein Isothiocyanate (FITC)-labelled Annexin V/Propidium iodide flow cytometry appeared to be the most sensitive, the most specific and the most user-friendly test for measurement of apoptosis of cells in culture conditions in suspension. The expression of 7A6 antigen on the mitochondrial membrane appeared to be not specific for apoptotic cell death.     

Effect of Fas+ and Fas− Target Cells on the Ability of NK Cells to Repeatedly Fragment DNA and Trigger Lysis via the Fas Lytic Pathway

We and others have recently shown that human NK cells express the Fas ligand (FasL) constitutively and that they can trigger the lysis of Fas positive (Fas+) target cells (TC) by apoptosis. We have also previously demonstrated that NK cells exposed to sensitive TC temporarily lose their ability to lyse sensitive TC via the granule-mediated pathway and that this loss is recovered when inactivated NK cells (NKi) are incubated in medium supplemented with IL-2, IL-12 or IL-15. In this study, we investigated the fate of the Fas-lytic pathway in NK cells exposed to either Fas+ or Fas– TC. To this end, we exposed NK cells to Jurkat (Fas–) or Jurkat (Fas+) TC for up to 6 h, separated NK cells from the TC and assessed the residual lytic activity against K562, a traditional human NK cell target, Jurkat Fas+ and Jurkat Fas– TC. Fas lytic activity was determined in calcium free medium, in the presence or absence of two distinct Fas-blocking monoclonal antibodies and a Fas.Fc fusion protein. In parallel experiments, the extent of DNA fragmentation in the three TCs was also assayed by the JAM test. Our results indicate that: (i) NK cells exposed to susceptible Fas+ TC temporarily lose most of their lytic potential due to the granule-mediated pathway, while only partially losing the Fas-lytic pathway. They also partially lose their ability to fragment DNA. (ii) NK cells exposed to Fas+ TC completely recover the Fas lytic pathway and the ability to fragment DNA via the Fas/Fas ligand when incubated in medium supplemented with IL-2 for 18 h.     

Clustering Timber Harvests and the Effect of Dynamic Forest Management Policy on Forest Fragmentation

To integrate multiple uses (mature forest and commodity production) better on forested lands, timber management strategies that cluster harvests have been proposed. One such approach clusters harvest activity in space and time, and rotates timber production zones across the landscape with a long temporal period (dynamic zoning). Dynamic zoning has been shown to increase timber production and reduce forest fragmentation by segregating uses in time without reducing the spatial extent of timber production. It is reasonable to wonder what the effect of periodic interruptions in the implementation of such as strategy might be, as would be expected in a dynamic political environment. To answer these questions, I used a timber harvest simulation model (HARVEST) to simulate a dynamic zoning harvest strategy that was periodically interrupted by changes in the spatial dispersion of harvests, by changes in timber production levels, or both. The temporal scale (period) of these interruptions had impacts related to the rate at which the forest achieved canopy closure after harvest. Spatial dynamics in harvest policies had a greater effect on the amount of forest interior and edge than did dynamics in harvest intensity. The periodically clustered scenarios always produced greater amounts of forest interior and less forest edge than did their never clustered counterparts. The results suggest that clustering of harvests produces less forest fragmentation than dispersed cutting alternatives, even in the face of a dynamic policy future. Although periodic episodes of dispersed cutting increased fragmentation, average and maximum fragmentation measures were less than if clustered harvest strategies were never implemented. Clustering may also be useful to mitigate the fragmentation effects of socially mandated increases in timber harvest levels. Implementation of spatial clustering during periods of high timber harvest rates reduced the variation in forest interior and edge through time, providing a more stable supply of forest interior habitat across the landscape. Received 19 September 1997; accepted 6 August 1998.     

Perspectives in the glycosciences – matrix-assisted laser desorption/ionization (MALDI) mass spectrometry of carbohydrates

The technique of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is reviewed with emphasis on the best type of matrix to use with carbohydrates, ways to obtain fragmentation and use of the technique for carbohydrate sequencing. © 1998 Rapid Science Ltd     

Landscape genetic structure of coastal tailed frogs (Ascaphus truei) in protected vs. managed forests

Habitat loss and fragmentation are the leading causes of species’ declines and extinctions. A key component of studying population response to habitat alteration is to understand how fragmentation affects population connectivity in disturbed landscapes. We used landscape genetic analyses to determine how habitat fragmentation due to timber harvest affects genetic population connectivity of the coastal tailed frog (Ascaphus truei), a forest-dwelling, stream-breeding amphibian. We compared rates of gene flow across old-growth (Olympic National Park) and logged landscapes (Olympic National Forest) and used spatial autoregression to estimate the effect of landscape variables on genetic structure. We detected higher overall genetic connectivity across the managed forest, although this was likely a historical signature of continuous forest before timber harvest began. Gene flow also occurred terrestrially, as connectivity was high across unconnected river basins. Autoregressive models demonstrated that closed forest and low solar radiation were correlated with increased gene flow. In addition, there was evidence for a temporal lag in the correlation of decreased gene flow with harvest, suggesting that the full genetic impact may not appear for several generations. Furthermore, we detected genetic evidence of population bottlenecks across the Olympic National Forest, including at sites that were within old-growth forest but surrounded by harvested patches. Collectively, this research suggests that absence of forest (whether due to natural or anthropogenic changes) is a key restrictor of genetic connectivity and that intact forested patches in the surrounding environment are necessary for continued gene flow and population connectivity.     

Identifying forest‐obligate birds in tropical moist cloud forest of Andean Ecuador

ABSTRACT Large‐scale transformation of forested landscapes is a major factor in loss of biological diversity in the American tropics. Investigators examining the responses of species to deforestation rarely control for variation in the amount of forest relative to other habitats at the landscape‐level. Bellavista Reserve on the western slope of the Andes in Ecuador is located between similar‐sized areas of pristine, protected forest, and deforested landscapes. We used strip‐transect counts and mist netting to evaluate habitat use by passerine birds in a habitat mosaic consisting of abandoned pastures, forest edges, forest fragments, and large blocks of interior tropical montane cloud forest (TMCF). During 3600 net hours, we had 1476 captures, including 346 recaptures. Of 78 species captured in mist nets, 30 had sufficient counts for Poison Rate Regression (PRR) modeling (a statistical method for comparing counts). Twelve species (40%) had capture patterns indicative of an affinity for mature TMCF, and 6 species (20%) had significantly higher counts in degraded areas (forest edge, forest fragment, and regenerating pastures) than in interior TMCF. The remaining 40% showed no significant bias in detection among habitats. Combined with strip‐count data, our results suggest that about 38% of the 119 species sampled at the Bellavista Reserve occur primarily in mature TMCF, avoiding edges and early second‐growth forest. Populations of these species may be vulnerable to further loss, fragmentation, and degradation of TMCF and, as such, deserve additional study and a place on lists of species of conservation concern.     

Effects of small‐scale grassland fragmentation and frequent mowing on population density and species diversity of orthopterans: a long‐term study

Abstract 1. Habitat fragmentation is considered one of the major threats to invertebrate diversity in semi‐natural grassland. However, the effects of habitat fragmentation through mowing on the rich insect fauna of these grasslands have not been sufficiently investigated and experiments are especially rare. 2. We studied the impact of small‐scale grassland fragmentation on orthopterans over 7 years in an experiment which allowed us to additionally investigate the effect of frequent mowing on Orthoptera communities. 3. Overall, Orthoptera density and species richness increased over time. This was likely a result of increased small‐scale habitat heterogeneity and the provision of a short‐turf habitat suitable for xerophilous species. The fragmentation affected orthopteran density and species composition but not species richness whose response lagged behind the changes in abundance. 4. Responses differed between suborders. Ensifera density was higher in fragment than in control plots. Caelifera density did not differ between fragment and control plots. The mown matrix was an unsuitable habitat for most of the species, particularly within the Ensifera. 5. Our experiment shows that even small‐scale fragmentation can affect Orthopteran communities and that the effects became more pronounced over time. As the mown matrix was unsuitable for many Ensifera species, they may go locally extinct when large areas are mown simultaneously.     

Mate-locating behaviour, habitat-use, and flight morphology relative to rainforest disturbance in an Afrotropical butterfly

To cope with environmental constraints, organisms can show variation in phenotype, either by genetic adaptation or phenotypic plasticity. These patterns are especially pronounced in ecosystems that are under anthropogenic influence. Due to human-induced disturbances such as logging and deforestation, tropical forests comprise such a system. To date, most studies have dealt with ecological responses at the community level relative to forest disturbance or degradation. However, the evolutionary consequences of tropical forest deterioration on behaviour and functional morphology have received far less attention compared to temporal regions. From a resource-point of view, light conditions are essential for heliotherms such as butterflies. Because degradation of tropical cloud forests in the Taita Hills (Kenya) is very pronounced, the present study tested whether this induced changes in mate-location strategies, habitat-use, and functional flight morphology in a forest butterfly, Salamis parhassus. According to predictions from temperate regions, it was hypothesized that the species would change its mate location strategy from perching to patrolling in more disturbed forests, that this higher mobility results in a faster occupancy of light gaps, and that it accords with a higher wing loading within populations from undisturbed forests. These hypotheses were confirmed by field surveys and experiments. The present study demonstrates that degradation of tropical forests does not only affect communities (e.g. species richness), but also the behaviour and functional morphology of individual species.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 830–839.