Cell Death In Higher Plants Research Articles

Domain swap between two type-II metacaspases defines key elements for their biochemical properties.

Type-II metacaspases are conserved cysteine proteases found in eukaryotes with oxygenic photosynthesis, including green plants and some algae, such as Chlamydomonas and Volvox. Genetic and biochemical studies showed that some members in this protease family could be involved in oxidative stress-induced cell death in higher plants, but their regulatory mechanisms remain unclear. Biochemically, two distinct classes of type-II metacaspases are exemplified by AtMC4 and AtMC9 from Arabidopsis, with AtMC4 activation dependent on calcium under neutral pH, whereas AtMC9 is active only under mildly acidic pH, regardless of the availability of calcium. Here, we constructed all six possible combinations between the p20, linker, and p10 domains from AtMC4 and AtMC9. Our results show that calcium stimulation of AtMC4 activity is associated with essential amino acids located in its p20 domain. In contrast, the acidic pH optimum trait is lost from AtMC9 if one or two of its domains are replaced by that from AtMC4, suggesting that multiple interactions between domains in AtMC9 may be responsible for this property. Consistent with this, we found conserved 'signature' residues in each of the three domains that distinguish AtMC4- and AtMC9-like classes of metacaspases. Tracing the origin of the AtMC9 class, we found evidence for its appearance between lycophytes and gymnosperms, coincident with the evolution of more complex root archetypes in terrestrial plants. Our work suggests that the distinctive properties of the AtMC9-like protease could be associated with special cellular physiology in the roots of gymnosperms and angiosperms that are distinct from lycophytes.

Programmed cell death is induced by hydrogen peroxide but not by excessive ionic stress of sodium chloride in the unicellular green alga Chlamydomonas reinhardtii

Eukaryotic microalgae serve as indicators of environmental change when exposed to severe seasonal fluctuations. Several environmental stress conditions are known to produce reactive oxygen species in cellular compartments, resulting in oxidative damage and apoptosis. The study of cell death in higher plants and animals has revealed the existence of an active ‘programmed cell death’ (PCD) process and similarities between such processes suggest an evolutionary origin. A study was undertaken to examine the morphological, biochemical and molecular responses of the unicellular green alga Chlamydomonas reinhardtii after exposure to oxidative (10 mM H2O2) and osmotic (200 mM NaCl and 360 mM sorbitol) stress. Concentrations of H2O2 (2–50 mM), NaCl and sorbitol (100–800 mM) were negatively correlated with growth. Biochemical analyses showed an increase in intracellular H2O2 production (2.2-fold with H2O2 and ~1.2–1.4-fold with NaCl and sorbitol) and activities of some antioxidant enzymes [super oxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX)]. Alteration of mitochondrial membrane potential (MMP) was observed upon treatment with H2O2 and NaCl, but not with sorbitol, indicating that the ionic stress component of NaCl altered the MMP. In addition, H2O2 led to the activation of a caspase-3-like protein, increase in the cleavage of a poly(ADP) ribose polymerase-1 (PARP-1)-like enzyme and formation of DNA nicks and laddering. With NaCl and sorbitol, no caspase activation, nor oligonucleosomal DNA laddering was observed, indicating non-apoptotic death. However, genomic DNA of NaCl (800 mM)-stressed cells, but not those of sorbitol-treated cells showed complete shearing. We conclude that the ionic rather than the osmotic component of NaCl leads to necrosis. These results unequivocally suggest that the vegetative cells of C. reinhardtii respond differentially to various stress agents, leading to different death types in the same organism. Moreover, unlike most other organisms, when exposed to NaCl this alga does not undergo PCD.

Singlet oxygen-dependent translational control in the tigrina-d.12 mutant of barley

The tigrina (tig)-d.12 mutant of barley is impaired in the negative control limiting excess protochlorophyllide (Pchlide) accumulation in the dark. Upon illumination, Pchlide operates as photosensitizer and triggers singlet oxygen production and cell death. Here, we show that both Pchlide and singlet oxygen operate as signals that control gene expression and metabolite accumulation in tig-d.12 plants. In vivo labeling, Northern blotting, polysome profiling, and protein gel blot analyses revealed a selective suppression of synthesis of the small and large subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase (RBCSs and RBCLs), the major light-harvesting chlorophyll a/b-binding protein of photosystem II (LHCB2), as well as other chlorophyll-binding proteins, in response to singlet oxygen. In part, these effects were caused by an arrest in translation initiation of photosynthetic transcripts at 80S cytoplasmic ribosomes. The observed changes in translation correlated with a decline in the phosphorylation level of ribosomal protein S6. At later stages, ribosome dissociation occurred. Together, our results identify translation as a major target of singlet oxygen-dependent growth control and cell death in higher plants.

Depletion of UDP-d-apiose/UDP-d-xylose Synthases Results in Rhamnogalacturonan-II Deficiency, Cell Wall Thickening, and Cell Death in Higher Plants

D-apiose serves as the binding site for borate cross-linking of rhamnogalacturonan II (RG-II) in the plant cell wall, and biosynthesis of D-apiose involves UDP-D-apiose/UDP-D-xylose synthase catalyzing the conversion of UDP-D-glucuronate to a mixture of UDP-D-apiose and UDP-D-xylose. In this study we have analyzed the cellular effects of depletion of UDP-D-apiose/UDP-D-xylose synthases in plants by using virus-induced gene silencing (VIGS) of NbAXS1 in Nicotiana benthamiana. The recombinant NbAXS1 protein exhibited UDP-D-apiose/UDP-D-xylose synthase activity in vitro. The NbAXS1 gene was expressed in all major plant organs, and an NbAXS1-green fluorescent protein fusion protein was mostly localized in the cytosol. VIGS of NbAXS1 resulted in growth arrest and leaf yellowing. Microscopic studies of the leaf cells of the NbAXS1 VIGS lines revealed cell death symptoms including cell lysis and disintegration of cellular organelles and compartments. The cell death was accompanied by excessive formation of reactive oxygen species and by induction of various protease genes. Furthermore, abnormal wall structure of the affected cells was evident including excessive cell wall thickening and wall gaps. The mutant cell walls contained significantly reduced levels of D-apiose as well as 2-O-methyl-L-fucose and 2-O-methyl-D-xylose, which serve as markers for the RG-II side chains B and A, respectively. These results suggest that VIGS of NbAXS1 caused a severe deficiency in the major side chains of RG-II and that the growth defect and cell death was likely caused by structural alterations in RG-II due to a D-apiose deficiency.

Lam, E., Fukuda, H., Greenberg, J., eds. Programmed cell death in higher plants

Lam, E., Fukuda, H., Greenberg, J., eds. Programmed cell death in higher plants

E. Lam, H. Fukuda & J. Greenberg (eds): Programmed Cell Death in Higher Plants.

E. Lam, H. Fukuda & J. Greenberg (eds): Programmed Cell Death in Higher Plants.

A Genetic System Involving Superoxide Causes F1 Necrosis in Wheat (T. aestivumL.)

A genetic system in wheat is described in which F1 produced by crossing a drought tolerant cultivar C306 and high yielding cultivar WL711 exhibits leaf necrosis leading to the death of the plant. The mechanism underlying hybrid necrosis is not yet known. The hybrid exhibited a higher level of superoxide anion compared to the healthy leaves of parents at similar developmental stages. This increase in superoxide generation preceded necrotic lesion formation and displayed a gradient from the leaf tip to base. The leaf tip where necrotic lesions make their first appearance exhibited a higher level of superoxide compared to the base. Superoxide anion thus appears to play a vital role in necrosis of leaves in F1 hybrid. This genetic system can be a model system for understanding cell death in higher plants.

Programmed cell death during vascular system formation

Vascular plants have vessels and tracheids composed of dead tracheary elements. Differentiation of procambial or cambial cells to tracheary elements is a typical example of programmed cell death in higher plants. Recent studies on tracheary element differentiation, in particular with an in vitro differentiation system, have revealed a unique cell death process which differs from apoptosis. Herein I summarize the present knowledge about the induction of cell death, the morphological features of cell death, and the mechanism of autolysis, including the involvement of a DNase and cysteine proteases, during tracheary element differentiation.

Tracheary Element Differentiation.

Vascular plants, which are adapted for life on land, first appeared in the late Silurian period, some 400 million years ago. Since then they have evolved to fill a diverse range of habitats all over the earth. The vascular systems of land plants are composed of specialized conducting tissues, the xylem and the phloem, which provide both a pathway for water and nutrient transport and mechanical support for slender plants. The vascular system is also an important conduit for signal-transducing molecules. Tracheary elements (TEs), which are the distinctive cells of the xylem, are characterized by the formation of a secondary cell wall with annular, spiral, reticulate, or pitted wall thickenings. In the primary xylem, TEs differentiate from procambial cells, whereas in the secondary xylem, they arise from cells produced by the vascular cambium. As they mature, TEs lose their nuclei and cell contents, leaving hollow dead cells that form vessels or tracheids. The final stage of TE differentiation represents a typical example of programmed cell death in higher plants (see Pennell and Lamb, 1997, in this issue). TEs can also be induced to form in vitro from various types of cells, including cells of the phloem parenchyma and the cortex in roots, the pith parenchyma in shoots, the tuber parenchyma, and the mesophyll and epidermis in leaves (Roberts et al., 1988; Fukuda, 1992). In Zinnia elegans cell cultures, single mesophyll cells transdifferentiate directly into TEs without cell division in response to phytohormones (Fukuda and Komamine, 1980). The Zinnia system has proven to be particularly useful for studies of the sequence of events during TE differentiation. This is largely because differentiation occurs at a high frequency in Zinnia cultures and because the process can be followed in single cells (Chasan, 1994; Fukuda, 1994, 1996). Recently, I presented a general overview of xylogenesis (Fukuda, 1996). In this article, I focus on efforts to elucidate the molecular mechanisms underlying the in vitro differentiation of parenchyma cells into TEs.

Programmed cell death: a way of life for plants.

Cell death in higher plants has been widely observed in predictable patterns throughout development and in response to pathogenic infection. Genetic, biochemical, and morphological evidence suggests that these cell deaths occur as active processes and can be defined formally as examples of programmed cell death (PCD). Intriguingly, plants have at least two types of PCD, an observation that is also true of PCD in animals [Schwartz, L. M., Smith, W.W., Jones, M. E. E. & Osborne, B. A. (1993) Proc. Natl. Acad. Sci. USA 90, 980-984]. Thus, in plants, PCD resembles either a common form of PCD seen in animals called apoptosis or it resembles a morphologically distinct form of cell death. The ubiquitous occurrence and necessity of PCD for plant development and defense suggest that the underlying mechanisms of regulation and execution of these processes merit further examination.

高等植物におけるプログラム細胞死

高等植物におけるプログラム細胞死

Coordinated Activation of Programmed Cell Death and Defense Mechanisms in Transgenic Tobacco Plants Expressing a Bacterial Proton Pump.

In plants, programmed cell death is thought to be activated during the hypersensitive response to certain avirulent pathogens and in the course of several differentiation processes. We describe a transgenic model system that mimics the activation of programmed cell death in higher plants. In this system, expression of a bacterial proton pump in transgenic tobacco plants activates a cell death pathway that may be similar to that triggered by recognition of an incompatible pathogen. Thus, spontaneous lesions that resemble hypersensitive response lesions are formed, multiple defense mechanisms are apparently activated, and systemic resistance is induced in the absence of a pathogen. Interestingly, mutation of a single amino acid in the putative channel of this proton pump renders it inactive with respect to lesion formation and induction of resistance to pathogen challenge. This transgenic model system may provide insights into the mechanisms involved in mediating cell death in higher plants. In addition, it may also be used as a general agronomic tool to enhance disease protection.

高等植物のプログラムされた細胞死 道管の分化

高等植物のプログラムされた細胞死 道管の分化