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In the field, pathogens exist in a diverse and dynamic ecological environment with other organisms (Blakeman, 1993; Preece & Dickinson, 1971). We tend to assume that these other organisms are ‘benign’ or perhaps in competition with the pathogen to occupy a particular niche. However, bacteria are often observed to be in close association with pathogens, as witnessed by the difficulty of eliminating them during isolation of pathogens involved in plant diseases. We now have clear evidence (Dewey et al., pp. 489–497 in this issue) that they are sometimes neither benign, nor in competition, but actually helping the pathogen successfully infect its host even when the bacteria themselves are not pathogens of that host.

The arbuscular mycorrhiza (AM) is one of the most important symbioses on earth, formed between about 80% of vascular plants and AM fungi (Smith & Read, 1997), which are placed in the order Glomales (Morton & Benny, 1990). A discussion has recently arisen about the rDNA-gene and general genome organization of this fungal group, which, as far as is known, does not possess sexuality (Sanders, 1999). This discussion mainly arose as a result of the assumption that one spore of an AM fungus (Scutellospora castanea) contains small subunit (SSU) rRNA genes that belong, phylogenetically, to different families within the Glomales, namely the Gigasporaceae and Glomaceae (Hosny et al., 1999). Questions about genetic drift, infrequent recombination, concerted evolution and the validity of phylogenetic techniques have arisen (Hosny et al., 1999; Sanders, 1999). However, this assumption might well be based on the sequence of an Ascomycete contaminant.

CONTENTS

Summary 389

I. INTRODUCTION 389

II. 1. The SNARE hypothesis 393

2. SNAREs and associates 395

(a) NSF 395

(b) SNAP 395

(c) SNARE 395

(d) Rab and Rab-associated proteins 397

(e) Other interacting proteins 398

(f) Clostridial neurotoxins 399

3. New perspectives on SNARE function 400

(a) Revisiting the SNARE hypothesis 400

(b) Roles for SNARE proteins 401

III. VESICLE TRAFFICKING IN PLANTS 402

1. Identifying secretory proteins in plants 402

2. Placing syntaxins in order 403

3. Functional analysis of plant SNAREs in vesicle trafficking 404

4. Small GTPases 405

5. Searching for a vesicle–fusion complex 405

IV. REGULATION OF VESICLE TRAFFICKING IN PLANT CELLS 406

1. Hormonal control of secretion 406

(a) Auxin-evoked secretion and cell growth 406

(b) Gibberellic acid and secretion by barley aleurone 407

(c) Abscisic acid control of stomatal aperture 407

2. Coupling secretion to cytosolic-free Ca+ concentration 408

3. Stress-evoked secretion 409

(a) Mechanical stress 409

(b) Wounding and pathogen attack 409

V. CONCLUSION 410

Acknowledgements 411

References 411

Eukaryotic cells share a set of secretory pathways for the flux of membrane and protein material. In 1993, ideas about the functioning of three major proteins of the neurosecretory complex were consolidated in the SNARE hypothesis, which proposed that the interaction of these proteins provides both the specificity for vesicle targeting and the molecular machinery for fusion between vesicle and target membranes. Subsequetly, the organization, molecular mechanics and control of vesicle trafficking have become topics of intense research, and the hypothesis has evolved to accommodate new discoveries from the analysis of secretion in yeast and mammals. It is likely to be challenged again as more information comes to light about secretory processes in plants. New tools for measuring and manipulating vesicle traffic and secretion are now being used, drawing on in vivo fluorescence and capacitance recording as well as genetic engineering. These new technologies have already begun to yield details wholly unexpected from past studies. Here we focus on recent findings relating to the mechanisms of vesicle trafficking and the background to these developments, highlighting both current understanding of the molecular events of secretion and the gaps therein, as well as discussing emerging themes from work with plants.

Summary 3

I. introduction 4

II. the cyanobacteria 7

III. the heterocyst 9

1. Function and metabolism 9

2. Heterocyst structure 12

(a) Overview 12

(b) The polysaccharide (homogeneous) layer 12

(c) The glycolipid (laminated) layer 12

(d) The septum and microplasmodesmata 12

3. Nitrogen regulation and heterocyst development 12

4. Heterocyst development 13

(a) The proheterocyst 13

(b) Proteolysis associated with heterocyst development 14

(c) RNA polymerase sigma factors 14

(d) Developmental regulation of heterocyst cell wall and nitrogenase gene expression 14

(e) Genome rearrangements associated with heterocyst development 15

5. Genes essential for heterocyst development 15

(a) hetR 15

(b) Protein phosphorylation and the regulation of hetR activity 16

(c) hetR in nonheterocystous cyanobacteria 16

(d) Other heterocyst-specific genes 16

6. Heterocyst spacing 18

(a) Patterns of heterocyst differentiation 18

(b) Genes involved in heterocyst spacing 18

(c) Disruption of heterocyst pattern 18

7. Filament fragmentation and the regression of developing heterocysts 20

8. The nature of the heterocyst inhibitor 20

9. Cell selection during differentiation and pattern formation 20

(a) Cell division 20

(b) DNA replication and the cell cycle 21

(c) Competition 21

10. Models for heterocyst differentiation and pattern control 21

IV. the akinete 23

1. Properties of akinetes 23

2. Structure, composition and metabolism 24

3. Relationship to heterocysts 24

4. Factors that influence akinete differentiation 24

5. Extracellular signals 25

6. Akinete germination 25

7. Genes involved in akinete differentiation 26

V. conclusion 26

Acknowledgements 27

References 28

Cyanobacteria are an ancient and morphologically diverse group of photosynthetic prokaryotes. They were the first organisms to evolve oxygenic photosynthesis, and so changed the Earth's atmosphere from anoxic to oxic. As a consequence, many nitrogen-fixing bacteria became confined to suitable anoxic environmental niches, because the enzyme nitrogenase is highly sensitive to oxygen. However, in the cyanobacteria a number of strategies evolved that protected nitrogenase from oxygen, including a temporal separation of oxygenic photosynthesis and nitrogen fixation and, in some filamentous strains, the differentiation of a specialized cell, the heterocyst, which provided a suitable microaerobic environment for the functioning of nitrogenase. The evolution of a spore-like cell, the akinete, almost certainly preceded that of the heterocyst and, indeed, the akinete may have been the ancestor of the heterocyst. Cyanobacteria have the capacity to differentiate several additional cell and filament types, but this review will concentrate on the heterocyst and the akinete, emphasizing the differentiation and spacing of these specialized cells.

The relationships of foliage assimilation capacity per unit area (PPmax) with leaf dry mass per unit area (LMA) and nitrogen content per unit area (NP) differ between species and within species grown in different habitats. To gain a more mechanistic insight into the dependencies of PPmax on LMA and NP, this literature study based on 597 species from a wide range of earth biomes with woody vegetation examines the relations between leaf photosynthetic capacity and the components of LMA (leaf density (D, dry mass per volume) and thickness (T)), and also the correlations of D and T with leaf nitrogen content and fractional leaf volumes in different tissues. Across all species, PPmax varied 12-fold and photosynthetic capacity per unit dry mass (Pmmax) 16-fold, NP 12-fold, and nitrogen per unit dry mass (Nm) 13-fold, LMA 46-fold, D 13-fold, and T 35-fold, indicating that foliar morphology was more plastic than foliar chemistry and assimilation rates. Although there were strong positive correlations between PPmax and NP, and between Pmmax and Nm, leaf structure was a more important determinant of leaf assimilation capacities. PPmax increased with increasing LMA and T, but was independent of D. By contrast, Pmmax scaled negatively with LMA because of a negative correlation between Pmmax and D, and was poorly related to T. Analysis of leaf nitrogen and tissue composition data indicated that the negative relationship between D and Pmmax resulted from negative correlations between D and Nm, D and volumetric fraction of leaf internal air space, and D and symplasmic leaf fraction. Thus, increases in leaf density bring about (1) decreases in assimilative leaf compounds, and (2) extensive modifications in leaf anatomy that may result in increases in intercellular transfer resistance to CO2. Collectively, (1) and (2) lead to decreased Pmmax, and also modify PPmax versus LMA relationships.

Cryptogamous plants in the forest communities of Bialowieza National Park. Functional groups analysis and general synthesis. Ed. by J. B. FALINSKI and W. MULENKO. 30×21 cm. Pp. 224 with 4 plates and numerous text-figures. Phytocoenosis (N. S.) 8, Archivum Geobotanicum 6. 1996. P/b.

Cryptogamous plants in the forest communities of Bialowieza National Park. Ecological atlas of seminal and cryptogamous plants. Ed. by J. B. FALINSKI and W. MULENKO. 30×21 cm. Pp. 522 with 2000 maps. Phytocoenosis (N. S.) 9, Supplementum Cartographicae Geobotanicae 7. 1997. H/b.

Deciduous woody pioneer species (Juniperus communis, Populus tremula, Salix sp. div.) in the secondary succession and regeneration. By J. B. FALINSKI. 30×21 cm. Pp. 156 with 8 plates, 2 end-paper charts and numerous text-figures. Phytocoenosis (N. S.) 10, Supplementum Cartographicae Geobotanicae 8. 1998. P/b.

Synanthropization of plant cover in new Polish research. Ed. by J. B. FALINSKI, W. ADAMOWSKI and B. JACKOWIAK. 30×21 cm. Pp. 279 with 2 plates, 2 end-paper maps and numerous text-figures. Phytocoenosis (N. S.) 10, Supplementum Cartographicae Geobotanicae 9. 1998. P/b.

The relative ozone sensitivities of 25 German native herbaceous plant species, representative of arable field margins or disturbed habitats, were examined over three consecutive growing seasons. Plants were grown from seed, potted into natural soils and exposed in open-top chambers for the entire season to different ozone-exposure regimes covering a range of concentrations from <5 to 48 ppb (seasonal 8 h daily mean). The assessment of ozone effects was carried out by recording the first day of visible symptom appearance and the percentage of injured leaves at the end of vegetative growth. Species exhibited contrasting patterns of symptom expression under ozone stress, with either ozone-specific symptoms or ozone-enhanced foliar pigmentation and senescence. Classifications of species according to their ozone susceptibility varied depending on whether measurement was of the total extent of visible injury, ozone threshold doses for the incidence of symptoms, or modelled exposure–response relationships. The most sensitive species exhibiting ozone-specific symptoms were Cirsium arvense and Sonchus asper, which responded to accumulated ozone exposures <1500 ppb.h (AOT40). For these and three other species, an AOT40 peak of a single day was found to be responsible for the incidence of ozone-specific symptoms, i.e. injury occurred rapidly within a few days of the day with the highest AOT40, while other species responded only to longer-term ozone exposures. The relative ozone sensitivity of the species was calculated by combining the different sensitivity criteria, and possible systematic trends (taxonomic or evolutionary features) are pointed out. The results suggest it may be possible to use a particular group of native herbaceous plant species with contrasting patterns of ozone sensitivity as a biomonitoring system in the field. This allows plant responses to be related either to peak values or to prolonged ozone exposure, making it possible to distinguish between short- and long-term effects of ozone.