Ernst-Detlef Schulze Erwin Beck Klaus Mller-Hohenstein

Plant Ecology
With 506 Figures, most of them in colour, and 101 Tables

12

Professor Dr. Ernst-Detlef Schulze Max-Planck-Institute for Biogeochemistry P.O. Box 10 01 64 07701 Jena Germany Professor Dr. Erwin Beck Department of Plant Physiology University of Bayreuth 95440 Bayreuth Germany Professor Dr. Klaus Mller-Hohenstein Department of Biogeography University of Bayreuth 95440 Bayreuth Germany

Translated by: Gudrun Lawlor, FIL Dr. Kirsten Lawlor Dr. David Lawlor 9 Burywick Harpenden Hertfordshire AL5 2AQ UK

Original title: Ernst-Detlef Schulze/Erwin Beck/Klaus Mller-Hohenstein, Pflanzenkologie Copyright 2002 Spektrum Akademischer Verlag GmbH, Heidelberg

ISBN 3-540-20833-X Springer-Verlag Berlin Heidelberg New York

Library of Congress Control Number: 2004107209 This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilm or in any other way, and storage in data banks. Duplication of this publication or parts thereof is permitted only under the provisions of the German Copyright Law of September 9, 1965, in its current version, and permission for use must always be obtained from Springer-Verlag. Violations are liable for prosecution under the German Copyright Law. Springer is a part of Springer Science+Business Media springeronline.com Springer Berlin Heidelberg 2005 Printed in Germany The use of general descriptive names, registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. Editor: Dr. Dieter Czeschlik, Heidelberg Desk editor: Dr. Andrea Schlitzberger, Heidelberg Production: Karl-Heinz Winter, Heidelberg Cover design: design & production GmbH, Heidelberg, Germany Cover illustration: Claus Diercks, Willy Giltmann, Hamburg, Germany Typsetting: K & V Fotosatz GmbH, Beerfelden 31/3150 5 4 3 2 1 0 Printed on acid-free paper

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nomical importance with regard to the production of secondary plant metabolites, because more carbohydrates (and nitrogen) are then available for growth. In an interesting experiment, the cost-benefit relation of jasmonate-induced defence was analysed (Baldwin 1998). Seed production of the annual Nicotiana attenuata was measured after some plants had been treated with methyl jasmonate to induce the defence reactions while others were left untreated. Both populations were then exposed to different intensities of herbivory by the natural spectrum of herbivores and phytophagous insects. Under herbivory pressure the (previously) induced plants produced significantly more seeds than the non-induced unprotected control plants. With little or no herbivory, however, the non-induced plants were superior. In nature, herbivory is the normal case, and therefore species capable of jasmonate-induced defence are favoured by selection. Jasmonate plays the role of transmitter not only in the signal cascade after wounding, but also after infestation by pathogens. The reaction to both stimuli may be very different, not only regarding the metabolic end products: If the signal cascade is triggered by a fungal elicitor and if arachidonic acid instead of, or in addition to, linolenic acid is released, a different isozyme of the HMGR is induced, compared to the reaction in which jasmonate is the elicitor (hmg1 vs. hmg2). The biological sense behind this differentiation is still unknown. A jasmonate-independent signal pathway was identified, beside the jasmonate-dependent transduction of the wound signal (Titarenko et al. 1997). However, the jasmonate-independent signal pathway does not lead to a systemic induction, but only to a local induction of defensive genes as in the previously discussed systemin-independent signal pathway. This path is shown in Fig. 1.10.5 by the broken arrow. It is interesting that the isoprenoids which are released function as kairomones 1 and particularly attract parasitoids, which then parasitise phytophages. Ichneumon wasps are especially attracted by the volatile terpenoids (Par and Tumlinson 1999; Kessler and Baldwin 2001). Isoprenoids thus serve as signals with very high specificity regarding the host plant of the phy1

tophage, but the signal does not give much information about the type of phytophage. This is understandable, as the signal originates from the plant and not from the phytophage. It might be a kind of co-evolution that the inducible signals are produced almost exclusively during the day, when parasitoids are active. Induction of HMGR can thus be interpreted as an active defence strategy of plants against herbivory.

1.10.2
Pathogen Attack and Defence Recommended Literature
Durrant WE, Dong X (2004) Systemic acquired resistance. Annu Rev Phytopathol 42:185209 Hammond-Kosack K, Jones JDG (2000) Responses to plant pathogens. In: Buchanan BB, Gruissem W, Jones RL (eds) Biochemistry and molecular biology of plants. American Society of Plant Physiologists, Rockville, MD, pp 11021156 Heil M, Baldwin IT (2002) Fitness costs of induced resistance: emerging experimental support for a slippery concept. Trends Plant Sci 7:6167 In addition to phytophagous insects, plants may also suffer from infestation by pathogens like viruses, bacteria and fungi. Pathogens often cause considerable damage to the high-yielding cultivars of our crop plants and therefore their interaction with the host plant has been the topic of extensive research in the scientific area of phytopathology. Within the scope of this book, the reaction spectrum of plants to pathogen attacks can only be described in a cursory fashion. We distinguish between compatible and incompatible interactions. In compatible interactions host and pathogen get along with each other, i.e. infestation is strong, the pathogen is virulent and the plant is susceptible. In incompatible interactions, the infested plant can cope with the attack of pathogens by establishing defence reactions, i.e. it is resistant and the pathogens are called avirulent. In compatible interactions, pathogens are able to circumvent, inactivate or tolerate the plant defences. Generally, however, the resistance of plants to the broad

Kairomones are (usually) volatile signal compounds, which are useful for the detecting organism: attractants, repellents (escape), stimulants, warning signals.

a spectrum of potential, more or less omnipresent pathogens, is very high. Fungi and bacteria proliferate and expand within the extracellular space of the plant, therefore cell walls are one of the preformed barriers these pathogens have to overcome. Often cell walls are very difficult to penetrate because of coating with or insertion of cutin, or because of lignification.

Pathogen Attack and Defence

247

serving to support growth of the fungus. Other fungi start infection of a plant similarly as described, but instead of forming haustoria they invade the plant cell with secondary hyphae destroying the plant's plasma membrane. The plant cells die and the fungi live from the contents of dead plant cells. Without appropriate plant defence, fungi overgrow the plant tissue and their virulence is often increased by formation of exo- or zoospores.

1.10.2.1

Infection by Fungi A fungal infection starts from a spore adhering to the cuticle by an adhesion pad on the surface of a plant. A germ tube develops from the spore and an appressorium is formed (see Fig. 1.10.12). Within this appressorium a high pressure is built up exerting a mechanical force and with the additional help of cutinases, cellulases and pectinases the fungus enzymatically disrupts cuticle and cell wall, without damaging the plant plasma membrane. Primary hyphae expand in the extracellular space and haustoria are formed. Haustoria are nutrient-absorbing cells which invade plant cells by infolding the plant plasma membrane. The plant cell therefore remains intact, but their nutrients are consumed

1.10.2.2

Induced Defence Resistant plants are able to prevent proliferation and expansion of pathogens. They do so by induction of defence responses which include (see Fig. 1.10.10) stiffening of their cell walls by synthesis of callose, a b-1,3-glucan, synthesis of hydroxyproline-rich glycoproteins, which cross-link cell wall components, and increased lignification (see Fig. 1.10.12, formation of a papilla), synthesis of so-called PR proteins (pathogenesis-related proteins), some of which are chitinases and glucanases which hydrolyse fungal and bacterial cell walls, others have anti-oxidative capacities,

Fungus (pathogen) Exogenous elicitors Receptors Toxins Enzymes Endogenous elicitors

? Secondary messengers PGIPs Defense response PR-proteins Phytoalexins Proteinase inhibitors Structural components

Gene activation

Cell wall

Plasma membrane

Nucleus

n Fig. 1.10.10. Model of the interaction between pathogen and host in the hypersensitive reaction. The fungus releases cell wall lysing enzymes and elicitors from its hyphae, and in some cases also toxins. Via a signal cascade, the fungal elicitors, and the elicitors produced by hydrolysis of the plant cell wall, induce the expression of genes in the nucleus, which are involved in the synthesis of defensive enzymes that either directly attack the fungus (for instance PR proteins) or produce defensive compounds (phytoalexins and enzyme inhibitors). The polygalacturonase inhibitor (PGIP) highlighted in the model prevents degradation of oligogalacturonides, products of the action of a fungal polygalacturonase, which act as elicitors and thus increase the defence reaction. While the defence reactions are induced in cells surrounding the infection site during the hypersensitive response, the infected cell itself is sacrificed by programmed cell death. (After He 1999)

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Biotic Stress: Herbivory, Infection, Allelopathy

Box 1.10.3

Salicylic acid
icylic acid is the glucoside salicin. C4H Cinnamate-4-hydroxylase; PAL phenylalanine ammonia lyase; GTase glucosyl-transferase (after Chong et al. 2001).

Salicylic acid can be produced via several biosynthetic pathways, either via cinnamic acid coenzyme A thioester or via cinnamic acid glucose ester. Free benzoic acid is probably only a by-product. The storage form of sal-

O O C S-CoA L-Phenylalanine CoA-ligase PAL O C OH trans-Cinnamic acid C4H Cinnamoyl-glucose Benzoyl-glucose Lignin Phytoalexins GTase C O-Glucose O C O-Glucose OH C OH Free benzoic acid O O C O-Glucose C OH O-Glucose Salicin O OH Salicylic acid O Cinnamoyl-CoA Benzoyl-CoA OH Salicyloyl-CoA C OH O C S-CoA C S-CoA O

p-Coumaric acid

Salicyloyl-glucose ester

synthesis of phytoalexins (Greek alekein = to fend off ), a diverse group of low molecular weight secondary metabolites, predominantly compounds of the classes isoprenoids, flavonoids and stilbenes, which exert an antibiotic effect against a broad spectrum of fungi and bacteria. The defence responses are induced in cells surrounding the infested cell. One mediator of the induction of PR proteins is salicylic acid 2 (see Box 1.10.3 and Table 1.10.1). In resistant plants, the infested cell itself often dies by programmed cell death, the final result of the so-called hypersensitive response (HR). During an HR, reactive oxygen species (ROS, see Chap. 1.3.5.3) are produced, leading to an oxidative burst (Lamb and Dixon 1997). ROS are toxic for the pathogen
2

Acetylsalicylic acid, aspirin, is the most produced medicine worldwide. It is better tolerated by humans than free salicylic acid, the actual agent. This substance from the bark of the willow tree (Salix) was used in ancient times in western Europe and by North American Indians to relieve pain and fever. In plants, it increases resistance against pathogens, and is also used as a spray to prevent mildew. Many amateur gardeners water their plants from time-to-time with aspirin solutions.

and the host and, therefore, synthesis of antioxidants in cells around the infection site is plausible. Why are the defence responses induced during incompatible but not during compatible interactions? In an incompatible interaction, signals are transmitted between pathogen and host, which trigger the defence responses. The basis of this signal transmittance is a certain genetic background of both partners. Plants contain R genes (resistance genes), while pathogens contain avr genes (avirulence genes) or vir genes (virulence genes). For an interaction to be incompatible, host and pathogen must have corresponding R and avr genes (Hammond-Kosack and Jones 2000; Staskawicz 2001). The products of R genes are often integral plasma membrane proteins which function as receptors. Binding of a proper elicitor to the receptor then induces via a signal transduction chain (similar as during herbivory) the defence responses. The products of avr genes may be the elicitors per se or, in most cases, avr genes encode proteins which are involved in the synthesis of elicitors. Upon a fungal infestation, elicitors are often degradation products of cell walls, either of plant or fungal origin. They are very specific oligosac-

Pathogen Attack and Defence

249

n Fig. 1.10.11. Systemic acquired resistance of cucumber plants to the Anthracnose fungus Colletotrichum lagenarium. One cotyledon of the plant on the left was infected with spores of the fungus. Obviously, it is suffering severely from necrosis. The plant on the right was left untreated. One week later, 20 drops of a spore suspension were placed on a main leaf of both plants. The leaf of the plant on the left remained fully healthy as it had become resistant. The leaf of the plant on the right, which experienced no immunisation , exhibited necrotic areas at the positions of every drop of spore suspension. (Conrath and Kauss 2000)

charides. In one case, this elicitor is produced by the combined action of a fungal polygalacturonase (hydrolysing pectin of the plant cell wall) and a polygalacturonase inhibitor protein (PGIP), which is synthesised by the plant after infection, leading to oligogalacturonides of the appropriate length to function as elicitor (see Fig. 1.10.10). This is probably the result of a co-evolution between a former virulent fungus and a susceptible plant cultivar. Elicitors may also be secreted glycoproteins which bind to plant receptors. Regarding bacteria, avr-gene products are proteins which often are transferred into the plant's cytoplasm interacting with intracellular receptor proteins which guard the defence response. In the presence of a corresponding avr-gene product, the defence responses are induced. The difference between resistant and susceptible plants is not that susceptible plants were unable to produce PR proteins or phytoalexins, but that their production is not triggered or not triggered in time because a proper signal (from the pathogen) or a proper receptor (in the plant) is missing.

A S

C CW

PM

A A S CW PM C

1.10.2.3

Systemic Acquired Resistance The hypersensitive response often includes the establishment of a systemic acquired resistance (SAR) against pathogens. SAR is reminiscent of the systemic defence against phytophagous insects. It means that the defence response (mainly synthesis of PR proteins) is induced in non-in-

n Fig. 1.10.12. Compatible (A) and incompatible (B) interactions during infestation by a fungus. A A germinating spore (S) develops a germ tube from which an appressorium (A) is formed. Within the appressorium a high pressure is built up and by secretion of cell wall lysing enzymes a primary hypha (P) is growing through the cuticle (C) and cell wall (CW). Later secondary hyphae (H) enter the cell by destroying the plasma membrane (PM). B Defence against a fungal infestation in a (systemically) resistant plant. The fungus is unable to penetrate the outer wall of the epidermal cell, as a papillose wall thickening develops under the appressorium. This papilla is also impregnated with a strong layer of lignin. (After Conrath and Kauss 2000)

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Biotic Stress: Herbivory, Infection, Allelopathy

fested plant organs remote from the infection site (Fig. 1.10.11). When pathogens later try to infect these plant organs they fail because of the defence mechanisms already preformed (Fig. 1.10.12). As during the local defence response, salicylic acid is part of the signal transduction chain leading to the expression of PR proteins in the non-infested tissue. Interestingly, the SAR is directed against a broad spectrum of pathogens, not only against the pathogen of the primary infection. Establishment of a SAR requires a mobile signal probably moving in the phloem. Salicylic acid has been found to be translocated in the phloem, but despite this fact and though it is necessary for the induction of PR-protein synthesis in the remote site, it does not seem to be the mobile signal responsible for SAR induction. This still remains elusive.

Kalancho, where root exudates from the mother plant inhibit growth of the daughter plants which are formed at the edge of the leaf (Fig. 1.10.13). The main components of the root exudate (in decreasing amounts) are gallic acid, caffeic acid, p-hydroxybenzoic acid, protocatechuic acid and p-coumaric acid, again all phenolic substances (Br et al. 2000). It is not yet known how these phenolic compounds affect growth of competitors; inhibition of germination often plays an important role.

Summary
1. Poisonous plant compounds, which are constitutively formed and are usually products of secondary metabolism, protect plants from herbivory, however, never completely. Some phytophagous insects take up such compounds while feeding on plants and accumulate them to protect themselves from their enemies. During the course of evolution, however, counteractions developed again. Volatile secondary plant metabolites often act as messengers which either frighten off or entice. 2. In addition to the constitutively expressed defence system of plants, reactions are also induced after wounding or infestation by pathogens. These reactions are triggered by pressure waves in the case of wounding, and by elicitors in the case of pathogen infestation. 3. If a plant is wounded, the vascular bundles are almost always affected. If air enters into the xylem, which is under negative pressure, a positive pressure wave is triggered across the whole plant. A depolarisation wave, decreasing with increasing distance from the damaged site, follows the change of pressure. The depolarisation wave activates intracellular signal chains, initially the formation of systemin from prosystemin, followed by activation of a lipase which releases linolenic acid from biomembranes. The free linolenic acid undergoes many metabolic reactions, initiated by lipoxygenase. One of the products is jasmonate, a stress hormone, but traumatin, a wound stress hormone, which activates cell division in the damaged tissue, may also be produced. 4. Jasmonate, like many phytohormones, has pleiotropic effects; it induces the expression of various genes, which, insofar as they are known, encode proteins serving to counteract stress. These genes are active even in those

1.10.3
Allelopathy
Many interactions between plants and herbivores and plants and pathogens are known. Less well known is that plants affect the growth of other plants. Such a phenomenon, allelopathy (Greek allelos, another; pathos, suffering), cannot always be unequivocally demonstrated, and was often said to be responsible for reduced plant growth without good evidence for such an interpretation. The classical example for allelopathy is the inhibitory effect on germination by leaves of the walnut trees ( Juglans regia, J. nigra). The inhibition is caused by juglone (1-hydroxynaphthalene), which is formed from 5-glucosyl-1,4,5-trihydroxynaphthalene under the influence of bacteria. Another classical example is the inhibition of grasses in the garigue of southern California, in the presence of the shrubs Salvia leucophylla and Artemisia californica. They produce a broad spectrum of possibly toxic, volatile terpenoids which impair growth of grasses. However, recently, doubts have been expressed about this interpretation; it is now thought that rabbits shelter under the bushes and graze the grasses causing their low biomass production. In most cases known, compounds which act allelopathically are phenolics which are metabolised by soil microorganisms and are thus activated. A good example of intraspecific allelopathy is shown by several representatives of the genus

Summary

251

n Fig. 1.10.13. Intraspecific alle-

lopathy in Kalancho (Kalancho daigremontiana). A Same age daughter plantlets were placed in a sand soil at regular intervals from the mother plant. After 150 days the height of the daughter plants clearly shows the inhibitory effect of the mother plant on their growth. B Quantification shows that the inhibition of the young plants depends on the distance from the mother plant. (After Br et al. 2000)

25 20
Stem height (cm)

15 10 5 0 24 21 18 Dist anc 15 e fro 12 mm othe 9 6 r pla nt (c 3 m) 4 1 0

M

5 2
on th

parts of the plant that have not been exposed to stress or develop after the stress. Because of this systemic reaction, this phenomenon is also called immunisation. 5. Plants distinguish in their reaction between simple mechanical wounding and damage because of herbivory. In damage caused by herbivory, the regurgitant of phytophagous insects or their larvae is very important. This digestive fluid contains fatty acids conjugated to amino acids as elicitors. Synthesis of proteinase inhibitors, as well as synthesis of volatile signal compounds of the group of terpenes, is predominantly induced. Additionally, non-volatile defensive substances are formed after induction by jasmonate, e.g. the alkaloid

nicotine, which is toxic for many insects and warm-blooded animals. 6. In addition to the transduction of the wound signal by jasmonates, there is also a jasmonate-independent path which activates the defence reactions. 7. Compatible and incompatible reactions are distinguished in pathogen attacks. In compatible reactions, there is a strong infestation, whilst the incompatible reactions indicate resistance. Resistance may be constitutive, directly acquired or acquired by immunisation, the so-called systemic acquired resistance. Salicylic acid derived from phenolic metabolism is the signal substance inducing the defence system in infected or in non-infected plant parts.

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Biotic Stress: Herbivory, Infection, Allelopathy tomato leaves in response to wounding. Plant Physiol 111:797803 Conrath U, Kauss H (2000) Systemisch erworbene Resistenz Das Immunsystem der Pflanze. Biologie unserer Zeit 30:202208 Creelman RA, Mullet JE (1997) Biosynthesis and action of jasmonates in plants. Annu Rev Plant Physiol Plant Mol Biol 48:355381 Feussner I, Khn H, Wasternack C (2001) Lipoxygenase-dependent degradation of storage lipids. Trends Plant Sci 6:268273 Frey M, Chomet P, Glawischnig E, Stetter C, Grun S, Winklmair A, Eisenreich W, Bacher A, Meeley RB, Briggs SP, Simcox K, Gierl A (1997) Analysis of a chemical plant defense mechanism in grasses. Science 227:696699 He D (1999) Pflanzenphysiologie: molekulare und biochemische Grundlagen von Stoffwechsel und Entwicklung der Pflanzen. UTB Ulmer Stuttgart, 608 pp Kessler A, Baldwin IT (2001) Defensive function of herbivore-induced plant volatile emissions in nature. Science 291:21412144 Korth KL, Dixon RA (1997) Evidence for chewing insectspecific molecular events distinct from a general wound response in leaves. Plant Physiol 115:12991305 Lamb C, Dixon RA (1997) The oxidative burst in plant disease resistance. Annu Rev Plant Physiol Plant Mol Biol 48:251276 Meindl T, Boller T, Felix G (1998) The plant wound hormone systemin binds with the N-terminal part to its receptor but needs the C-terminal part to activate it. Plant Cell 10:15611570 Mitchell-Olds T, Gershenzon J, Baldwin IT, Boland W (1998) Chemical ecology in the molecular era. Trends Plant Sci 3:362365 Par PW, Tumlinson JH (1999) Plant volatiles as a defense against insect herbivores. Plant Physiol 121:325331 Rouster J, Leah R, Mundy J, Cameron-Mills V (1997) Identification of a methyl-jasmonate responsive region in the promoter of a lipoxygenase gene expressed in barley grain. Plant J 11:513523 Sharkey TD (1996) Emission of low molecular mass hydrocarbons from plants. Trends Plant Sci 3:7882 Stahlberg R, Cosgrove DJ (1997) The propagation of slow wave potentials in pea epicotyls. Plant Physiol 113:209 217 B, Davies E (1998) The wound response in tomato Stankovic involves rapid growth and electrical responses, systemically up-regulated transcription of proteinase inhibitor and calmodulin and down-regulated translation. Plant Cell Physiol 39:268274 Staskawicz BJ (2001) Genetics of plant-pathogen interactions specifying plant disease resistance. Plant Physiol 125:7376 Stout MJ, Workman J, Duffey SS (1994) Differential induction of tomato foliar proteins by arthropod herbivores. J Chem Ecol 20:25752594 Titarenko E, Rojo E, Len J, Snchez-Serrano JJ (1997) Jasmonic acid-dependent and -independent signaling pathways control wound-induced gene activation in Arabidopsis thaliana. Plant Physiol 115: 817826 Wasternack C, Parthier B (1997) Jasmonate-signalled plant gene expression. Trends Plant Sci 2:302307

8. The most frequently occurring form of an incompatible reaction is the hypersensitive response: In addition to the synthesis of PR proteins (pathogenesis-related proteins, mainly lytic enzymes) and low molecular weight antibiotically active phytoalexins, the hypersensitive response comprises the programmed cell death of the infected cell, thereby depriving the pathogen from its resources. The hypersensitive reaction can be recognised by small, well-defined lesions. 9. Hypersensitive reactions are triggered by socalled elicitors, which may be derived from the pathogen, but also from the plant cell wall attacked by the pathogen. They bind specifically to the receptors which are encoded by R(esistance) genes. The specificity of elicitors is based on the avr genes (avirulence genes) of the pathogen. 10. Inhibition of growth and development of plants by neighbouring plants is called allelopathy, if it is based on excretion of incompatible chemical substances and not on simple competition. Classical examples are inhibition of germination and growth caused by rotting leaves of walnut trees, and the inhibition of growth of grasses by volatile compounds from Lamiaceae and Asteraceae. Intraspecific allelopathy is demonstrated by Kalanchoe daigremontiana, where the mother plant inhibits growth of plantlets developing at the edges of its leaves by secretion of a broad spectrum of phenolic compounds from the roots.

References
Baier M, Dietz K-J (1999) Alkyl hydroperoxide reductases: the way out of the oxidative breakdown of lipids in chloroplasts. Trends Plant Sci 4:166168 Baldwin IT (1998) Jasmonate-induced responses are costly but benefit plants under attack in native populations. Proc Natl Acad Sci USA 95:81138118 Br W, Pfeifer P, Dettner K (2000) Biochemische Interaktionen zwischen Kalanchoe-Pflanzen. Biol Unserer Zeit 30:228234 Chong J, Pierrel M-A, Atanassova R, Werck-Reichhart D, Fritig B, Saindrenan P (2001) Free and conjugated benzoic acid in tobacco plants and cell cultures. Induced accumulation upon elicitation of defense responses and role as salicylic acid precursors. Plant Physiol 125:318328 Conconi A, Miquel M, Browse JA, Ryan CA (1996) Intracellular levels of free linolenic and linoleic acids increase in