Phylogeny and Ecological Evolution of Gall-Inducing Sawflies (Hymenoptera: Tenthredinidae)
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University of Joensuu, PhD Dissertations in Biology No:6 Phylogeny and ecological evolution of gall-inducing sawflies (Hymenoptera: Tenthredinidae) by Tommi Nyman Joensuu 2000 Nyman, Tommi Phylogeny and ecological evolution of gall-inducing sawflies (Hymenoptera: Tenthredini- dae). – University of Joensuu, 2000, 92 pp. University of Joensuu, PhD Dissertations in Biology, n:o 6. ISSN 1457-2486 ISBN 951-708-962-7 Key words: coevolution, convergent evolution, extended phenotypes, gall morphology, gradualism, insect-plant interactions, Nematinae, phylogeny, Salix, sawfly, speciation The nematine sawflies (Hymenoptera: Tenthredinidae) that induce galls on willows (Salix spp.) form one of the most abundant and speciose herbivore groups in the Holarctic region. The purpose of this thesis was to study the evolutionary history of the nematine gallers. The three main questions addressed were: (1) what is the sequence in which different gall types have evolved in the nematines; (2) how has the utilization of host plants evolved in the group; and (3) why has gall induction evolved in the nematines? The first question was studied by reconstructing the phylogeny of representative nema- tine species by using enzyme electrophoresis and DNA sequencing. According to the results, species that induce true closed galls evolved from species that induce leaf folds or rolls. Thereafter, several different gall types evolved gradually, and the evolution of each gall type was followed by a radiation to new host species. The phylogenies show that gall morphology is mainly determined by the insects, i.e., the gall represents an extended phenotype of the galler. The two phylogenies were also used to study the second question. The results show that parallel cladogenesis between willows and the gallers is not a sufficient explanation for the current host associations, since many willow species have been colonized multiple times by nematine species representing different gall types. Likewise, it seems that the overall chemi- cal similarity of hosts does not determine the direction of host shifts. Consequently, other explanations, such as host phenology, geographical distribution, or habitat, are probably needed for explaining the patterns of host use in the nematine sawflies. Similar results were obtained on a smaller scale in an enzyme electrophoretic study of the species that induce bud galls. The third question was studied by comparing the chemical properties of sawfly galls to the chemical properties of the ungalled tissues of the willow hosts. The galls were found to contain comparatively low levels of most phenolic defense compounds that are found in other willow tissues. Thus, it appears that the galls are nutritionally beneficial for the sawfly larvae. The changes in chemistry occur in a highly coordinated pattern in all studied galler- host pairs, which suggests that the sawflies are able to manipulate the phenolic biosynthesis in their hosts. The end result is a striking convergence of the chemical properties of the galls both within and between host species. Tommi Nyman, Department of Biology, University of Joensuu, P.O. Box 111, FIN-80101 Joensuu, Finland CONTENTS LIST OF ORIGINAL PUBLICATIONS 1. INTRODUCTION 7 1.1. Gall-inducing insects 7 1.2. Purpose of this study 8 2. THE STUDY SYSTEM 8 2.1. Willows 8 2.2. The nematine gallers 9 3. THE EVOLUTION OF DIFFERENT GALL TYPES 11 3.1. Hypotheses 11 3.2. Allozyme results 12 3.3. DNA results 12 3.4. Combined evidence 14 3.5. Discussion 15 4. THE EVOLUTION OF HOST PLANT USE 16 4.1. Hypotheses 16 4.2. Host-plant relationships in the nematines 17 4.3. Euura mucronata: a polyphage or a sibling species complex? 18 4.4. Discussion 19 5. THE CHEMICAL ECOLOGY OF NEMATINE SAWFLIES 22 5.1. The Nutrition Hypothesis and previous studies 22 5.2. The chemical properties of Pontania galls on willows 23 5.3. Discussion 23 6. CONCLUDING REMARKS 26 ACKNOWLEDGEMENTS 28 REFERENCES 29 LIST OF ORIGINAL PUBLICATIONS This thesis is based on the following articles, referred to in the text by their Roman numerals. I Nyman, T., Roininen, H. & Vuorinen, J. A. 1998. Evolution of different gall types in willow-feeding sawflies (Hymenoptera: Tenthredinidae). Evolution 52: 465-474. II Nyman, T., Widmer, A. & Roininen, H. 2000. Evolution of gall morphology and host- plant relationships in willow-feeding sawflies (Hymenoptera: Tenthredinidae). Evolu- tion 54: 526-533. III Nyman, T. The willow bud galler Euura mucronata Hartig (Hymenoptera: Tenthredini- dae): one polyphage or many monophages? Manuscript. IV Nyman, T. & Julkunen-Tiitto, R. 2000. Manipulation of the phenolic chemistry of wil- lows by gall-inducing sawflies. Proc. Natl. Acad. Sci. USA 97: 13184-13187. Some unpublished results are also presented. 7 1. INTRODUCTION nisms include mechanical damage, plant hormone analogs, and genetic manipulation 1.1. Gall-inducing insects (Hori 1992, Price 1992). Genetic manipula- tion is involved in the process of gall induc- Meyer (1987) defines galls as “all manifes- tion by the bacterium Agrobacterium tume- tations of growth, whether positive or nega- faciens. The bacterium inserts a portion of a tive, and of abnormal differentiation induced Ti plasmid into the genome of some of the on a plant by animal and plant parasites”. cells of its host, which eventually leads to Usually galls can be seen as abnormal abnormal growth (Davey et al. 1994). Al- growth caused by an increase in the number though it has been suggested that such ma- (hyperplasia) or size (hypertrophy) of plant nipulation may also occur in the case of in- cells at the site of the gall (Meyer 1987). sect gallers (Cornell 1983), no evidence for Galls can be induced on virtually all parts of such a mechanism has been found (Price plants, and their complexity varies from 1992). It is likely that the mechanism of gall simple pouches to highly structured organs induction varies between galler taxa, but with specifically differentiated tissues (Dre- currently it would seem that the production ger-Jauffret & Shorthouse 1992). of plant hormone analogs represents the The ability to induce galls has evolved most likely explanation (Hori 1992, Roskam convergently in many taxa, ranging from 1992). For example, McCalla et al. (1962) microbes and fungi to nematodes and arthro- found uric acid, glutamic acid, and adenine pods (Meyer 1987). In insects alone, gall derivatives in the female accessory glands of induction has evolved in at least the follow- the sawfly galler Pontania pacifica Marlatt. ing seven orders: Thysanoptera, Hemiptera, These compounds are closely related to cy- Homoptera, Lepidoptera, Coleoptera, Dip- tokinins, which are natural growth- tera, and Hymenoptera (Meyer 1987, Dre- promoting hormones in plants (Hartley ger-Jauffret & Shorthouse 1992). The num- 1992). High concentrations of cytokinins ber of independent origins of gall induction have been found in galls induced by Pon- in insects is, however, considerably higher, tania proxima Lepeletier, but apparently since within most of these orders there have they are produced by the plant (Leitch 1994). been several separate lineages leading to the Furthermore, studies on other galler/plant galling habit (Dreger-Jauffret & Shorthouse combinations have found that the relative 1992, Roskam 1992). concentrations of cytokinins are not neces- The fossil record of plant galls shows sarily higher in all galls (Mapes & Davies that unknown gall-inducing insects were 1998). Mapes and Davies (1998) found cy- already present during the Late Carbonifer- tokinins and IAA in larvae of the dipteran ous period, over 300 million years ago (La- stem galler Eurosta solidaginis Fitch, but it bandeira & Phillips 1996). Numerous galls is not clear whether the larvae produce these have been found in younger plant fossils, but hormones de novo, or sequester them from in most cases the identity of the gall inducer their diets. remains unknown (Larew 1992, Scott et al. Different galler taxa tend to specialize in 1994). However, it is evident that several different plant taxa, but some groups of gall-inducing insect groups were already in plants are attacked more than others (Ro- existence at the time of the initial radiation skam 1992). For example, relatively few of angiosperms during the mid-Cretaceous; insect species induce galls on ferns and possible taxa include aphids, midges, and gymnosperms (Roskam 1992). In contrast, cynipid wasps (Scott et al. 1994). over 90% of galls occur on dicots, and Although gallers are common, the within dicots the vast majority of galls occur mechanism by which insects induce galls on Fagaceae, Asteraceae, and Rosaceae remains largely unknown. Possible mecha- (Abrahamson & Weis 1987). In Britain, 8 Salix (Salicaceae) and Quercus (Fagaceae) two questions were studied by reconstructing species support the largest numbers of insect the phylogeny of representative nematines gallers (Kennedy & Southwood 1984). using allozyme electrophoresis and DNA A common observation is that gall- sequencing, and the last one was studied by inducing insect species are strictly host- analyzing the chemical properties of sawfly specific (Meyer 1987, Dreger-Jauffret & galls using high-performance liquid chro- Shorthouse 1992). This is probably a result matography. of the tight link between the gallers and their hosts, because gall induction may be possi- ble only during a short period of time during 2. THE STUDY SYSTEM the growing season, and the available plant species (or even individuals or clones within 2.1. Willows species) may differ in their reactions to the gall-inducing stimuli (Smith 1970, Price Willows (Salix spp.) form one of the taxo- 1992, Whitham 1992, Craig et al. 1994, nomically and ecologically most diverse Peries 1994, Csóka 1994, Kokkonen 2000). plant genera in the Northern Hemisphere It is important to note that the degree of host (Argus 1973). The oldest willow fossils have specificity may have been underestimated, been found from North American deposits since galler species are often extremely dif- dating from the early Eocene, ca. 55-65 mil- ficult to identify by morphological methods, lion years ago (Collinson 1992).