doi:10.1111/j.1420-9101.2004.00808.

TARGET REVIEW

Costs and benets of genetic heterogeneity within organisms

M. PINEDA-KRCH * & K. LEHTILA
*Department of Zoology, 6270 University Boulevard, University of British Columbia, Vancouver, BC, Canada Department of Botany, Stockholm University, Stockholm, Sweden

Keywords:
chimera; clonality; intraclonal variation; modular organisms; mosaic; unitary organisms.

Abstract
An increasing number of studies have recently detected within-organism genetic heterogeneity suggesting that genetically homogeneous organisms may be rare. In this review, we examine the potential costs and benets of such intraorganismal genetic heterogeneity (IGH) on the tness of the individual. The costs of IGH include cancerous growth, parasitism, competitive interactions and developmental instability, all of which threaten the integrity of the individual while the potential benets are increased genetic variability, size-specic processes, and synergistic interactions between genetic variants. The particular cost or benet of IGH in a specic case depends on the organism type and the origin of the IGH. While mosaicism easily arise by genetic changes in an individual, and will be the more common type of IGH, chimerism originates by the fusion of genetically distinct entities, and is expected to be substantially rare in most organisms. Potential conicts and synergistic effects between different genetic lineages within an individual provide an interesting example for theoretical and empirical studies of multilevel selection. documented from all branches of life suggesting that it is much more common than previously believed. Traditionally IGH is viewed as an aberration resulting in a multitude of deleterious effects, for instance tumour formations. Advantages conferred by IGH are considered rare. One notable exception is the genetic diversity of the immune system (Youssouan & Pyeritz, 2002). Studies evaluating the signicance of IGH are predominantly concerned with short-term consequences, such as potential developmental costs. In the light of the accumulating number of studies identifying natural IGH (Table 1) the importance of considering long-term ecological and evolutionary signicance of IGH is increasing. In this review, we examine the potential costs and benets of genetically heterogeneous organisms and set the stage for further inquiries into its ecological and evolutionary signicance.

Introduction
Individuality plays a central role in ecology and evolution where concepts such as population, birth, reproduction, and death are dened at the level of the individual organism. In spite of this, the concept of individuality is lacking a clear denition and used ambiguously in the literature. The key attributes of individuality has traditionally involved the simultaneous and invariable presence of physiological unity and autonomy, genetic uniqueness, and genetic homogeneity (Santelices, 1999). The invariability of the rst two attributes have over the years been disproved for many organism groups. Physiological unity and autonomy is absent in social colonies and genetic uniqueness is lacking in clonally derived organisms (see Santelices, 1999, for details). Genetic homogeneity is still, however, commonly assumed to hold true. Over the last few years, however, studies have accumulated where naturally occurring intraorganismal genetic heterogeneity (IGH) has been
Correspondence: Mario Pineda-Krch, Department of Zoology, 6270 University Boulevard, University of British Columbia, Vancouver, BC V6T 1Z4, Canada. Fax: +1 604 822 2416; e-mail: pineda@zoology.ubc.ca

Intraorganismal genetic heterogeneity

An individual is traditionally dened as reproductive, physiologically united, autonomous, genetically homogeneous and unique entity, which is also the main unit of selection (Tuomi & Vuorisalo, 1989; Fagerstro m et al., 1998; Michod, 1999; Santelices, 1999). The denition

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Table 1 Distribution of effects resulting from IGH among various extant taxa with a modular (m) or unitary structure (u). IGH type refers to chimerism (c) or mosaicism (m) and the net phenotypic effects of the IGH at the individual level are detrimental ()), advantageous (+) or neutral (n).
Taxa Bacteria Myxobacteria Protists Cellular slime moulds Algae Species Structure IGH type Effect Comments References

Myxococcus xanthus Dictyostelium spp. Chondrus canaliculatus, Gracilaria chilensis, Mazzaella laminarioides, Sarcothalia crispata Armillaria spp., myccorhizal fungi, Scutellospora castanea Ficus burtt-davyii, Juniperus spp., Nicotiana spp., Pelagonium spp., Rhizospora mangle, Zea mays

m m m

c c c

) ) +

Cell parasitism Cell parasitism, cheating Synergism resulting in a larger, more branched (Healthier) phenotype, increased survival

1 2 3

Fungi Plants

m m

c, m c, m ),+, n Developmental instabilities, GMH, size-specic advantage.

4 5

Animals Invertebrates Cnidaria

Tunicata Vertebrates Horse Cat Marmosets Humans

Clavularia hamra, Heteroxenia fuscescens, Hydrozoa spp., Nephthea spp., Parerythropium fulvum fulvum Diplosoma listerianum, Botryllus schlosseri Equus caballus Felix domesticus Callithrix jacchus Homo sapiens

m, u

), n

Developmental instability: disruption of polyp formation, slower growth

Cell parasitism

u u u u

c m c c, m

n n n ), n

Blood chimerism Tortoiseshell coat Blood chimerism Developmental instability. Blood chimerism, gynandromorphism, mosaic disorders of the skin, autoimmune diseases, genetic disorders, metabolic derangements, skeletal dysplasias, muscular dystrophy, clotting disorders, tumour-suppression disorders

8 9 10 11

l & Papp (2000); Velicer et al. (2000). Buss (1982); Pa l & Papp (2000); Strassmann et al. (2000). Filosa (1962); Buss (1982, 1999); Ennis et al. (2000); Pa 3 Meneses & Santelices (1999); Meneses et al. (1999); Santelices et al. (1999); Poore & Fagerstro m (2000, 2001). 4 Sanders (1999); Money (2000); Peabody et al. (2000); Stenlid (2000); Kuhn et al. (2001). 5 Whitham & Slobodchikoff (1981); Jerling (1985); Ruth et al. (1985); Klekowski (1988); Edwards et al. (1990); Thomson et al. (1991); Gill et al. (1995); Szymkowiak & Sussex (1996); Marcotrigiano (1997, 2000); Korn (2002). 6 Grosberg (1988); Hughes (1989); Rinkevich & Shapira (1999); Barki et al. (2002). 7 Tilney-Bassett (1986); Rinkevich & Weissman (1987); Grosberg (1988); Hughes (1989); Rinkevich & Weissman (1992); Szymkowiak & Sussex (1992); Pancer et al. (1995); Stoner & Weissman (1996); Rinkevich & Shapira (1999); Sommerfeldt & Bishop (1999); Stoner et al. (1999); Ben-Shlomo et al. (2001); Sommerfeldt et al. (2003). 8 Vandeplassche (1993). 9 Benirschke (1981). 10 Signer et al. (2000). 11 zc Opitz & Utkus (2001); Paller (2001); Barinaga (2002); Nelson (2002); Pearson (2002); Youssouan & Pyeritz (2002); O elik (2002).
2

can be broadened by releasing each of these conditions (Santelices, 1999). If permanent physiological integration is not required, organisms such as cellular slime moulds count as multicellular individuals. The individual cells of cellular slime moulds are able to act independently with respect to their environment but have a multicellular fruiting stage (Strassmann et al., 2000). It has also been suggested that a colony of eusocial insects is an individual organism where working individuals are physiologically

autonomous but cannot reproduce independently (Wilson & Sober, 1989; Santelices, 1999). In this paper, we deal with cases where individuals are at least temporarily physiologically united and discuss the implications of genetic heterogeneity within such entities. The majority of our knowledge on the prevalence of naturally occurring IGH and its effects mainly come from the elds of horticulture, animal husbandry, and medicine. There are two main types of IGH, mosaicism and

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chimerism, which refer to the functional origin of the heterogeneity. In a mosaic individual the IGH originates from various forms of genetic change, e.g. somatic mutations, mitotic recombination, mitotic gene conversion (Otto & Hastings, 1998; Youssouan & Pyeritz, 2002), genome duplications, and changes in ploidy level (Santelices, 1999). Chimeric individuals, in contrast, originate through a fusion process, either by fusion of two individuals or exchange of genetically distinct parts which can be single-celled or multicellular units (Fig. 1). Nuclear transfer such as heterokaryosis in fungal hyphae (Wu & Glass, 2001) and in red algae (Poore & Fagerstro m, 2000) is also categorized as a chimera formation. Formally a chimera is dened as the presence of two or more cell lines that are derived from different zygotes. A clear distinction between chimeras and mosaics is not always available in the reported cases, often the two types are used indiscriminantly or interchangeably. Unless indicated otherwise we will assume the above denitions, which are used in much of the recent literature (Rinkevich, 2000; Pearson, 2002; Sommerfeldt et al., 2003). The generation of natural chimeras requires

the coalescence of two or more individuals and is, hence, substantially more difcult and risky than the formation of a mosaic which is a slower and more progressive process. The fusion of two unrelated individuals gives rise to disruptive internal conicts compromising the function and integrity of the individual. Normally, cellular self-recognition systems prevent fusion with all nonself cells except close kin (Buss, 1982; Grosberg & Strathmann, 1998). Thus it seem likely that the majority of genetically heterogeneous individuals will be mosaics rather than chimeras. A notable exception are cellular slime moulds and myxobacteria where the relationship is reversed due to the ease by which fusion occurs and the ephemeral nature of the multicellular entity. While we may expect chimeras to be rare, evidence is accumulating indicating that genetic mosaics are probably the rule while genetic homogeneity, one of the conventional hallmarks of individuality, is the exception (Santelices, 1999; Rinkevich, 2000; Loxdale & Lushai, 2003). We will here use the general terms genetic heterogeneity or IGH for any organism composed of a heterogeneous assemblage of modules.

Precursors

Ontogeny

Sequestration

Singlecelled

and 1 or 2 6 3 or or 7 8 9 10

Multicelled

11 4 and 5 or

12

13

14

15

Fig. 1 The origin and fate of IGH in a hypothetical life cycle where precursor cells are sequestered late in ontogeny (i.e. lacking an early sequestered germ line). IGH may arise either through the coalescence of autonomous single (1) or multi-celled (4) precursors creating a chimeric organism, through genetic change taking place during ontogeny creating a mosaic organism (2 and 5), or from a genetically heterogeneous multicellular precursor (3). The circles surrounding the lower-level units (cells, modules, or groups of modules) represent the boundaries dening the higher-level units, usually dened as an individual. During the course of ontogeny (6) IGH may prevail or be eliminated. In the upper part of the gure the life cycle is recurrently contracted to a single-celled stage (1, 2, and 710) which may be meiotically (i.e. meiocytes) or mitotically derived (e.g. stem cells). In the bottom part of the gure the precursor as well as the sequestration of subsequent precursors is multi-celled (35 and 1115), i.e. mitotically derived. The multi-celled mitotic lineage corresponds to the meristem meristem cycle by Fagerstro m (1992), where new organisms arise vegetatively. During clonal propagation IGH may be eliminated through single-celled (8 and 9) or multi-celled sequestration (12 and 14) or persist through a multi-celled sequestration (13) although most likely with an altered composition.

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High degrees of IGH, i.e. many different genetic lineages within an individual, have been detected in a number of different types of organisms. Thomson et al. (1991) sampled leaves from strangler g trees (211 per tree) from several branches in 14 individuals of six species of gs. Thirteen of the individual trees (all species) showed multiple genotypes (28 genotypes), including the trees where only two leaves had been sampled. All in all the 14 trees represented at least 45 genetic individuals. Peabody et al. (2000) analysed the genetic composition of the fungus Armillaria gallica. A 10-cell isolate from a single basidome produced nine different genotypes when analysed for variation at six nuclear loci. Arbuscular mycorrhizal fungi have multi-nuclear spores where the nuclei may differ in their genotypes (Stenlid, 2000). In many red algae multi-chimeras are regularly formed through sporeling coalescence between two or more genetically distinct spores. Santelices et al. (1999) showed that a larger number of coalescing spores altered the morphology and increased the survival of the developing algae. Meneses et al. (1999) reported rapid within-individual genetic divergence of the red alga Gracilaria chilensis during growth (they do not, however, report on measures taken to avoid contamination of the material by foreign DNA). In the tunicate Botryllus schlosseri multi-chimerism commonly occurs naturally and outperforms bichimeras (Rinkevich & Shapira, 1999). Ben-Shlomo et al. (2001) found IGH on average in 8% (maximum of 14%) of the sampled colonies of B. schlosseri. It is likely that this is an underestimate of the level of IGH in the population as they did not collect colonies showing morphological evidence of IGH and the genetic evidence was based on only one microsatellite loci. Sommerfeldt & Bishop (1999) found that one-third of sampled colonies in the compound ascidian Diplosoma listerianum possessed multiple genotypes, some colonies with up to four different genotypes. Multi-chimeras have also been documented in corals and hydroids (Rinkevich & Shapira, 1999). In other organisms, studies detecting multi-chimerism are lacking. Nevertheless, patterns of phenotypic variability in several studies indicate that multi-chimerism may be common in organisms such as the fungal parts of lichens (Fahselt, 1996; Honegger, 1996), plants (Edmunds & Alstad, 1978; Edwards et al., 1990; Dawson & Bliss, 1993), and invertebrate animals (Gill et al., 1995).

comprise, in fact, a large fraction of multicellular life (Table 1). Organisms belonging to this group range across many of lifes extremes from the oldest and largest to the smallest and most ephemeral creatures. Among these organisms are plants, fungi, and many benthic invertebrates, e.g. hydroids, corals, bryozoans and colonial ascidians (Vuorisalo & Mutikainen, 1999). Unitary organisms have a closed developmental programme starting from a single-celled zygote and ending with a determinate adult organism. There is a xed number of constituent parts (arms, legs, organ systems) at predetermined positions (the head is always situated on top of the spinal column), which are established early during ontogeny. Modular organisms, on the other hand, have an open-ended developmental programme starting from a single cell (zygote or stem cell) or multicellular stage (various vegetative propagules) (Fagerstro m et al., 1998) and developing by the iteration of basic structural units of construction referred to as modules (Tuomi & Vuorisalo, 1989). The result is an adult organism with a variable number of parts at various positions. The low level of terminal differentiation, plastic development, the absence of an early sequestered germ line, and the often long life span predisposes modular organisms towards becoming genetically heterogeneous over time (Fig. 1) (Klekowski & Godfrey, 1989; Otto & Hastings, 1998; Marcotrigiano, 2000).

Costs and benets of IGH

The presence of genetically distinct replicating units in a multicellular organism can lead to several different kinds of interactions among the genetic lineages resulting in the manifestation of costs or benets at various level of the organization (Grosberg & Strathmann, 1998). This is particularly true in organisms with a modular construction where any of the levels of organization can act as a causal level of selection (Tuomi & Vuorisalo, 1989). If phenotypic traits of a module modify its survival and reproduction, modules and individuals may both work as levels of selection (Tuomi & Vuorisalo, 1989; Vuorisalo et al., 1997). For simplicity, we assume here that the costs and benets of IGH are primarily manifested at the level of the individual. There are no conceptual difculties in extending this framework to encompass all types of modular organisms. On the contrary, the hierarchical nature of modular organisms, where the units of selection may be dened at arbitrary levels of construction, suggests that it may be an appropriate extension (Gill et al., 1995). Presence of genetically heterogeneous lineages within an individual may confer both advantages and disadvantages. For instance, the fusion of two individuals may result in an increased size, which is advantageous, but may at the same time result in intraorganismal conicts. We rst outline the possible net effects of costs and benets of IGH on tness of an individual and then discuss specic costs and benets in more detail.

IGH in modular organisms

One of the most important steps in the evolution of morphological complexity and diversity has been the advent of modular construction, i.e. a body plan made up of repeated structural units modules (Carroll, 2001). Although modularity per se is rarely appreciated in ecological and evolutionary studies and remains somewhat enigmatic to many biologists, modular organisms

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General net effects IGH is usually assumed to convey a cost to the individual resulting in aberrant phenotypes and lowering the tness of the individual. In unitary organisms, with preformistic development (Buss, 1983), IGH in the somatic lineage is often associated with cancerous growth (Youssouan & Pyeritz, 2002), germ cell parasitism (Buss, 1982; Stoner & Weissman, 1996), somatic cell parasitism (Rinkevich & Weissman, 1992),andother anomalies(Benirschke,1981). The coexistence of multiple genotypic lineages within an organism often results in intraorganismal conicts threatening the harmony and well-being of the organism. In the case of somatic mutations this could lead to a selsh (defecting) cell lineage that could either parasitize the wild type lineage (Buss, 1982) or potentially replace it (Klekowski, 1988; Otto & Hastings, 1998). Other potential conicts are, e.g. developmental instabilities (Stebbins, 1986; Mller & Swaddle, 1997) or intra-individual competition (Otto & Orive, 1995). The IGH conferring benets to the individual by increasing its tness is a scenario that rarely gets attention. In unitary organisms, with a high degree of integration among its parts, this situation is assumed to be rare. In modular organisms, the open-ended development with a low degree of integration allows advantageous lineages be selected through intraorganismal selection. Examples of such advantages are increased genetic variability, synergistic interactions between the variants (e.g. division of labour and ry, 1999), mate complementation of function; Szathma location (Buss, 1982), size-specic attributes enhancing tness (e.g. growth rates, reproductive success and survivorship) (Buss, 1982; Foster et al., 2002), and selection through heterosis (Rinkevich & Weissman, 1992). A genetically heterogeneous soma may also not have any effect on individual tness. There are two aspects of this; all lineages may be neutral with respect to the higher-level phenotype or a potentially detrimental effect of one lineage may be neutralized or suppressed in the presence of another genetic lineage. Specic deleterious effects on individual tness

Tragedy of the commons

Both individual cells and a group of cells (an individual organism) achieve best when there is low competitiveness and prudent use of resources. This cooperative state is, however, only a stable strategy when there is a strong coupling between successes at the two levels (Frank, 1998). The coexistence of multiple genetic lineages within an organism creates a breeding ground for conicts and cheating where lineages pursuing their own interests increase their returns relative to other lineages while decreasing the tness of the organism. Problems of this type of group-benecial behaviour has

been referred to as the tragedy of the commons (Hardin, 1968; Frank, 1998; Michod, 1999) and is central in many instances of IGH. Based on the functional relationship between the component lineages in a genetically heterogeneous organism two scenarios of are possible, (i) cell parasitism, where at least one lineage depends on another for its existence and (ii) intraorganismal competition, where none of the lineages depend on each other for their independent existence. A genetically heterogeneous organism that cannot control the proliferation of its lower-level lineages will become parasitized by selsh lineages. In a worst case scenario the integrity of the organism may be compromised such as in cancerous growth while in other cases it may lead to a lowered tness of the individual. Because the parasitizing lineage is incapable of maintaining itself in the absence of the original form, it may adopt a subordinate role in the organism (Buss, 1982; Rinkevich & Shapira, 1999). Such lineage will then persist together with the supportive lineage and may parasitize the soma (somatic cell parasitism) or germ line (germ line parasitism; Hastings, 1989). The parasitic variant is at an advantage when rare, when the probability of it reaching xation through random drift is low and its presence does not detract too much from the tness of the organism. Cell parasitism has been demonstrated, among others, in cellular slime moulds, in myxobacteria, and in ascidians (Table 1). In these organisms independent cells aggregate to form a multicellular reproductive structure. Only a limited number of cells will have the opportunity to reproduce while the rest will form a nonreproductive stalk in slime moulds and autolyse in myxobacteria. Several studies have shown that in reproductive structures composed of a heterogeneous assemblage of cells certain lineages cheat by avoiding the nonreproductive fate (Strassmann et al., 2000; Velicer et al., 2000). The cost to the higher-level unit, the group in this particular case, is the formation of an aberrant fruiting structure. In slime moulds the aggregated cells may fail to produce the stalk of the fruiting structure or will produce a shorter stalk making the group more susceptible to predation and less efcient in dispersal. The cheating lineage, however, benets by gaining access to the germ line and is maintained at a low equilibrium frequency of 5% while the reproductive output of the group is reduced with about 20%. Even lineages with a normal stalk formation may work as cheaters in genetically heterogeneous fruiting structures by disproportional stalk/spore allocation (Strassmann et al., 2000). In colonial tunicates, conspecics coalesce forming a chimeric organism often resulting in stem cells of the soma or germ line becoming parasitized by one of the genetic lineages (Stoner et al., 1999). The frequency by which cell parasitism is detected suggests that this may be a common outcome of IGH.

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In intraorganismal competition any of the lineages can outcompete the other lineages and go to xation, as opposed to somatic cell parasitism where the parasitizing lineage is contingent on a supportive lineage. Intraorganismal competition between genetically heterogeneous lineages may incur a cost to the individual and lower its tness. The deleterious lineage may reach xation in the organism (Pineda-Krch & Fagerstro m, 1999) or become incorporated into the germ line, or both. Rinkevich & Weissman (1992) observed that fusion of two colonies of the tunicate Botryllus schlosseri did not increase the tness (growth, fecundity, and survivorship) of the chimeric colony compared with the summed performance of two separate colonies. The study does not, however, state whether tness decreased. IGH was especially harmful for the partner losing the competition, but even the winning partner could suffer from cell parasitism of the other partner (Rinkevich & Weissman, 1987). Interestingly enough, IGH consisting of more than two distinct lineages, i.e. a multi-chimera was more stable having a smaller reduction in tness (Rinkevich & Shapira, 1999).

mosaic meristems of owering maple Abutilon produce either phenotypically normal leaves or deformed leaves, depending on the meristem layer where the different variants occur (Szymkowiak & Sussex, 1996). Specic advantageous effects on individual tness

Synergism
When several genetically distinct lineages interact so that their combined phenotype is different from the sum of their separate phenotypes, a novel phenotype emerges the phenotype of the individual cannot simply be derived from the phenotypes of its component lineages. Attributes of biochemical pathways such as anthocyanin synthesis, cellular pH, cell size, and fragrance have been shown to be autonomously expressed in mosaic plants (Sussex, 1985) and are likely candidates for synergism to give rise to emergent phenotypes. An extreme case of synergism takes place between lineages that are not able to produce normal structures when occurring by themselves but are able to form a functional entity when brought together. Apart from diploidy, laboratory evidence of this form of synergism, where all the lineages are aberrant when alone, is common (Buss, 1982), but we found no reports on naturally occurring synergism of this form. Normally functioning genetically homogeneous organisms may also benet from being associated with genetically different lineages, e.g. from an increase in the developmental stability, speed of growth, and reduced propensity to fragment (Rinkevich & Shapira, 1999). Santelices et al. (1999) showed that formation of chimeras with more than two genotypes through sporeling coalescence in various red algae (Table 1) resulted in better survival by decreasing grazing, mechanical damage and interspecic competition. Rinkevich & Shapira (1999) reported that heterogeneous individuals composed of more than two genetically distinct lineages of the tunicate Botryllus schlosseri grew faster, reached larger sizes, and did not fragment, as opposed to individuals consisting only of two lineages. It remains to be studied what the relative tnesses of the various genotypes within the individual are, i.e. whether the lower-level units also benet from coalescence.

Developmental instability
Developmental stability refers to the capacity for developmental pathways to resist perturbations during ontogeny while developmental instability is viewed as deviations from an intended phenotype (Mller & Swaddle, 1997). IGH may interfere with the normal developmental pathways by disrupting intercellular communication, developmental coordination and differentiation (Steward, 1978; Becraft & Freeling, 1991) resulting in an aberrant phenotype or arresting ontogeny altogether. Although these effects commonly occur in unitary organisms, where even minor developmental disruptions can have fatal consequences (Benirschke, 1981), the developmental plasticity of modular organisms enables them to resist even severe interference in the ontogenetic program resulting in a phenotype that may be aberrant but nevertheless functioning. Many examples of developmental instabilities in plants are known from horticulture where both mosaic and chimeric plants are abundant (Szymkowiak & Sussex, 1996; Otto & Hastings, 1998). In plants, where developmental patterning largely depends on positional information (Wolpert, 1998), the spatial arrangement of cell lineages within a meristem is important in determining the phenotype of an organism. Szymkowiak & Sussex (1992) and Szymkowiak (1996) studied mosaic (refered to as chimeras) forms of tomatoes Lycopersicon esculentum and L. peruvianum holding two lineages mutant and wild type that affected attributes of oral phenotype (size of oral meristem and carpal numbers). They found that it is the genotype of the internal meristem layers that determines the phenotype. Similarly, phenotype depends on the spatial arrangement of the lineages in genetically heterogeneous owering maples. Some

The genetic mosaicism hypothesis

The genetic mosaicism hypothesis (GMH) was rst proposed to explain patterns of coevolution between long-lived plants and their short-lived, and rapidly evolving, herbivores and pests. According to the GMH, long-lived arborescent plants accumulate somatic mutations and become genetic mosaics as they grow. The GMH predicted that selection can act on this new genetic material during the somatic stage of ontogeny (Gill et al., 1995). Despite the name, the Genetic Mosaicism Hypothesis, the hypothesis does not require the IGH to be of a mosaic type. It is equally applicable to chimeric

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heterogeneity and perhaps a more suitable name would be the Genetic Heterogeneity Hypothesis, GHH. Under the GMH the phenotypic variability of the higher-level unit increases conferring a greater phenotypic repertoire when dealing with environmental contingencies such as herbivory, pests, and micro-evolutionary adaptations (Silander, 1985; Rinkevich & Shapira, 1999). Herbivores, for example, do not cope well with varying food quality (Suomela & Nilson, 1994; Karban et al., 1997). In long-lived modular organisms, variable module quality entails both a short-term and long-term edge in the interspecic arms race with pests and herbivores. Genetic mosaicism may be one way of producing such variation in quality (Whitham & Slobodchikoff, 1981; Gill & Halversson, 1984). Edwards et al. (1990) observed that Eucalyptus melliodora trees, which normally suffer from high levels of herbivory by Christmas beetle, Anoplognathus montanus, sometimes carry uninfested branches. It turned out that the content of volatile oils between damaged and undamaged leaves differed, which was possibly caused by newly arisen mutations in meristematic tissue. In its simplest form, novel benecial variants are selected for, as in the case of E. melliodora, with the potential of replacing the original variant. In an alternative scenario, however, the resistant leaves may protect vulnerable parts of the plant and thus benet the whole individual. The GMH is not limited to interactions with pests, conceptually it is equally applicable to any benet to the individual IGH may have. In chimeric strangler gs (Ficus spp.), e.g. IGH may result in owering asynchrony among genetically distinct branches that have originated by fusion of separate trees (Thomson et al., 1991). Although the benets of this asynchrony have not been investigated, one potential benet could be that the gs, which are spaced far apart and depend on wasps for their pollination, could more easily attract and maintain a large enough wasp population for pollination if they ower continuously. Synchronous owering, on the other hand, may also be benecial as outcrossing could take place within the crown of a single tree.

(Thomson et al., 1991). The net benets of coalescence depend on the absolute size and the relative size difference between the coalescing units. Large differences in sizes may only be advantageous to the smaller unit while the large unit may receive little benet (Thomson et al., 1991). Evidence for size-specic benets of fusion has been found in cellular slime moulds (Foster et al., 2002), in algae (Santelices et al., 1999), in tunicates (Rinkevich & Shapira, 1999), and in trees (Thomson et al., 1991).

The fate of IGH

Selection can work at many stages of development at various levels of organization, all inuencing the fate of the heterogeneity. Theoretical models have shown that selection within the individual may drive one of the lineages to xation (Otto & Orive, 1995; Pineda-Krch & Fagerstro m, 1999; Pineda-Krch & Lehtila , 2002). Withinindividual selection may, however, also maintain a persistent IGH. The maintenance and propagation of genetic mosaicism originating late in development, for example, would be difcult to explain in the absence of a selective advantage at the cell level (Youssouan & Pyeritz, 2002). Numerous theoretical models of cell-lineage selection have been developed making various assumptions regarding the developmental program and types of selective pressures. For example, cell-lineage dynamics in plants has been studied several times (Antolin & Strobeck, 1985; Slatkin, 1985; Otto & Orive, 1995; Pineda-Krch & Fagerstro m, 1999; Orive, 2001; PinedaKrch & Lehtila , 2002). The reader is referred to the excellent review by Otto & Hastings (1998) for further details regarding some of these models. All of these models have in common that explicit selection takes place only at the cell level. Assuming that selection at different levels acts concordantly (Otto & Orive, 1995; Otto & Hastings, 1998) enables the interpretation of these models from multilevel selection scenario. Multilevel selection acting in opposite directions would be costly to both levels. In these cases, cells that proliferate fastest are those that are deleterious at the individual level, e.g. cancerous cells, or alternatively, cells that proliferate slower would benet the individual by cooperating more efciently (Michod & Roze, 2000). Studies of such opposing selective pressures will be instrumental for our understanding of the evolution and maintenance of cellular self-recognition mechanisms and conict mediating systems necessary to maintain the integrity of genetically heterogeneous units (Pineda-Krch & Fagerstro m, 1999; Michod & Roze, 2000). We have little knowledge, however, about the relationship between selection acting at the individual level to selection at the cellular level (Tuomi & Vuorisalo, 1989; Otto & Hastings, 1998) and many questions are waiting to be addressed both theoretically and empirically.

Size-specic processes
Demographic parameters, e.g. fecundity and mortality, are often size-specic in modular organisms rather than age-specic as in unitary organisms (Otto & Orive, 1995; Hall & Hughes, 1996; Pedersen, 1999; Meesters et al., 2001) where the size of the organism often directly affects survival probability and reproductive output (Roff, 1992; Hutchings, 1997; Marfenin, 1997). Coalescence, i.e. the fusion and integration of separate individuals, results in a rapid size increase and can potentially increase the tness of the newly formed individual, e.g. by increasing herbivore or pest resistance, enhancing mechanical stability, increasing resource acquisition (light, nutrients and water) and/or giving organism an edge in competitive interactions with conspecics

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Currently there is a paucity of models considering such multilevel selection (for exceptions, see Pedersen & Tuomi, 1995; Michod & Roze, 2000; Hadany, 2001). Some theoretical works have dealt with synergy among genetically distinct lineages. In general, models of group selection and kin selection (e.g. Queller, 1992; Wilson, 1997) can be applied to the theoretical analysis of IGH. One example is the model of Hudson et al. (2002), who studied the evolutionary dynamics in the reproduction of cellular slime moulds. They suggest that the proliferation of selsh lineages may be counteracted by anticheater adaptations and processes of kin selection, which would maintain the synergistic features of the multicellular structure. The model of Hudson et al. (2002) does not directly analyse the benets of chimeric vs. genetically homogeneous fruiting bodies, whereas the ry, 1989; Maynard stochastic corrector model (Szathma ry, 1997) explicitely deals with the Smith & Szathma maintenance of genetic heterogeneity. The stochastic corrector model was originally devised to explain the origin of compartmentalized templates in a prebiological setting (e.g. RNA molecules in cells) and it assumes synergism among RNA templates conferring advantages to the compartment. The templates are analogous to suborganismal units such as modules or groups of modules while the compartment is equivalent to the organism. Although the templates compete against each other, or one is altruistic and the other is parasitic, both lineages are required for the proper functioning of the compartment. The tness of the compartment (measured as rate of replication) is zero if it is genetically homogeneous and maximal if it contains equal numbers of the two types. Although stochastic, the model shows that the probability distribution of the various compositions is constant and IGH persists. Whether such a stable IGH will occur in modular organism where the component lineages act synergistically remains an open question. When IGH is benecial to the individual the question arises under what conditions can the genetically heterogeneous individual replicate itself rather than its subunits independently, i.e. how can the heritability of a genetically heterogeneous organism be maintained? A requirement for IGH to be passed on to an offspring is that the sequestration of the precursor cells of the next generation is multicellular, in other words, propagation has to be clonal using multicellular propagules (Fig. 1). The actual composition of lineages in a genetically heterogeneous organism (composition is here dened as the number of coexisting genotypes, their proportions, and spatial arrangement) is a complex function of processes taking place at several levels of organization. These include the frequency of mutational events and coalescence formation, details of the ontogeny, intraorganismal selection, stochasticity (Pineda-Krch & Fagerstro m, 1999), spatial constraints, and the complexity of the life histories of the organism. As Gill et al. (1995) assert, the composition will vary through the differential growth and survival of

modules. The composition of multicellular propagules will most likely differ from the composition of the parent, and the sequestration will potentially result in the elimination of one or several genetic lineages (see paths 12 and 14 in Fig. 1). For example, an individual where the lineages are compartmentalized, as when one lineage occurs in the supportive structures while the other in the tissue giving rise to runners, may not be able to transfer its genetic assemblage to its clonally generated offspring. In a stochastically ideal scenario, i.e. in the absence of spatial constraints (e.g. meristem stratication) and where the genetic lineages are well mixed, such as the hypothetical organism in Fig. 1, the composition of the sequestered propagules will have a hypergeometric probability distribution (Pineda-Krch & Fagerstro m, 1999). How can IGH be eliminated when it is detrimental to the individual? Coalescence between unrelated individuals is likely to give rise to severe intraorganismal conicts threatening to eliminate one or both of the component lineages. The obvious way of controlling such conicts is to avoid them in the rst place by restricting fusion to closely related individuals (Maldonado, 1998; Rinkevich & Shapira, 1999) by employing various incompatibility barriers (Stoner et al., 1999), e.g. histocompatibility systems in animals (Rinkevich & Shapira, 1999) and heterokaryon incompatibility in fungi (Wu & Glass, 2001). An alternative strategy is to allow structural fusion but not physiological integration which has been found in trees (Thomson et al., 1991). IGH that arises through genetic change can be maintained or revert to genetic uniformity by a single-celled sequestration (paths 8 and 9 in Fig. 1), intraorganismal selection (Otto & Orive, 1995; Pineda-Krch & Fagerstro m, 1999), intraorganismal structural compartmentalization (e.g. the stratication of apical meristems with a limited exchange between the layers; Pineda-Krch & Lehtila , 2002), restricted cell movement (e.g. the xed cell walls in plants), or by simply shedding parts (Marfenin, 1997).

Future directions
Our hope is that this review of the literature pertaining to IGH and the discussion of the potential costs and benet has set the stage for further studies in this eld. Future studies will increase our understanding of the biology of genetically heterogeneous organisms and the processes underlying their ecology and evolution. Some of the key questions are: 1 Assessing the rates at which useful functional variation arise (Sunnucks et al., 1998; Bataillon, 2000), e.g. rates of somatic mutation and other genetic changes as well as frequency of successful coalescence events similar to the work by Bishop & Sommerfeldt (1999) and Rinkevich & Shapira (1999). 2 Investigate the frequency and pattern of naturally occurring IGH (Slatkin, 1985) such as in the work by Peabody et al. (2000) on the fungi Armillaria gallica and

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Thomson et al. (1991) on gs. The functional relationships between lower-level composition and the phenotype of the individual will be of particular importance to study. Part of this work will involve the development and standardizing of new methods for screening natural populations for IGH. 3 Elucidating attributes of the ontogeny that affects the longevity of the IGH (e.g. meristem structure and sequestration patterns during branching and vegetative propagation). 4 Embracing a multilevel approach to establish the degree of cooperation among heterogeneous lineages required to form a stable phenotype. 5 Further investigations, similar to Pineda-Krch & Fagerstro m (1999), Poore & Fagerstro m (2000) and Pineda-Krch & Lehtila (2002), into the evolutionary potential of persistent mitotic lineages in modular organisms, e.g. the probability of mutant lineages being purged or reaching xation at the cellular level.

References
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Conclusions
The number of studies detecting naturally occurring IGH has recently begun to accumulate suggesting that IGH may be substantially more common than previously believed. The long held paradigm that individuals for obvious reasons must be genetically homogeneous, usually despite a lack of supporting empirical evidence, is breached. The assumption of intraorganismal genetic homogeneity is proximately an issue of genotype resolution dependent upon the resolving power of molecular markers but ultimately it is a scientic simplication for the sake of convenience (Loxdale & Lushai, 2003). When dealing with short-lived, sexually reproducing organisms with an early sequestered germ line, i.e. the majority of model organisms, this simplication is valid. A large proportion of multicellular life does not, however, fall into this category. Although it has been acknowledged that modular organisms are different (Jerling, 1985; Stearns, 1992; Vuorisalo & Mutikainen, 1999) and that IGH in modular organisms will have major implications for many aspects of their ecology and evolution, the eld remains largely unexplored.

Acknowledgments
We are grateful to Torbjo rn Fagerstro m, Sally Otto, Alistair Poore, and Dilara Ally for fruitful discussions and two anonymous referees for comments who substantially improved the manuscript. MP-K was partially nanced by the Royal Physiographical Society, the Wallenberg Foundation, The European Science Foundation, The Swedish Foundation for International Cooperation in Research and Education, and by grants from the Swedish Natural Science Research Council to Torbjo rn Fagerstro m. KL was partially nanced by Swedish Natural Science Research Council.

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