Miscel.lania Zoologica 20.1 (1997) 9

The value of cytogenetics for the taxonomy and evolution of Leaf Beetles (Coleoptera, Chrysornel idae)

E. Petitpierre

Petitpierre, E., 1997. The value of cytogenetics for the taxonomy and evolution of Leaf Beetles (Coleoptera, Chrysomelidae). Misc. Zool., 20.1: 9-18.

The value of cytogenet~csfor the taxonomy and evolut~on of Leaf Beetles (Coleoptera, Chrysomebdae) - The advantages and pitfalls of cytogenetics for the taxonomy and evolution of Leaf Beetles are discussed Karyology may provide clues for distinguishing cryptic sibling species as demonstrated in Chrysol~naaur~chalcea and Casslda v~r~d~sThe phylogenetic value of karyotypes can only be substantiated on the grounds of three general rules extensive sampling to determine the most widespread karyotype or meioformula, the criterion of parsimony, and parallel evolution of other characters A modal karyotype cannot be equalized a prior1 to the most primitive karyotype Hence, for simple effects of sampling, in a few Leaf Beetle subfamilies only the ancestral karyotype can be reasonably assessed The Chrysomelinae subfamily is studied in significantly greater detall, and the probable interrelationships of their higher taxa based on the chromosomal findings and their correlation with other characters of phylogenetic interest is discussed The presumed effects of deme size and specialized phytophagy on the karyological evolution of Chrysomelinae genera are also dealt with

Key words: Cytogenetics, Leaf Beetles, Chrysomelinae, Taxonomy, Evolution.

(Rebut: 10 1 97; Acceptació definitiva: 11 111 97)

Eduard Petltplerre, Lab de Genetlca, Dept de Blolog~aAmb~ental, Unlv de les llles Balears- UIB, 07071 Palma de Mallorca, Espanya (Spa~n)

This work was supported by project DGICYT PB93-0419, Ministry of Education and Culture, Spain.

O 1997 Museu de Zoologia Petitpierre lntroduction species is its karyotype, defined by chromo- some number, size and morphology of each The Chrysomelidae or Leaf Beetles are one of individual chromosorne, and when pre- the largest farnilies of Coleoptera with some sent, such as in most animals, the type of 35,000 described species, and probably a sex-chromosome system. The karyotype of great number as yet undescribed (LAWRENCE,a species is usually constant and can there- 1982; JOLIVET & COX, 1996). This is the most fore be considered as a definite element, irnportant taxon of the superfarnily Chryso- like any morphological character, for taxo- meloidea, where Cerambycidae and Bruchi- nomic purposes. dae are also included among a total of five A simple manner to determine the rough families (CROWSON,1981). karyotype of a beetle species is by the The recent publication of three books rneioformula or karyotypic formula, the dealing with the biology of Chryso- number of autosomal bivalent plus the type melidae, in the broadest sense (JOLIVET of sex-chromosome system, usually obtained et al., 1988, 1994; JOLIVET & COX, 1996), from males, at rneiotic metaphase I (SMITH& and specially several of its chapters dedi- VIRKKI,1978). Thus, a rneioformula of 9+Xyp, cated to the phylogeny and evolution of means nine autosomal bivalent and a sex- the group, has given rise to considerable chromosome bivalent made up of a large X interest in these subjects from a multidis- and a small y, noted by a capital letter and ciplinary view. Two authors specialized in a srnall letter respectively, held together by Leaf Beetles recently questioned the use- a 'parachute' (p) type of association, be- fulness of cytogenetics as a suitable tool cause it resernbles this configuration. Equal- for phylogenetic and evolutionary studies. ity of meioformulas between two species CROWSON(1994) stated that 'chromo- does not mean they have identical karyo- somal patterns do not usually provide reli- types. If this were true the huge nurnber able markers for higher taxa ,... marked of beetles with the modal 9+Xyn rneioformula karyotypic differences may be found be- would be indistinct in theii karyotypes, tween species which in other respects seem which is clearly nonsense. In fact, alrnost al1 closely allied, and very similar karyotypes sisecies can be differentiated in their can persist in extensively varied groups', to karyotypes provided they are surveyed conclude 'the main systematic (and biologi- through a high resolution analysis, narnely cal value) of karyotypic studies is likely to by chromosome banding, in situ hybridiza- be around the level of species, as demon- tion, and a deep study of meiosis. The only strated e.g. by Virkki's studies in Alticinae'. known exceptions to this rule in animals In addition, REID (1995) did not use (just over 1%), are the homosequential spe- karyology among the 71 characters used in cies of Drosophila, whose polytenic chro- his long and detailed cladistic analysis of rnosomes enabled a high resolution analy- Leaf Beetles, because 'karyology tends to sis. They have identical patterns of bands consider common to primitive, but this is and are therefore impossible to distinguish unacceptable for several reasons, including (WHITE,1978). sample bias and failure to distinguish be- Specifically regarding Leaf Beetles, tween an ancestral state frorn one shared by closely related species can differ or not in a recent radiation of species', adding finally their meioformulas, but when they are co- that 'at present levels of study, kariology is incident even a conventional morphometric useless for chrysomelid phylogeny except at analysis of mitotic metaphase chromoso- trivial levels, as already noted by CROWSON mes, which is not an extremely power- (1981)'. Nevertheless the topic is worthy of ful technique, can demonstrate differences, more detailed study and discussion. as shown in two lndian species of Cassida (YADAV& PILLAI, 1975), the allied species of Chrysolina feeding on Labiatae plants The value of karyotype for the taxonomy (PETITPIERRE,1983), and 13 North American of Leaf Beetles Leptinotarsa (HSIAO& HSIAO,1983). Both CROWSON(1994) and REID (1995) One characteristic of the phenotype of a agree in the acceptance of the karyotype Miscel.lania Zoologica 20.1 (1997) as a valuable character to species tax- The karyotype as a phylogenetic character onomy. Nevertheless, and contrary to the latter author, the species level is not at al1 Phylogeny implies recognition of polarity trivial for taxonomy and phylogeny, ei- and the modern and extensive cladistic ther in basic or in applied research, and analyses in Leaf Beetles assume ances- several examples could be quoted to sup- tral (plesiomorphous) and derived (apo- port this view. morphous) states in many characters to Two recent findings illustrate the value build up the most parsimonious trees, of karyotype for taxonomy and phylo- phenograms or cladograms (LEE,1993; REID, genetic interrelationships. The first is rep- 1995). Likewise, cytogenetic data may be resented by the complex of taxa known as used in phylogenetic studies if polarity of Chrysolina aurichalcea from Japan and Korea, change is reasonably inferred. Before dis- whose existence was proved by karyotypic cussing how to answer the question of analysis showing distinct chromosomal races, polarity in evolutionary cytogenetics, sev- with n=32, 2n=42 and 2n=46 diploid num- eral important premises should be clearly bers, that should be rightly consider- settled. First of all, there is not a prior; ed as sibling species (PETITPIERRE,1981; FUJI- reason for assuming that extant species YAMA, 1989). The spermatids of siblings with with primitive morphology, behaviour and n=16 and n=23 chromosomes had no sig- life history should also have the most primi- nificant differences in their DNA content, tive karyotypes. This may occur or not, be- so, the chromosomal shifts involved in cause evolution proceeds in a mosaic way the origin of the latter from the former, with some characters being derived and 0th- according to the common trend found for ers conserved with respect to the ancestral the genus, have not changed this param- state. A correspondence between karyotype eter (PETITPIERREet al., 1991). A second and evolutionary ancestry could be true if example of siblings was found in the tor- the cytogenetic characters determine, at toise beetle Cassida viridis, the spe- least to some degree, the states of the cimens from Switzerland and from Catalo- non-cytogenetic characters, andlor if the nia (NE Spain), dsiplayed 2n=24, while those rates of karyotypic change and those of from two geographical sources in Andalucia other characters are correlated across (S Spain) had 2n=30 chromosomes (PETITPIERREevolving lineages. Consequently, the et al., 1988, in prep.). Whereas the adults cytogenetic characters cannot be taken of Chrysolina aurichalcea siblings show alone in phylogenetic analyses, especially slight differences in male genitalia and eco- in taxonomic categories higher than ge- logical niches (FUJIYAMA,1989; FUJIYAMAet al., nus. They can serve to test phylogenetic 1991; FUJIYAMA & TAKANASHI,1994), those of hypotheses principally based on exo- Cassida viridis siblings are identical but phenotypic characters, that is morphologic their larvae have not as yet been' studied and behavioural traits, and this is their and might provide some clues for distinc- main evolutionary usefulness. tion. But how can we determine the direc- Greater efforts should be directed to tion of karyotypic change between two the karyological characterization of Leaf states A and B: A -> B or B -> A? A Beetle species by using conventional common practice, followed by several evo- techniques of staining to extend the lutionary cytogeneticists, has been to sample analyses, and also high resolu- equalize the modal karyotype to the an- tion techniques, especially in the taxa cestral state. This presumed equality is where conservative chromosome num- obviously false because it is strongly de- bers and meioformulas are prevalent. pendent on the biases and extension Nevertheless, the karyotype is only a part of sampling (STEVENS,1980). Even when of the genetic knowledge needed for an these conditions of unbiased and sufficient accurate definition of a species, along with sampling are met, a modal karyotype (in a the classic exophenotypic characters of wide sense) for a group, does not neces- morphology, life history, ecology and bio- sarily mean the most widespread karyotype geography. among the supraspecific taxa of the group. 12 Petitpierre

If taxon X is constituted, for example, of interrelationships among them is true. five tribes, M, N, O, P and Q, and karyotype There is considerable controversy re- I appears in 60% of the whole sam- garding ancestrallprimitive state as either pled species but is restricted to the tribe commonality or ancestral, as most wide- M, while the karyotype Il appears in only spread. With regard to the subfamilies of 30% of the whole but in each of the five Chrysomelidae, and assuming the neces- tribes, karyotype I is the modal and sary premise of an adequate and repre- karyotype II is the most widespread in the sentative sampling of karyotypes in each higher taxa of X. This difference between subfamily, the most widespread karyotype modal and most widespread, together with and meioformula, not the modal one by the inclusion of a right outgroup and analy- itself, can presumably be taken as the an- sis of parsimony, is crucial to understanding cestral karyotype if it is the most parsi- any presumed polarity in evolutionary cyto- monious. This is also the opinion of genetics. MADDISON(1985) in his large study on the In an overview on the applications of cytogenetics of Bembidion ground Bee- molecular systematics HILLIS& MORITZ(1990) tles (Carabidae). claim that cytogenetics is an appropriate Among the different chromosomally and effective method to infer phylogenies examined subfamilies of Leaf Beetles, the among taxa whose divergences range amount of cytogenetic data is not suffi- from five to 50 million years (rnya), but it cient to ascertain the most widespread is marginally appropriate or appropriate karyotype in Eumolpinae since only four of only under limited circumstances for lower the fourteen tribes (SEENO& WILCOX,1982) time frames of divergence (O to 5 mya) or have been surveyed. In Cryptocephalinae higher (50 to 500 rnya). two of the five tribes (SEENO& WILCOX,1982) Therefore, only closely related species are known, but each for only one genus. diverged within the past five rnya and Their show very different modal numbers, those that diverged since more than 50 Cryptocephalus (Cryptocephalini) 2n = 30, mya, cannot be generally subjected to a and Pachybrachis (Pachybrachini) 2n = 16 cytogenetic analysis for estimating their which precludes any phylogenetic assess- phylogenies. Furthermore, SANTIAGO-BLAYment. This may be due to either the poor (1994) reports that the fossil record for tribal screening or to the striking heteroge- eight extant subfamilies of the Leaf neity between the two previous genera. Beetles goes back to Mesozoic (245 to 66.4 Three other subfamilies, Galerucinae, rnya), and most of them first came in the Alticinae and Cassidinae, are particularly Jurassic period (195 to 135 rnya). Assum- diverse in higher taxonomic groups. Moreo- ing these times of origin for a good ver, in Alticinae, the current tribes are so number of Chrysomelidae subfamilies ei- misleading (SEENO& WILCOX,1982) that ther in Jurassic or in Cretaceous pe- they also prevent any reliable attempt to riods, they fall apart from the range of determine their presumed ancestral karyo- time where cytogenetics is appropriate to types. Nevertheless, a valuable approach to deduce phylogenies. Four subfamilies or the cytotaxonomy, but not the whole tribes of Leaf Beetles share a modal chro- phylogeny of Alticinae, was published by mosome number of 2n = 16, Criocerinae, VIRKKI(1 988). Pachybrachini (Cryptocephalinae), Eumol- On the contrary, two additional sub- pinae and Hispinae (s. str.), although families, Donaciinae and Criocerinae, with in agreement with most phylogenetic pro- fewer species and higher taxa, have been posals there are no grounds to assume suff iciently surveyed (PETITPIERREet al., 1988, any direct phylogenetic relatedness among in prep.) to suggest their presumed them, except maybe for Criocerinae and most widespread karyotypes, 2n = 30 and Hispinae (MANN& CROWSON,1981; REID, 2n = 16, respectively. Finally, the subfamily 1995). Very likely, their common modal Chrysomelinae, the best relatively well- number should be explained as a simple known higher taxon of Leaf Beetles will convergence, at least as far as striking be discussed in greater depth on continu- substantial evidence of allied phylogenetic Miscel.lania Zoologica 20.1 (1997) 13

The phylogenetic significance of karyotype Chrysomelina is n = 17 (48% of species). in Chrysomelinae he- modal value for the whole Chryso- melinae, n = 12, cannot be assumed as Of the roughly 3,000 described species the ancestral value because it appears in (DACCORDI,1996). the meioformulas andlor only one species of Timarchini, the most karyotypes of Chrysomelinae have been de- primitive taxon of the subfamily in adult termined for 203 species (PETITPIERREet al. and larval morphology, life history and be- 1988; PETITPIERRE& JUAN, 1994; PETITPIERRE,in haviour, and does not occur in Ento- prep.), which corresponds to 6.8% of the moscelina. total group. Although the genus screening On the contrary, Chrysomelina, the most is not very large, in 30 of the 132 present evolved subtribe in larval morphology, life genera (23%) (DACCORDI,1994, 1996) most history and behaviour (PATERSON,1931; current tribes and subtribes have been KIMOTO,1962; TAKIZAWA,1976), has the high- checked in a rather good sampling. On the est modal number, n = 17, among the five other hand, a general agreement has been subtribes of Chrysomelinae, which would reached about the monophyly of Chryso- probably imply the most advanced phylo- melinae (REID,1995; DACCORDI,1996). a genetic position. In summary, n = 10 is the favourable condition to perform their most widespread chromosome number in phylogenetic analysis. Timarchini, and in the Cerambycidae, the SEENO & WILCOX(1982) followed the divi- right outgroup of Leaf Beetles, and due sion of the subfamily in two tribes, also to the correspondence with the remar- Timarchini and Chrysomelini. The latter was kable primitiveness of the Timarcha, it can further split into twelve subtribes, but the reasonably be taken as the ancestral value present taxonomic views based on recon- for the whole tribe Timarchini. However, sideration of adult characters and, pricipally, as this number of n=10 has further been on a much better knowledge of the larval found in two species of Chrysolinina, one and pupal morphology, have reduced their Cosmogramma from South America and one number to only five subtribes: Timarchina Oreina from Europe, it might also be the (Timarchini), Entomoscelina, Paropsina, ancestral number for Chrysolinina too. Nev- Chrysolinina and Chrysomelina (DACCORDI, ertheless in accordance with the parsimony 1994, 1996). However, STEINHAUSEN (1996) rules and assuming n = 12 as the ancestral goes even further in joining Entomoscelina number for Chrysolinina, it is much easier with Paropsina (=Gonioctenina), based on to explain the origin of the previous n = 10 their larval and pupal characteristics, which chromosome species by one centric fusion does not seem substantiated until a much in each, rather than the reverse, that is, the larger screening of immature stages in gen- derivation of 59 species with n = 12 chro- era of Entomoscelina becomes available. mosomes from those by a clearly higher The diversity of haploid chromosome figure of centric fissions. The same reason- numbers for these five subtribes of Chryso- ing could be applied in trying to ascertain melinae is given in table 1. The modal the presumed ancestral number for Chry- number for the subfamily is n = 12 chromo- somelina, with n = 17 chromosomes the somes, a value found in 84 (41.3%) of the most widespread, and in agreement with total 203 checked species. This is the most their derived morphological and beha- frequent number in Chrysolinina (51.7% of vioural features, it can be rightly taken as species) and almost the only number dis- the ancestral karyotypic value for the played in Paropsina (96% of species), but it subtribe. is very scarcely represented in Timarchina (3% of species), or in Chrysomelina (8% of Phenotypic correlates of karyotypes in species), and has not been observed to date Chrysomelinae in Entomoscelina.The Timarchina have n=10 as modal value (57% of taxa), the Ento- The larval morphology of Chrysomeli- moscelina have a mode at n=13 (83% nae offers excellent characters to set up of species) although the number of sam- groups of species more clearly differenti- pled species is still scanty, while that of ated than their corresponding adult stages. 14 Petitpierre

Table 1. Distribution of haploid chromosome numbers among higher taxa of Chrysomelinae Leaf Beetles: T. Timarchini; Chrl. Chrysolinina (including six polyploid parthenotes of Calligrapha); P. Paropsina; E. Entomoscelina; Chrm. Chrysomelina; gen. Genera; sp. Species Distribución de los números de cromosomas haploides en los grandes taxones de Chrysomelinae. (Para abreviaturas ver arriba.)

Examined Haploid numbers gen. sp 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

T 135-- -1202 1541------1 .- Chrl 12 114 1 - - - 2 359 16 13 515 210 3 - 2 1

P 6 23 ------22 ------1.- --

E 3 6 ------51 -- ---.----

Chrm 8 25 ------2 -24512 --.----

Total 30 203 1 - - 122 5 841113 6 8 1715 211 3 1 2 1

Three large groups of larvae are generally (PETITPIERRE,1995). The three main types recognized by several authors (PATERSON, of larvae and their defensive allomones 193 1; HENNIG,1938; KIMOTO,1962; TAKIZAWA, correspond with the three basic modal 1976; Cox, 1982): 1. Non-tuberculate larvae karyotypes of Timarchini (n = lo), Chryso- (Timarchini); 2. Tuberculate but 'non- linina and Gonioctenina (n = 12), and glanduliferous' larvae, having only one pair Chrysomelina (n = 17). Another interest- of defensive glands (Chrysolinina, Entomos- ing insight of cytotaxonomic value comes celina and Paropsina, including the former from genome size, since the species of Gonioctenina); 3. Tuberculate and glandu- Chrysomelina have significantly smaller liferous larvae, with nine pairs of glands genomes than those of the other subtribes, (Chrysomelina, including the former Phyllo- very likely an apomorphic condition of these dectina). These three types of larvae are most advanced chrysomelines (PETITPIERRE& rather well correlated with the recent re- JUAN, 1994). sults of chemotaxonomy which have been All these features of phylogenetic value discussed and summarized by PASTEELS& have also been validated by the analyses ROWELL-RAHIER(1989) and PASTEELS(1993). of nucleotide sequences of mitochondrial Thus, in the subtribe Chrysomelina, al1 the DNA (HSIAO, 1994a, 1994b). This has al- studied genera and species, except for one lowed some phylogenetic trees to be built which is misclassified, secrete nitropro- up, where the genera of Chrysomelina panoic acid and isoxazolinone glucosides, clearly branched off from those of while Chrysolinina-Doryphorina-Gonioc- Chrysolinina-Doryphorina. Nevertheless, in tenina produce cardenolides, ethanolamine the second and more extensive paper or amino acid derivatives, with the excep- (HSIAO, 1994b), only one species of tion of the Chrysolina subgenus Hypericia, Timarchini was included as an out- which is characterized by secreting polyo- group, and the few checked Paropsina (3 xygenated steroid glycosides as well as spp.) and Entornoscelina (1 sp.), were not ethanolamine. Finally, the ancestral tribe clearly clustered. A rnuch larger screen- Timarchini is strikingly separated from ing of these three higher taxa is therefore the previous taxa by its probable secretion necessary for a more conclusive picture of of anthraquinones as defensive substances their genetic relatedness. . - Miscel.lania Zooloqica 20.1 (1997)

Some clues to explain the karyological reduced in the latter. Furthermore, the diversity of Leaf Beetles oligophagous or monophagous versus polyphagous feeding selection is very likely All chromosomal shifts which may be related to the deme size, being much larger detected either inter- or intraspecifica- in the latter. One very interesting study on Ily depend on mutation as a primary factor. insect numbers has demonstrated increased This means that any new mutant is opportunities for speciation in the specializ- heterozygous for such a chromosomal ed phytophagous insects respect to the non- change, and assuming that this mutation phytophagous insect taxa (MITTERet al., does not negatively affect the survival of 1988). In our particular frame of Chrysome- the mutant individual, it can only be kept linae Leaf Beetles, we chose twelve genera and spread if it becomes fixed. Hence it with a minimum number of four chromo- becomes a homozygous condition. Several somally examined species to test the previ- factors can account for the fixation of those ous hypotheses. A significant correlation chromosomal shifts without deleterious was found between the rate of chromo- effects causing their elimination by natural somal evolution, given as standard devia- selection. BENGTSSON(1980) has detailed the tion of diploid numbers, and the species five most important models leading to richness per genus (r = 0.53). A much higher fixation of chromosome mutations: 1. Seg- correlation was obtained between the rate regation distortion (or meiotic drive in a of chromosomal evolution per genus and more frequently used name); 2. Selective the number of host plant families selected advantage; 3. Recombination modification; by these congeneric species (r = 0.84), but 4. lnbreeding and homozygote advantage; there were no clear-cut differences in the 5. Random genetic drift. Among these five chromosomal evolution between flying and models the last two have attracted more flightless species genera (PETITPIERREet al., interest and attention than the others. 1993). Therefore, the oligophagous or CHESSER& BAKER (1986) have developed monophagous feeding preferences increase computer simulation models to determine the chance for chromosomal evolution and the conditions for the stochastic fixation of speciation, because the conditions sug- chromosomal mutations in small isolated gested by CHESSER& BAKER (1986) are prob- demes. These models predict fixation un- ably fulfilled, although the interrelation- der the conditions of: a. A small number ship between chrornosomal evolution and of initial founders (5 or 10); b. A relatively speciation is a matter of debate, and indeed small reduction in fecundity due to meiotic not necessarily univocal. Furthermore, if ex- problems; and c. A high number of off- amples of meiotic drive in Leaf Beetles are spring per mating. Moreover, LANDE(1985) reported in the next years, we will estimated the capacity of fixation of a new have more valuable information to under- chromosomal change through colonization stand their karyological evolution and the of further demes by the mutant individuals. processes leading to fixation of new These conditions can be applied to the chromosomal variants. Leaf Beetles as an approach to understand- ing their chromosomal evolution. Effec- tive population size is a crucial factor in Resumen order to explain thefixation of chromo- somal changes due to inbreeding and El valor de la citogenética para la taxo- genetic drift. Although direct measures of nomía y la evolucíón de los crisornélidos population sizes in Leaf Beetles are lacking, (Coleoptera, Chrysornelidae) some indirect features may provide valuable information in this regard. The fly- Se discuten en detalle las ventajas e incon- ing capacity versus flightlessness is a char- venientes de la citogenética para la taxono- acter with a presumed immediate effect on mía y estudio de la evolución de los criso- the dispersa1 potentiality of a species. In a mélidos. La cariologia puede suministrar broad sense, the local population or deme medios para distinguir especies gemelas size is probably enlarged in the former and cripticas como se demuestra en Chrysolina aurichalcea y Cassida viridis. El valor pects of the biology of Chrysomelidae: filogenético de los cariotipos solo puede XIX-XXIII (P. H. Jolivet, M. L. Cox & E. invocarse en base a tres reglas generales: un Petitpierre, Eds.). Kluwer Acad. Publ., amplio muestre0 para conocer el cariotipo Dordrecht (Netherlands). o meiofórmula más extendida, el criterio de DACCORDI,M., 1994. Notes for phylogenetic parsimonia, y la evolución paralela en study of Chrysomelinae, with descrip- otros caracteres. Desde luego, un cariotipo tions on new taxa and a list of al1 the modal no puede ser considerado a priori known genera (Coleoptera: Chrysomeli- como el más primitivo, portanto, por simples dae, Chrysomelinae). In: Proceedings razones de muestreo, sólo puede reconocer- of the Third lnternational Symposium on se el cariotipo ancestral en unas pocas the Chrysomelidae, Beijing 1992: 60-84 (D. subfamilias. Entre ellas los Donaciinae, con G. Furth, Ed.). Backhuys Publ., Leiden 2n = 30 (Xy ) y los Criocerinae, con 2n = (Netherlands). 16 (Xy ), parLcen estar razonablemente es- - 1996. Notes on the distribution of tableAdos, pero no los Cryptocepha- the Chrysomelinae and their possible ori- linae, Eumolpinae, Galerucinae, Alticinae, gin. In: Chrysomelidae Biology, vol. Hispinae y Cassidinae, porque aunque se 1: 399-412 (P. H. A. Jolivet & M. L. Cox, han estudiado bastantes especies de todas Eds.), SPB Academic Publ., Amsterdam. estas subfamilias, en todas ellas quedan FUJIYAMA, S., 1989. Species problems in todavía muchas tribus o grupos de géneros, Chrysolina aurichalcea (Mannerheim) como sucede en los Alticinae, sobre las cua- with special reference to chromosome les no hay datos cromosómicos. La sub- numbers. Entomography, 6: 443-452. familia Chrysomelinae, con casi un 7% de FUJIYAMA,S., SASAWAKI, A. & NAGAO,M. 1991. sus 3000 especies caracterizadas cromoso- Geographic distribution of the two types micamente, permite un análisis más profun- of chromosome number and frequency of do, para discutir las posibles relaciones de the colour forms in Chrysolina sus tribus y subtribus basadas en los datos aurichalcea complex in Nagano Prefec- citogenéticos y las claras correspondencias ture, central Honshu, Japan, with special observadas con otros caracteres de va- reference to natural environment. Ann. lor filogenético. También se tratan los Environ. Sci., Shinshu Univ., 13: 21-31. probables efectos del tamaño de las pobla- FUJIYAMA,S. & TAKANASHI,H., 1994. 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