Cytogenetic studies of three Lycosidae species from Argentina (Arachnida, Araneae)

ABSTRACT

GENETICS

Cytogenetic studies of three Lycosidae species from Argentina (Arachnida, Araneae)

María A. Chemisquy¹

Sergio G. Rodríguez Gil²

Cristina L. Scioscia³

Liliana M. Mola^2,4

¹Instituto de Botánica Darwinion, San Isidro, Buenos Aires, Argentina

²Laboratorio de Citogenética y Evolución, Departamento de Ecología, Genética y Evolución, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina

³División Aracnología, Museo Argentino de Ciencias Naturales "Bernardino Rivadavia", Ciudad Autónoma de Buenos Aires, Argentina

⁴Consejo Nacional de Investigaciones Científicas y Técnicas, Ciudad Autónoma de Buenos Aires, Argentina

Cytogenetic studies of the family Lycosidae (Arachnida: Araneae) are scarce. Less than 4% of the described species have been analyzed and the male haploid chromosome numbers ranged from 8+X₁ X ₂ to 13+X₁ X ₂. Species formerly classified as Lycosa were the most studied ones. Our aim in this work was to perform a comparative analysis of the meiosis in "Lycosa" erythrognatha Lucas, "Lycosa" pampeana Holmberg and Schizocosa malitiosa (Tullgren). We also compared male and female karyotypes and characterized the heterochromatin of "L." erythrognatha. The males of the three species had 2n = 22, n = 10+X₁X₂, all the chromosomes were telocentric and there was generally a single chiasma per bivalent. In "Lycosa" pampeana, which is described cytogenetically for the first time herein, the bivalents and sex chromosomes showed a clustered arrangement at prometaphase I. The comparison of the male/female karyotypes (2n = 22/24) of "Lycosa" erythrognatha revealed that the sex chromosomes were the largest of the complement and that the autosomes decreased gradually in size. The analysis of the amount, composition and distribution of heterochromatin with C-banding and staining with DAPI- and CMA₃ - showed that "Lycosa" erythrognatha had little GC-rich heterochromatin in the pericentromeric region of all chromosomes. In addition, the actual occurrence of the genus Lycosa in the Southern Hemisphere is discussed.

Cytogenetic studies of the family Lycosidae (Arachnida, Araneae) are scarce and were performed in less than 4% of the 2324 known species (Platnick, 2008). Most of the analyzed species had only telocentric or acrocentric chromosomes, which ranged from 2n = 18, n = 8+X₁X₂ (male) to 2n = 28/30 (male/female), n = 13+X₁X₂ (male). The 2n = 28/30, which is present in 50% of the analyzed species, is probably the modal diploid number of the family. The sex chromosome mechanism X₁X₂/X₁X₁X₂X₂ (male/female) occurs in 94% of the lycosid species and is considered as an ancestral trait in spiders. The derived systems are: X0 in Lycosabarnesi, L. nordenskjoldi, Wadicosa quadrifera and an unidentified species of Schizocosa (Schizocosa sp. 2 in Table 1); X₁X₂X₃ in an unidentified species of Lycosa (Lycosa sp. 8 in Table 1), and a neo-XY system with multiple X chromosomes in Pardosa morosa (^{Král, 2004}). Most cytological studies have been performed in species formerly classified as Lycosa and the most common male haploid chromosome number was n = 13+X₁X₂ in the Northern Hemisphere species. The Southern Hemisphere species presented complements with n = 10+X₁X₂ (males) or derived from it (Table 1).

The content, distribution and composition of the constitutive heterochromatin in spiders have been poorly analyzed (^{Brum-Zorrilla and Cazenave, 1974}; ^{Brum-Zorrilla and Postiglioni, 1980}; ^{Rowell, 1985}; ^{Datta and Chatterjee, 1988}; ^{Rowell, 1991}; ^{Gorlova et al., 1997}; ^{Silva et al., 2002}; ^{Araujo et al., 2005a}, 2005b; Rodríguez Gil et al., 2007). The first characterization of heterochromatin in spiders was performed in Schizocosa malitiosa using C-banding. All chromosomes showed small pericentromeric heterochromatic bands in this species and in all the other Lycosidae analyzed (^{Brum-Zorrilla and Cazenave, 1974}; ^{Brum-Zorrilla and Postiglioni, 1980}; ^{Gorlova et al., 1997}).

Morphological and molecular phylogenetic studies questioned the taxonomic position of a number of species currently placed in Lycosa (sensu lato) (including Lycosa erythrognatha), since they appear not to be closely affiliated with Lycosa tarantula (Linnaeus 1758), the type species of the genus (^{Murphy et al., 2006}; ^{Álvares and Brescovit, 2007}). In view of this uncertainty, we named the species with its original combination, but with the generic name inside inverted commas.

In this work we analyzed and compared the meiotic behavior of “Lycosa” erythrognatha Lucas 1836, “Lycosa” pampeana Holmberg 1876 and Schizocosamalitiosa (Tullgren 1905). We used C-banding, DAPI- and CMA₃-staining to analyze the male and female karyotypes and the amount, composition and distribution of the heterochromatin in “L.” erythrognatha. We compared our results with those reported for other lycosid species. A literature review of the cytogenetics Lycosidae and a discussion on the actual occurrence of the genus Lycosa in Southern Hemisphere are also included.

Figure 2 Meiosis in “Lycosa” erythrognatha (2n = 22, n = 10+X1X2 and n = 10): (a) spermatogonial prometaphase; (b) zygotene; (c) pachytene; (d) early diplotene; (e) late diplotene with two univalents (arrows) and a bivalent with two chiasmata (arrowhead); (f) diakinesis; (g) metaphase I; (h) anaphase I; (i) telophase I; (j) metaphase II with sex chromosomes; (k) anaphase II with sex chromosomes; (l) telophase II without sex chromosomes. The arrowheads point to the sex chromosomes. Bar = 10 μm.

Figure 3 Testicular cells of “Lycosa” erythrognatha (2n = 22 = 20+X1X2) after staining with CMA3 (a, c, e, f) and with DAPI (b, d): (a-b) spermatogonial prometaphase; (c-d) pachytene; (e) diakinesis; (f) metaphase I. The arrowheads point to the sex chromosomes; the V points to CMA3 bright bands and the asterisk marks the DAPI dull regions. Bar = 10 μm.

Figure 4 Meiosis of “Lycosa” pampeana male (n = 10+X1X2 and n = 10): (a) pachytene; (b) diplotene; (c) diakinesis; (d-f) prometaphase I; (g) telophase I; (h) prometaphase II with sex chromosomes; (i) telophase II without sex chromosomes. The arrowheads point to the sex chromosomes. Bar = 10 μm.

Figure 5 Meiosis in Schizocosa malitiosa (2n = 22, n = 10+X1X2 and n = 10): (a) spermatogonial prometaphase; (b) pachytene; (c) diplotene with a bivalent with two chiasmata; (d) diakinesis; (e) metaphase I; (f) anaphase I; (g) telophase I; (h) metaphase II with sex chromosomes; (i) metaphase II without sex chromosomes; (j) telophase II with sex chromosomes. The arrowheads point to the sex chromosomes. Bar = 10 μm.

We analyzed 21 males and nine females of “Lycosa” erythrognatha from Buenos Aires City and surroundings (34°48' S - 58°41' W) (17 males, five females), Martín García Island Natural Preserve (34°18' S - 58°24' W, Buenos Aires Province) (two males), Magdalena (35°08' S - 57°51' W, Buenos Aires Province) (one female), Parque Nacional “El Palmar” (24°08' S - 64°58' W, Entre Ríos Province) (one female), Malargüe (35°48' S - 69°59' W, Mendoza Province) (one male, two females) and Posadas (27°40' S - 55°93' W, Misiones Province) (one male); four males of “Lycosa” pampeana from Buenos Aires City and surroundings; and seven males of Schizocosa malitiosa from Martín García Island Natural Preserve (one male), Gualeguaychú (33°04' S - 58°43' W, Entre Ríos Province) (three males, one subadult male), San Juan Poriahú (27°61' S - 56°98' W, Corrientes Province) (one male) and Embalse de Río Tercero (32°17' S - 64°25' W, Córdoba Province) (one male).

Adult males and females were collected in the field and reared at the Arachnology Division of the Museo Argentino de Ciencias Naturales “Bernardino Rivadavia” (MACN). Voucher specimens were deposited in the National Collection of Arachnology (MACN-Ar, Cristina Scioscia).

For male meiotic analyses, testes were dissected out and kept in 3:1 ethanol:acetic acid at 4 °C. Preparations were obtained by squashing in iron propionic haematoxylin.

Fluorescent staining with 4'-6-diamidino-2-phenylindole (DAPI) and chromomycin A₃ (CMA₃) was carried out on unstained chromosomes. After squashing a piece of testis in 45% acetic acid, the coverslip was removed with the dry-ice method and slides were air-dried. The sequential DAPI-CMA₃ staining was performed according to ^{Rebagliati et al. (2003)}.

For the mitotic analysis specimens of “Lycosa” erythrognatha were injected with 0.1 mL of a 0.01% colchicine solution. After 1.25 h, several drops of haemolymph were removed from the coxal joints and the gonads together with some digestive tissues were dissected. Each sample was suspended in 2 mL of hypotonic solution (KCl 0.56%) for 15 min, centrifuged at 800 rpm for 5 min, and fixed in 1 mL of 3:1 ethanol:acetic acid. The cell suspension was dropped onto clean slides, air-dried and stained with Giemsa for chromosome counting and karyotyping. C-banding was carried out according to ^{Sumner (1972)}.

Chromosome measurements were performed in twelve well-spread mitotic metaphases using the MicroMeasure version 3.3 software (Reeves and Tear, 2000). The total haploid complement length (TCL) in females was calculated by adding the mean value of each chromosome pair (in arbitrary units). In males, the TCL was calculated after the analysis of the relative length of all chromosomes, which was used to identify those having no homologues (sex chromosomes). The male and female idiograms were drawn based on the length of each chromosome pair in relation to the TCL. Chromosomes were also measured with a vernier caliper in order to estimate the TCL in microns.

The diploid number in somatic cells was 22 in males and 24 in females. All the chromosomes were telocentric (Figures ^{1 a and b}). The X₁ and X₂ were the largest chromosomes of the complement, with 12.83% and 11.69% of the TCL, respectively, whereas the autosomes decreased gradually in size, with the largest and smallest pairs representing 9.54% and 5.47% of the TCL, respectively (Figures ^{1 a, b and d}). The total haploid complement length (TCL) was 43.8 μm, the sex chromosomes were 5.03 + 0.04 μm and 4.78 + 0.1 μm long and the autosomes ranged between 4.13 + 0.25 μm and 2.64 + 0.09 μm. C-banding revealed the presence of small positive bands in the pericentromeric region of all chromosomes (^{Figure 1c}).

“Lycosa” erythrognatha (2n = 22, n = 10+X₁X₂ and n = 10)

At spermatogonial prometaphases and metaphases the sex chromosomes and the autosomes were isopycnotic (Figure 2a). At prophase I, up to pachytene, the sex chromosomes were positively heteropycnotic and closely associated (Figures 2 b and c). The sex chromosomes were usually isopycnotic from diakinesis onwards, but appeared negatively heteropycnotic in some cells; in both cases they remained associated and differed in size (Figure 2d-f). Bivalents had a single proximal or interstitial chiasma (Figure 2d-g; 3e-f), but two chiasmata could occasionally be observed in one of the largest bivalents (Figure 2e). In six individuals, a pair of medium-sized autosomal univalents were seen in a low frequency at diakinesis (less than 10% of the cells) and two univalent pairs were seen in a single cell (Figure 2e). The sex chromosomes were located apart from the bivalents at metaphase I (Figures 2g; 3f) and precociously migrated together towards the same pole at anaphase I (Figure 2h-i). This resulted in two types of metaphase II, one with ten autosomes and the other with ten autosomes plus the X₁X₂ chromosomes (Figure 2j). The sister chromatids of each sex chromosome were always closely associated, whereas the autosomal chromatids were only associated by the centromeric region (Figure 2j). The sex chromosomes and autosomes migrated simultaneously and were positioned slightly apart at anaphase II (Figure 2k) resulting in cells with ten autosomes (Figure 2l) and with ten chromosomes plus X₁X₂ in telophase II. An atypical meiosis was observed in some cells of all the males (unpublished data).

Sequential DAPI-CMA₃-staining of spermatogonial prometaphases and metaphases revealed that all the pericentromeric C-positive bands were bright after CMA₃-staining and showed no differential fluorescence with DAPI (Figure 3a-b). The sex chromosomes were brightly fluorescent after DAPI- and CMA₃-staining at early prophase I. The CMA₃-bright bands observed in mitotic chromosomes were composed of several smaller CMA₃-fluorescent bands at pachytene. The same bands were generally dull after DAPI-staining (Figure 3c-d). A single pericentromeric CMA₃-bright band could be observed in the autosomes and sex chromosomes from diplotene onwards (Figure 3e-f).

“Lycosa” pampeana (2n = 22, n = 10+X₁X₂, n = 10)

The sex chromosomes were positively heteropycnotic and closely associated at early prophase I (Figure 4a) and turned isopycnotic at diakinesis, when they remained associated and showed different sizes. All autosomal bivalents presented a single proximal or distal chiasma and bivalents with two chiasmata were never found (Figure 4b-c). Bivalents adopted a particular disposition during prometaphase I, with some of them (from one to four) lining up on the equatorial plate and the others located near the poles. The sex chromosomes were either in the cell equator or at one pole (Figure 4d-f). The bivalents and sex chromosomes lined up on the equatorial plane at metaphase I and the sex chromosomes migrated together to the same pole at anaphase I (Figure 4g). Two types of prometaphases II and metaphases II could be distinguished, one with ten and the other with 12 chromosomes (Figure 4h). Chromosomes with a telocentric morphology were clearly seen at anaphase I and II (Figures 4 g and i).

Schizocosa malitiosa (2n = 22, n = 10+X₁X₂, n = 10)

Twenty-two isopycnotic chromosomes were seen in spermatogonial prometaphases and metaphases (Figure 5a). The sex chromosomes were closely associated and positively heteropycnotic at early prophase I (Figure 5b) and turned isopycnotic from diakinesis onwards, when they were close to each other and showed different sizes (Figure 5c-d). Most of the bivalents had a single interstitial or distal chiasma, and less frequently a proximal one, as could be seen at metaphase I (Figure 5e). Some cells also presented one bivalent with two distal chismata (Figure 5c). The sex chromosomes were not lined up on the equatorial plate at metaphase I, but closer to one pole (Figure 5e), and they migrated together to the same pole at anaphase I (Figure 5f). The sex chromosomes remained condensed and positively heteropycnotic at prophase II (Figure 5g) and were similar in size to the largest autosomes at metaphase II. The sister chromatids of the X₁ and X₂ chromosomes were always associated, whereas autosomal chromatids were only associated by their centromeric region (Figure 5h-i). The chromosomes of this species were also telocentric (Figure 5h-j).

Only ten species of Lycosidae from South America have been cytogenetically studied. Two of them belonged to the genus Schizocosa Chamberlin 1904 and the remaining eight to the genus Lycosa Latreille 1804 (Table 1). They were collected in Uruguay, Brazil and Argentina.

In this work, we found a 2n = 22 (20+X₁X₂, male) in Schizocosamalitiosa. This species had a karyotype with all telocentric chromosomes, with the sex chromosomes being the largest of the complement and chiasmata mainly at interstitial or distal positions. In populations of S. malitiosa from Uruguay, ^{Brum-Zorrilla and Cazenave (1974)} and ^{Brum-Zorrilla and Postiglioni (1980)} described 2n = 22 in males and 2n = 24 in females, telocentric chromosomes, sex chromosomes that were the smallest of the complement and bivalents with proximal chiasmata in the males. These results suggest that S. malitiosa is polytypic for the size of the sex chromosomes and chiasma position. In the males of an unidentified Schizocosa species (Schizocosa sp. 2 in Table 1) included within the “malitiosa group”, ^{Postiglioni and Brum-Zorrilla (1981)} found 2n = 23 (22+X), with a metacentric X chromosome probably resulting from the fusion of two telocentric X chromosomes. The five species from the USA already studied (S. communis, S. crassipes, S. ocreata, S. rovneri, S. stridulans) also had n = 10+X₁X₂, whereas an unidentified species from India (Schizocosa sp. 1 in Table 1) had n = 13+X₁X₂ (^{Painter, 1914}; ^{Hard, 1939}; ^{Mittal, 1960}, 1963; ^{Stratton, 1997}). These results allowed us to conclude that the modal chromosome number for the genus is 2n = 22/24 (male/female) and that the sex chromosome determination system is of the X₁X₂/ X₁X₁X₂X₂ type.

The present work represents the first cytogenetic study conducted in “Lycosa” pampeana. This species had 2n = 22 (20+X₁X₂, male), with all telocentric chromosomes. At prometaphase I, the bivalents and the sex chromosomes were peculiarly arranged into three groups, one group being located on the equatorial plane and the remaining two at the cell poles. This very unusual chromosome disposition in spiders was formerly described for three species of Tegenaria (Agelenidae) by ^{Revell (1947)}, who considered that it resulted from the primary polarization of the bivalents at early prophase I due to the attraction of the heterochromatic regions by the centrosome. When the centrosome began to split, each new centrosome acted as a polarization centre for bivalents, which gradually became aligned on the metaphase plate. This stage, named “transitional metaphase” by ^{Revell (1947)}, resembles the prometaphases observed in “Lycosa” pampeana.

“Lycosa” erythrognatha had 2n = 22/24 (male/female), n = 10+X₁X₂, n = 10 in males, all telocentric chromosomes and the sex chromosomes were the largest of the complement. Bivalents usually had one chiasma, although bivalents with two chiasmata and univalents were occasionally seen and probably resulted from desynapsis. The number and location of the chiasmata during meiosis varied largely among cells. Such variation may be determined not only genetically, but also by environmental factors, both internal and external to the individual (^{John and Lewis, 1965}; ^{Jones, 1987}; ^{Appels et al., 1998}). The presence of univalents and bivalents with two chiasmata in the same individual and even in the same cell may be a consequence of changes in the mechanisms regulating chiasma frequency and distribution. The chromosome number, some karyotypic features and the meiotic behavior herein observed in “Lycosa” erythrognatha are consistent with results obtained in specimens of the same species from Uruguay and Brazil (Díaz and Sáez, ^1966a, ^1966b; ^{Giroti et al., 2007}).

“Lycosa” erythrognatha is characterized by scanty GC-rich heterochromatin located in the pericentromeric region of all chromosomes. The DAPI- and CMA₃-bright fluorescence of the sex chromosomes during early prophase I is consistent with the allocycly of these chromosomes during male meiosis and probably reflects different degrees of chromatin condensation rather than differences in base composition. The heterochromatin content has only been characterized in other four species of the family. C-banding of Alopecosa albofasciata showed small blocks of pericentromeric heterochromatin in the autosomes and uniformly heterochromatic sex chromosomes during male meiosis (^{Gorlova et al., 1997}). In Schizocosa malitiosa, Lycosa thorelli and in an unidentified species of Lycosa (Lycosa sp. 7 in Table 1), small AT-rich (Hoechst 33258 positive) C-positive bands were observed in the pericentromeric regions of all chromosomes. A few Hoechst-positive fluorescent bands found in the telomeric regions of some chromosomes of Lycosa sp. 7 (Table 1) were not C-positive (^{Brum-Zorrilla and Postiglioni, 1980}). In the male meiosis of Schizocosa malitiosa, the sex chromosomes were strongly positively heteropycnotic (^{Brum-Zorrilla and Cazenave, 1974}). The analysis of the heterochromatin content revealed that all the studied species of Lycosidae are characterized by a small amount of tandem repeated DNA sequences. On the other hand, there is some heterogeneity in heterochromatin composition, which can be summarized as follows: a) C-positive, AT-rich pericentromeric heterochromatin; b) C-positive, GC-rich pericentromeric heterochromatin, and c) AT-rich telomeric heterochromatin, undetectable with C-banding.

In a molecular phylogenetic reconstruction of the wolf spiders at the subfamily level, ^{Murphy et al. (2006)} stated that a number of species currently placed in Lycosa (sensu lato) (including “Lycosa” erythrognatha) do not form a clade and none of them appear to be closely affiliated with Lycosa tarantula (Linnaeus 1758), the type species of the genus; in contrast, “clades within the Lycosinae appear to reflect geographic regions rather than existing recognised morphological parameters”. The authors claimed that Australasian wolf spiders do not possess the defining features of Lycosa and that a critical study is necessary to determine their true taxonomic position.

The systematics of South American Lycosidae is poorly known and the taxonomic status of many of the species is far from being resolved. Most of the species originally described under the genus Lycosa were transferred to other genera on the basis of poor diagnostic morphological characters. Our karyological study supports previous morphological and behavioral evidence (unpublished data, two thesis works and congress presentations by several authors) indicating that “Lycosa” erythrognatha, “Lycosa” pampeana and a group of species from South America should be transferred to another genus (e.g.Schizocosa) or that they would belong to a new, still undescribed genus. ^{Álvares and Brescovit (2007)}, based exclusively on morphological characters, proposed that these species, as well as Schizocosa malitiosa, should be transferred to Hogna Simon 1885.

It is outstanding that all the South American species so far analyzed had 2n = 22/24 (male/female) or complements almost certainly derived from it. Even though the number of cytogenetically analyzed Schizocosa and Hogna species is not representative for the group, all the Schizocosa species from the USA already studied presented n = 10+X₁X₂, while Hogna species from the USA had n = 13+X₁X₂ and n = 11+X₁X₂ (Table 1).

Cytogenetic analyses of other South American species currently classified within Lycosinae are needed. The information obtained will provide baseline data on the karyotypic evolution within each genus. It will be particularly relevant to formulate a new revision of the taxonomy and phylogeny of the group taking into account morphological, cytogenetical and molecular data. This revision could confirm, as was the case for Australasian wolf spiders (^{Murphy et al., 2006}), that Lycosa does not occur in South America.

Acknowledgments

This study was supported by grants from the Buenos Aires University (UBA) to Dr L. Poggio and Dr L. M. Mola (Ex 317) and from the National Council of Scientific and Technological Research (CONICET) (PIP 5927 Poggio-Mola), and (PIP 5654 González-Scioscia). The authors thank Mr. Hernán Dinapoli for technical assistance and Prof. Gustavo Gagna for offering his house for specimen collections.

References

Álvares E.E.S., Brescovit A.D. (2007) The Lycosinae wolf spiders from Brazil with notes on species occurring in neighboring countries (Araneae, Lycosidae). 17th International Congress of Arachnology Abstract 64:São Pedro, SPhttp://www.ib.usp.br/~ricrocha/ISA17/CONGRESSOCOMPLETO.pdf

Araujo D., Brescovit A.D., Rheims C.A., Cella D.M. (2005a) Chromosomal data of two pholcids (Araneae, Haplogynae): A new diploid number and the first cytogenetical record for the new world clade. J Arachnol 33:591-596.

Araujo D., Cella D.M., Brescovit A.D. (2005b) Cytogenetic analysis of the neotropical spider Nephilengys cruentata (Araneomorphae, Tetragnathidae): Standard staining, NORs, C-bands and base-specific fluorochromes. Braz J Biol 65:193-202.

Bole-Gowda B.N. (1953) Chromosome study of fifteen species of Indian spiders. Proc 40 Indian Sci Congr, Lucknow 3:179-180.

Bole-Gowda B.N. (1958) A study of the chromosomes during meiosis in twenty-two species of Indian spiders. Proc Zool Soc Bengal 11:69-108.

Brum-Zorrilla N., Cazenave A.M. (1974) Heterochromatin localization in the chromosomes of Lycosa malitiosa (Arachnida). Experientia 30:94-95.

Brum-Zorrilla N., Postiglioni A. (1980) Karyological studies from Uruguayan spiders. I. Banding pattern in chromosomes of Lycosa species (Araneae-Lycosidae). Genetica 54:149-153.

Datta S.N., Chatterjee K. (1988) Chromosome and sex determination in 13 araneid spiders of North-Eastern India. Genetica 76:91-99.

Datta S.N., Chatterjee K. (1989) Study of meiotic chromosomes of four hunting spiders of north eastern India. Perspect Cytol Genet 6:417-424.

Giroti A.M., Araujo D., Oliveira E.G., Brescovit A.D., Cella D.M. (2007) Cytogenetics of some true Lycosoid spiders (Araneomorphae):Chromosomes of two Lycosa species (Lycosidae)and possible occurrence of B-chromosomes in Trechalea sp. (Trechaleidae). 17th International Congress of Arachnology Abstract 245:São Pedro, SP http://www.ib.usp.br/~ricrocha/ISA17/CONGRESSOCOMPLETO.pdf

Gorlov I.P., Gorlova O.Y., Logunov D.V. (1995) Cytogenetic studies on Siberian spiders. Hereditas 122:211-220.

Gorlova O.Y., Gorlov I.P., Nevo E., Logunov D.V. (1997) Cytogenetic studies on seventeen spiders species from Israel. Bull Br Arachnol Soc 10:249-252.

Hackman W. (1948) Chromosomenstudien an Araneen mit besonderer Berücksichtigung der Geschlechtschromosomen. Acta Zool Fennica 54:1-101.

Hard W.L. (1939) The spermatogenesis on the lycosid spider Schizocosa crassipes (Walckenaer). J Morph 65:121-154.

John B., Lewis K.R. (1965) The meiotic system. Protoplasmatologia 6:1-335.

Jones G.H. (1987) Chiasmata. MeiosisMoens P.B.New YorkAcademic Press213-244.

Kageyama A., Seto T., Inoue H. (1978) Chromosomes of Japanese lycosid spiders. Chromosome Information Service 25:26-27.

Král J. (2004) Evolution of the neo-sex chromosome system in spiders: Karyotype analysis of Tegenaria ferruginea (Agelenidae)and Pardosa morosa (Lycosidae). 16th International Congress of Arachnology Abstract 91:Ghent, Belgium http://users.ugent.be/~jpmaelfa/Abstracts%20Lezingen%20(all).pdf

Matsumoto S. (1977) An observation of somatic chromosomes from spider embryo-cells. Acta Arachnol 27:167-172.

Mittal O.P. (1960) Chromosome number and sex mechanism in twenty species of the Indian spiders. Res Bull Panjab Univ ns Sci 11:245-247.

Mittal O.P. (1961) Chromosome number and sex mechanism in twenty-one species of the India spiders. Res Bull Panjab Univ ns Sci 12:71-273.

Mital O. P. (1962) An analysis of the chromosome complement infive of the Indian spiders belonging to the subfamily Lycosinae. Proc 49 Indian Sci Congr Abstracts Part III section VII:349-350.

Mittal O.P. (1963) Karyological studies on the Indian spiders. I. A comparative study of the chromosomes and sex determinating mechanism in the family Lycosidae. Res Bull Panjab Univ ns Sci 14:59-86.

Montgomery T.H. (1905) The spermatogenesis of Syrbula and Lycosa, with general consideration upon chromosome reduction and the heterochromosomes. Proc Acad Nat Sci Phil 57:162-205.

Murphy N.P., Framenau V.W., Donnellan S.C., Harvey M.S., Park Y.C., Austin A.D. (2006) Phylogenetic reconstruction of the wolf spiders (Araneae, Lycosidae) using sequences from the 12S rRNA, 28S rRNA, and NADH1 genes: Implications for classification, biogeography, and the evolution of web building behaviour. Mol Phylogenet Evol 38:583-602.

Painter S. (1914) Spermatogenesis in spiders. Zool Jahrb Abt Anat Ontog Tiere 38:1-101.

Parida B.B., Sharma G.P. (1987a) Cytological studies on Indian spiders. I. Meiosis in three species of wolf spiders (Lycosidae, Arachnida). Caryologia 40:89-97.

Parida B.B., Sharma G.P. (1987b) Chromosome number, sex mechanism and genome size in twenty seven species of Indian spiders. Chromosome Information Service 43:11-13.

Parida B.B., Mohanty P.K., Sahoo P., Mohapatra A. (1986) Studies on spermatocytic chromosomes of an acuatic wolf spider Hippasa madhuae Tikader and Malhotra (Lycosidae, Araneae). Curr Sci 55:997-998.

Postiglioni A., Brum-Zorrilla N. (1981) Karyological studies on Uruguayan spiders. II. Sex chromosomes in spiders of the genus Lycosa (Araneae-Lycosidae). Genetica 56:47-53.

Rebagliati P.J., Papeschi A.G., Mola L.M. (2003) Meiosis and fluorescent banding in Edessa meditabunda and Edessa rufomarginata (Heteroptera, Pentatomidae, Edessinae). Eur J Entomol 100:11-18.

Revell S.H. (1947) Controlled X-segregation in Tegenaria. Heredity 1:337-347.

Rodríguez Gil S.G., Merani M.S., Scioscia C.L., Mola L.M. (2007) Cytogenetics in three species of Polybetes Simon 1897 from Argentina (Araneae, Sparassidae) I. Karyotype and chromosome banding pattern. J Arachnol 35:227-237.

Rowell D.M. (1985) Complex sex-linked translocation heterozygosity and its role in the evolution of social behaviour. J Genet Cytol 28:168-170.

Rowell D.M. (1991) Chromosomal fusion and meiotic behaviour in Delena cancerides (Araneae, Sparassidae). II. Chiasma position and its implications for speciation. Genome 34:567-573.

Sharma G.P. (1961) A study on the chromosomes of two lycosid spiders. Proc Zool Soc Calcuta 14:33-38.

Sharma G.P., Gupta B.L. (1956) Cytological studies on the male germ cells of the spider, Pardosa sp. , with observations under the phase contrast microscope. Res Bull Panjab Univ ns Sci 84:5-19.

Sharma G.P., Jande M.S., Tandon K.K. (1959) Cytological studies on Indian spiders. IV. Chromosome complement and meiosis in Selenops radiatus Latr. (Selenopidae) and Leucage decorata (Blackw.) (Tetragnathidae), with special reference to XXXO-type of male sex-determining mechanisms. Res Bull Panjab Univ ns Sci 10:73-80.

Sharma G.P., Jande M.S., Grewal M.S., Chopra R.N. (1958) Cytological sudies on the Indian spiders. II. Chromosome complement and male meiosis in seven species of the family Lycosidae. Res Bull Panjab Univ 156:255-269.

Sharma N., Parida B.B. (1987) Study of chromosomes in spiders from Orissa. Pranikee 8:71-76.

Silva R.W., Klisiowicz D.D.R., Cella D.M., Mangili O.C., Sbalqueiro I.J. (2002) Differential distribution of constitutive heterochromatin in two species of brown spider: Loxosceles intermedia and L. laeta (Araneae, Sicariidae), from the metropolitan region of Curitiba, PR, Brazil. Acta Biol Par Curitiba 31:123-136.

Sokolov I.I. (1960) Studies on nuclear structures in Araneina. I. Karyological peculiarities in spermatogenesis. Voprosy Cytologii i Protistologii160-186.

Srivastava S.C., Shukla S. (1986) Chromosome number and sex determining mechanism in forty-seven species of Indian spiders. Chromosome Information Service 41:23-26.

Stratton G.E. (1997) Investigation of species divergence and reproductive isolation of Schizocosa stridulans (Araneae, Lycosidae) from Illinois. Bull Br Arachnol Soc 10:313-321.

Sumner A.T. (1972) A simple technique for demostrating centromeric heterochromatin. Exp Cell Res 75:304-305.

Suzuki S. (1954) Cytological studies in Spiders. III. Studies on the chromosomes of fifty-seven species of spiders belonging to seventeen families, with general considerations on chromosomal evolution. J Sci Hiroshima Univ, B. 1 15:24-150.

Tugmon C.R., Brown J.D., Horner N.V. (1990) Karyotypes of seventeen USA spider species (Araneae, Araneidae, Gnaphosidae, Loxoscelidae, Lycosidae, Oxyopidae, Philodromidae, Salticidae and Theridiidae). J Arachnol 18:41-48.

Wise D.A. (1983) An electron microscope study of the karyotypes of two wolf spiders. Can J Genet Cytol 25:161-168.

Wise D.A. (1984) The ultraestructure of an intraspindle membrane system in meiosis of spider spermatocytes. Chromosoma 90:50-56.

Wise D.A., Shaw R.G. (1984) The mechanism of non-random chromosome segregation in lycosid spiders. J Cell Biol 99:246

Internet Resources

Platnick N.I.The World Spider Catalog, v. 8.5. American Museum of Natural History, online (2008) Available from: http://research.amnh.org/entomology/spiders/catalog/index.html

Reeves A., Tear J.Micromeasure for Windows (2000) Available from: http://www.colostate.edu/Depts/Biology/Micromeasure.[3.3]

Table 1
- Chromosome data of Lycosidae species.

Specie		n (male)	Origin	References
Allocosa georgicola (Walckenaer 1837)	28	13+X1X2	USA	Wise, 1983; Wise, 1984 (sub Lycosa georgicola); Wise and Shaw, 1984 (sub Lycosa georgicola)
Alopecosa aculeata (Clerck 1757)		13+X1X2	Finland	Hackman, 1948 (sub Tarentula aculeata)
A. albofasciata (Brullé 1832)		13+X1X2	Israel	Gorlova et al., 1997
A. pulverulenta (Clerck 1757)		13+X1X2	Finland	Hackman, 1948 (sub Tarentula pulverulenta)
Anomalomma sp.	28	13+X1X2	India	Sharma et al., 1959
Anomalosa harishi (Dyal 1935)	28	13+X1X2	India	Mittal, 1961; Mittal, 1963 (sub Anomalomma harishi)
Arctosa alpigena (Doleschall 1852)	26	12+X1X2	Finland	Hackman, 1948
A. leopardus (Sundevall 1833)	26	12+X1X2	Finland	Hackman, 1948
Arctosa sp.	28 30 f	13+X1X2	India	Mittal, 1960, 1963
Crocodilosa leucostigma (Simon 1885)	28	13+X1X2	India	Srivastava and Shukla, 1986 (sub Lycosa leucostigma)
Evippa praelongipes (O.P.-Cambridge 1870)	26	12+X1X2	India Israel	Sharma et al., 1958; Gorlova et al., 1997
Gladicosa pulchra (Keyserling 1877)	28	13+X1X2	USA	Montgomery, 1905 (sub Lycosa insopita); Gowan, 1985 (sub Lycosa insopita) (1)
Hippasa agelenoides (Simon 1884)	28 24	13+X1X2 11+X1X2	India	Bole-Gowda, 1953; Sharma et al., 1958
H. madhuae Tikader and Malhotra 1980	28	13+X1X2	India	Parida et al., 1986
H. olivacea (Thorell 1887)	28	13+X1X2	India	Parida and Sharma, 1987a; Parida and Sharma, 1987b; Sharma and Parida, 1987
H. pisaurina Pocock 1900	26 28	12+X1X2 13+X1X2	India India	Mittal 1960, 1963 Srivastava and Shukla, 1986
Hippasa sp.	22	10+X1X2	India	Parida and Sharma, 1987b; Sharma and Parida, 1987
Hogna ammophila (Wallace 1942)		11+X1X2	USA	Gowan, 1985 (sub Lycosa ammophila)(1)
H. helluo (Walckneaer 1837)		13+X1X2	USA	Gowan, 1985 (sub Lycosa helluo)(1)
H. himalayensis (Gravely 1924)	28	13+X1X2	India	Mittal, 1962, 1963 (sub Lycosa himalayensis)
H. lenta (Hentz 1844)		11+X1X2	USA	Gowan, 1985 (sub Lycosa lenta)(1)
Hygrolycosa rubrofasciata (Ohlert 1865)	20 22 f		Russia India	Gorlov et al., 1995
Lycosa barnesi Gravely 1924	27	13+X	India	Srivastava and Shukla, 1986
L. bistriata Gravely 1924	28	13+X1X2	India	Bole-Gowda, 1953; Bole-Gowda, 1958
L. carmichaeli Gravely 1924	28 22	13+X1X2 10+X1X2	India	Mittal, 1961, 1963 Srivastava and Shukla, 1986
L. cf. praegrandis C. L. Koch 1878		10+X1X2	Israel	Gorlova et al., 1997 (sub Lycosa cf. nordmanni)
L. chaperi Simon 1885	22	10+X1X2	India	Mittal, 1962, 1963
L. coelestis L. Koch 1878	26	12+X1X2	Japan	Suzuki, 1954
“Lycosa” erythrognatha Lucas 1836	22 24 f	10+X1X2	Uruguay Brazil India	Díaz and Sáez, 1966a(2), 1966b(2); Giroti et al., 2007 This work
L. madani Pocock 1901	24	11+X1X2	India	Mittal, 1962, 1963
L. nigrotibialis Simon 1884	28 24	13+X1X2 11+X1X2	India	Mittal, 1961, 1963 Srivastava and Shukla, 1986
L. nordenskjoldi Tullgren 1905		9+X	Uruguay	Díaz and Sáez, 1966a, 1966b (sub Lycosa nordenskölii)
“Lycosa” pampeana Holmberg 1876		10+X1X2	Argentina	This work
L. sericovittata Mello-Leitao 1939	22	10+X1X2	Brazil	Giroti et al., 2007
L. thorelli (Keyserling 1877)	22	10+X1X2	Uruguay	Brum-Zorrilla and Postiglioni, 1980; Postiglioni and Brum-Zorrilla, 1981
Lycosa sp. 1	28	13+X1X2	India	Bole-Gowda, 1953, 1958
Lycosa sp. 2	28	13+X1X2	Japan	Suzuki, 1954
Lycosa sp. 3	28	13+X1X2	India	Sharma et al., 1958
Lycosa sp. 4		13+X1X2		Sokolov, 1960
Lycosa sp. 5		13+X1X2	India	Mittal, 1961 (sub Lycosa sp. nov.)
Lycosa sp. 6	28	13+X1X2	India	Mittal, 1962, 1963
Lycosa sp. 7	22 24 f	10+X1X2	Uruguay	Brum-Zorrilla and Postiglioni, 1980
Lycosa sp. 8	23	10+X1X2X3	Uruguay	Postiglioni and Brum-Zorrilla, 1981 (sub Lycosa sp.2 (thorelli group))
Lycosa sp. 9	22	10+X1X2	Uruguay	Postiglioni and Brum-Zorrilla, 1981 (sub Lycosa sp.3)
Lycosa sp. 10	18	8+X1X2	India	Srivastava and Shukla, 1986 (sub Lycosa sp.I)
Lycosa sp. 11	28	13+X1X2	India	Srivastava and Shukla, 1986 (sub Lycosa sp.II)
Lycosa sp. 12	22	10+X1X2	India	Sharma and Parida, 1987; Parida and Sharma, 1987a; Parida and Sharma, 1987b (sub Lycosa sp.1)
Lycosa sp. 13	28	13+X1X2	India	Parida and Sharma, 1987b, Sharma and Parida, 1987 (sub Lycosa sp.2)
Margonia himalayensis (Gravely 1924)	28	13+X1X2	India	Mittal, 1961, 1963 (sub Venonia himalayensis)
Ocyale kumari Dyal 1935	28	10+X1X2	India	Sharma et al., 1958
Pardosa agrestis (Westring 1861)	28	13+X1X2	Russia	Gorlov et al., 1995
P. agricola (Thorell 1856)	28	13+X1X2 15(MII) h	Finland	Hackman, 1948 (sub Lycosafluviatilis)
P. amentata (Clerck 1757)	28	13+X1X2	Finland	Hackman, 1948 (sub Lycosapaludicola, sub Lycosasaccata); Sokolov, 1960
P. astrigera L. Koch 1878	28 30 f	13+X1X2	Japan	Suzuki, 1954 (sub Lycosacinereofusca, sub LycosaT-insignita); Matsumoto, 1977 (sub PardosaT-insignita); Kageyama et al., 1978
P. basiri (Dyal 1935)	22	10+X1X2	India	Mittal, 1960, 1963
P. birmanica Simon 1884	28	13+X1X2	India	Bole-Gowda, 1953, 1958 (sub Lycosabirmanica); Srivastava and Shukla, 1986 (sub Lycosabirmanica); Parida and Sharma, 1987a, 1987b; Sharma and Parida, 1987; Datta and Chatterjee, 1989
P. fletcheri (Gravely 1924)	28	13+X1X2	India	Srivastava and Shukla, 1986 (sub Lycosafletcheri)
P. lahorensis Dyal 1935	28	13+X1X2	India	Sharma et al., 1958
P. laura Karsch 1879	28 30 f	13+X1X2	Japan	Kageyama et al., 1978
P. leucopalpis Gravely 1924	28 24	13+X1X2 11+X1X2	India India	Bole-Gowda, 1953, 1958; Mittal, 1963, Srivastava and Shukla, 1986
P. lugubris (Walckenaer 1802)	28	13+X1X2	Russia	Gorlov et al.,1995
P. monticola (Clerck 1757)	28	13+X1X2	Finland	Hackman, 1948 (sub Lycosamonticola)
P. mulani (Dyal 1935)	28	13+X1X2	India	Sharma et al., 1958
P. oakleyi Gravely 1924	26	12+X1X2	India	Srivastava and Shukla, 1986
P. palustris (Linnaeus 1758)	24 28	12+X1X2 13+X1X2	Finland Russia	Hackman, 1948 (sub Lycosatarsalis) Gorlov et al., 1995
P. plumipes (Thorell 1875)	28	13+X1X2	Russia	Gorlov et al., 1995
P. pseudoannulata (Bösenberg and Strand 1906)	28 30 f	13+X1X2	Japan India	Suzuki, 1954 (sub Lycosapseudoannulatus); Kageyama et al., 1978 (sub Lycosapseudoannulata) Bole-Gowda, 1953, 1958 (sub Lycosaannandalei); Srivastava and Shukla, 1986 (sub Lycosaannandalei)
P. pullata (Clerck 1757)	28	13+X1X2	Finland	Hackman, 1948 (sub Lycosapullata)
P. sumatrana (Thorell 1890)	24	11+X1X2	India	Sharma, 1961 (sub Lycosasumatrana); Srivastava and Shukla, 1986 (sub Lycosasumatrana)
Pardosa sp. 1		13+X1X2	India	Bole-Gowda, 1953, 1958
Pardosa sp. 2	28	13+X1X2	India	Sharma and Gupta, 1956
Pardosa sp. 3	28	13+X1X2	India	Mittal, 1960
Pirata latitans (Blackwall 1841)	24	11+X1X2	India	Mittal, 1962, 1963
P. piraticus (Clerck 1757)	26	12+X1X2	Finland	Hackman, 1948
P. procurvus (Bösenberg and Strand 1906)	26 28 f	12+X1X2	Japan	Kageyama et al., 1978
P. subpiraticus (Bösenberg and Strand 1906)	26 28 f	12+X1X2	Japan	Kageyama et al., 1978
P. uliginosus (Thorell 1856)	24	11+X1X2	Finland	Hackman, 1948
Pirata sp.		12+X1X2		Sokolov, 1960
Rabidosa punctulata (Hentz 1844)		13+X1X2	USA	Gowan, 1985(1)
R. rabida (Walckenaer 1837)	28 30 f	13+X1X2	USA	Wise, 1983, 1984 (sub Lycosarabida); Wise and Shaw, 1984 (sub Lycosarabida); Tugmon et al., 1990 (sub Lycosarabida)
Schizocosa communis (Emerton 1885)	22	10+X1X2	USA	Painter, 1914 (sub Lycosacommunis)
S. crassipes (Walckenaer 1837)	22	10+X1X2	USA	Hard, 1939
S. malitiosa (Tullgren 1905)	22 24 f	10+X1X2	Uruguay Argentina	Brum-Zorrilla and Cazenave, 1974; Brum-Zorrilla and Postiglioni, 1980 (sub Lycosamalitiosa) This work
S. ocreata (Hentz 1844)	22	10+X1X2	USA	Stratton, 1997
S. rovneri Uetz and Dondale 1979		10+X1X2	USA	Stratton, 1997
S. stridulans Stratton 1984		10+X1X2	USA	Stratton, 1997
Schizocosa sp. 1	28	13+X1X2	India	Mittal, 1960, 1963
Schizocosa sp. 2	23	11+X	Uruguay	Postiglioni and Brum-Zorrilla, 1981 (sub Lycosa sp.1 (malitiosa group))
Trochosa punctipes (Gravely 1924)	28	13+X1X2	India	Sharma, 1961 (sub Lycosapunctipes)
T. ruricola (De Geer 1778)	26 28 f	12+X1X2 14(MI)	Finland	Hackman, 1948
T. spinipalpis (F.O.P.-Cambridge 1895)	26	12+X1X2	Finland	Hackman, 1948
Venonia sp.	26	12+X1X2	India	Mittal, 1963
Wadicosa quadrifera (Gravely 1924)	27	13+X	India	Srivastava and Shukla, 1986 (sub Lycosaquadrifer)
Xerolycosa miniata (C.L. Koch 1834)	22	10+X1X2	Finland Russia	Hackman, 1948 Gorlov et al., 1995
X. nemoralis (Westring 1861)	26 22	12+X1X2	Finland Russia	Hackman, 1948 Gorlov et al., 1995

⁽¹⁾Gowan TD (1985). The life history and reproduction of the wolf spider, Lycosa lenta Hentz. PhD. Thesis. University of Florida. 259 pp. ⁽²⁾Díaz and Sáez (1966b) handwrote on each reprint of their paper that the spiders they originally classified as two separate species having n = 10+X₁X_2, actually belonged to “Lycosa” erythrognatha.

Received: January 25, 2008; Accepted: May 8, 2008

Liliana María Mola. Laboratorio de Citogenética y Evolución, Departamento de Ecología, Genética y Evolución, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Güiraldes y Costanera Norte, C1428EHA, Ciudad Autónoma de Buenos Aires, Argentina. E-mail: [e-mail].

Appels et al. , 1998

Introduction

Figure 1 “Lycosa” erythrognatha chromosomes (2n = 22): male (a) and female (b) karyograms; C-banded male metaphase (c); relative chromosome sizes in the male (d). Bar = 10 μm.

Materials and Methods

Results

Karyotype of “Lycosa” erythrognatha

Male meiosis

Discussion

Díaz M., Sáez F.A. (1966a) Investigaciones citogenéticas sobre algunas especies de araneidos uruguayos. Anales II Congreso Latinoamericano de Zoología 1:São Paulo3-9.

Díaz M., Sáez F.A. (1966b) Karyotypes of South America Araneida. Mem Inst Butantan 33:153-154.

Tables

2

n

Associate Editor: Yatiyo Yonenaga-Yassuda

Publication Dates

• Publication in this collection
  19 Nov 2008
• Date of issue
  2008

History

• Received
  2501
• Accepted
  8052