Chromosome number evolution in skippers (Lepidoptera, Hesperiidae).

Lepidoptera (butterflies and moths), as many other groups of animals and plants, simultaneously represent preservation of ancestral karyotype in the majority of families with a high degree of chromosome number instability in numerous independently evolved phylogenetic lineages. However, the pattern and trends of karyotype evolution in some Lepidoptera families are poorly studied. Here I provide a survey of chromosome numbers in skippers (family Hesperiidae) based on intensive search and analysis of published data. I demonstrate that the majority of skippers preserve the haploid chromosome number n=31 that seems to be an ancestral number for the Hesperiidae and the order Lepidoptera at whole. However, in the tribe Baorini the derived number n=16 is the most typical state which can be used as a (syn)apomorphic character in further phylogenetic investigations. Several groups of skippers display extreme chromosome number variations on within-species (e.g. the representatives of the genus Carcharodus Hübner, [1819]) and between-species (e.g. the genus Agathymus Freeman, 1959) levels. Thus, these groups can be used as model systems for future analysis of the phenomenon of chromosome instability. Interspecific chromosomal differences are also shown to be useful for discovering and describing new cryptic species of Hesperiidae representing in such a way a powerful tool in biodiversity research. Generally, the skipper butterflies promise to be an exciting group that will significantly contribute to the growing knowledge of patterns and processes of chromosome evolution.

Introduction

The main karyotypic features of organisms, particularly the number of chromosomes, tend to be stable within species (White 1973, King 1993). New chromosomal rearrangements usually originate as heterozygotes and are often – although not always (Lukhtanov et al. 2011) – associated with heterozygote disadvantage. The spread of such rearrangements to fixation within a large population has low probability (King 1993). Therefore, many organisms are characterized by chromosomal conservatism, a situation in which all closely related taxa demonstrate the same chromosome number.

In contrast to chromosomal conservatism, chromosomal instability characterizes situations where multiple closely related taxa (populations, subspecies and/or species) belonging to a single phylogenetic lineage differ drastically from each other by major chromosomal rearrangements, sometimes resulting in high variability in chromosome number.

Both phenomena - chromosomal conservatism and chromosomal instability - are clearly expressed in insects of the order (butterflies and moths). The modal haploid number of chromosomes (n) of n = 31 or n = 30 (Suomalainen 1969, Lukhtanov 2000) is preserved in the majority of lepidopteran families (Robinson 1971). At the same time, numerous cases of chromosomal instability have been discovered in the butterfly families, e.g. in (Emmel et al. 1995), (Lukhtanov 1991, Lukhtanov et al. 2011, Dinca et al. 2011), (Brown et al. 1992, 2004, 2007a, 2007b) and (Brown et al. 2012). This phenomenon was analyzed in more detail in the family (Kandul et al. 2004, 2007, Lukhtanov et al. 2005, 2006, 2008, Vershinina and Lukhtanov 2010, 2013, Vila et al. 2010, Talavera et al. 2013, Przybyłowicz et al. 2014).

Skippers (the family ) are studied to a lesser extent with the respect of karyotype evolution than the other butterfly families mentioned above (but see: Emmel and Trew 1973, Saura et al. 2013). This family includes about 4000 species under 567 genera and is a globally distributed group found in all continents except Antarctica (Warren et al. 2008). The tribal level classification of skippers, based on combined analysis of molecular and morphological data, was recently elaborated by Warren and colleagues (Warren et al. 2008, 2009).

Here I provide a first world-wide survey of chromosome numbers in skippers based on intensive search and analysis of published data.

Results

The results of literature search are presented in the Table below. It includes all the discovered chromosome counts except n=13 for PageBreak (Bremer et Grey, 1853), noted by Bigger (1960) as “”. The name was long used for the species of Europe, but it actually refers to its Far Eastern sister species, and the European taxon is now called (Esper, 1777) (ICZN 2000). Both European and Far Eastern species have the same chromosome number n=29 (Federley 1938, Lorković 1941, Abe et al. 2006), not n=13 as indicated by Bigger (1960). Thus, the species name used by Bigger (1960) was probably misidentification.

The classification of skippers accepted in this paper follows Warren and colleagues (Warren et al. 2008, 2009).

Discussion

Modal chromosomal numbers

The table gives the chromosome numbers of 205 species of skippers, i.e. about 5% of the species of the world fauna. This number is not enough to infer any final statements about peculiarities of chromosome numbers distribution within the . However, several tentative conclusions can be made. The haploid chromosome number n=31 was found in 50 studied species of skippers and, thus, it is a clear modal number for the family at whole. Interestingly, n=31 was found in representatives of all investigated subfamilies, except for . However, in the last subfamily only one species was karyologically studied until now, and discovery of n=31 in is not excluded in future. The next most common numbers are n=29 (43 species), n=30 (33 species) and n=28 (13 species).

Subfamilies and have a sharp peak at n=31. In the subfamily n=31 was also found (only one species studied).

Within the subfamily , the modal number n=31 is found in the tribe . The tribe is characterized by the most common n=28. The modal number in the tribe is n=30. The tribe has peaks at n=30 and n=31. In the tribe , n=29, n=30 and n=31 were found as the most common numbers.

In the family n=29 was found (only one species studied).

Within the subfamily , the tribes , , , and are characterized by the most common n=29. Very variable chromosome numbers (from n=5 to n=50) were found in the tribe Aeromachini. It is difficult to infer the modal number for the last tribe. However, it should be noted that one species, , has n=31 as the majority of other skippers. The tribe (subfamily ) has a clear peak at n=16, so it is exceptional with respect to the modal number of chromosomes.

The overall evidence indicates that chromosome numbers of , , , and conform to the lepidopteran modal of n=31 (Robinson 1971). This number seems to be an ancestral one for the as for the order at whole (Suomalainen 1969, Lukhtanov 2000). This modal number (or its deviation to n=30, n=29 and 28) were preserved in the majority of skippers. However, in the tribe the number n=16 was evolved and, thus, represents a derived trait which can be used as a (syn)apomorphic character in further phylogenetic studies of the family .

Between- and within-species variations in chromosome number

Several groups of skippers display extreme chromosome number variations at the within-species level (Table). The most extreme variations in number of chromosome elements were observed in first meiotic metaphase of , and (Table, de Lesse 1960). The nature of these variations remains unknown, and there are two plausible explanations for this phenomenon. First, this variation can be explained by presence of so-called B-chromosomes (=additional chromosomes, =supernumerary chromosomes) (de Lesse 1960). B-chromosomes consist mainly of repetitive DNA and can sometimes accumulate through processes of mitotic or meiotic drive (Jones et al. 2008). B-chromosomes can be distinguished from normal A-chromosomes because they are usually smaller and can be seen as additional chromosomes present in only some of the individuals in a population (Camacho et al. 2000, Jones et al. 2008).

Second, this kind of variation can be caused by violations in meiotic chromosome pairing resulting in appearance of univalents (instead of bivalents) in meiotic prophase (Lorković 1990). This type of variation was studied in detail by Maeki and Ae (1979) in butterfly genus and is expected if regular or irregular interspecific mating occurs in nature. Anyway, the nature of intraspecific variations observed in is different from that discovered in the Wood White butterfly (Linnaeus, 1758) (). In the last species the compared range of intraspecific variation in chromosome number (from n=28 to n=53) was caused by multiple chromosome fusions/fissions accumulated within the species (Lukhtanov et al. 2011, Dinca et al. 2011).

Between-species variation exists in numerous genera of skippers (Table 1) and is especially expressed in the Nearctic genus Freeman, 1959, in which the range of haploid numbers was discovered from n =5 in to n=38 in (Freeman 1969). This range is comparable of even exceeds the range found in chromosomally diverse genera from other butterfly families (Lorković 1990, Lukhtanov et al. 2005, Talavera et al. 2013). Thus, the genera of can be used as model systems for future analysis of the phenomenon of chromosome instability.

Detecting cryptic species using analysis of chromosomal differences

Recent years karyological data have been widely used in studies of butterfly taxonomy and in biodiversity research as main or additional chracters for detecting cryptic species (e.g. Dinca et al. 2011) and for synonymizing biological entities that were incorrectly described as distinct species (e.g. Vila et al. 2010). The family is not excluded in this respect. In the genus Hübner, [1819], two cryptic species (n=24) and (n=41) were discovered through extensive chromosome analysis of different populations (de Lesse 1960, 1967b, Larsen 1982, Saitoh 1984).

In the genus Hübner, [1819], our unpublished chromosome data (see Table) were used to recognize and then to describe two morphologically similar species, and (De Prins and van der Poorten 1995).

Thus, interspecific chromosomal differences are useful for discovering and describing new cryptic species of representing in such a way a powerful tool in biodiversity research.

Table 1.

Chromosome number of skippers (, ) of the world fauna (Us are univalents; 2n is diploid chromosome number).
Years of the species descriptions are given square brackets in cases where they were not stated in the original sources but were inferred from reliable external evidence.

#	Species	Haploid chromosome number	Country	Reference
Subfamily Coeliadinae
1	Bibasis aquilina (Speyer, 1879)	29	Japan	Maeki 1953
	Bibasis aquilina chrysaeglia (Butler, 1881)	31 (2n=62)	Japan	Abe et al. 2006
2	Bibasis jaina formosana Fruhstorfer, 1911	31	Taiwan	Maeki and Ae 1968b
3	Choaspes benjaminii (Guérin-Méneville, 1843)	31	Japan	Maeki 1953
	Choaspes benjaminii japonica (Murray, 1875)	31	Japan	Saitoh et al. 1978
4	Coeliades anchises jucunda (Butler, 1881)	30	Oman	Saitoh 1982
5	Coeliades ernesti (Grandidier, 1867)	31	Madagascar	de Lesse 1972
6	Coeliades fervida (Butler, 1880)	23	Madagascar	de Lesse 1972
7	Coeliades forestan arbogastes (Guenee, 1863)	31	Madagascar	de Lesse 1972
8	Coeliades ramanatek (Boisduval, 1833)	31	Madagascar	de Lesse 1972
Subfamily Euschemoninae no chromosomal data available
Subfamily Eudaminae
9	Achalarus casica (Herrich-Schäffer, 1869)	29	USA (Texas)	Emmel and Trew 1973
10	Achalarus lyciades (Geyer, 1832)	31	USA (Connecticut)	Maeki 1961
11	Achalarus toxeus (Plötz, 1882)	16	Mexico	Maeki and Remington 1960
12	Astraptes anaphus (Godman et Salvin, 1896)	31	Bolivia	de Lesse 1967a
13	Astraptes fulgerator (Walch, 1775)	31	Peru	Kumagai et al. 2010
14	Astraptes naxos (Hewitson, 1867)	31	Brazil	Saura et al. 2013
15	Astraptes phalaecus (Godman et Salvin, 1893)	25	Guatemala	de Lesse 1967a
16	Astraptes longipennis (Plötz, 1882)	31	Costa Rica	Kumagai et al. 2010
		31	Peru	Kumagai et al. 2010
		31	Brazil	Kumagai et al. 2010
17	Autochton sp.	20, 21	Brazil	Kumagai et al. 2010
18	Chioides albofasciatus (Hewitson, 1867)	31	Mexico	de Lesse 1970a
	Chioides albofasciatus (Hewitson, 1867) (as Chioides catillus)	31	Mexico	Maeki and Remington 1960
	Chioides albofasciatus (Hewitson, 1867)	31	USA (Texas)	Emmel and Trew 1973
19	Entheus priassus pralina Evans, 1952	22	Brazil	Saura et al. 2013
20	Epargyreus barisses (Hewitson, 1874)	31	Argentina	de Lesse 1967
21	Epargyreus clarus (Cramer, 1775)	31	USA (Florida)	Maeki 1961
22	Epargyreus clavicornis tenda Evans, 1955	ca 29–30	Guatemala	de Lesse 1970a
23	Oechydrus chersis (Herrich-Schäffer, 1869)	31	Bolivia	de Lesse 1967a
24	Phocides polybius phanias (Burmeister, 1880)	16	Brazil	Saura et al. 2013
25	Tarsoctenus praecia plutia (Hewitson, 1857)	15	Brazil	Saura et al. 2013
26	Thorybes pylades pylades (Scudder, 1870)	31	USA (Connecticut)	Maeki 1961
27	Udranomia spitzi (Hayward, 1942)	29	Brazil	de Lesse and Brown 1971
28	Urbanus dorantes dorantes (Stoll, 1790)	31	Mexico	de Lesse 1970a
29	Urbanus doryssus doryssus (Swainson, 1831)	14	Costa Rica	Kumagai et al. 2010
30	Urbanus proteus (Linnaeus, 1758)	31	Bolivia	de Lesse 1967a
		31	Mexico	de Lesse 1970a
		31	USA (Florida)	Maeki 1961
31	Urbanus simplicius (Stoll, 1790)	31	Argentina	de Lesse 1967a
32	Urbanus teleus (Hübner, 1821)	31	Argentina	de Lesse 1967a
Subfamily Pyrginae
Tribe Pyrrhopygini
33	Elbella lamprus (Hopffer, 1874)	40	Brazil	de Lesse 1970a
34	(?) Jemadia sp.	32(?)	Brazil	Saura et al. 2013
35	Mimoniades montana J. Zikán, 1938	27	Brazil	Saura et al. 2013
36	Mimoniades nurscia (Swainson, 1821)	28	Ecuador	de Lesse 1967a
	Mimoniades nurscia malis (Godman et Salvin, 1879)	28	Colombia	Saura et al. 2013
37	Mimoniades sp.	21	Colombia	Saura et al. 2013
38	Mimoniades sp.	28	Colombia	Saura et al. 2013
39	Mimoniades versicolor (Latreille, [1824])	28	Brazil	de Lesse and Brown 1971
40	Pyrrhopyge charybdis Westwood, 1852	14(?)	Brazil	Saura et al. 2013
41	Pyrrhopyge pelota Plötz, 1879	28	Argentina	de Lesse 1967a
42	Pyrrhopyge sp.	15	Brazil	Saura et al. 2013
43	Sarbia sp.	30	Brazil	Saura et al. 2013
Tribe Tagiadini
44	Daimio tethys (Ménétriés, 1857)	30	Japan	Maeki 1953, Maeki and Makino 1953
45	Daimio tethys moorei Mabille, 1876	30	Taiwan	Maeki and Ae 1968b
46	Eagris lucetia (Hewitson, 1876)	30	Uganda	de Lesse 1968
47	Eagris sabadius astoria Holland, 1896	30	Kenya	de Lesse 1968
48	Eretis lugens (Rogenhofer, 1891	28	Kenya	de Lesse 1968
Tribe Celaenorrhinini
49	Sarangesa phidyle (Walker, 1870)	29	Senegal	de Lesse and Condamin 1962
Tribe Carcharodini
50	Carcharodus alceae (Esper, [1780])	31	Croatia	Lorković 1941
51	Carcharodus boeticus Reverdin, 1913	43–47	Spain	de Lesse 1960
	Carcharodus boeticus Reverdin, 1913	40–52	France	de Lesse 1960
	Carcharodus boeticus Reverdin, 1913	38–46	Italy	de Lesse 1960
52	Carcharodus dravira (Moore, 1874)	37–48 (with Us)	Iran	de Lesse 1960
53	Carcharodus flocciferus (Zeller, 1847)	32–41 (with Us)	France (Cauterets)	de Lesse 1960
54	Carcharodus flocciferus (Zeller, 1847)	42–58 (with Us)	Italy	de Lesse 1960
55	Carcharodus lavatherae (Esper, [1783])	30	France (Salau, Ariege)	de Lesse 1960
56	Carcharodus orientalis Reverdin, 1913	31–32	Lebanon	de Lesse 1960
		30	Turkey (Van)	de Lesse 1960
		30–37 (with Us)	Turkey (Amasya)	de Lesse 1960
57	Carcharodus stauderi ambiguus Verity, 1925	30	Lebanon	de Lesse 1960
		30	Turkey	de Lesse 1960
58	Hesperopsis alpheus (W. H. Edwards, 1876) (as Pholisora)	34	USA (Texas)	Emmel and Trew 1973
59	Muschampia nomas (Lederer, 1855)	30	Lebanon	de Lesse 1960
60	Muschampia proteides (Wagner, 1929)	30	Lebanon	Larsen 1975
61	Muschampia proto (Ochsenheimer, 1808)	30	Spain	de Lesse 1960
		30	Lebanon	Larsen 1975
62	Pholisora catullus (Fabricius, 1793)	29	?USA	Lorković in Robinson 1971
63	Spialia orbifer (Hübner, [1823])	30	Croatia	Lorković 1941
		31	Turkey	de Lesse 1960
64	Spialia phlomidis (Herrich-Schäffer, [1845])	31	Turkey	de Lesse 1960
65	Spialia sertorius (Hoffmannsegg, 1804)	31	Slovenia	Lorković 1941
Tribe Erynnini
66	Chiomara asychis georgina (Reakirt, 1868)	31	Mexico	de Lesse 1970a
	Chiomara asychis georgina (Reakirt, 1868)	32	USA (Texas)	Emmel and Trew 1973
67	Chiomara sp.	31	Trinidad	Wesley and Emmel 1975
68	Ebrietas anacreon (Staudinger, 1876)	31	Argentina	de Lesse 1967a
69	Ebrietas osyris (Staudinger, 1876)	31	Argentina	de Lesse 1967a
70	Erynnis baptisiae (W. Forbes, 1936)	31	USA (Connecticut)	Maeki 1961
71	Erynnis funeralis (Scudder et Burgess, 1870)	31	Argentina	de Lesse 1967a
72	Erynnis horatius (Scudder et Burgess, 1870)	31	USA (Florida)	Maeki 1961
73	Erynnis icelus (Scudder et Burgess, 1870)	30	USA (Connecticut)	Maeki 1961
74	Erynnis juvenalis juvenalis (Fabricius, 1793)	30	USA (Connecticut)	Maeki 1961
75	Erynnis lucilius (Scudder et Burgess, 1870)	31	USA (Connecticut)	Maeki and Remington 1960a
76	Erynnis marloyi (Boisduval, [1834])	31	Lebanon	de Lesse 1960
77	Erynnis montanus (Bremer, 1861)	31 (2n=62)	Japan	Abe et al. 2006
	Erynnis montanus (Bremer, 1861)	31	Japan	Maeki 1953
78	E. persius (Scudder, 1863)	31	USA (Connecticut)	Maeki 1961
79	Erynnis tages (Linnaeus, 1758)	31	Croatia	Lorković 1941
		31	France	de Lesse 1960
		31	England	Bigger 1960
80	Erynnis tristis tatius (W. H. Edwards, 1883)	31	USA (Texas)	Emmel and Trew 1973
81	Gesta gesta (Herrich-Schäffer, 1863)	32	Tobago	Wesley and Emmel 1975
82	Grais stigmaticus (Mabille, 1883)	31	Mexico	Maeki and Remington 1960a
83	Theagenes albiplaga (C. Felder et R. Felder, 1867)	31	Bolivia	de Lesse 1967a
Tribe Achlyodidini
84	Achlyodes pallida (R. Felder, 1869) (as Achlyodes selva)	15	Bolivia	de Lesse 1967a
		15	Mexico	de Lesse 1970a
85	Zera zera zera (Butler, 1870)	34	Brazil	de Lesse and Brown 1971
Tribe Pyrgini
86	Anisochoria sublimbata Mabille, 1883	31	Argentina	de Lesse 1967a
87	Antigonus erosus (Hübner, [1812])	31	Mexico	de Lesse 1970a
88	Antigonus liborius Plötz, 1884	31	Argentina	de Lesse 1967a
89	Celotes nessus (W. H. Edwards, 1877)	14, 13	USA (Texas)	Emmel and Trew 1973
90	Heliopetes arsalte (Linnaeus, 1758)	30	Bolivia	de Lesse 1967a
	Heliopetes arsalte (Linnaeus, 1758)	30	Mexico	de Lesse 1970a
91	Heliopetes laviana (Hewitson, 1868)	29	USA (Texas)	Emmel and Trew 1973
92	Heliopetes macaira (Reakirt, [1867])	29	USA (Texas)	Emmel and Trew 1973
93	Heliopetes omrina (Butler, 1870)	30	Argentina	de Lesse 1967a
94	Heliopyrgus americanus (Blanchard, 1852)	30	Chile	de Lesse 1967a
95	Paches loxus (Westwood, [1852])	31	Guatemala	de Lesse 1970a
96	Pyrgus aladaghensis De Prins et van der Poorten, 1995	ca 18–21	Turkey	Lukhtanov and Kandul 1995 (in Hesselbarth et al. 1995)
97	Pyrgus albescens Plötz, 1884	30 (2n=60)	USA (Texas)	Goodpasture 1976
	Pyrgus albescens Plötz, 1884	28	USA (Texas)	Emmel and Trew 1973
98	Pyrgus alveus (Hübner, [1803])	24	Finland	Federley 1938
		24	Croatia	Lorković 1941
		24	Turkey	Lukhtanov and Kandul 1995 (in Hesselbarth et al. 1995)
99	Pyrgus bellieri (Oberthür, 1910)	27	France	de Lesse 1960
100	Pyrgus bocchoris (Hewitson, 1874)	30	Argentina	de Lesse 1967a
101	Pyrgus bolkariensis De Prins et van der Poorten, 1995	30	Turkey	Lukhtanov and Kandul 1995 (in Hesselbarth et al. 1995)
102	Pyrgus cacaliae (Rambur, 1839)	30	Italy	de Lesse 1960
103	Pyrgus carlinae (Rambur, [1839])	30	Italy	de Lesse 1960
104	Pyrgus carthami (Hübner, [1813])	29	Italy	de Lesse 1960
105	Pyrgus cirsii (Rambur, [1839])	30	France (Peyreleau, Aveyron)	de Lesse 1960
106	Pyrgus fides Hayward, 1940	30	Chile	de Lesse 1967a
107	Pyrgus maculates (Bremer et Grey, 1852)	31 (2n=62)	Japan	Abe et al. 2006
108	Pyrgus malvae (Linnaeus, 1758)	31	Finland	Federley 1938
		33	England	Bigger 1960
109	Pyrgus oileus (Linnaeus, 1767)	30 (2n=60)	USA (Texas)	Goodpasture 1976
		32	USA (Texas)	Emmel and Trew 1973
110	Pyrgus onopordi (Rambur, [1839])	30	France	Lorković 1941
111	Pyrgus serratulae (Rambur, [1839])	30	France	Lorković 1941
112	Trina geometrina geometrina (C. Felder et R. Felder, 1867)	31	Brazil	de Lesse and Brown 1971
Subfamily Heteropterinae
113	Butleria quilla Evans, 1939	29	Chile	de Lesse 1967a
Subfamily Trapezitinae
114	Trapezites eliena Hewitson, 1868	31	Australia	Maeki and Ogata 1971
Subfamily Hesperiinae
Tribe Aeromachini
115	Aegiale hesperiaris (Walker, 1856)	24	Mexico	Freeman 1969
116	Agathymus alliae (Stallings et Turner, 1957)	38	USA (Arizona)	Freeman 1969
117	Agathymus aryxna (Dyar, 1905)	5	Mexico	Freeman 1969
118	Agathymus baueri (Stallings et Turner, 1954)	15	USA (Arizona)	Freeman 1969
119	Agathymus chisosensis (Freeman, 1952)	18	USA (Texas)	Freeman 1969
120	Agathymus estelleae valverdiensis Freeman, 1966	9	USA (Texas)	Freeman 1969
	Agathymus estelleae estelleae (Stallings et Turner, 1958)	9	Mexico	Freeman 1969
121	Agathymus freemani Stallings, Turner et Stallings, 1960	15	USA (Arizona)	Freeman 1969
122	Agathymus gilberti Freeman, 1964	21	USA (Texas)	Freeman 1969
123	Agathymus mariae chinatiensis Freeman, 1964	22	USA (Texas)	Freeman 1969
	Agathymus mariae lajitaensis Freeman, 1964	22	USA (Texas)	Freeman 1969
	Agathymus mariae mariae (Barnes et Benjamin, 1924)	22	USA or Mexico	Freeman 1969
	Agathymus mariae rindgei Freeman, 1964	22	USA (Texas)	Freeman 1969
124	Agathymus micheneri Stallings, Turner et Stallings, 1961	20	Mexico	Freeman 1969
125	Agathymus neumoegeni florenceae (Stallings et Turner, 1957)	10	USA (Texas)	Freeman 1969
	Agathymus neumoegeni macalpinei (Freeman, 1955)	10	USA (Texas)	Freeman 1969
126	Agathymus polingi (Skinner, 1905)	10	USA (Arizona)	Freeman 1969
127	Agathymus remingtoni (Stallings et Turner, 1958)	9	Mexico	Freeman 1969
128	Alera vulpina (C. Felder et R. Felder, 1867)	ca27	Ecuador	de Lesse 1967a
129	Ankola fan (Holland, 1844)	10	Uganda	De Lesse 1968
130	Arotis derasa (Herrich-Schäffer, 1870) (as Euphyes)	28	Brazil	de Lesse and Brown 1971
131	Erionota thrax thrax (Linnaeus, 1767)	29	Malaysia	Saitoh and Kumagai 1974
132	Euphyes leptosema Mabille, 1891	ca28	Argentina	de Lesse 1967a
133	Megathymus coloradensis coloradensis Riley, 1877	27	USA	Freeman 1969
134	Megathymus coloradensis kendalli Freeman, 1965	27	USA (South central Texas)	Freeman 1969
	Megathymus coloradensis louiseae Freeman, 1963	27	USA (Western Texas)	Freeman 1969
	Megathymus coloradensis navajo Skinner, 1911	27	USA	Freeman 1969
	Megathymus coloradensis reinthali Freeman, 1963	27	USA (Texas)	Freeman 1969
	Megathymus coloradensis reubeni Stallings, Turner et Stallings, 1963	27	USA (Texas)	Freeman 1969
	Megathymus coloradensis stallingsi Freeman, 1943	27	USA	Freeman 1969
	Megathymus coloradensis wilsonorum Stallings et Turner, 1958	27	?Mexico	Freeman 1969
135	Megathymus violae Stallings et Turner, 1956	27	USA	Maeki 1961, Freeman 1969
136	Megathymus yuccae buchholzi Freeman, 1952	26	USA (Florida)	Freeman 1969
137	Pardaleodes incerta (Snellen, 1872)	17	Uganda	de Lesse 1968
138	Stallingsia maculosus (Freeman, 1955)	50	USA (Texas)	Maeki 1961, Freeman 1969
139	Suastus gremius (Fabricius, 1798)	23	Taiwan	Maeki and Ae 1968b
140	Thoressa varia (Murray, 1875)	31 (2n=62)	Japan	Abe et al. 2006
141	Thoressa varia (Murray, 1875)	31	Japan	Maeki 1953
Tribe Baorini
142	Gegenes gambica (Mabille, 1878)	41	Yemen	Saitoh 1984
		41	Turkey	de Lesse 1960
		41	Lebanon	Larsen 1982
143	Gegenes nostrodamus (Fabricius, 1793)	15	Egypt	Larsen 1982
		15	Israel	Saitoh 1979, Larsen 1982
144	Gegenes pumilio (Hoffmansegg, 1804)	24	France	de Lesse 1960
		24	Alger	de Lesse 1967b
145	Parnara guttata (Bremer et Grey, 1852)	16	Japan	Maeki 1953, Maeki and Makino 1953
		16	China	Saitoh and Abe 1981
146	Pelopidas conjucta conjucta (Herrich-Schäffer, 1869)	16	Hong Kong	Maeki and Ae 1968a
147	Pelopidas jansonis (Butler, 1878)	16 (2n=32)	Japan	Abe et al. 2006
148	Pelopidas mathias (Fabricius, 1798)	16	Japan	Maeki and Remington 1960
149	Pelopidas thrax (Hübner, [1821])	16	Lebanon	Larsen 1975
150	Polytremis lubricans (Herrich-Schäffer, 1869)	16	Taiwan	Maeki and Ae 1968b
151	Polytremis pellucida (Murray, 1875)	16, 17, 18 (2n=32, 33)	Japan	Abe et al. 2006
		16	Japan	Maeki and Remington 1960
152	Zenonia zeno (Trimen, 1864)	16	Uganda	de Lesse 1968
Tribe Taractrocerini
153	Ocybadistes walkeri sothis Waterhouse, 1933	28	Australia	Maeki and Ogata 1971
154	Potanthus flavus (Murray, 1875)	29 (2n=58)	Japan	Abe et al. 2006
155	Telicota ancilla horisha Evans, 1934	29	Taiwan	Maeki and Ae 1968b
156	Telicota colon stinga Evans, 1949	29	Japan (Okinava)	Abe et al. 2006
157	Telicota ohara formosana Fruhstorfer, 1911	29 (2n=58)	Taiwan	Abe et al. 2006
Tribe Thymelicini
158	Copaeodes minima (W.H. Edwards, 1870)	29	USA (Florida)	Maeki 1961
159	Thymelicus sylvestris (Poda, 1761)	27	England	Bigger 1960
160	Thymelicus sylvaticus (Bremer, 1861)	10 (2n=20)	Japan	Abe et al. 2006
161	Thymelicus acteon (Rottemburg, 1775)	28	Spain	de Lesse 1970c
162	Thymelicus hyrax (Lederer, 1861)	29	Lebanon	Larsen 1975
163	Thymelicus leoninus (Butler, 1878)	9 (2n=18)	Japan	Abe et al. 2006
164	Thymelicus lineola (Ochsenheimer, 1808)	29	Finland	Federley 1938
		29	Lebanon	Larsen 1975
Tribe Calpodini
165	Ebusus ebusus (Cramer, [1780])	29	Mexico	de Lesse 1970a
166	Lychnuchus celsus (Fabricius, 1793)	30	Brazil	de Lesse and Brown 1971
167	Panoquina hecebolus (Scudder, 1872)	29	USA (Texas)	Emmel and Trew 1973
168	Panoquina ocola (W. H. Edwards, 1863)	29	USA (Texas)	Emmel and Trew 1973
169	Panoquina panoquin (Scudder, 1863)	29	USA (Florida)	Maeki 1961
170	Panoquina panoquinoides (Skinner, 1891)	29	USA (Texas)	Emmel and Trew 1973
Tribe Anthoptini no chromosomal data available
Tribe Moncini
171	Amblyscirtes aenus W.H. Edwards, 1878	28, 29	USA (Texas)	Emmel and Trew 1973
172	Amblyscirtes cassus W. H. Edwards, 1883	29	USA (Texas)	Emmel and Trew 1973
173	Amblyscirtes celia (Skinner, 1895)	29	USA (Texas)	Emmel and Trew 1973
174	Amblyscirtes phylace W.H. Edwards, 1878	29	USA (Texas)	Emmel and Trew 1973
175	Amblyscirtes texanae Bell, 1927	29	USA (Texas)	Emmel and Trew 1973
176	Amblyscirtes vialis (W. H. Edwards, 1862)	29	USA (Connecticut)	Maeki 1961
177	Cymaenes sp.	31	Tobago	Wesley and Emmel 1975
178	Enosis immaculata immaculata (Hewitson, 1868)	29	Ecuador	Kumagai et al. 2010
179	Lerema accius (Smith, 1797)	29 (2n=58)	USA (Texas)	Goodpasture 1976
		29	USA (Texas)	Emmel and Trew 1973
180	Moeris vopiscus (Herrich-Schäffer, 1869)	27	Peru	Kumagai et al. 2010
181	Nastra lherminier (Latreille, [1824])	30	USA (Connecticut)	Maeki 1961
182	Thargella caura (Plötz, 1882)	25	Brazil	de Lesse and Brown 1971
183	Vettius coryna (Hewitson, [1866])	31, ca32	Ecuador	de Lesse 1967a
184	Vettius phyllus prona Evans, 1955	26	Brazil	de Lesse and Brown 1971
185	Vettius triangularis (Hübner, [1831])	26	Brazil	Kumagai et al. 2010
Tribe Hesperiini
186	Asbolis capucinus (Lucas, 1857)	48	USA (Florida)	Maeki 1961
187	Cynea iquita (Bell, 1941)	29	Argentina	de Lesse 1967a
188	Hesperia comma (Linnaeus, 1758)	28	Italy	de Lesse 1970c
		28	Lebanon	Larsen 1975
189	Hesperia florinda Butler, 1878	28 (2n=56)	Japan	Abe et al. 2006
190	Hylephila fasciolata (Blanchard, 1852)	29	Argentina	de Lesse 1967a
191	Hesperia phyleus (Drury, 1773)	29	Argentia	de Lesse 1967a
		29	USA (Florida)	Maeki 1961
192	Hesperia signata (Blanchard, 1852)	29	Chile	de Lesse 1967a
193	Ochlodes ochraceus (Bremer, 1861)	29 (2n=58)	Japan	Abe et al. 2006
		24	Japan	Maeki and Remington 1960
194	Ochlodes sylvanoides (Boisduval, 1852)	29	USA	Maeki 1961
195	Ochlodes sylvanus (Esper, 1777)	29	Finland	Federley 1938
		29	Croatia	Lorković 1941
196	Ochlodes venatus (Bremer et Grey, 1853) (as sylvanus Esper, 1777)	29 (2n=58)	Japan	Abe et al. 2006
197	Oligoria maculata (W. H. Edwards, 1865)	29	USA (Florida)	Maeki 1961
198	Poanes hobomok hobomok (Harris, 1862)	29	?USA	Lorković in Robinson 1971
199	Poanes taxiles (W. H. Edwards, 1881)	29	USA	Maeki 1961
200	Poanes zabulon (Boisduval et Le Conte, [1837]) (as Polites zabulon)	29	USA (Connecticut)	Maeki 1961
201	Polites themistocles (Latreille, [1824])	29	USA (Florida)	Maeki 1961
202	Poanes vibex catilina (Plötz, 1886)	29	Argentina	de Lesse 1967a
	Poanes vibex praeceps (Scudder, 1872)	27	USA (Texas)	Emmel and Trew 1973
	Poanes vibex vibex (Geyer, 1832)	29	USA (Florida)	Maeki 1961
203	Wallengrenia egeremet (Scudder, 1863)	28	USA (Texas)	Emmel and Trew 1973
204	Wallengrenia otho curassavica (Snellen, 1887)	28–30	USA (Texas)	Emmel and Trew 1973
205	Wallengrenia premnas (Wallengren, 1860)	27	Argentina	de Lesse 1967