Karyotaxonomy of Greek Serpentine Angiosperms

Botanical Journal of the Linnean Society, 2002, 139, 109–124. With 19 ﬁgures

Karyotaxonomy of Greek serpentine angiosperms

THEOPHANIS CONSTANTINIDIS1*, ELEFTHERIA-PERDIKO BAREKA2 and GEORGIA KAMARI2 Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 1Institute of Systematic Botany, Department of Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, GR – 118 55 Athens, Greece 2Section of Plant Biology, Department of Biology, University of Patras, GR – 265 00 Patras, Greece

Received October 2001; accepted for publication February 2002

The chromosome numbers are given for 20 angiosperm taxa, most of which grow predominately or exclusively on serpentine in Greece, and the karyotype morphology is illustrated in all cases but one. Chromosome data are provided for the first time for ten taxa (Alyssum pogonocarpum, Centaurea charrellii, C. vlachorum, Cephalaria fanourii, Matricaria tempskyana, Onosma stridii, Scorzonera doriae, Silene fabarioides, S. salamandra, Trinia glauca ssp. pindica) and for the endemic monospecific genus Leptoplax. A new number is reported for Thymus teucrioides ssp. candilicus. An unexpected dysploid number is recorded for a population of Leontodon hispidus ssp. hispidus. Chromosome number and karyotype details from Greek populations are presented for the rest of the taxa. Chromosomal evidence supports close relationships among members of Onosma subsect. Asterotricha. The unusual chromosome number of Centaurea vlachorum supports its placement either in C. sect. Jacea or C. sect. Cyanus. The evolution of taxa in sections Vierhapperia, Pulvinares, and Nervosae of Scorzonera appears to be connected with particular chromosomal rearrangements and dysploidy. Leptoplax does not share the same chromosome number with Peltaria but with Bornmuellera instead, something that facilitates infrageneric hybridization. Further issues of taxonomy, distribution, evolution and conservation of serpentine species are briefly discussed where appropriate. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124.

ADDITIONAL KEYWORDS: chromosome numbers – distribution – endemism – Flora Hellenica – karyotype morphology – ultramaﬁc substrate.

INTRODUCTION when their relatives are not obvious or are distantly distributed (Wild & Bradshaw, 1977; Brooks, 1987). Rich in silica, magnesium and iron, the serpentine Palaeoendemics may be restricted to small areas as a rocks and soils (also sometimes called ophiolitic or result of severe range contraction, leaving them often ultramaﬁc) have long been known to support a spe- with a disjunct distribution. cialized ﬂora and to affect the form of vegetation Certain cases of differentiation and evolution, occa- (Brooks, 1987; Baker, Proctor & Reeves, 1992; Roberts sionally with karyological data included, have been & Proctor, 1992). Plant species growing on serpentine investigated in serpentine endemics or species occu- may be morphologically distinct and in some cases pying both serpentine and nonserpentine substrates restricted to such geoedaphics, i.e. serpentine ende- (Westerbergh & Saura, 1992; Mayer, Soltis & Soltis, mics. Endemic taxa whose taxonomically closely 1994; Dolan, 1995; Sˇtepánková, 1996; Krahulcová & related congener is easily recognized, often growing in Sˇtepánková, 1998). Evidence derived from chromo- proximity to the serpentine endemic and occasionally some number and morphology may be useful in on a different rock type, are usually called neoen- tracing taxonomic and evolutionary relationships of demics. A neoendemic may have undergone minimal serpentine species, especially when information on range expansion (Mayer & Soltis, 1994). Palaeoen- related species is also available. However, beyond demics can also been found on serpentine, especially taxonomic description of certain taxa, very little work has been devoted to the study of chromosome number, *Corresponding author. E-mail: [email protected] karyotype morphology and possible evolutionary

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 109 110 T. CONSTANTINIDIS ET AL. consequences of taxa with particular preference to cultivated in the Experimental Botanical Garden, serpentine in Greece. University of Patras. Taxa names, provenances and This paper aims to investigate chromosome number chromosome numbers are summarized in Table 1. and karyotype morphology of angiosperm taxa, most The chromosome counts were made from root of which grow predominately or exclusively on serp- tip metaphases. In all cases root tips were subject entine in Greece. It forms part of our studies on the to a chemical pretreatment (0.03% v/w aqueous 8- karyology of the Greek flora (Constantinidis, Kamari hydroxyquinoline) and/or chilling (4°C) for 3–24h & Phitos, 1997; Constantinidis & Kamari, 2000), and before being processed further. Details of the squash on the various peculiarities of plant life on serpentine technique used are reported in Constantinidis et al. (e.g. edaphic endemism and speciation, disjunct dis- (1997). Chromosome terminology principally follows Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 tributions, metal hyperaccumulation; Constantinidis, Levan, Fredga & Sandberg (1964) and Stebbins (1971). 1999; Psaras et al., 2000). The karyological informa- Nomenclature for the taxa examined mostly follows tion obtained is used in conjunction with published evi- the published volumes of Med-Checklist (Greuter, dence on taxa related to those growing on serpentine Burdet & Long, 1984, 1986, 1989) or, in genera not to address various taxonomic or evolutionary issues. appearing in them or revised more recently, Tutin Eleven of our chromosome reports are published for et al. (1976, 1993), Strid (1986a), Strid & Tan (1991a) the first time, new chromosome numbers are given for and Greuter (1997). Investigated taxa are listed by two taxa, karyotype morphology is given for the first their families, in alphabetical order. time in 13 taxa, and novel karyological records based on Greek populations are provided for three taxa. Details on the distribution and conservation of ser- RESULTS AND DISCUSSION pentine taxa have been added where appropriate. BORAGINACEAE Onosma elegantissima Rech. fil. & Goulimy MATERIAL AND METHODS Nomos Grevenon, Eparchia Grevenon: Lower slopes Living plant material of perennial taxa and seeds of of Mt. Vourinos, c. 2.5–3.3km after Dafneron vil- annual species was collected in various serpentine lage along road to Exarchos. Serpentine slopes with areas of Greece. Individual plants (3–8 per taxon) were Juniperus oxycedrus and Buxus sempervirens. Alt.

Table 1. Alphabetical list of the taxa investigated (E = endemic to Greece) with provenance and chromosome number found: = first report; = first record based on Greek material; = new chromosome number; +=first report of karyotype morphology

Taxon Provenance 2n

Alyssum pogonocarpum Rodos island 16 Centaurea charrellii (E) Domokos area, Sterea Ellas 36 Centaurea thracica Mt. Katachloron area, Sterea Ellas 18 Centaurea vlachorum (E) Mts. Aftia/Flega, Ipiros 24 + Cephalaria fanourii (E) Mt. Gramos, Ipiros/Makedonia 18 + Crepis merxmuelleri (E) Mt. Smolikas, Ipiros 8 + Hesperis laciniata ssp. laciniata Mt. Smolikas, Ipiros 12 Leontodon hispidus ssp. hispidus Mt. Smolikas, Ipiros 12 Leptoplax emarginata (E) Domokos area, Sterea Ellas 16 + Matricaria tempskyana (E) Mt. Zigos, Ipiros 36 + Onosma elegantissina (E) Mt. Vourinos, Makedonia 14 + Onosma stridii (E) Mt. Kallidromon, Sterea Ellas 14 + Ranunculus pedatus Domokos & Mt. Kallidromon, Sterea Ellas 16 Scorzonera doriae Eptachori area, Makedonia 12 + Silene fabaria ssp. domokina (E) Domokos area & Mt. Gerania, Sterea Ellas 24 + Silene fabarioides Mt. Mavrovouni, Ipiros 24 + Silene haussknechtii (E) Mt. Smolikas, Ipiros 24 + Silene salamandra (E) Rodos island 24 + Thymus teucrioides ssp. candilicus (E) Domokos area, Sterea Ellas 28 Trinia glauca ssp. pindica Mt. Aftia/Flega, Ipiros 18 +

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 KARYOTAXONOMY OF GREEK SERPENTINE ANGIOSPERMS 111 c. 720–750m, Lat. 40°10¢ N, Long. 21°36¢ E, 30.iv.2000, distributed quite differently. Karyological information Constantinidis 9010 (UPA). 2n = 14 (Fig. 1). on many Asiatic species is still lacking. Known only from a single mountain in north central Greece (Vourinos) and growing from c. 700 up to Onosma stridii Teppner 1800m, this attractive species has been found in Nomos Fthiotidos, Eparchia Lokridos: Mt. several localities forming multiple rosettes on rather Kallidromon, c. 2.7km after the village of Modion, shallow, serpentine ground. We have not observed it along road to Reginion. Slopes with scrub and a dry growing on limestone but Teppner (1991a) gave it as streambed. Vegetation composed mainly of Cistus such. This being true, Onosma elegantissima should

salviifolius, Quercus coccifera, Pistacia lentiscus, Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 not be considered as an obligate serpentine endemic. Juniperus oxycedrus, etc. Ophiolithic substrate. Alt. c. In agreement with Strid (1995) and Strid & Tan (2000) 580–620 m, Lat. 38°40 N, Long. 22°41 E, 22.iv.1998, we considered it as hardly threatened. Most popula- ¢ ¢ Constantinidis 7522 (UPA). 2n 14 (Fig. 2). tions encountered appeared to be healthy, untouched = Onosma stridii, a very local species known from a by browsing animals (except perhaps for some missing small part of the Mt. Kallidromon serpentine area, flowering shoots), and with a good number of juvenile should be considered as an obligate serpentine ste- individuals. noendemic. It was described relatively recently by A report of the O. elegantissima chromosome Teppner (1988b) from a single collection made in 1987. number is given by Teppner (1991a, 1991b) as 2n = 14, Our search on Mt. Kallidromon located only two without further details. Our investigation confirmed populations: the one mentioned above (locus classi- the same diploid number. The chromosomes are cus) and a second to the north-north-west of it that mostly metacentric. Satellites were observed on two forms part of the understorey of locally sparse Pinus of the smallest pairs. The chromosome formula of halepensis woodland. This second population is com- the population examined is given as 2n = 10m + posed of individuals with the usual white or whitish 2m-SAT + 2m/sm-SAT = 14. Examination of mitotic flowers and others with pink, pinkish or tinged pink plates at metaphase and especially at late prophase flowers, a colour that is unique among Greek Onosma revealed clear heterochromatic bands, mostly situated species. at centromeric and distal parts of the longer chromo- Karyological examination of the locus classicus some arms. The number and size of chromosomes and population revealed a diploid karyotype with 2n 14. the rather late condensing euchromatin support place- = The chromosomes are mostly metacentric, with at ment of O. elegantissima complement with the O. least two very obvious satellites situated on the short echioides L. cytotype, characteristic of asterotrichous arm of the smallest chromosome pair. Another pair of Onosma species with x = 7 (Teppner, 1971, 1991b). satellites is sometimes observed on a metacentric When originally described (Rechinger, 1957), chromosome pair. Therefore, the karyotype formula of Onosma elegantissima was compared to the Albanian the population examined is given as 2n 10m endemic O. mattirolii Bald. Judging from the pub- = + 2m-SAT 2m/sm-SAT 14. This is the first report on lished karyological information on the latter species + = the chromosome number and karyotype morphology of (Baltisberger & Baltisberger, 1995) some differences O. stridii. from C. elegantissima are noted. In particular, clear Together with O. elegantissima, O. stridii belongs submetacentric chromosomes exist that we did not to the asterotrichous Onosma species characterized find in O. elegantissima and do not match an O. by a base number of x 7, a diploid cytotype and a echioides cytotype. = clear separation of euchromatin and heterochro- Several karyotypes of members of O. subsect. Aster- matin at prophase or even early metaphase stages otricha (Boiss.) Gürke studied so far with 2n = 14 (echioides type). The taxonomic affinity between the present a remarkable similarity, which is of proba- two species that was assumed by Teppner (1988b) is ble taxonomic importance, especially if the diversity further supported by karyological evidence. Although of chromosome number and morphology observed O. stridii has pollen larger than that of O. elegantis- in Onosma is taken into account. Indeed, the karyo- sima, they are both diploid with very similar kary- type of O. elegantissima, a single mountain endemic, otype morphology. appears almost identical to those of O. echioides from Italy (Teppner, 1971; Raimondo, Rossitto & Ottonello, 1983), the Greek endemic O. erecta Sm. ssp. erecta CARYOPHYLLACEAE (Teppner, 1988a), the single mountain endemic O. stridii (see below), and O. inexspectata Teppner, an Silene fabaria (L.) Sm. ssp. domokina Greuter endemic species of SW Anatolia (Teppner, 1974). This Nomos Attikis/Viotias, Eparchia Megaridos/ stable and conservative karyotype feature may actu- Korinthias: northern foothills of Mt. Gerania, between ally represent a symplesiomorphy shared by species Pefkogiali and Mavrolimni settlements. Ophiolitic

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 112 T. CONSTANTINIDIS ET AL. Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 1 2

3 4

5 6

7 8

Figures 1–8. Photomicrographs of mitotic metaphases. Scale bars = 10 mm. Fig. 1. Onosma elegantissima (2n = 14). Fig. 2. O. stridii (2n = 14). Fig. 3. Silene fabaria ssp. domokina (2n = 24). Fig. 4. S. fabarioides (2n = 24). Fig. 5. S. haussknechtii (2n = 24). Fig. 6. S. salamandra (2n = 24). Fig. 7. Centaurea thracica (2n = 18). Fig. 8. C. charrellii (2n = 36).

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 KARYOTAXONOMY OF GREEK SERPENTINE ANGIOSPERMS 113 substrate. Alt. 5–10m, Lat. 38°03¢N, Long. 23°07¢E, with 2n = 24, the most common number in Silene. Most 09.ix.1994, Constantinidis 3238 (UPA). 2n = 24 of the chromosomes are metacentric, with only a (Fig. 3). few being metacentric/submetacentric. Three pairs of Nomos Fthiotidos, Eparchia Domokou: c. 3.8km satellites were observed, two on the short arms and south of Domokos, along a secondary road to an aban- one on the long arms of metacentric chromosomes. doned quarry. Rocky slopes. Ophiolithic substrate. Alt. c. 510m. Lat. 39°05¢ N, Long. 22°18¢ E, 28.vi.1998, Silene haussknechtii Hausskn. Constantinidis (seeds collected). 2n 24. = Nomos Ioanninon, Eparchia Konitsis: Mt. Smolikas, Described very recently by Greuter (1995), this

c. 6.2km along forest road starting just south-west Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 neoendemic taxon shows a disjunct distribution of the village of Pades and leading to the upper parts between the serpentine parts around the town of of the mountain. Openings of Pinus forest and road Domokos and the north-west serpentine slopes of Mt. sides, serpentine. Alt. c. 1580m, Lat. 40°03¢ N, Long. Gerania. Although it occurs predominately on serpen- 20°54¢ E, 01.ix.1997, Phitos, Kamari & Constantinidis tine, it was also observed growing on flysch, usually in 25608 (UPA). 2n = 24 (Fig. 5). secondary, man-made habitats such as scree along Earlier literature records, specimens examined road embankments and disturbed alluvia. and personal observations indicate that Silene Its chromosome number, 2n 24, was reported by = haussknechtii is a serpentine endemic. It has been Melzheimer (1987) under the name S. fabaria ssp. considered to be restricted to serpentine of north-west thebana (Orph. ex Boiss.) Melzh. Of the two popu- Greece but its occurrence in serpentine areas of lations examined by Melzheimer only that from adjacent Albania cannot be ruled out (Greuter, 1997). Domokos is considered to represent the true ssp. Its chromosome number has been reported as domokina, while the population from Thiva (Thive) 2n = probably 24, ranging between 22 and 25 was later reclassified as ssp. fabaria (Greuter, 1995). (Oxelman, 1995) in material from the foothills of Mt. The same chromosome number of 2n = 24 is confirmed Vasilitsa. Our count confirms the number of 2n = 24 here, based on material from two disjunct populations. and at the same time the karyotype morphology of Chromosomes are mostly metacentric and submeta- the species is presented. Most of the chromosomes centric, with at least three pairs of clearly visible are metacentric, with two pairs of satellites on the satellites situated on the short arm of the longest short arms of the longest submetacentric pair and a metacentric/submetacentric chromosome pair and on medium-sized metacentric chromosome pair. In some the long arms of two metacentric pairs. metaphase plates a spherical formation has been Based on the provided locality data, all previous found, almost equalling the short arm of some chro- chromosome numbers for Silene fabaria s.l. that mosome pairs in size. It might be a B-chromosome, appear in the literature (Damboldt & Phitos, 1968; possibly partly responsible for the earlier difficulties Degraeve, 1980; Baltisberger & Aeschimann, 1988) in estimating the exact chromosome number of the should be attributed to S. fabaria ssp. fabaria. This species. last taxon is distributed in east Greece, Aegean islands and Turkey, mostly in coastal, rocky places and on a variety of substrates (Melzheimer & Ulrich, 1994; Silene salamandra Pamp. Greuter, 1997). Nomos Dodekanisou, Eparchia Rodou: Rodos island, c. 4.4km north-east of the village of Laerma. Sparse Pinus brutia forest, scree and dry stream beds, ultra- Silene fabarioides Hausskn. mafic substrate. Alt. c. 320m, Lat. 36°11¢ N, Long. Nomos Ioanninon, Eparchia Metsovou: Mt. 27°55¢ E, 18.iv.1998, Constantinidis K214 (UPA). Mavrovouni, c. 15.5km along forest road on the 2n = 24 (Fig. 6). mountain. The road starts c. 2.5km after (north-west) A rare and local endemic of Rodos island, Silene Metsovon (turnoff to Milea). Rocky slopes with Pinus salamandra is an attractive annual species of S. close to a stream. Mainly ophiolite. Alt. 1350–1400 m, sect. Atocion Otth. Its taxonomic position has been Lat. 39°51¢ N, Long. 21°05¢ E, 18.vi.2000, Constanti- elucidated by Carlström (1986a), who simultaneously nidis 9136 (UPA). 2n = 24 (Fig. 4). recorded it as an exclusive endemic of the ophioli- A Balkan endemic species, Silene fabarioides is thic areas of Rodos (Carlström, 1986b). However, often found on serpentine in Greece but is not the altitudinal range of the species (0–500m) given by confined to it (Melzheimer, 1980; T. Constantinidis, Greuter (1997) indicates that it should also grow by pers. observ.). Its distribution includes Albania, F.Y.R. the sea, where the ophiolithic substrate does not exist. Macedonia, Bulgaria and Greece (Greuter, 1997). Ultramafic rocks are found in the central parts Its chromosome number and morphology are pre- of Rodos island, presumably having chemical and/ sented here for the first time. The species is diploid or petrological synthesis differing from the similar-

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 114 T. CONSTANTINIDIS ET AL. looking formations on the Greek mainland and Evvia. species is able of reproducing itself sexually. All the They are rather sparsely covered with Pinus brutia capitula of the plants examined contained viable forest. During our visits on the island, S. salamandra achenes that germinated under cultivation, 2 months was observed growing on gravelly and rather steep after being collected, without presenting any signs of hill slopes, unstable scree, and dry streambeds with dormancy. As, however, breeding systems and repro- coarse ultramafic sand. The species achieves a luxuri- ductive ecology of C. charrellii are virtually unknown, ous growth under cultivation in normal soil, and there is always the possibility that seed setting is pre- increases its size up to six times compared to wild vented in very small populations, especially in cases specimens. in which the species is self-incompatible. Normal Chromosome number and karyotype morphology of sexual reproduction, viable seeds, progeny that are Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 S. salamandra are reported here for the first time. All identical to their parents, and lack of related species chromosomes appear to be metacentric. Satellites are in Greece make the assumption of Georgiadis (1995) not usually observable, with the exception of a dis- that C. charrellii might be of hybrid origin improba- tinct, spherical one on a chromosome of average size. ble, unless it is an ancient allotetraploid derived from The same chromosome number of 2n = 24 has been the doubling of the chromosome number of a hybrid found in S. insularis Barbey (Damboldt & Phitos, between two diploid species. 1966), a close relative of S. salamanda endemic to the The chromosome number and a microphotograph of island of Karpathos. The diploid number of 24 is, a metaphase plate of C. charrellii are presented here however, a very common and conservative feature in for the first time. The species is tetraploid, with Silene (although not in S. subg. Conoimorpha), and 2n = 36. The somewhat solid appearance of the chro- uniformity of karyotype does not permit conclusions of mosomes does not permit a detailed morphological karyotaxonomic importance to be made. description, but at least metacentric and submetacentric chromosome types occur in the complement. Four satellites were also observed. COMPOSITAE The chromosome number of 2n = 36 is clearly based on x = 9, a common basic number in Centaurea found Centaurea charrellii Halácsy & Dörfler in several sections (e.g. Hellwig, 1994; Garcia-Jacas, Nomos Fthiotidos, Eparchia Domokou: Road Lamia- Susanna & Mozaffarian, 1998a; Garcia-Jacas et al., Domokos, very close to the crossing towards Omvriaki 1998b). As far as C. sect. Cynaroides is concerned, two village. Margins of cultivated land and among bushes, other members have been investigated karyologically, ophiolithic substrate. Alt. c. 500m, Lat. 39°06¢ N, C. amanicola Hub.-Mor. from Turkey (Gardou & Long. 22°18¢ E, 26.vi.1997, Constantinidis 6965 (UPA). Tchehrehgocha, 1975) and C. imperialis Boiss. ex 2n = 36 (Fig. 8). Walp. from Iran (Garcia-Jacas et al., 1998a). They are One of the rarest endemic species of Centaurea in both diploid with 2n = 18. As examination of relevant Greece, C. charrellii is currently known from two very literature did not reveal chromosome numbers in any small and severely threatened populations in the area other species of the more than 22 members of C. sect. of Domokos. Both populations do not exceed 40 indi- Cynaroides, ours is the first report of tetraploidy in the viduals in total and the species has justifiably been section. characterized as Endangered (Georgiadis, 1995). Since its original collection in the locus classicus close to Vodena, now the city of Edessa (north Greece), it has Centaurea thracica (Janka) Hayek not been reported again from the same area, nor any Nomos Karditsis, Eparchia Karditsis: Mt. Katachloron, other area of Greece besides Domokos. along road from Kedros village to Loutra Smokovou, Owing to its tall habit, bright yellow florets (super- c. 2.6km south of Kedros. Meadow, rocky slopes and ficially resembling the much more common C. saloni- road sides. Ophiolithic substrate. Alt. c. 220 m, Lat. tana Vis. from a distance) and taxonomic/chorological 39°11¢ N, Long. 22°02¢ E, 27.vi.1997, Constantinidis position, i.e. the only representative of the Asiatic C. 7061 (UPA). 2n = 18 (Fig. 7). sect. Cynaroides Boiss. ex Walp. in Greece and Europe, By analogy to Centaurea charrellii, C. thracica is the without any close relatives, and growing more than only representative of the Asiatic sect. Microlophus 1200km from any other member of the same section (Cass.) DC. in Europe, but it is not as rare as C. char- (Wagenitz, 1975a) the species itself is distinct and rellii. We have always collected or observed C. thracica unique. While collecting around Domokos we found C. on serpentine in the area of Domokos, the foothills of charrellii twice, growing on serpentine or on a mixture Mt. Katachloron (the population examined karyologi- or serpentine and flysch, usually under the protection cally), the medium-altitude parts of Mt. Kratsovon of grazed Quercus coccifera scrub. Contrary to previ- and between the villages of Kakoplevri and Oxinia, ous reports (Georgiadis, 1995), we found that the north-west of Kalambaka. A record from Evvia island

(Rechinger, 1961) also strengthens this correlation. evident by its banding effect in metaphase plates. However, exclusive conjunction of this species with Differentiation of staining may reflect heterochro- ultramafic substrates still has to be proved. matin rich areas on this particular submetacentric The diploid chromosome number found, 2n = 18, is chromosome pair. The karyotype formula of the popu- a confirmation of an earlier report by Damboldt & lation examined is given as 2n = 18m + 2m- Matthäs (1975), on material obtained from the botani- SAT + 2sm + 2sm-SAT. cal garden of Sofia, Bulgaria. This is the first time The diploid karyotype of C. vlachorum is based on that the chromosome number of a Greek population is x = 12, a number intermediate between the more pri- examined. Most of the chromosomes are metacentric mitive x > 12 and the more advanced x < 12 groups of or submetacentric, with two satellites visible on the Centaureinae (Garcia-Jacas, Susanna & Ilarslan, Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 short arm of a submetacentric chromosome pair and 1996). Within Centaurea, x = 12 is confirmed in three two on the shorter arms of a metacentric chromosome sections, namely Mesocentron (Cass.) DC. (species of pair. This is generally in close agreement with the sect. Hymenocentron (Cass.) DC. are often included karyotype drawing provided by Damboldt & Matthäs here), Cyanus, Jacea and Hyalaea DC. Based on the (1975). available chromosome evidence, no relationship As already noted by Garcia-Jacas et al. (1998b) between C. vlachorum and C. sect. Acrocentron is C. sect. Microlophus is characterized by two basic supported. All members of sect. Acrocentron so far numbers, x = 8 and x = 9. It is not clear yet to what examined karyologically have the chromosome base extent those different numbers are related to the tax- of x = 10 or x = 11 (Garcia Jacas & Susanna de la onomy and phylogeny of its species, as to our knowl- Serna, 1992; Routsi & Georgiadis, 1999), with x = 12 edge, chromosome numbers of only three out of more essentially unknown in the section. Furthermore, than ten species comprising the section are known morphological characters clearly exclude any close currently. relationship between C. vlachorum and members of sections Mesocentron/Hymenocentron and of the small section Hyalaea. In sect. Cyanus, x = 12 is known in Centaurea vlachorum Hartvig C. cyanus L. and C. montana L. (several counts in Nomos Ioanninon, Eparchia Dodonis/Metsovou: Mts. Fedorov, 1969) while in sect. Jacea it is found in, e.g. Aftia/Flega, c. 16.5km after turn to Milea of the road C. phrygia L. ssp. nigriceps (Dobrocz.) Dostál (see Ioannina-Trikala and along a secondary road that Fedorov, 1969), in which confirmation of the count leads to the upper parts of the mountain. Mixed would be welcome. Fagus-Pinus forest and small clearings. Ophiolite. Alt. To conclude, karyological evidence indicates that the c. 1650 m, Lat. 39°52¢ N, Long. 21°06¢ E, 28.vii.1999, isolated placement of C. vlachorum in C. sect. Jacea Constantinidis 8669 (UPA). 2n = 24 (Fig. 9). (contradicted by bract morphology) is in accordance Centaurea vlachorum is a rare endemic species with its unusual chromosome number. The same known from only two mountains, Milea and Aftia of number exists in sect. Cyanus, but the placement of Northern Pindos (Hartvig, 1981). It has always been C. vlachorum in this section is not in agreement with found associated with serpentine and can therefore be its pollen morphology. considered as a serpentine endemic. Since its description, the systematic placement of the species has been intriguing. C. vlachorum is considered to be a distinct Crepis merxmuelleri Kamari & Hartvig species without close relatives and an isolated member Nomos Ioanninon, Eparchia Metsovou: Mt. Smolikas, of C. sect. Jacea (Mill.) DC. (in its widely ascribed southern slopes of the mountain, c. 5.7km along sense, including sect. Lepteranthus DC.; see Wagenitz, road from Pades village to Armata village. Rocky 1975b, 1980), although morphological features of places by the road, scree and road sides. Ophiolitic bracts are reminiscent of sect. Cyanus (Mill.) DC. and substrate. Alt. c. 1050 m, Lat. 40°02¢ N, Long. 20°56¢ E, Acrocentron (Cass.) DC. (Gamal-Eldin & Wagenitz, 03.ix.1997, Constantinidis 8634 (UPA) and Phitos, 1991). The chromosome number of the species, previ- Kamari & Constantinidis 26819 (UPA). 2n = 8 ously unknown, would elucidate its sectional place- (Fig. 10). ment, given that several groups of Centaurea have Crepis merxmuelleri is a rare species, described quite characteristic base numbers. recently from the serpentine belt of Mt. Smolikas C. vlachorum is diploid with 2n = 24. Chromosomes (Kamari & Hartvig, 1988). The description was based are of average size and mostly metacentric. Satellites on two specimens collected on the north-east parts of are found on the shorter arms of a metacentric and the mountain, at an altitude of c. 1250–1550m. Our a submetacentric pair. It is characteristic that one collections, found on the south-east part of the same pair of chromosomes often has a condensation rate mountain and at a slightly lower altitude, extend differing from that of the rest of the complement, somewhat the known distribution of the species.

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 116 T. CONSTANTINIDIS ET AL. Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 9 10

11 12

Figures 9–13. Photomicrographs of mitotic metaphases. Scale bars = 10 mm. Fig. 9. Centaurea vlachorum (2n = 24). Fig. 10. Crepis merxmuelleri (2n = 8). Fig. 11. Leontodon hispidus ssp. hispidus (2n = 12). Fig. 12. Scorzonera doriae (2n = 12); arrowheads indicate secondary constrictions of the longest chromosome pair. Fig. 13. Matricaria tempskyana (2n = 36).

Compared to the isotype (Hartvig & Seberg 5104, The chromosome number of 2n = 8 has already been UPA!), the original description and a recent collection reported for C. merxmuelleri (Kamari, 1991). The (Constantinidis & Kouki 9876, ACA!, UPA!), our speci- number is confirmed here, and a detailed karyotype mens present a significant difference: they completely analysis is given for the first time. The species is lack the yellowish-brown glandular indumentum, diploid with submetacentric, acrocentric, and subtelo- which has been attributed to C. merxmuelleri as a key centric chromosomes. Small satellites are observed on character (Kamari, 1991). the small arm of the subtelocentric chromosome pair.

The karyotype formula is given as 2n = 2sm + 4st + Rousi (1973) is greater (two–three pairs), compared 2t-SAT = 8. with our results (one pair). Overall morphology, chromosome number, and kary- The dysploid reduction of chromosome number from otype analysis of C. merxmuelleri are strongly remi- 2n = 14 to 2n = 12 seems to be related to the loss of a niscent of C. turcica Degen & Bald. (see Constantinidis chromosome pair, nevertheless additional chromoso- & Kamari, 2000; for a presentation of the karyotype mal translocations cannot be ruled out. Relative size of C. turcica and for references). The two species of chromosomes in the serpentine population, when resemble each other in that they both have glandular compared with published information from different and eglandular forms in different populations. It countries, does not support chromosome fusion. As Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 seems that they are closely related species differing in Arabidopsis-type sequence repeats (TTTAGGG)n have their substrate preferences. C. turcica grows on lime- been traced only at telomeric positions in L. hispidus stone and rarely on flysch, while C. merxmuelleri has with 2n = 14 (Fuchs, Brandes & Schubert, 1995), it been found so far only on serpentine. would be of interest to check the 2n = 12 population using fluorescent in situ hybridization (FISH) for similar signals, in an effort to trace possible chromo- Leontodon hispidus L. ssp. hispidus some rearrangements. It is still unknown whether or Nomos Ioanninon, Eparchia Konitsis: Mt. Smolikas, not such a chromosome change was induced by the close to the small lake known as Drakolimni, along serpentine soil in which L. hispidus grows. road from a pen situated c. 10km north of Pades village. Stony slopes, scree and rocky places. Ophiolitic substrate. Alt. c. 2000–2200m. Lat. 40°05¢ N, Long. Matricaria tempskyana (Freyn. & Sint.) Rauschert 20°54¢ E, 22.vi.1998, Phitos, Kamari & Constantinidis [Syn. Tripleurospermum tempskyanum (Freyn & cult. 9137 (UPA). 2n = 12 (Fig. 11). Sint.) Hayek] Although Leontodon hispidus ssp. hispidus is a Nomos Ioanninon, Eparchia Metsovou: Mt. Zigos, widespread taxon having broad ecological preferences c. 1.2km after ‘Rachi Kataras’ along road to the with respect to the geological substrate, the karyotype telecommunication station. Wooded slopes and road- examination of a population growing on the serpentine sides. Ophiolithic substrate. Alt. c. 1780m, Lat. parts of Mt. Smolikas is included here because of its 39°46¢ N, Long. 21°13¢ E, 07.vii.1999, Constantinidis unexpected chromosome number. All five plants exam- 8634 (UPA). 2n = 36 (Fig. 13). ined and 13 microphotographs of metaphase plates This Greek endemic species is found in montane taken confirm the chromosome number of 2n = 12. areas of north-west Greece, where it grows mostly This differs from the many previous counts of 2n = 14 on serpentine (Strid & Tan, 1991b). In two popula- for L. hispidus, reported from different countries, e.g. tions examined morphologically, the one cited above Bulgaria (Kuzmanov & Georgieva, 1976), the British and another growing in Pinus forest margins close to Isles (Morton, 1977), Finland (Uotila & Pellinen, the village of Milea, the vast majority of individuals 1985), Spain (Izuzquiza & Nieto Feliner, 1991) and were lacking outer ligulate florets but in some cases Greece (Strid & Franzén, 1981). There are also some these were present in a much-reduced, rudimentary rare reports of triploid plants and some aneuploid form. cases with 2n = 16 or 18 (De Groot, 1977) but, to our The tetraploid chromosome number of 2n = 36 and knowledge, the number 2n = 12 is new for this taxon. the karyotype morphology for Matricaria tempskyana Given the unusual chromosome number counted, the are given here for the first time. Three chromosome specimens were again checked morphologically but no types were observed in the complement: metacentric significant deviation from other forms of L. hispidus and submetacentric chromosomes are dominating, was found, bearing in mind the variability observed in while acrocentric ones are limited to eight pairs. this taxon. Four small satellites mark the short arm of a sub- The karyotype formula of the Smolikas population metacentric and an acrocentric chromosome pair. The is found as 2n = 2m + 6sm + 2sm-SAT + 2st-SAT = 12. karyotype formula of the population examined is given In some plates one of the metacentric chromosomes, as 2n = 14m + 2m/sm + 10sm + 2sm-SAT + 6st + the smallest in size in the complement, tends to be 2st-SAT = 36. submetacentric. The acrocentric satellited chromosome pair is the longest in the complement, while the submetacentric one is the shortest. Our formula is Scorzonera doriae Degen & Bald. close to the general classification of chromosomes Nomos Kastorias, Eparchia Kastorias: c. 5.0–5.8km reported by Rousi (1973) and Pittoni (1974), but the south of Eptachori village. Serpentine slopes, rocks incidence of acrocentric (st) chromosomes given by and scree by the road. Alt. c. 840m, Lat. 40°13¢N,

Long. 20°58¢E, 09.vii.1999, Constantinidis 8683 Pinus brutia forest, scree and dry stream beds, ultra- (UPA). 2n = 12 (Fig. 12). mafic substrate. Alt. c. 320m, Lat. 36°11¢N, Long. Scorzonera doriae is endemic to the western Balkan 27°55¢E, 18.iv.1998, Constantinidis K 219 (UPA). Peninsula. In Greece, it is known from a few localities, 2n = 16. all on serpentine of the North Pindos area (see Lack Endemic to Rodos island and adjacent Turkish & Kilian, 1991 for mountain reports). mainland (Carlström, 1987), Alyssum pogonocarpum The chromosome number and karyotype morphology was found growing on open ultramafic scree and of S. doriae are presented here for the first time. The gravel of central Rodos, together with other interest- species is diploid with 2n = 12. Five out of six chro- ing species such as Aethionema arabicum (L.) O. E. mosome pairs in the complement are metacentric, Schulz, Cleome iberica DC., Linum virgultorum Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 while the longest sixth pair is submetacentric, charac- Planchon, and Silene salamandra. Its chromosome terized by a satellite on the short arm. The karyotype number of 2n = 16 is reported here for the first time. formula is given as 2n = 10m + 2sm-SAT = 12. The somewhat blurred appearance of chromosomes The longest chromosome pair of S. doriae is of in metaphase plates, however, did not permit their particular interest. A wide, poorly stained centromeric illustration. area separates the long from the short arm in each Alyssum pogonocarpum was compared with A. chromosome, and one or two secondary constrictions hirsutum, considered to be its closest ally (Carlström, may be visible on each long arm (Fig. 12). A closer 1984). The two species, in addition to morphologi- examination of other members in S. sect. Vierhappe- cal divergence, also exhibit different chromosome ria Lipsch., to which S. doriae belongs, reveals that S. numbers. Alyssum pogonocarpum is diploid with hirsuta L. from Spain (Díaz de la Guardia & Blanca, 2n = 16, while A. hirsutum is polyploid with 2n = 46 1987) and probably S. biebersteinii Lipsch. from the or 48 (see Ancˇev & Dudley, 1981, and reference in Caucasus area (Nazarova, 1997) have a karyotype Goldblatt & Johnson, 1994). morphology similar to that of S. doriae. Long, submetacentric chromosome pairs are not usually found in Scorzonera members with 2n = 14, where chromo- Hesperis laciniata All. ssp. laciniata somes bearing satellites are among the smallest in Nomos Ioanninon, Eparchia Konitsis: Mt. Smolikas, the complement. A comparison of the chromosome the south-east lower parts of the mountain, c. 3.4km idiograms given by Nazarova (1997) is even more after the village of Distraton along road to Samarina. informative. Long, submetacentric chromosome pairs Ophiolithic scree. Alt. c. 1050m. Lat. 40°02¢ N, Long. bearing a satellite are also found in n = 6 karyotypes 21°00¢E, 11.viii.1998, Constantinidis & Garofalo of sections Pulvinares (Boiss.) Lipsch. and Nervosae (seeds collected, voucher: K242, UPA). 2n = 12 Lipsch. Remarkable similarities in chromosome struc- (Fig. 14). ture between S. hirsuta and S. hispanica L. have Most populations of Hesperis laciniata are found already urged Díaz de la Guardia & Blanca (1987) to on calcareous soils, usually in rocky places, gravel and propose that each long submetacentric chromosome in scree. Populations growing on serpentine occur more S. hirsuta may be the result of a reciprocal transloca- rarely, at least in Greece. Morphological comparison of tion combining two smaller, metacentric chromo- serpentine and limestone collections of H. laciniata somes. Here, the same procedure of metacentric ssp. laciniata did not reveal any noteworthy (usually marked with satellites) chromosome fusion in differences. Scorzonera members with n = 7 to yield long, sub- The chromosome number of 2n = 12, found in metacentric chromosomes in Scorzonera members three plants of H. laciniata ssp. laciniata from Mt. with n = 6 is given a more general perspective. If n = 7 Smolikas, is the result of the first karyological in Scorzonera is accepted as ancestral (Nazarova, examination of a Greek population. The karyotype 1997), then n = 6 is derived and secondary. Chromo- analysis revealed the formula of 2n = 6m + somal rearrangements such as chromosome fusion 2m-SAT + 2sm + 2sm/st-SAT = 12, with satellites should have played an important role in the evolution usually difficult to observe on the contracted chromo- of particular Scorzonera sections. Obviously, they com- somes of metaphase plates. This diploid number is plement the general evolutionary scheme proposed by in accordance with the reports given by Ancˇev & Nazarova (1997). Goranova (1997) for H. laciniata ssp. laciniata and by Constantinidis & Kamari (1994) for H. laciniata ssp. CRUCIFERAE secundiflora (Boiss. & Spruner) Breistr. Small differences between the above-cited reports and this study Alyssum pogonocarpum A. Carlström centre on the relative position of the satellites and the Nomos Dodekanisou, Eparchia Rodou: Rodos island, classification of one chromosome pair as submetacen- c. 4.4km north-east of the village of Laerma. Sparse tric or subtelocentric. The gametic number of n = 6,

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 KARYOTAXONOMY OF GREEK SERPENTINE ANGIOSPERMS 119 Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 14 15

16 17

18 19

Figures 14–19. Photomicrographs of mitotic metaphases. Scale bars = 10 mm. Fig. 14. Hesperis laciniata ssp. laciniata (2n = 12). Fig. 15. Leptoplax emarginata (2n = 16). Fig. 16. Cephalaria fanourii (2n = 18). Fig. 17. Thymus teucrioides ssp. candilicus (2n = 28). Figure 18. Ranunculus pedatus (2n = 16). Fig. 19. Trinia glauca ssp. pindica (2n = 18). given by Ruíz de Clavijo (1994) for material from although its sole species may occasionally be found in Spain should be better attributed to H. l. ssp. the bibliography as belonging to Peltaria Jacq., under spectabilis (Jord.) Rouy & Foucaud and not to H. l. ssp. the name of P. emarginata (Boiss.) Hausskn. Leptoplax laciniata, especially if the work on the genus by is endemic to Greece, distributed from the area of Dvorˇák, 1971) is taken into account. North Pindos to the north, down to Mt. Gerania in the south. The genus is also found close to the border with Albania, where is has not yet been reported (see Leptoplax emarginata (Boiss.) O. E. Schulz Paparisto et al., 1988), so it should be sought there. Nomos Fthiotidos, Eparchia Domokou: c. 1km south Leptoplax is always associated with serpentine and of Ekkara village along road to Ano Agoriani village. can be considered as a serpentine endemic. Stony places by a small stream. Ophiolithic substrate. The chromosome number of 2n = 16 and a micropho- Alt. c. 200 m, Lat. 39°08¢ N, Long. 22°11¢ E, 26.vi.1997, tograph of the karyotype of Leptoplax are reported Constantinidis 7020 (UPA). 2n = 16 (Fig. 15). here for the ﬁrst time. The species is diploid. Chro- Leptoplax O. E. Schulz is usually considered an mosomes are small and appear to be either meta- independent, monotypic genus (e.g. Hartvig, 1986a), centric or submetacentric but clear identiﬁcation is

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 120 T. CONSTANTINIDIS ET AL. difﬁcult. A satellite appears on one metacentric chro- match any of the Cephalaria karyotype formulae given mosome but its homologue is not usually visible. by Verlaque (1985) and adds to the karyotypic hetero- Leptoplax does not have the same chromosome geneity of the genus. number as Peltaria, in which several counts have given the stable number of 2n = 14 or the tetraploid 2n = 28 (see Fedorov, 1969; Van Loon, 1987 for LABIATAE references). This difference in base chromosome Thymus teucrioides Boiss. & Spruner ssp. candilicus number supports the splitting of Leptoplax from (Beauv.) Hartvig Peltaria, although there are cases in Cruciferae

Nomos Fthiotidos, Eparchia Domokou: c. 5km south Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 where more than one chromosome base-number of Ekkara village along road to Ano Agoriani village. exists in the same genus. In contrast, Leptoplax has Stony slopes, field margins and road sides. Ophiolithic the same chromosome number as Bornmuellera substrate. Alt. c. 460m, Lat. 39°07 N, Long. 22°12 E, Hausskn., something that may facilitate hybridiza- ¢ ¢ 26.vi.1997, Constantinidis 7005 (UPA). 2n 28 tion, pollen fertility in hybrids and introgression = (Fig. 17). between species of the two genera, a process that This is the second chromosome count of Thymus has already been observed in nature and reported by teucrioides ssp. candilicus, a Greek neoendemic taxon Hartvig (1986a). apparently found exclusively on serpentine. The first count, originating from a population growing on the serpentine parts of Mt. Gerania, gave the number of DIPSACACEAE 2n = 30 (Constantinidis et al., 1997). The new count of Cephalaria fanourii Perdetzoglou & Kit Tan 2n = 28 adds a different chromosome number to this taxon and simultaneously addresses the case of dys- Nomos Ioanninon/Kastorias, Eparchia Konitsis/ ploidy in Thymus. Unfortunately, karyotype morphol- Kastorias: Mt. Gramos, upper parts of Bouchetsi ogy is not possible to determine from the Domokos summit, c. 100–400 m north-north-west of the highest population. Lack of chromosome details is due mostly summit. Steep, east-facing, treeless serpentine slopes to the small size of chromosomes in Thymus, and does and scree. Alt. c. 1680m, Lat. 40°14 N, Long. 20°57 E, ¢ ¢ not allow any direct karyotype comparison among 13.ix.2000, Constantinidis & Kyparissis 9150 (UPA). populations. 2n 18 (Fig. 16). = To our knowledge, no other chromosome counts A rare serpentine endemic, Cephalaria fanourii is are known in Th. sect. Teucrioides besides those of known, up to now, from a single locality, the serpen- Thymus teucrioides ssp. candilicus. Different chromo- tine top area of Mt. Bouchetsi, where it has been col- some numbers in the same Thymus species, similar lected independently by P. Hartvig and colleagues, and to the case reported here, are not exceptional. For by D. Perdetzoglou (Perdetzoglou & Tan, 1995). example, Th. praecox Opiz has been found to have According to Perdetzoglou & Tan (1995), C. fanourii 2n 54, 56–58 (Jalas & Pohjo, 1965 as T. humifusus belongs to C. sect. Leucocephalae. This section is a = Bernh.; Trela-Sawicka, 1972) and Th. glabrescens member of C. subgen. Denticarpus, which supposedly Willd. ( Th. odoratissimus Miller) has 2n 28, 32, 56, includes species whose achenes have eight apical = = 58 (Trela-Sawicka, 1968). A cytogeographical investi- teeth (‘achenium apice dentibus 8 minoribus vel lon- gation of Th. sect. Teucrioides in Greece, where it is gioribus’, see Szabó, 1940). This feature obviously does largely distributed, might be of interest. not apply to C. fanourii or to its relatives (as defined by Perdetzoglou & Tan, 1995) C. laevigata (Waldst. & Kit.) Schrader and C. coriacea (Willd.) Steud. In C. RANUNCULACEAE fanourii there are four small, apical teeth on the achene, alternating with four minute ones that may Ranunculus pedatus Waldst. & Kit. even be absent. C. laevigata achenes have no apical Nomos Fthiotidos, Eparchia Domokou: About 5.2km teeth at all, while those of C. coriacea also lack teeth south-south-east of Ekkara village, along road to or may rarely have minute ones. Thus, the character Ano Agoriani. Slopes with Quercus coccifera, Junipe- of apical teeth on the achene allows reliable distinc- rus oxycedrus and Cistus spp., small meadows and tion between C. fanourii and its related taxa. margins of fields. Ophiolithic substrate. Alt. c. The chromosome number of C. fanourii, reported 580–620m, Lat. 39°07¢ N, Long. 22°12¢ E, 18.iii.1998, here for the first time, is 2n = 18, the most common Constantinidis 7386 (UPA). 2n = 16 (Fig. 18). number in Cephalaria (Verlaque, 1985). Karyotype Nokos Fthiotidos, Eparchia Lokridos: Mt. morphology is given as 2n = 8m + 2m-SAT + 2sm + Kallidromon, c. 8.3km after the village of Elatia, along 2sm-SAT + 4st = 18. The satellites are large and forest road to Reginion. Slopes with Quercus coccifera, clearly visible. This chromosome formula does not Arbutus andrachne, Pistacia lentiscus, etc. Ophiolithic

© 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 2002, 139, 109–124 KARYOTAXONOMY OF GREEK SERPENTINE ANGIOSPERMS 121 substrate. Alt. c. 630m, Lat. 38°40¢N, Long. 22°46¢E, Trinia glauca is not usually given or, when given, 23.iv.1998, Constantinidis 7595 (UPA). does not permit an unambiguous classiﬁcation of A rare species in Greece, Ranunculus pedatus was chromosomes. known with certainty until now from a single collection on the ophiolithic summit area of Mt. Kratsovon in the North Pindos area (Strid, 1986b). Our collec- CONCLUSIONS tions from serpentine areas extend the distribution In certain cases, karyological information did not of the species in Greece further south, and to a lower provide evidence of taxonomic importance between altitude. It is expected that more populations will

serpentine species and their relatives, as, e.g. in sec- Downloaded from https://academic.oup.com/botlinnean/article-abstract/139/1/109/2433713 by guest on 24 November 2019 be found if serpentine areas are carefully searched in tions of Silene, where chromosome number and kary- early spring. otype morphology tend to be conservative features. The chromosome number of 2n = 16 is counted on Differences were found in other groups of taxa, and Greek material of R. pedatus for the first time. The were referring to ploidy level (diploid Alyssum pogono- two populations examined present a similar kary- carpum vs. polyploid A. hirsutum), chromosome num- otype, which has the following formula: 2n = 6m + ber (between the genera Leptoplax and Peltaria) 2sm + 2sm-SAT + 2st + 2st-SAT + 2t-SAT = 16. The or karyotype morphology (Onosma elegantissima vs. satellites are small. The chromosome number is in O. mattirolii). Differences also exist among popula- agreement with reports from other countries, e.g. tions of the same taxon (dysploidy in a serpentine Slovakia (Májovsky´ et al., 1978). The tetraploid population of Leontodon hispidus ssp. hispidus). These number of 2n = 32 is also reported for this species (see are indications of karyotype differentiation and may Goldblatt, 1988 for reference). The karyotype formula reflect taxonomic relationships between serpentine of R. pedatus given here conforms to the general species and their allies that are more distant than karyotype scheme of R. sect. Ranunculastrum pre- were considered previously. For Onosma, Crepis and sented by Goepfert (1974). Leptoplax in particular, new taxonomic relationships are proposed, which are in concordance with karyological data and observations in the field. UMBELLIFERAE In other examples, chromosome number and Trinia glauca (L.) Dumort ssp. pindica Hartvig karyotype morphology of serpentine taxa are related to evolutionary modifications characterizing supra- Nomos Ioanninon, Eparchia Dodonis/Metsovou: Mts. specific taxonomic units (e.g. common karyotype Aftia/Flega. c. 16.5km after turn to Milea of the road morphology of Onosma subsect. Asterotricha, chro- Ioannina-Trikala and along a secondary road that mosomal rearrangements in Scorzonera sects. Vier- leads to the upper parts of the mountain. Mixed happeria, Pulvinares and Nervosae). These shared Fagus-Pinus forest and small clearings. Ophiolite. Alt. evolutionary features may be irrelevant to the geologi- c. 1650m, Lat. 39°52¢ N, Long. 21°06¢E, 28.vii.1999, cal substrate on which plant taxa grow today, or may Constantinidis 8663 (UPA). 2n = 18 (Fig. 19). precede adaptation and specialization on serpentine. Originally described as a distinct taxon by Hartvig Our results so far indicate that karyotype changes (1986b) and distributed in Greece and Albania, Trinia in serpentine taxa, compared to their close allies, glauca ssp. pindica is known to grow both on ser- follow no strict modification patterns. The reaction of pentine and limestone substrates of several Greek plants to the serpentine factor may vary, and concen- mountains. Its chromosome number and karyotype tration on particular taxonomic groups in the future morphology are presented here for the first time. will afford more precise information on karyotypic This taxon is diploid with 2n = 18. The karyotype differentiation enhanced by serpentine. consists mostly of metacentric chromosomes, with a pair of very clear satellites marking a metacentric/ submetacentric chromosome pair. The karyotype ACKNOWLEDGEMENTS formula can be described as 2n = 12m + 2m/sm + 2m/sm-SAT + 2sm = 18. This work was partially supported by a grant from the The chromosome number of 2n = 18 in Trinia glauca Greek Ministry for Development, General Secretariat ssp. pindica agrees with several chromosome reports for Research and Technology (Project EPET II, no. on T. glauca ssp. glauca from various European coun- 9513514). tries (Kieft & Van Loon, 1978; Váchová & Lhotská, 1978; Váchová & Lhotska, 1980; Van Loon, 1980; REFERENCES Bellomaria & Hrusˇka, 1983; Nikolov, 1991) or Trinia glauca s.l. (Van Loon & Van Setten, 1982; Castroviejo Ancˇev ME, Dudley TR. 1981. Reports. In: Löve Á, ed. IOPB & Nieto Feliner, 1986). Karyotype illustration in chromosome number reports LXXIII. Taxon 30: 856.

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