Chromosome number. 2n = 22 (Fig. 1)

Voucher specimen. ITALY. Toscana. Cerbaie (Pisa) (WGS84: 43.751228 N, 10.719234 E), 55 m, 7 May 2015, D. Dolci (PI n. 000455, 000456, 000457, 000458). Lazio. Torano (Rieti) (WGS84: 42.157098 N, 13.270760 E), 760 m, 30 April 2015, D. Dolci (PI n. 000453, 000454); Vallerotonda (Frosinone) (WGS84: 41.588942 N, 14.007237 E), 765 m, 3 May 2015, D. Dolci (PI n. 000459, 000460).

Figure 1. 

Polygala flavescens DC. subsp. flavescens, 2n = 22 from Cerbaie, Tuscany (a), Torano, Lazio (b), and Vallerotonda, Lazio (c). Scale bar: 10 μm.

Method. Squash preparations were made on root-tips taken from plants cultivated in the Botanical Garden of Pisa and obtained from germinating seeds collected in the field. Root-tips were pre-treated with 0.4% colchicine for 3 hours and then fixed in Carnoy fixative solution for 1 hour. After hydrolysis in HCl 1N at 60° C, the tips were stained in leuco-basic fuchsine.

Observations.Polygala flavescens subsp. flavescens is an Italian endemic taxon, described from central Italy and currently recorded from Emilia Romagna to Basilicata (Conti et al. 2005, Del Guacchio 2010). These are the first chromosome counts for this species (Bedini et al. 2010 onwards), and they also represent the first records of 2n = 22 cytotypes in the genus Polygala L. (Rice et al. 2014).

Chromosome number. 2n = 22 (Fig. 2)

Voucher specimen. ITALY. Toscana. Monte Argentario (Grosseto) (WGS84: 42.421952 N, 11.140779 E), 130 m, 6 May 2015, D. Dolci (PI n. 000466, 000467, 000468, 000469).

Figure 2. 

Polygala flavescens subsp. maremmana (Fiori) Arrigoni, 2n = 22. Scale bar: 10 μm.

Method. Squash preparations were made on root-tips taken from plants cultivated in the Botanical Garden of Pisa and obtained from germinating seeds collected in the field. Root-tips were pre-treated with 0.4% colchicine for 3 hours and then fixed in Carnoy fixative solution for 1 hour. After hydrolysis in HCl 1N at 60° C, the tips were stained in leuco-basic fuchsine.

Observations.Polygala flavescens subsp. maremana is an Italian endemic, originally described as a form based on plants from Mt. Argentario (Peruzzi et al. 2015). Currently, it is recorded from the coasts of southern Tuscany, from San Vincenzo (Leghorn) to Capalbio (Grosseto) (Arrigoni 2014). Our chromosome count, performed on plants from the locus classicus, is the first for this subspecies (Bedini et al. 2010 onwards), and it agrees with the chromosome number reported above for P. flavescens subsp. flavescens.

Chromosome number. 2n = 22 (Fig. 3)

Voucher specimen. ITALY. Marche. Fano (Pesaro e Urbino) (WGS84: 43.864231 N, 12.984113 E), 25 m, 24 April 2015, D. Dolci (PI n. 000461, 000462).

Figure 3. 

Polygala flavescens subsp. pisaurensis (Caldesi) Arcang., 2n = 22. Scale bar: 10 μm.

Method. Squash preparations were made on root-tips taken from plants cultivated in the Botanical Garden of Pisa and obtained from germinating seeds collected in the field. Root-tips were pre-treated with 0.4% colchicine for 3 hours and then fixed in Carnoy fixative solution for 1 hour. After hydrolysis in HCl 1N at 60° C, the tips were stained in leuco-basic fuchsine.

Observations.Polygala flavescens subsp. pisaurensis is an Italian endemic, originally described as a species based on plants from Pesaro (Peruzzi et al. 2015). Currently, it is recorded for coastal hills of Emilia-Romagna and Marche (Arrigoni 2014). Our chromosome count, performed on plants from the locus classicus area, is the first for this subspecies (Bedini et al. 2010 onwards), and it agrees with the chromosome numbers reported above for P. flavescens subsp. flavescens and P. flavescens subsp. maremmana.

F. Roma-Marzio, D. Dolci, L. Peruzzi

Chromosome number. 2n = 30 (Fig. 4)

Voucher specimen. ITALY. Calabria. Aspromonte, Roccaforte del Greco (Reggio Calabria), near the Menta’s artificial basin (WGS84: 38.12427 N, 15.90080 E), metamorphic rock outcrops along the road, 1400 m, 21 August 2017, L. Peruzzi & K.F. Caparelli (seeds collected in the field and stored in the Germplasm Bank of Department of Biology, Pisa, under acc. n. PI-20170073).

Figure 4. 

Dianthus brutius Brullo, Scelsi & Spamp. subsp. brutius, 2n = 30. Scale bar: 10 μm.

Method. Squash preparations were made on root-tips obtained from germinating seeds collected in the field. Root-tips were pre-treated with 0.4% colchicine for 3 hours and then fixed in Carnoy fixative solution for 1 hour. After hydrolysis in 1N HCl at 60° C, the tips were stained in leuco-basic fuchsine.

Observations.Dianthus brutius subsp. brutius is endemic to Calabria (Peruzzi et al. 2014), occurring in the mountain part of the Aspromonte area, and it belongs to D. vulturius Guss. & Ten. group (Brullo et al. 2000). This is the first chromosome count for this species, and the number is consistent with the basic chromosome number (x = 15) typical for the genus Dianthus L. (Bedini et al. 2010 onwards; Rice et al. 2014).

Chromosome number. 2n = 30 (Fig. 5)

Voucher specimen. ITALY. Calabria. Melito di Porto Salvo (Reggio Calabria), Pentidattilo (WGS84: 37.95946 N; 15.76115 E), conglomerate cliffs, 300 m, 21 August 2017, L. Peruzzi & K.F. Caparelli (seeds collected in the field and stored in the Germplasm Bank of Department of Biology, Pisa, under acc. n. PI-20170072).

Figure 5. 

Dianthus brutius subsp. pentadactyli Brullo, Scelsi & Spamp., 2n = 30. Scale bar: 10 μm.

Method. Squash preparations were made on root-tips obtained from germinating seeds collected in the field. Root-tips were pre-treated with 0.4% colchicine for 3 hours and then fixed in Carnoy fixative solution for 1 hour. After hydrolysis in 1N HCl at 60° C, the tips were stained in leuco-basic fuchsine.

Observations.Dianthus brutius subsp. pentadactyli is endemic to Calabria (Peruzzi et al. 2014), occurring in the lowest part of the Ionian slope of Aspromonte area (Brullo et al. 2000). This is the first chromosome count for this subspecies (Bedini et al. 2010 onwards), and it agrees with the chromosome number reported above for D. brutius subsp. brutius.

Chromosome number. 2n = 30 (Fig. 6)

Voucher specimen. ITALY. Calabria. Montebello Jonico (Reggio Calabria), along the road to Masella (WGS84: 37.97780 N, 15.75978 E), cliffs, 320 m a.s.l., 21 August 2017, L. Peruzzi & K.F. Caparelli (PI).

Figure 6. 

Dianthus vulturius Guss. & Ten. subsp. aspromontanus Brullo, Scelsi & Spamp., 2n = 30. Scale bar: 10 μm.

Method. Squash preparations were made on root-tips obtained from germinating seeds collected in the field. Root-tips were pre-treated with 0.4% colchicine for 3 hours and then fixed in Carnoy fixative solution for 1 hour. After hydrolysis in 1N HCl at 60° C, the tips were stained in leuco-basic fuchsine.

Observations.Dianthus vulturius subsp. aspromontanus is endemic to Calabria (Peruzzi et al. 2014). It is distributed only in a small area of southern Aspromonte, growing as a chasmophyte (Brullo et al. 2000). This is the first chromosome count for this species, and it is consistent with chromosome numbers reported above for the related D. brutius subsp. brutius and D. brutius subsp. pentadactyli.

Chromosome number. 2n = 24 (Fig. 7)

Voucher specimen. ITALY. Calabria. Melito di Porto Salvo (Reggio Calabria), Pentidattilo (WGS84: 37.95278 N, 15.76156 E), cliffs along the road, 255 m a.s.l., 21 August 2017, Lorenzo Peruzzi & K.F. Caparelli (seeds collected in the field and stored in the Germplasm Bank of Department of Biology, Pisa, under acc. n. PI-20170071).

Figure 7. 

Silene calabra Brullo, Scelsi & Spamp., 2n = 24. Scale bar: 10 μm.

Method. Squash preparations were made on root-tips obtained from germinating seeds collected in the field. Root-tips were pre-treated with 0.4% colchicine for 3 hours and then fixed in Carnoy fixative solution for 1 hour. After hydrolysis in 1N HCl at 60° C, the tips were stained in leuco-basic fuchsine.

Observations. Silene calabra is endemic to Calabria (Peruzzi et al. 2014). According to Brullo et al. (1997), this species belongs to S. sect. Siphonomorpha Otth., and it shows a close morphological relationship mainly with taxa of the S. mollissima (L.) Pers. group. This is the first chromosome count reported for this species, and it agrees with that reported for the closely related S. oenotriae Brullo (Peruzzi et al. 2007), and with those reported for other taxa within S. sect. Siphonomorpha (Naciri et al. 2017, and literature cited therein).

R.B. Chiavegatto, M. D’Antraccoli, F. Roma-Marzio, L. Peruzzi