Chromosome numbers for the Italian flora: 9

Abstract

In this contribution, new chromosome data obtained on material collected in Italy are presented. It includes counts from six populations of three taxa within the genus Pulmonaria , two of which are endemic to Italy (P. vallarsae subsp. apennina and P. vallarsae subsp. vallarsae); the other is the widespread European P. officinalis . In addition, two counts from Potentilla detommasii and Stachys thirkei , two eastern Mediterranean species, are also reported.

Full text

Chromosome numbers for the Italian ora 101
Chromosome numbers for the Italian flora: 9
Giovanni Astuti1, Gianni Bedini1, Daniela Ciccarelli1,
Lijuan Liu1, Manuel Tiburtini1, Lorenzo Peruzzi1
1 Dipartimento di Biologia, Università di Pisa, Via Derna 1, 56126 Pisa, Italy
Corresponding author: Giovanni Astuti (gastuti@biologia.unipi.it)
Academic editor: G. Domina  |  Received 2 June 2020 |  Accepted 12 June 2020  |  Published 8 July 2020
Citation: Astuti G, Bedini G, Ciccarelli D, Liu L, Tiburtini M, Peruzzi L (2020) Chromosome numbers for the Italian
ora: 9. Italian Botanist 9: 101–110. https://doi.org/10.3897/italianbotanist.9.54973
Abstract
In this contribution, new chromosome data obtained on material collected in Italy are presented. It in-
cludes counts from six populations of three taxa within the genus Pulmonaria, two of which are endemic
to Italy (P. vallarsae subsp. apennina and P. vallarsae subsp. vallarsae); the other is the widespread European
P. ocinalis. In addition, two counts from Potentilla detommasii and Stachys thirkei, two eastern Mediter-
ranean species, are also reported.
Keywords
cytogeography, cytotaxonomy, Emilia-Romagna, Friuli Venezia Giulia, Marche, Toscana, Trentino-Alto Adige
How to contribute
Texts concerning new chromosome data should be submitted electronically to Gio-
vanni Astuti (gastuti@biologia.unipi.it), including indications on voucher specimens
and methods used.
Italian Botanist 9: 101–110 (2020)
doi: 10.3897/italianbotanist.9.54973
http://italianbotanist.pensoft.net
Copyright Giovanni Astuti et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC
BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
RESEARCH ARTICLE

Giovanni Astuti et al. / Italian Botanist 9: 101–110 (2020)
102
Chromosome counts
Boraginaceae
Pulmonaria vallarsae A.Kern. subsp. apennina (Cristof. & Puppi) Cecchi & Selvi
Chromosome number. 2n = 22 (Figs 1a, 2)
Voucher specimen. Italy. Toscana. Molino del Pallone (Sambuca Pistoiese, Pis-
toia), sulla sponda toscana del ume Reno (WGS84: 44.100658N, 10.962573E),
margine di bosco, ca. 500 m s.l.m., 5 April 2019, L. Liu & G. Astuti (PI n° 034194–
034202); Passo del Muraglione (San Godenzo, Firenze), sulla SS67 scendendo verso
San Benedetto in Alpe (WG84: 43.935302N, 11.658088E), ca. 900 m s.l.m., 9 April
2019, L. Liu & G. Astuti (PI n° 034203–034214).
Chromosome number. 2n = 28 (Fig. 1b)
Voucher specimen. Italy. Toscana. Molino del Pallone (Sambuca Pistoiese, Pis-
toia), sulla sponda toscana del ume Reno (WGS84: 44.100658N, 10.962573E),
margine di bosco, ca. 500 m s.l.m., May 2015, P. Pupillo (plant only temporarily cul-
tivated at the Botanic Garden of Pisa University).
Method. Squash preparations were made on root tips obtained from potted plants.
Root tips were pre-treated with 0.4% colchicine for 3 hours and then xed in Carnoy
xative solution for 1 hour. After hydrolysis in HCl 1N at 60 °C, the tips were stained
in leuco-basic fuchsine.
Observations. e overall morphology of these plants is congruent with P. vallar-
sae subsp. apennina, a subspecies endemic to peninsular Italy (Cecchi and Selvi 2015).
In the population from Molino del Pallone we found ten individuals showing 2n = 22
(Fig. 1a) and one individual with 2n = 28 chromosomes (Fig. 1b). Vosa and Pistolesi
(2004) found a few individuals with an intermediate chromosome number (2n = 26)
Figure 1. Pulmonaria vallarsae A.Kern. subsp. apennina (Cristof. & Puppi) L.Cecchi & Selvi from Moli-
no del Pallone (Sambuca Pistoiese, Pistoia), 2n = 22 (a) and 2n = 28 (b). Scale bar: 10 μm.

Chromosome numbers for the Italian ora 103
among plants showing 2n = 22 chromosomes at Passo del Muraglione. However, in the
present study, we found there only plants with 2n = 22 chromosomes (12 individuals
studied). In both cases, we highlight the rarity of chromosome numbers dierent from
2n = 22, which is typical of P. vallarsae subsp. apennina, although a population showing
2n = 26 chromosomes has been recently recorded from Abruzzo (Astuti et al. 2019).
Pulmonaria vallarsae A.Kern. subsp. vallarsae
Chromosome number. 2n = 22 (Figs 3, 4)
Voucher specimen. Italy. Trentino-Alto Adige. Pian delle Fugazze (Vallarsa,
Trento) (WGS84: 45.760208N, 11.171882E), ca. 1160 m s.l.m., 17 April 2019, L.
Liu & G. Astuti (PI n° 034215–034219); Alla sorgente del Cop, Bellaria di Cei (Villa
Lagarina, Trento) (WGS84: 45.96022N, 11.04163E), ca. 900 m s.l.m., 17 July 2019,
L. Liu & G. Astuti (PI n° 034220–034222).
Method. Squash preparations were made on root tips obtained from potted plants.
Root tips were pre-treated with 0.4% colchicine for 3 hours and then xed in Carnoy
xative solution for 1 hour. After hydrolysis in HCl 1N at 60 °C, the tips were stained
in leuco-basic fuchsine.
Observations. Pulmonaria vallarsae subsp. vallarsae is endemic to Trentino-Alto
Adige and Veneto (Cecchi and Selvi 2015), and it was described by Kerner (1878) on
plants occurring in Vallarsa, the valley of the river Leno situated southeast of Rovereto,
which is the locus classicus for this species (Puppi and Cristofolini 1996). Pian delle Fu-
gazze, the northern summit of the valley, is among the localities cited in the protologue.
Figure 2. Pulmonaria vallarsae A.Kern. subsp. apennina (Cristof. & Puppi) L.Cecchi & Selvi from Passo
del Muraglione (San Godenzo, Firenze), 2n = 22. Scale bar: 10 μm.

Giovanni Astuti et al. / Italian Botanist 9: 101–110 (2020)
104
Figure 3. Pulmonaria vallarsae A.Kern. subsp. vallarsae from Pian delle Fugazze (Vallarsa, Trento),
2n=22. Scale bar: 10 μm.
Figure 4. Pulmonaria vallarsae A.Kern. subsp. vallarsae from Bellaria di Cei (Villa Lagarina, Trento),
2n=22. Scale bar: 10 μm.
All the ve individuals sampled here displayed the typical chromosome number of P.
vallarsae (including P. vallarsae subsp. apennina), 2n = 22, without any variation. is
chromosome number was also found in the three individuals sampled in the other pop-
ulation at Bellaria di Cei, which is ca. 40 km far from the locus classicus of this species.

Chromosome numbers for the Italian ora 105
Pulmonaria hirta L.
Chromosome number. 2n = 28 (Fig. 5)
Voucher specimen. Italy. Emilia-Romagna. Parco di Monte Paderno (Bologna)
(WGS84: 44.452272N, 11.320769E), ca. 280 m s.l.m., 3 April 2019, L. Liu & G.
Astuti (PI n° 034223–034226).
Chromosome number. 2n = 22 (Fig. 6)
Voucher specimen. Italy. Emilia-Romagna. Parco di Monte Paderno (Bologna)
(WGS84: 44.452272N, 11.320769E), ca. 280 m s.l.m., 3 April 2019, L. Liu & G.
Astuti (PI n° 034227–034228).
Method. Squash preparations were made on root tips obtained from potted plants.
Root tips were pre-treated with 0.4% colchicine for 3 hours and then xed in Carnoy
xative solution for 1 hour. After hydrolysis in HCl 1N at 60 °C, the tips were stained
in leuco-basic fuchsine.
Observations. ese plants display intermediate morphological features between
the typical P. hirta and the typical P. vallarsae subsp. apennina (Puppi and Cristofolini
1996; Cecchi and Selvi 2015), although showing a closer resemblance to the former
species, whose range spreads from SE France to C Italy (Cecchi and Selvi 2015). Four
out of six samples in this population were found to have 2n = 28 chromosomes, which
is typical for P. hirta (Astuti et al. 2019; Pupillo et al. 2019), whereas the remaining
two individuals were found to have 2n = 22 chromosomes. Plants with 2n =22 and 28
chromosomes grow together and do not show any pattern relating chromosome num-
ber to a specic morphological syndrome, as already observed by Vosa and Pistolesi
(2004) for other populations.
Figure 5. Pulmonaria hirta L. from Parco di Monte Paderno (Bologna), 2n = 28. Scale bar: 10 μm.

Giovanni Astuti et al. / Italian Botanist 9: 101–110 (2020)
106
Figure 6. Pulmonaria hirta L. from Parco di Monte Paderno (Bologna), 2n = 22. Scale bar: 10 μm.
Pulmonaria ocinalis L. subsp. ocinalis
Chromosome number. 2n = 16 (Fig. 7)
Voucher specimen. Italy. Friuli Venezia Giulia. Lungo la SP31 sotto il santuario
di Castelmonte (Prepotto, Udine), (WGS84: 46.092828N, 13.516041E), ca. 580 m
s.l.m., 21 March 2019, L. Liu & G. Astuti (PI n° 034229–034237).
Observations. Plants in this population show some morphological features that
are intermediate between P. ocinalis and P. vallarsae, especially in terms of leaf shape
and maculation. However, the cordate leaf base and the presence of aculeoli (very short
Figure 7. Pulmonaria ocinalis L. subsp. ocinalis from Castelmonte (Prepotto, Udine), 2n = 16.
Scale bar: 10 μm.

Chromosome numbers for the Italian ora 107
conical hairs) clearly place these plants within P. ocinalis subsp. ocinalis (Bolliger
1982), a taxon that is widespread in Europe. is is also conrmed by the chromo-
some number 2n = 16, found in all nine individuals studied, which is typical of P.
ocinalis (Sauer 1975; Bolliger 1982). Because of the faint maculation of most of the
plants, these plants resemble P. ocinalis subsp. marzolae G.Astuti, Peruzzi, Cristof.
& P.Pupillo, known so far for Trentino-Alto Adige only. However, the presence of ac-
uleoli in the population studied here does not match with their absence in P. ocinalis
subsp. marzolae (Astuti et al. 2014). erefore, these plants could be safely ascribed to
a faintly spotted form of P. ocinalis.
L. Liu, G. Astuti, G. Bedini, D. Ciccarelli, L. Peruzzi
Rosaceae
Potentilla detommasii Ten.
Chromosome number. 2n = 14 (Fig. 8)
Voucher specimen. Italy. Basilicata. Belvedere del Malvento (Viggianello, Po-
tenza), (WGS84: 39.902987N, 16.136537E), ca. 1620 m s.l.m., 13 August 2018, L.
Peruzzi (seeds collected in the eld and stored at the Seed Bank of the Department of
Biology, University of Pisa).
Method. Squash preparations were made on root tips obtained from germinating
seeds. Root tips were pre-treated with 0.4% colchicine for 3 hours and then xed in
Carnoy xative solution for 1 hour. After hydrolysis in HCl 1N at 60 °C, the tips were
stained in leuco-basic fuchsine.
Observations. is species spreads from peninsular Italy to Turkey, but it is not
recorded for Croatia and Montenegro (Kurtto 2009). Here we report the rst chromo-
some count for P. detommasii from Italy, which agrees with the only other count avail-
able for this species (Markowa 1971), conrming its diploid status.
G. Astuti, L. Peruzzi
Figure 8. Potentilla detommasii Ten. from Belvedere del Malvento (Viggianello, Potenza), 2n = 14.
Scale bar: 10 μm.

Giovanni Astuti et al. / Italian Botanist 9: 101–110 (2020)
108
Lamiaceae
Stachys thirkei K.Koch
Chromosome number. 2n = 30 (Fig. 9)
Voucher specimen. Italy. Marche. Monte Rosato (Fermo) (WGS84: 43.129556N,
13.702611E), prato arido a sud del bosco su substrato argilloso, 88 m, 6 June 2015,
M. Tiburtini (FI).
Method. Squash preparations were made on root tips obtained from cultivated
plants. Root tips were pre-treated with 0.4% colchicine for 4 h and then xed in Car-
noy solution for 1 h. After hydrolysis in 1N HCl at 60 °C, the tips were stained with
leuco-basic fuchsine.
Observations. is species grows in the eastern Mediterranean, showing a main
distribution range in the Balkans, but extending eastwards to Turkey and westwards to
Italy (Ball 1972). is species has been recorded for Marche (Bartolucci et al. 2019),
where it has been found in only two localities. e specimen investigated here comes
from one of these localities, and our count agrees with a previous report for Italy
(Falciani and Fiorini 1996). is chromosome number is also typical of all the species
belonging to Salvia sect. Eriostomum (Homans. & Link) Dumort. (Falciani 1997).
G. Astuti, M. Tiburtini
Acknowledgements
Many thanks are due to the following people for their help in locating populations of
Pulmonaria: Paolo Pupillo and Giancarlo Marconi (Bologna) for Molino del Pallone,
Parco di Monte Paderno and Pian delle Fugazze; Filippo Prosser (Rovereto) for Bellaria
di Cei; Giuseppe Pistolesi (Pisa) for Passo del Muraglione. We are also indebted to Sil-
via Zublena and Francesco Roma-Marzio at the Botanic Garden and Museum of Pisa
Figure 9. Stachys thirkei K.Koch, 2n = 30 from Monte Rosato (Fermo). Scale bar: 10 μm.

Chromosome numbers for the Italian ora 109
for their help in collecting root tips of Stachys thirkei. e authors gratefully acknowl-
edge the nancial support of “Progetto di Ricerca di Ateneo” (PRA) of the University
of Pisa, grant number PRA_2018_15, for the chromosome counts of Pulmonaria.
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