This is the accepted version of the journal article: Sanjuan, Josep; Matamoros, Damià; Casanovas i Vilar, Isaac; [et al.]. «Palaeoe- cology of Middle Miocene charophytes from the Vallès[U+2012]Penedès and Vilanova basins (Catalonia, Spain)». Historical biology, (September 2022). DOI 10.1080/08912963.2022.2106861 This version is available at https://ddd.uab.cat/record/265986 under the terms of the license 1 Palaeoecology of Middle Miocene charophytes from the Vallès‒Penedès 2 and Vilanova basins (Catalonia, Spain) 3 Josep Sanjuan1,2*, Damià Matamoros3, Isaac Casanovas-Vilar4, Alba Vicente 1,2,5, Josep 4 Anton Moreno-Bedmar6, Jonathan Holmes7, Carles Martín-Closas1,2 5 1 Departament de Dinàmica de la Terra i de l’Oceà, Facultat de Ciències de la Terra, 6 Universitat de Barcelona-UB, 08028 Barcelona, Catalonia (Spain) 7 2 Institut de Recerca de la Biodiversitat (IRBio). Universitat de Barcelona-UB, 08028 8 Barcelona, Catalonia (Spain) 9 3 Dianastrasse 10a, 9200 Gossau SG, Switzerland 10 4 Institut Català de Paleontologia Miquel Crusafont, Universitat Autònoma de 11 Barcelona, Edifici ICTA-ICP, c/Columnes s/n, Campus de la UAB, 08193 Cerdanyola 12 del Vallès, Barcelona, Catalonia (Spain) 13 5 Instituto Politécnico Nacional. Av. IPN s/N, Col. Playa Palo de Santa Rita. C.P. 14 23096, La Paz, BCS, Mexico 15 6 Instituto de Geología, Universidad Nacional Autónoma de México, Ciudad 16 Universitaria, Coyoacán, 04510, Ciudad de México, Mexico 17 7 Environmental Change Research Centre, Department of Geography, University 18 College London, London WC1E 6BT, United Kingdom 19 20 *Corresponding author: josepsanjuan@ub.edu 21 22 Abstract 23 A diverse charophyte assemblage from the Early-Middle Miocene of the Vallès‒Penedès 24 and Vilanova basins (Catalonia, NE Spain) is here described and illustrated for the first 25 time. This flora has been recovered from three localities: els Casots (Subirats), Vilobí del 26 Penedès, and Mas de l´Alonso‒el Pi Gros (Vilanova i La Geltrú). The charophyte 27 assemblages comprise ten different species distributed among three distinctive aquatic 28 environments, and are approximately simultaneous to major fossil vertebrate sites of these 29 basins. Sphaerochara ulmensis, Chara cf. vulgaris, Chara molassica var. notata, 1 30 Lychnothamnus barbatus var. antiquus, Lychnothamnus sp., Nitellopsis (Tectochara) 31 merianii, and Nitellopsis sp. occur associated to abraded benthic foraminifera in organic- 32 rich claystones related to palustrine and shallow freshwater coastal lakes. 33 Lamprothamnium papulosum forms monospecific assemblages in gypsum-claystone 34 alternations attributed to a coastal brackish water salina. Chara cf. hispida and Chara sp. 35 are found associated with ostracods in marls and limestones related to a permanent 36 oligohaline and alkaline lake. The discovery of this aquatic flora sheds new light on the 37 palaeoenvironmental conditions that prevailed in the Vallès‒Penedès Basin during the 38 Langhian in the context of the Mid-Miocene Climatic Optimum and in the Vilanova Basin 39 during the Serravalian. 40 Keywords: Charophyta; Neogene; Aragonian; palaeolimnology; paleolake; 41 palaeoecology 42 Introduction 43 In this work we describe three Miocene charophyte assemblages from the Vallès- 44 Penedès and Vilanova i la Geltrú basins (Catalonia), which are near the northeastern coast 45 of Spain. 46 The Vallès‒Penedès Basin is one of the best known Miocene basins of eastern Iberia, 47 mostly famous by its dense and continuous land mammal record that comprises hundreds 48 of localities and thousands of specimens (see Casanovas-Vilar et al. 2016a and references 49 therein). A relatively rich fossil pollen and leaf flora has also been documented from this 50 basin (Bessedik 1985; Sanz de Siria Catalán 1993, 1994, respectively), allowing for a 51 comparison with other Southern European floras. Despite its rich continental record, quite 52 surprisingly fossil charophytes have yet to be reported from this area. 53 In the Iberian Peninsula Miocene charophytes have been described in detail from several 54 basins, including the Ebro Basin (e.g., Feist et al. 1994), the Loranca Basin (e.g., Julià de 2 55 Agar 1991), the Teruel Basin in the Iberian Chain (e.g., Ludwig 1987) and the Portuguese 56 part of the Tagus Basin (Soulié-Märsche 1978). Less remarkable records include the 57 Intermediary Depression in Central Spain (Ortíz et al. 1998), La Cerdanya Basin in the 58 Eastern Pyrenees (e.g., Soulié-Märsche and Martín-Closas 2003) and from the Balearic 59 Islands (Martín-Closas and Ramos 2005). The state of the art about Iberian Miocene 60 charophytes surprisingly contrasts with the lack of charophytes studies from the Vallès‒ 61 Penedès Basin, and more considering that both land vertebrates and marine mollusks have 62 been thoroughly studied since the late 19th Century. 63 In this work we describe the first fossil charophyte assemblages from the Vallès‒Penedès 64 Basin and use them to provide accurate palaeoenvironmental reconstructions of Miocene 65 continental aquatic environments. These derive from three distinct localities, els Casots 66 and Vilobí del Penedès, located in the southern sector (Penedès) of the Basin, and Mas 67 de l´Alonso‒el Pi Gros, located in the western sector of the Vilanova i La Geltrú sub- 68 basin (Fig. 1). The els Casots and Vilobí del Penedès localities have been accurately dated 69 by means of bio- and magnetostratigraphy to about 16 Ma (see following section), that is 70 coinciding with the onset of the Miocene Climatic Optimum (MCO). The MCO 71 represented a brief greenhouse interval between ~17 and 15 Ma characterized by an 72 interruption of the Antartic icesheet build up (Zachos et al. 2001, 2008; Foster et al. 2012) 73 and mid-latitude temperatures as much as 3 ⁰C (You et al. 2009) or even 7 ⁰C 74 (Steinthorsdottir et al. 2020) higher than present. Fossil charophytes from both localities 75 (els Casots and Vilobí del Penedès) not only represent the first charophyte assemblages 76 from the Vallès-Penedès, but also provide new palaeoenvironmental information on 77 continental aquatic environments during the MCO in the western Mediterranean. The 78 sites from the Vilanova Basin i.e. Mas de l’Alonso and el Pi Gros date back to the 3 79 Serravallian (see below), that is, they postdate the MCO and coincides with a time of 80 gradual cooling (see Zachos et al., 2001, 2008; Steinthorsdottir et al., 2020). 81 The three outcrops studied, els Casots, Vilobí del Penedès and Mas de l’Alonso‒el Pi 82 Gros correspond to three different palaeoenvironments, ranging from the late Burdigalian 83 to the Serravalian. They record a diverse charophyte flora and show the potential interest 84 that the study of fossil charophytes has for the understanding of the Vallès‒Penedès non- 85 marine palaeoenvironments. The presence of other microfossils (foraminifers and 86 ostracods) provide further information on the palaeoenvironments associated to the 87 charophyte flora. 88 Geological setting 89 The Vallés‒Penedès Basin is part of a large system of Neogene grabens that mark the 90 present eastern Iberian coastline, continuing further offshore (Roca and Guimerà 1992; 91 Roca et al. 1999; Cabrera et al. 2004). These basins are part of the so-called València 92 Trough which originated because of the opening of the western Mediterranean during the 93 late Oligocene and Miocene (Roca et al. 1999; Roca 2001). 94 The Vallès‒Penedès Basin is bounded by NE-SW oriented normal faults that formed after 95 the tectonic inversion of ancient alpine compressional structures during the late Oligocene 96 to Early Miocene (Fontboté 1954; Roca and Guimerà 1992). These limit the basin, which 97 is 100 km long by 12-14 km wide, with the surrounding horsts of the Catalan Litoral 98 Ranges. The Vallès-Penedès main fault is located in its northwestern margin, where the 99 basement is attained at up to 4000 m depth (Bartrina et al. 1992; Roca et al. 1999). Basin 100 basement, and corresponding sediment thickness, gently grades far away from the active 101 margin towards the southeastern margin of the basin to just a few meters (Cabrera et al. 102 1991). The southwestern margin of the basin is only affected by minor faults which were 103 overlapped by Early to Middle Miocene sedimentary sequences (Cabrera 1981a; Cabrera 4 104 and Calvet 1996). These were later fractured as result of minor fault reactivations 105 (Cabrera 1981a; Cabrera and Calvet 1996). The basin is divided into two distinct sectors 106 or sub-basins, Vallès and Penedès, by the NW-SE oriented Llobregat fault (Fontboté 107 1954; Cabrera and Calvet 1996). The semi-graben also shows a strong internal 108 compartmentation due to the occurrence of small horsts and grabens, broadly parallel to 109 the main fault (Permanyer 1990; Cabrera and Calvet 1996). 110 The Miocene record of the Vallès-Penedès basin has been subdivided into four main 111 lithostratigraphic units which have been dated combining bio- and magnetostratigraphy 112 (Agustí et al. 1985; Cabrera and Calvet 1996; Cabrera et al. 1991; Casanovas-Vilar et al. 113 2016b; de Gibert and Casanovas-Vilar 2011): 114 (1) Basal Breccia Unit. This consists of monogenic conglomerates and breccias and crops 115 out only at a few points in the Garraf-Montnegre horst. It dates back to the Ramblian 116 mammal age (early Burdigalian) in the Vallès sector and it is younger in the Penedès, 117 dating back to the early Aragonian (late Burdigalian and earliest Langhian; de Gibert and 118 Casanovas-Vilar 2011; Casanovas-Vilar et al. 2022). 119 (2) Lower Continental Units. Consist of alluvial fan red beds, including conglomerates, 120 sandstones, and siltstones. Older alluvial fan deposits within this unit were sourced from 121 the southeastern basin margin, while younger ones were also sourced from the 122 northwestern reliefs and expanded in the lowlands covering wider areas. Shallow 123 carbonate and evaporite lacustrine systems developed in zones of the southeastern part of 124 the basin (Cabrera et al. 1979; Cabrera 1981a, b), such as the Subirats detritic-carbonatic 125 system, which includes the site of els Casots (see Casanovas-Vilar et al. 2022). This unit 126 mainly covers the Burdigalian (Early Miocene) and coincides with the rifting phase in the 127 Vallès‒Penedès Basin (Cabrera et al. 2004). The els Casots site has been dated to the 128 earliest Langhian (15.9 Ma). 5 129 (3) The Marine and Transitional Units. These developed during the late Burdigalian to 130 the early Serravallian and were related to a global sea-level rise that took place in the 131 context of the MCO between 17 and 15 Ma (Miller et al. 2020). These also coincide with 132 the beginning of the post-rift phase, when the faults that bounded the southeastern margin 133 of the basin became inactive and were onlapped and overlapped by the sedimentary infill 134 (Cabrera and Calvet 1996; Cabrera et al. 1991, 2004; Roca et al. 1999). At least three 135 episodes of marine transgression and regression are recorded in the basin: late 136 Burdigalian, Langhian and early Serravallian (Cabrera et al. 1991; Cabrera and Calvet 137 1996; Roca et al. 1999; de Gibert and Casanovas-Vilar 2011; Casanovas-Vilar et al. 138 2016b). Coastal saline lakes (sabkhas) developed just below the marine units in the area 139 of Vilobí del Penedès, which represents a lifted block of the basin basement (Permanyer 140 1990). A diverse set of gypsum deposits up to 60 m thick developed in these sabkhas (Ortí 141 and Pueyo 1976; Agustí et al. 1990). These gypsum deposits are overlain by claystones, 142 which have yielded various charophyte assemblages, that are ultimately covered by 143 oyster-rich greyish claystones and bioclastic limestones corresponding to the Langhian 144 transgression. The gypsum units have been generally correlated to the late Burdigalian 145 (Ortí and Pueyo 1976; Agustí et al. 1990; de Gibert and Casanovas-Vilar 2011), while the 146 overlying claystones, which include small mammal fossils, indicate a younger, earliest 147 Langhian, age (Jovells-Vaqué, 2020). 148 (3) Upper Continental Units. These cover the Serravalian and Tortonian and represent the 149 late post-rift phase in the basin (Cabrera et al. 2004). They consist of alluvial fan deposits 150 that were sourced from the reliefs in the northwestern margin while the Vallès-Penedès 151 main fault remained active (Agustí et al. 1985, 1997; Cabrera and Calvet 1996; Roca et 152 al. 1999; Casanovas-Vilar et al. 2016a, b; de Gibert and Casanovas-Vilar 2011). Most of 153 the fossil vertebrate sites of the Vallès-Penedès Basin are located in the distal to marginal 6 154 facies of alluvial fan systems (Casanovas-Vilar et al. 2016 a, b), but this unit has not 155 yielded charophyte localities thus far. 156 There are several smaller grabens next to the coast, south of the Garraf-Montnegre horst 157 of the Catalan Coastal Ranges. These include the Pla de Barcelona, Baix Llobregat, Sant 158 Andreu de la Barca, Olesa de Bonesvalls and Vilanova basins, and all formed in the 159 context of the same extensional processes that originated the Vallès-Penedès Basin. 160 The Vilanova basin is about 5 km wide by 11 km long (Fig. 1). Same as the Vallès- 161 Penedès, its northwestern margin is limited by a major normal fault affecting the pre- 162 Miocene basement, while the southeastern margin is defined by minor normal faults that 163 are overlapped by Quaternary sediments or are currently under sea in most of the basin 164 (Ramos-Guerrero et al. 1996). Ramos-Guerrero et al. (1996) defined up to four 165 litostratigraphic units attaining a maximum thickness of about 400 m. The oldest unit 166 (M0) is represented by the marginal complex (breccias) associated and genetically related 167 to the main faults that bound the basin. The other three units show a vertical evolution 168 from continental alluvial environments at the base (M1), freshwater and brackishwater 169 lacustrine facies (M2), to marine (littoral and restricted shelf-bay) at the top (M3). The 170 charophyte assemblages studied here (localities of Mas de l’Alonso and el Pi Gros) have 171 been recovered from the M2 unit (Intermediate Detrital-Carbonate Unit) which is mainly 172 composed of lacustrine marls and limestones. These litostratigraphic units have been 173 correlated to the Serravallian, although the lowermost alluvial conglomeratic units may 174 correspond to Langhian (Cabrera et al. 2004). 175 -------------------------------------------Figure 1 near here----------------------------------------- 176 177 Materials and methods 7 178 The fossil flora herein studied has been recovered from soft rocks (claystones and marls) 179 from three distinct localities: els Casots, Vilobí del Penedès, in the Penedès sub-basin and 180 Mas de l´Alonso‒el Pi Gros, in the Vilanova i la Geltrú sub-basin (Fig. 1). The 181 coordinates of these localities are: els Casots (Subirats), 41°24'56.57"N 01°48'41.78"E; 182 Vilobí del Penedès, 41°23'15.24"N, 01°39'10.90"E; Mas de l´Alonso (Vilanova i la 183 Geltrú), 41°14'57.28"N, 1°43'39.59"E, and el Pí Gros (Vilanova i la Geltrú), 184 41°14'52.57"N, 01°42'50.09"E. 185 In total, 12 samples of 2 kg each one were treated to extract the microfossils. Samples 186 were disaggregated in water, hydroxide peroxide and sodium carbonate and later wet- 187 sieved using sieves with mesh apertures of 1, 0.5, and 0.3 mm. In case of Vilobí del 188 Penedès (sample V6/7), about 2000 kg of sediments were screen-washed during a field 189 campaign undertaken by the Institut Català de Paleontologia (ICP) in the 1990s. The field 190 campaign focused on the recovery of microvertebrate remains and the smallest mesh 191 aperture used during the sieving process was 0.5 mm. In April 2021, 2 kg of sediments 192 were taken from the same outcrop to recover a smaller microfossil fraction (200 – 500 193 µm), which was lost in the previous campaign because a larger mesh size was used. 194 Nine out of the twelve studied samples yielded microfossils including gyrogonites, 195 ostracod carapaces, skeletons of foraminifera, and fish teeth (Fig. 2). Microfossils were 196 sorted after inspecting the residue washed sediment under a light microscope. Selected 197 gyrogonites were measured using the software Motic Images Plus 2.0 ML in a 198 stereomicroscope Motic BA310 with a built-in digital camera housed in the Departament 199 de Dinàmica de la Terra i de l´Oceà (Facultat de Ciències de la Terra), Universitat de 200 Barcelona). The biometric parameters considered were the gyrogonite height (μm), the 201 gyrogonite width (μm), the number of spiral turns observed in lateral view and the 202 isopolarity index (gyrogonite height/gyrogonite width × 100). Well-preserved 8 203 microfossils were selected and photographed using the scanning electronic microscope 204 (SEM) QUANTA 200 located at the Centres Científics i Tecnològics, Universitat de 205 Barcelona (CCiTUB). Thin-sections, ca. 30 μm in thickness, were prepared from four 206 limestone beds from els Casots and Mas de l´Alonso-el Pi Gros localities (Fig. 2). The 207 microfossils illustrated herein are curated at the Institut Català de Paleontologia Miquel 208 Crusafont (ICP) at Sabadell (Catalonia, Spain) with catalogue numbers IPS122528, 209 IPS122530 and IPS127514 to IPS127517. 210 Results 211 Depositional setting and age 212 Els Casots (Subirats) 213 Els Casots is a key vertebrate fossil site situated in the municipality of Subirats, in the 214 south-western margin of the Penedès sub-basin (Fig. 1). The Miocene succession at els 215 Casots area directly overlies the basement, which consists of Mesozoic (mainly Early 216 Cretaceous) marine carbonates (Casanovas-Vilar et al. 2022). The Miocene succession 217 was studied by means of geological core sampling and is described in detail in Casanovas- 218 Vilar et al. (2022). The core records a basal breccia followed by a 25 m-thick, cyclically- 219 arranged mudstone/limestone succession that at the bottom includes thin (mm to dm 220 thick) sub-bituminous coal deposits. The upper part of the cycle is a carbonate term that 221 includes a variety of bioclastic- to micrite-dominated limestones that range from a few 222 centimetres to a maximum of a couple of meters thick. The carbonate facies include 223 bioclastic laminae alternated with very thin tufa-like carbonates (Cabrera 1979). Thin 224 ferruginous laminae are associated to some of the bioclastic facies. Both carbonates and 225 siliciclastic mudstones may include abundant plant fragments and freshwater gastropod 226 shells or casts. The stratigraphic succession indicates lacustrine to palustrine 227 environments with cyclically oscillating water level. This lacustrine/marshy succession 9 228 has yielded abundant and remarkably complete terrestrial vertebrate fossils in the 229 excavated area of the site. The vertebrate fauna comprises up to 75 different vertebrate 230 species including amphibians, reptiles, birds and mostly mammals. The finding of several 231 articulated partial skeletons indicate that the site records an autochthonous to 232 parautochthonous vertebrate assemblage (Casanovas-Vilar et al. 2022). The charophyte 233 flora studied here was recovered from a 3.5 m thick section laterally equivalent to the 234 vertebrate-bearing lacustrine/marshy layers (samples C1-C4 in Fig. 2). This consists of 235 organic-rich claystones alternated with marls and a limestone bed (Figs 2 and 3A‒B). 236 Claystone beds range from 10 to 50 cm thick and show strong color variation from 237 yellowish, gray to dark gray (Figs 2 and 3A‒B). Some organic-rich claystones have 238 yielded abundant plant remains such as leaf fragments of helophytic plants, e.g., Typha, 239 well-preserved charophyte gyrogonites, abraded benthic foraminifera, plus vertebrate 240 bones, sometimes in anatomical connection. Some claystone intervals present root marks. 241 In some horizons abundant and irregular carbonate nodules occur (Fig. 3A). A 20 cm 242 thick limestone bed with packstone texture includes mollusk shell fragments, 150 µm thin 243 carbonate planar crusts (Fig. 3C-H) and oncoids (Fig. 3G), both interpreted as of 244 cyanobacterial origin. 245 Claystones at els Casots are attributed to deposition in a quiet and shallow lake. The 246 presence of nodular carbonates, mottling and root marks suggests that the original 247 lacustrine sediments were subject to fluctuations of the water table leading eventually to 248 subaerial exposure with a development of pedogenetic features, similar to those described 249 elesewhere by Alonso-Zarza and Wright (2010) and references herein. Limited oxygen 250 levels and rapid burial at the lake margins favored the preservation of mammal remains 251 and helophytic plant leaves found in darker claystone intervals. Charophytes from these 252 claystones grew under shallow lake conditions, while abraded remains of foraminifera 10 253 were probably whased in from nearby marine settings. The limestone layer studied at els 254 Casots is interpreted as deposited in a shallow pond. The occurrence of oncoids and 255 abundant mollusk-shell fragments suggest that higher hydrodynamic conditions and 256 perhaps trampling prevailed during its deposition. Moreover, the abundance of 257 cyanobacterial crusts and absence of charophyte remains in the same limestone bed 258 suggest that the water was eutrophic (Martín-Closas, 1999). 259 The lacustrine succession at els Casots ends rather abruptly and is followed by 260 reddish and mottled mudstones and sandstones that record subaerial distal channelized 261 alluvial fan facies. The continental deposits are in turn overlain by an oyster coquina and 262 a bioclastic quartzarenite, which marks the early Langhian transgressive surface in this 263 sector of the Vallès-Penedès Basin. 264 The age of els Casots is well constrained thanks to bio- and magnetostratigraphic 265 data (see Casanovas-Vilar et al. 2022). For a long time, this site was correlated to the 266 latest Early Miocene European mammal Neogene zone MN4, but the recent data indicate 267 a correlation to Aragonian subzone Cb, which corresponds to the earliest MN5. 268 Magnetostratigraphic data from the 30-m-thick els Casots succession indicate reverse 269 polarity which, coupled with the correlation to Aragonian subzone Cb (16.15–15.94 Ma; 270 boundaries after Van der Meulen et al. 2012), results in an estimated age of ~15.9 Ma for 271 the site of els Casots. Therefore, els Casots would coincides with the onset of the Miocene 272 Climatic Optimum (Zachos et al. 2001, 2008; Holbourn et al. 2015). Overall, 273 sedimentological, palaeobotanical (scarce macroflora remains) and faunal data indicate 274 that els Casots corresponds to an ancient freshwater lacustrine or palustrine environment 275 that developed in a tropical-subtropical climate with rainfall seasonality (Casanovas-Vilar 276 et al. 2022). 277 11 278 -------------------------------------------Figure 2 near here----------------------------------------- 279 280 -------------------------------------------Figure 3 near here----------------------------------------- 281 282 283 -------------------------------------------Figure 4 near here----------------------------------------- 284 285 Vilobí del Penedès 286 The section studied at Vilobí del Penedès overlies unconformably the Cretaceous 287 limestone substrate (Fig. 1). The base of the sequence is represented by non-marine 288 breccias, detritic red beds and limestones of the Lower Continental System, which in turn 289 are overlain by a 60 m thick evaporitic sequence mainly composed of secondary gypsum 290 but showing primary gypsarenite at the top (Ortí and Pueyo 1976; Ortí 1990). The 291 succession follows with a 7-m-thick interval dominated by reddish and grey claystones 292 which have yielded the charophyte flora studied herein (samples V1-V6/7 in Fig. 2.2) and 293 micromammal remains that had been previously attributed to the early Aragonian, MN4 294 zone, corresponding to the latest Early Miocene (Agustí et al. 1985, 1990; Casanovas- 295 Vilar et al. 2016a). However, the rodent fauna includes the same cricetid species recorded 296 at els Casots (Jovells-Vaqué and Casanovas-Vilar 2021) and amongst the eomyids only 297 the species Ligerimys ellipticus is present (Jovells-Vaquè 2020). Consequently, this 298 favors a correlation with Aragonian subzone Cb and indicate an early Middle Miocene 299 age (MN5) for the site instead (Jovells-Vaqué 2020). Therefore, it would be 300 chronologically very close to other sites of the Vallès-Penedès Basin immediately before 12 301 the Langhian regional marine transgression, such as els Casots or Sant Mamet (Sant Cugat 302 del Vallès). 303 The upper part of the Vilobí del Penedès succession, which has been studied here in more 304 detail, can be subdivided into three intervals. The first one is 2.5 m thick and overlies a 305 laminated gypsarenite (Fig. 4A). It is composed of an alternation of primary gypsum and 306 greenish claystones rich in charophytes and ostracods. Up to 4 parasequences of gypsum- 307 claystone alternations can be distinguished (Fig. 4B). A semi covered greenish claystone 308 bed, 1.5 m thick, lays over the gypsum-claystone alternation and yielded a second, 309 species-rich charophyte assemblage (Fig. 4C). The second interval is represented by 2.5 310 m of reddish claystone covered by an oyster bank that marks the base of the third interval 311 (Fig. 4D), which is dominated by grey marls and bioclastic limestones rich in marine 312 fossils such as echinoderms and coralline rhodophytes. 313 The evaporitic succession at Vilobí del Penedès has been interpreted by Bitzer (2004) and 314 references therein as having been deposited in a restricted coastal lake or salina (sabkha). 315 The facies alternation of gypsum and claystone horizons with charophytes observed 316 above the evaporitic unit, suggests that sediments settled under fluctuating salinity 317 conditions within the coastal salina. Gypsum layers precipitated in hypersaline 318 conditions, of up to 110‰ according to Bitzer (2004), while claystone layers were 319 deposited in more diluted conditions, corresponding to brackish water, as shown herein. 320 Each gypsum-claystone parasequence can be interpreted as small transgressive-regressive 321 cycle within the salina (Fig. 2). The absence of evaporites and the presence of a diverse 322 charophyte assemblage in the overlying grey claystone beds indicate that diluted 323 conditions finally prevailed, corresponding probably to a coastal freshwater lake (Fig. 324 4C). Finally, the oyster bank and the bioclastic limestone located atop of the section marks 325 the onset of the Langhian regional marine transgression (Fig. 4D). 13 326 Mas de l´Alonso‒el Pi Gros (Vilanova i la Geltrú): 327 The studied succession is located at the western margin of the Vilanova i la Geltrú basin 328 (Fig. 1) and corresponds to the Intermediate Detrital-Carbonate Unit (M2) of Ramos- 329 Guerrero et al. (1996). The outcrop at Mas de l´Alonso consists of dark grey organic-rich 330 claystones with charophytes topped by a metric yellowish lacustrine limestone bed (Fig. 331 2.4). Two samples provided charophytes (samples MA1 and MA2 in Fig. 2.4). About 1 332 km westwards, at the el Pi Gros locality, a lateral equivalent of the same non-marine 333 sequence crops out and shows a 3.5 m succession of yellowish limestone alternated with 334 marls, claystones, and siltstones (Figs 2.3 and 5). Four horizons yielded charophytes, i.e., 335 samples PG1-PG4 in Fig. 2.3. Light grey to yellowish limestone beds at the base of the 336 el Pi Gros section range in thickness from 10 to 30 cm. They display wackstone-packstone 337 textures and contain abundant microfossils such as ostracods and gyrogonites (Fig. 5C‒ 338 H). These limestones are overlain by two parasequences of limestone beds with 339 stromatolithic lamination covered by reddish bioturbated claystones (Figs 2.3 and 5B). 340 The upper part of the section at el Pi Gros shows 0.5 m-thick cross-bedded calcarenite 341 and is topped by a 0.6 m limestone bed displaying packstone textures rich in gyrogonites, 342 charophyte thalli, dasycladaleans, ostracods, and potamidid gastropods (Fig. 2.3). Above 343 these materials marine facies of the M3 Unit of Ramos-Guerrero et al. (1996) crop out. 344 Despite some authors suggested that the M2 unit is Langhian in age (Batllori and Moreno 345 2003), the micromammals from fossil sites located within this unit indicate younger ages 346 (Serravalian). The site of los Viñedos (Sant Pere de Ribes), yielded small vertebrate 347 remains that are suscintly reported in Ramos-Guerrero et al. (1996). The revised faunal 348 list includes the cricetids Megacricetodon crusafonti and Hispanomys cf. decedens which 349 would indicate a correlation with the Megarcricetodon crusafonti + Democricetodon 350 larteti Concurrent Range Subzone of the Vallès-Penedès Basin (Casanovas-Vilar et al. 14 351 2016b), thus indicating an age closer to 12.4–12.5 Ma, that is late Aragonian 352 (chronologically equivalent to the late Serravallian). A second site, Sant Pere de Ribes 1, 353 located at the Can Mestre quarry (Ramos-Guerrero et al. 1996), has yielded additional 354 small mammal remains. These include just a few teeth that are assigned to 355 Democricetodon crusafonti (referred to as Renzimys bilobatus in Ramos-Guerrero et al. 356 1996) a species that is first recorded during the late Aragonian in the Vallès-Penedès at 357 11.88 Ma (Casanovas-Vilar et al. 2016b) and slightly earlier (12.4 Ma) in the Calatayud- 358 Montalbán Basin (Aragon; Van Dam et al. 2014). Therefore, unit M3 would belong to 359 the late Serravallian as well. 360 According to Ramos-Guerrero et al. (1996), the sequence integrating the M2 and M3 units 361 grades upwards from lacustrine to brackish and finally marine environments. The 362 lacustrine conditions of Unit M2 were inferred because of the presence of freshwater 363 gastropods, helophytic plant stems, and charophyte remains (Batllori and Moreno 2003 364 and references therein). The locality of el Pi Gros is well known by the diverse and rich 365 mollusk fauna preserved as molds, and the bioerosion ichnofossils, both found in the 366 overlying marine M3 Unit (Batllori and Moreno 2003; Belaústegui et al. 2018). In 367 contrast, the fossils of the M2 Unit are poorly known. 368 Microfossils recovered from the lower marls at Mas de l’Alonso (sample MA-1, Fig. 3 369 and Table 1) and el Pi Gros (sample PG-2, Fig. 3, Table 1) are well preserved and do not 370 show signs of fragmentation or abrasion. Moreover, ostracod carapaces frequently appear 371 with the two valves articulated suggesting that they were buried in situ. The absence of 372 charophyte thalli in these marls suggest that there was a certain selection of the original 373 plant remains, and that gyrogonites may have been laterally transported from the 374 shallower areas of the lake where they grew. The occurrence of rare and abraded 375 foraminifera at Mas de l’Alonso is interpreted as result of transport from a nearby marine 15 376 coastal area. Altogether these data suggest that the depositional environment of these 377 marls corresponded to a relatively deeper part of a freshwater lake. The overlying 378 charophyte-rich limestones were deposited in shallower freshwater environments, where 379 lime-producing organisms thrived. Wackestone–packstone textures suggest the 380 prevalence of sublittoral lacustrine conditions (e.g., Gierlowski-Kordesch 2010). Higher 381 in the series, the substitution of charophyte-rich marl and limestones by stromatolite 382 limestones and reddish claystone suggest that palaeoenvironmental conditions changed, 383 difficulting the growth of charophytes. The occurrence of the terrigenous red claystone 384 atop of the stromatolite, probably indicating deposition in a floodplain, suggest that the 385 nutrient input to the lake may have increased during the development of stromatolites, 386 hindering the development of charophytes. 387 388 -------------------------------------------Figure 5 near here----------------------------------------- 389 390 Systematic palaeobotany (Charophytes) 391 Division Charophyta Migula, 1897 392 Class Charophyceae Smith, 1938 393 Order Charales Lindley, 1836 394 Family Characeae Richard ex C. Agardh, 1824 395 Subfamily Nitelloideae A. Braun in Migula, 1897 396 Genus Sphaerochara (Mädler, 1952) emend. Soulié-Märsche, 1989 397 Sphaerochara ulmensis (Straub, 1952) comb. nov. Grambast, 1962 398 Fig. 6A‒I 16 399 1952. Chara ulmensis Straub, p. 470, pl. A, fîg. 19. 400 1962. Sphaerochara ulmensis Grambast, p. 77. 401 Material: This species occurs at Vilobí del Penedès and Mas de l´Alonso‒el Pi Gros 402 (Table 1). Twenty-three gyrogonites were measured in sample MA1 and nine specimens 403 in sample V6/7 (Table 2 in supplementary data). 404 Description: Gyrogonites from MA1 are small, 398‒477 μm high (mean 437 μm) and 405 330‒431 μm wide (mean 391 μm), prolate spheroidal in shape, with an isopolarity index 406 of 112 (mean). Nine to ten spiral turns are visible in lateral view. Spiral cells are 53 µm 407 high in average and ornamented with an irregular mid-cellular crest (Fig. 6F‒H). The 408 thickness of the mid-cellular crest can vary significantly, and it can stretch the complete 409 width of the spiral cell (Fig. 6B, C and C‒E). The apex is rounded in lateral view. The 410 apical end of each spiral cell is ornamented with a small, individualized coma-shaped 411 nodule (Fig. 6A). The basal plate is visible from the outside and it is located within a large 412 pentagonal basal pore of ~75 µm across (Fig. 6D and I). The external part of the basal 413 plate bears a central nodule (Fig. 6D and I). Gyrogonites from the sample V6/7 at Vilobí 414 del Penedés (Fig. 6A‒D) are ~175 μm larger and wider than specimens extracted from 415 Mas de l´Alonso. 416 Remarks: This species shows strong morphological similarities to other ornamented 417 Sphaerochara gyrogonites from the European Paleogene displaying mid-cellular crests 418 such as S. hirmeri (Rasky 1945) Mädler 1952 and S. labellata Feist and Ringeade 1977. 419 Gyrogonites of S. ulmensis are longer and show a higher ISI index than the two 420 aforementioned taxa. Moreover, this species displays a thicker and more prominent mid- 421 cellular crest. The subtle differences between these three coeval ornamented 422 Sphaerochara species suggest that a comprehensive taxonomic revision should be 423 conducted in future studies. 17 424 Fossil record and distribution: This taxon has been reported in several Paleogene and 425 Neogene basins worldwide i.e., Europe, Middle East, and China mainly in sedimentary 426 sequences ranging in age from the upper Rupelian (early Oligocene) to the Tortonian 427 (Late Miocene). In Europe it is reported from several basins such as the Rhine Graben in 428 North and Central Germany (Straub 1952; Schwarz 1997 and references therein); the 429 Alpine Molasse in South Germany (Mädler 1955; Horn af Rantzien,1959; Reichenbacher 430 and Schwarz 1997); the Provence (Feist-Castel 1977), and the Languedoc, both in France 431 (Chellaï et al. 1982); the Ebro Basin in Catalonia (Feist et al. 1994); and the Cluj Basin 432 in Romania (Baciu and Feist 1999). S. ulmensis has also been reported by Mädler and 433 Staesche (1979) from several basins of Central Anatolia (Turkey), and Knobloch (1979) 434 further recovered S. ulmensis in Neogene rocks from Iraq. In China, S. ulmensis is found 435 in Oligocene deposits from the Bohai coastal region (Xinlun 1978) as well as in several 436 Chinese provinces, such as Sichuan (Huang et al. 1988), Yunnan (Liu 1989), Xinjiang 437 (Lu and Luo 1990) and Qinghai (Tang and Di 1991). 438 Palaeoecology: S. ulmensis is found in limestones attributed to well-developed lacustrine 439 systems, e.g., the Trévaresse limestone Formation in Aix-en-Provence, France (Feist- 440 Castel 1977). Based on the associated microfossils, Reichenbacher and Schwarz (1997) 441 concluded that S. ulmensis was a halophobous taxon, adapted exclusively to freshwater 442 habitats. 443 Subfamily Charoideae A. Braun in Migula 1897 444 Genus Chara Linnæus, 1753 445 Chara cf. vulgaris Linnæus, 1753 446 Fig. 6 J‒N 447 1753. Chara vulgaris Linnæus, p. 1156 18 448 1989. Chara vulgaris var. vulgaris Soulié-Märsche, p. 97, pl. 22 449 Material: This taxon dominates the flora in the sample C4 from els Casots (Table 1). Fifty 450 gyrogonites were measured (Table 2 in supplementary data). 451 Description: Gyrogonites are of medium size, 378‒584 μm high (mean 466 μm) and 256‒ 452 430 μm wide (mean 352 μm), showing a prolate to perprolate shape and an isopolarity 453 index ranging between 112 and 166 (mean 133). Ten to thirteen (mainly eleven) 454 convolutions are visible in lateral view (Fig. 6K‒M). Spiral cells are flat to concave, non- 455 ornamented and ~44 μm in height. Spiral cells expand at the apical area (Fig. 6J) forming 456 a psilocharoid apex type (Feist et al. 2005). The base is rounded or slightly pointed 457 showing a small pentagonal basal pore of ~48 μm in diameter (Fig. 6N). 458 Remarks: The population described herein is remarkably similar to gyrogonites of living 459 C. vulgaris in morphlogical characters such as the apical and basal structure, gyrogonite 460 width and convolution number in lateral view (Soulié-Märsché 1989; Sanjuan et al. 461 2017). However, the studied fossil sample shows gyrogonite heights ~100‒150 μm 462 shorter than living populations. 463 Fossil record and distribution: Fossils of C. vulgaris are poorly known in Neogene 464 sedimentary sequences. Ghetti et al. (2002) reported gyrogonites of C. vulgaris from Late 465 Miocene (early Messinian) rocks of the Velona Basin (Siena, Central Italy). Bathia et al. 466 (1998) found this species in the Plio-Pleistocene intermountainous Himalayan basins 467 (Kashmir, India). Fan et al. (1996) also recovered gyrogonites attributed to this taxon in 468 upper Pleistocene lacustrine facies in the Bangong Co Basin (Western Tibet). Large 469 samples of C. vulgaris were recovered by Soulié-Märsche (1991) and Soulié-Märsche et 470 al. (2010) from Pleistocene/Holocene paleolakes from Chad, Algeria and Sudan (Africa). 19 471 Palaeoecology: Chara vulgaris thrives in many kinds of shallow non-marine waterbodies 472 worldwide (Corillion 1972). This taxon grows in temporary ponds or permanent lakes 473 from the depth of several centimeters to up to 1 m (Krause 1997). It can live in freshwater 474 to oligohaline conditions, but calcified gyrogonites are only formed at salinities below 5 475 ‰ and in alkaline water (Ghetti et al. 2002 and references therein). It has also a wide 476 tolerance to different trophic conditions including eutrophic water (Korsch 2016). 477 478 Chara molassica var. notata (Straub, 1952) comb. nov. Soulié-Märsche, 1989 479 Fig. 6O‒U 480 1952. Chara molassica forma a Straub, p. 467, pl. A, fig. 4‒5 481 1965. Chara notata Grambast and Paul, p. 245 482 1989. Chara molassica var. notata Soulié-Märsche, p. 146 483 Material: Fifty specimens were extracted from Vilobí del Penedès and fifteen gyrogonites 484 from els Casots (Table 1). Thirty-five specimens were measured from the sample V6/7 485 (Table 2 in supplementary data). 486 Description: Gyrogonites are of medium size, 498‒599 μm high (mean 520 μm) and 320‒ 487 416 μm wide (mean 377 μm). They are subprolate in shape and display a wide isopolarity 488 index value, ranging between 108 and 161 (mean 138). Eight to twelve (commonly ten) 489 convolutions are visible in lateral view. Mean high of spiral cells is ~59 μm and they are 490 concave and ornamented with isolated small tubercles regularly arranged in row (Fig. 6I, 491 Q and T). Gyrogonites display a prominent psilocharoid-type apex (Fig. 6O and S). The 492 apex is ornamented with enlarged apical tubercles (Fig. 6S). The base is rounded or 493 slightly pointed showing a shallow pentagonal basal pore of ~36 μm in diameter (Fig. 6R 494 and U). 20 495 Fossil record and distribution: Chara molassica var. notata has been recovered in latest 496 Oligocene (Chattian) to Late Miocene (Tortonian) sedimentary sequences in Europe. 497 Feist-Castel (1977) and Feist and Ringeade (1977) described populations of this taxon 498 from the latest Oligocene/Early Miocene (Aquitanian) of the Provence and Aquitaine 499 basins in France respectively. Large samples of C. molassica var. notata were later 500 recovered in Aquitanian-aged rocks of the Alpine Molasse Basin in Switzerland (Berger 501 1983; Schwarz and Reichenbacher 1989). This variety is also well represented in the latest 502 Oligocene/Early Miocene of the Rhine Graben, Germany (Schwarz 1997 and references 503 therein). Julià de Agar (1991) reported this taxon from the Loranca Basin (Tagus Basin, 504 Spain). In the Ebro Basin (NE Spain) C. molassica var. notata is well represented in 505 Chattian to Tortonian non-marine deposits (Feist et al. 1994; González-Pardo 2012). This 506 species was also reported from the Middle East by Knobloch (1979) who found it in the 507 Miocene Fars Formation (Iraq). 508 Palaeoecology: Previous studies report this species in freshwater lacustrine facies (e.g., 509 Feist and Ringeade 1977). 510 511 -------------------------------------------Figure 6 near here------------------------------------- 512 513 Chara cf. hispida Linnæus, 1753 514 Fig. 7A‒H 515 1753. Chara hispida Linnæus, p. 1156‒1157 516 1989. Chara hispida var. major Soulié-Märsche, p. 92, pl. 23 517 1999. Chara cf. hispida var. major García, p. 311, Fig. 4a‒e 21 518 Material: Hundreds of well-preserved gyrogonites have been recovered in the el Pi Gros 519 section (Table 1). One hundred specimens were measured from the sample PG2 (Table 2 520 in supplementary data). 521 Description: Gyrogonites are medium- to large-sized, ranging between 540‒891 μm in 522 height (mean 726 μm) and 352‒621 μm in width (mean 500 μm). The dominant 523 gyrogonite shape is prolate spheroidal to perprolate (Feist et al. 2005), with an isopolarity 524 index ranging between 106 and 206 (mean 145). Nine to twelve (commonly ten) spiral 525 cells are visible in lateral view (Fig. 7A‒D). Spiral cells are flat to convex, about 88 μm 526 wide and without ornamentation. They become thinner towards the periphery of the apex. 527 The apex is psilocharoid and prominent, generally pointed (Fig. 7E‒F). The base is also 528 pointed showing a small pentagonal basal pore of ~59 μm in diameter (Fig. 7G‒H). 529 Remarks: The population at el Pi Gros displays smaller gyrogonites (~180 µm in length 530 and ~85 μm in width) than recent populations of C. hispida studied by Soulié-Marshé 531 (1989) or Détriché et al. (2009). 532 Fossil record and distribution: Fossils of Chara hispida have been described from 533 Quaternary sedimentary sequences of Europe, Africa and South America. Anadón et al. 534 (1987) reported gyrogonites of this species from the Pleistocene of the Guadix-Baza 535 Basin (SE Spain). Fossil samples are also reported from the Holocene lacustrine 536 sediments from the Wadi Shaw (Sudan) and intermountainous basins in the Middle Atlas 537 of Morocco (Soulié-Marsche 1991; Détriché et al. 2009). García (1999 and references 538 therein) reported gyrogonites of C. hispida from several Quaternary localities from 539 Argentina, i.e., Quebrada del Zonda (Province of San Juan), Laguna Salada Grande 540 (Buenos Aires Province), and Laguna del Bebedero (San Luis Province). 541 Palaeoecology: Extant C. hispida is a subcosmopolitan species thriving in lakes from 542 nearly all states in Europe, and a few localities in North Africa, the Caucasus, Central 22 543 Asia, and Eastern Siberia (Corillion 1972; Barinova et al. 2014 and references therein). It 544 dwells in alkaline permanent and well-oxygenated lakes at depths of up to 15 m. 545 However, this species mostly occurs between 0.5 and 7 m depth (Krause 1997). In spite 546 that preferred habitats of C. hispida are freshwater lakes, it can tolerate brackish water 547 (Mannschreck 2003). In agreement with habitat requirements of extant C. hispida, the 548 gyrogonite assemblage studied here would also be indicative of permanent and stable 549 freshwater lake conditions. 550 Chara sp. 551 Fig. 7I‒M 552 Material: A few gyrogonites were found at el Pi Gros locality (Table 1). Three specimens 553 were measured in sample PG2 (Table 2 in supplementary data). 554 Description: Gyrogonites are very variable in size ranging between 612‒825 μm in height 555 and 350‒400 μm in width. The dominant gyrogonite shape is perprolate, with a very high 556 isopolarity index ranging between 161 and 206. Six to seven spiral cells are visible in 557 lateral view (Fig. 7J‒L). Spiral cells are convex, without ornamentation. The apex is 558 psilocharoid and very prominent, generally pointed (Fig. 7I). The base is also pointed 559 showing a very small basal pore (Fig. 7M). 560 Remarks: This material could belong to extreme polymorphs of of Chara hispida, with 561 which it cooccurs in the same samples but in a lower proportion (Table 1). Stressful 562 environmental conditions are well known to result in the development of extreme 563 polymorphs in extant Chara species (e.g., Pukacz et al. 2012). 564 Genus Lamprothamnium J. Groves, 1916 565 Lamprothamnium papulosum (Wallroth, 1833) comb. nov. Groves, 1916 566 Fig. 7N‒U 23 567 1833. Chara papulosa Wallroth, p. 107 568 1916. Lamprothamnium papulosum Groves, p. 336 569 1969. Lamprothamnium priscum Castel and Grambast, p. 940‒941, pl. XXXII, figs 4‒7 570 1989. Lamprothamnium papulosum Soulié-Märsche, p. 150‒153, pl. XXXII 571 572 Material: Many gyrogonites were recovered from several samples at the base of the 573 Vilobí del Penedès section (Table 1). Sixty specimens were measured from the sample 574 V5 (Table 2 in supplementary data). 575 Description: Girogonites are of medium size, 569‒711 μm high (mean 631 μm) and 348‒ 576 542 μm wide (mean 469 μm), cylindrical or prolate in shape (Fig. 7N‒Q), with an 577 isopolarity index ranging from 113 to 177 (mean 135). Gyrogonites show a prominent 578 apex of lamprothamnoid type (Fig. 7R‒S), i.e., displaying a well-marked periapical 579 depression (Feist et al. 2005). Seven to eleven (commonly 9) spiral cells are visible in 580 lateral view. Spiral cells are concave, flat, or convex depending on the calcification degree 581 and ~81 μm high. Gyrogonites are not ornamented. The base of the gyrogonite is rounded 582 or slightly pointed with a small pentagonal pore, ranging in diameter from 40 to 60 μm 583 (Fig. 7T‒U). 584 Remarks: Fossil gyrogonites of L. papulosum from Vilobí del Penedés are ~75 μm 585 widerand display a reduced number of spiral cells in lateral view than gyrogonites from 586 the living populations of the same species from brackish waters of southern France 587 studied by Soulié-Märsché (1989). The studied gyrogonites are morphologically closer to 588 Lamprothamnium priscum Castel and Grambast, 1969, a fossil species synonymized with 589 L. papulosum by Soulié-Märsche (1989). 24 590 Fossil record and distribution: Extant Lamprothamnium papulosum is a quasi- 591 cosmopolitan species distributed throughout the world except in North and Central 592 America (Corillion 1972; García and Chivas 2004). The oldest known L. papulosum was 593 reported by Castel and Grambast (1969) and referred to as L. priscum, from the early 594 Eocene deposits from the Corbières mountains (southern France) and further from the 595 early Eocene of Ksour Mountains in Algeria (Mennad et al. 2021). Lamprothamnium 596 papulosum has been found in several Quaternary deposits of North Africa (Soulié - 597 Märsche 1982, 1998, 2007). 598 Palaeoecology: L. papulosum is adapted to aquatic environments subject to salinity 599 fluctuations and can thrive in sub-saline to hyper-saline waters up to 68‰ in salinity 600 (Burne et al. 1980). Despite Lamprothamnium can live in hypersaline conditions, the 601 required salinity to produce oospores and gyrogonites should be 20‒40‰ (Soulié- 602 Märsche 1998 and references therein). However, short periods of salinity close to 10% o 603 are also tolerated for germination of oospores and gyrogonites (Dubois, 1968). 604 605 606 -------------------------------------------Figure 7 near here------------------------------------- 607 608 Genus Lychnothamnus (Ruprecht, 1845) von Leonhardi, 1863 emend. A. Braun in 609 Braun et Nordstedt, 1882 610 Lychnothamnus barbatus (Meyen, 1827) comb. nov. von Leonhardi, 1863 611 Lychnothamnus barbatus var. antiquus Soulié-Märsche, 1989 612 Fig. 8A‒C 25 613 1827. Chara barbata Meyen, p. 75, pl. e, figs 7 and 8. 614 1863. Lychnothamnus barbatus Leonhardi, p. 72. 615 1989. Lychnothamnus barbatus var. antiquus Soulié-Märsche, p. 155, pl. 37, figs 1‒6. 616 Material: A few specimens were recovered from the sample V6/7 at Vilobí del Penedès 617 (Table 1). Three gyrogonites were measured (Table 2 in supplementary data). 618 Description: Gyrogonites are large (878 μm high and 733 μm wide), ellipsoidal in shape 619 with an isopolarity index of 120. In the few gyrogonites recovered, spiral cells do not 620 display any modification in the width or thickness at the periphery of the apex (Fig. 8A). 621 The apex is flat, and the base is tapered showing a small basal pore. Nine or ten concave 622 spiral cells in lateral view separated by bicarinate sutures can be clearly observed near the 623 base (Fig. 8B). They are 120 μm high and devoid of ornamentation. 624 Fossil record and distribution: Lychnothamnus barbatus var. antiquus has been found in 625 several Miocene/Pliocene peri-Mediterranean lacustrine basins from Europe, North 626 Africa, and the Middle East. It has been reported from the Ebro and the Cerdanya basins 627 (NE Spain), Rio Maior (Portugal), Provence (France), Maoče (Montenegro), Çankırı 628 (Turkey) and Bekaa in Lebanon (Sanjuan and Alqudah 2018 and references therein). 629 Palaeoecology: The living species L. barbatus mainly thrives in permanent lakes of 630 northern Europe (Corillion 1972) where it forms dense meadows at depths ranging 631 between 2 and 8 m. These extant populations grow in cold and oligo-mesotrophic waters 632 of freshwater lakes, frequently associated to phreatic sources (Krause 1997; Brzozowski 633 et al. 2019). 634 Lychnothamnus sp. 635 Fig. 8D‒F 26 636 Material: Only one complete specimen has been found. This gyrogonite and other 637 fragments attributed to this taxon were recovered from sample MA1 at Mas de l´Alonso 638 (Table 1 and Table 2 in supplementary data). 639 Description: Gyrogonites are large, 1022 μm high and 777 μm wide, cylindrical in shape 640 with an isopolarity index of 131. Spiral cells are ornamented with a characteristic 641 midcelular crest (Fig. 8E). Spiral cells do not show any modification at the periphery of 642 the apex and show a constant width. The apex is flat and ornamented with coma-shaped 643 tubercles (Fig. 8D). The base is tapered showing a star-shaped basal funnel (Fig. 8F). 644 Eight concave spiral cells are visible laterally, being separated by bicarinate sutures. This 645 character can be clearly observed near the base. 646 Remarks: The small number of recovered specimens hinders their identification to the 647 species level, but the ornamentation of this gyrogonite suggests affinities with species 648 formerly attributed to the fossil genus Stephanochara Grambast, which was synonymized 649 with Lychnothamnus by Soulié-Märsche (1989). 650 Genus Nitellopsis Hy, 1889 651 Subgenus Tectochara Grambast and Grambast-Fessard, 1954 652 Nitellopsis (Tectochara) merianii (Al. Braun ex Unger, 1852) comb. nov. Grambast and 653 Soulié-Märsche, 1972 654 Fig. 8G‒N 655 1852. Chara merianii Unger, p. 82, pl. 25, figs 10‒12. 656 1954. Tectochara merianii Grambast et Grambast-Fessard, p. 668. 657 1972. Nitellopsis (Tectochara) merianii Grambast et Soulié-Märsche, p. 11. 27 658 Material: Fifty gyrogonites were recovered from sample V6/7 at Vilobí del Penedès and 659 few fragments attributed to this taxon were found in sample MA1 (Table 1). Thirty-five 660 specimens were measured (Table 2 in supplementary data). 661 Description: Gyrogonites are large, ovoidal (Fig. 8K‒N), 781‒1120 μm high (mean 958 662 μm) and 656‒957 m wide (mean 824 μm), with an isopolarity index ranging between 104 663 and 135 (mean 116). Spiral cells are flat to convex. Seven to nine (frequently eight) spiral 664 cells are visible in lateral view (116 μm high). Gyrogonites display a nitellopsidoid apex 665 (Fig. 8G‒H), i.e., spiral cells become thinner and narrower at the periapical zone (Horn 666 af Rantzien 1959). The apex is flat and ornamented with large apical nodules. The basal 667 pole is conical showing a basal pore within a wide pentagonal funnel, 292 μm in diameter 668 (Fig. 8I‒J), sometimes star-shaped (Fig. 8J). 669 Fossil record and distribution: This species has been widely reported from several 670 Paleogene/Neogene basins from Europe, North Africa, the Middle East and Asia (Sanjuan 671 and Alqudah 2018 and references therein). The oldest representatives of this species were 672 discovered in latest Eocene (late Priabonian) deposits of the Ebro Basin (Sanjuan and 673 Martín-Closas 2014). Later, this species expanded its biogeographic range following 4 674 distributional phases to become finally subcosmopolitan in the Northern Hemisphere 675 during the Miocene (Soulié-Märsche et al. 1997; Soulié- Märsche et al. 2002; Sanjuan 676 and Martín-Closas 2015). 677 Palaeoecology: Nitellopsis (T) merianii is considered the ancestor of the living species 678 Nitellopsis obtusa (Sanjuan and Martín-Closas 2015). This taxon thrives in permanent, 679 cold, and deep alkaline lakes at a depth range between 4‒12 m, in large lakes of northern 680 Europe, Asia and North America (Corillion 1972). This species cannot tolerate salinities 681 higher than 5‰ (Katsuhara and Tazawa 1986). 682 Nitellopsis sp. 28 683 Fig. 8O‒Q 684 Material: A few abraded gyrogonites were recovered from sample V6/7, Vilobí del 685 Penedès. Only two specimens were measured (Table 2 in supplementary data). 686 Description: Gyrogonites are very large (1289 μm high and 1051 μm wide), ovoidal, with 687 an isopolarity index of 123. Spiral cells are flat and ornamented with a nodular mid- 688 cellular crest (Fig. 8P). Eight or nine spiral cells are visible in lateral view (155 μm high). 689 The apex is flat and of nitellopsidoid type (Horn af Rantzien 1959), ornamented with 690 small apical nodules (Fig. 8O). The basal pole is conical showing a small basal pore, 97 691 μm within a star-shaped funnel (Fig. 8Q). 692 Remarks: The main difference between this species and N. (T) merianii is the size and the 693 ornamentation. However, the small number of recovered specimens hinders their 694 identification to the species level. 695 696 -------------------------------------------Figure 8 near here------------------------------------- 697 698 Discussion 699 Charophyte taphonomy and palaeoecology 700 The non-marine deposits from els Casots, Vilobí del Penedés, and Mas de l´Alonso‒el Pi 701 Gros studied in this work have yielded a diverse number of charophyte and two ostracode 702 species which have characteristic palaeoenvironmental distributions. Several of them 703 have living representatives, which allow for more accurated palaeolimnological 704 inferences for the wetlands developed in the Vallès-Penedès and Vilanova i la Geltrú 705 basins during the Langhian and Serravalian. 29 706 707 We distinguish three different assemblages in the depositional environments studied: 708 Charophytes from shallow lakes: The microfossil assemblage associated to shallow lakes 709 varies locally. Charophyte assemblages at els Casots are dominated by Chara cf. vulgaris. 710 This living species has wide ecological tolerance and lives in brackish or freshwater 711 environments (estuaries, rivers, springs, ponds and lakes). It mostly occurs at very 712 shallow depths, from several centimeters up to 1 m and can live in eutrophic water 713 (Krause 1997). This species can withstand oligohaline conditions and it only produces 714 gyrogonites at salinities up to 5 ‰ (Ghetti et al. 2002). Associated taxa include C. 715 molassica var. notata and abraded tests of Ammonia sp. rotaliid foraminifera. C. 716 molassica var. notata is an extinct species related to freshwater lakes. The dominance of 717 C. vulgaris suggests very shallow water and that water trophism may have been high in 718 the paleolake of els Casots. 719 The flora in the upper part of the Vilobí del Penedès section (samples V5 and V6/7) 720 corresponds to a shallow lake as well. In this case the charophyte assemblage is dominated 721 by C. molassica var. notata. Other, less abundant, species include Sphaerochara 722 ulmensis, Nitellopsis (Tectochara) merianii, and Lychnothamnus barbatus var. antiquus. 723 A few poorly-preserved skeletons of rotaliid foraminifera (Ammonia sp.) also occur and 724 are considered allochthonous to the lake. Charophytes indicate the prevalence of 725 freshwater conditions. The occurrence of Nitellopsis (Tectochara) merianii and 726 Lychnothamnus barbatus var. antiquus (ancestors of the extant species Nitellopsis obtusa 727 and Lychnothamnus barbatus, respectively) thrive today in permanent, relatively deep 728 (2–8m), oligotrophic lakes with salinities not exceeding 5‰. The ecological requirements 729 of the charophyte assemblage at Vilobí del Penedès suggest that the lake was more stable 730 and oligotrophic than the contemporary lake of els Casots. 30 731 Finally, the assemblage from Mas de l´Alonso (sample MA1) also corresponds to shallow 732 lacustrine conditions. In spite of the scarce material recovered the charophyte assemblage 733 is quite diverse and also includes rare Ammonia sp. foraminifera. The charophyte 734 assemblage is constituted by Sphaerochara ulmensis, Chara cf. hispida, Lychnothamnus 735 sp. and fragmented gyrogonites attributed to Nitellopsis (Tectochara) merianii. This 736 assemblage indicates the prevalence of shallow freshwater conditions. Ammonia sp. tests 737 show evident signs of abrasion and are considered allochthonous, being probably 738 transported from nearby marine settings. 739 Charophytes from coastal salinas: Assemblages from Vilobí del Penedès (lower part of 740 the section) are dominated by the charophyte species Lamprothamnium papulosum, 741 which occurs associated to abraded ostracod carapaces. L. papulosum is the most 742 halotolerant species among living charophytes, surviving at salinities up to 70‰ (Burne 743 et al. 1980). Despite withstanding such hypersaline conditions, living populations of L. 744 papulosum require salinities around 20‰ (maximum 40‰) to produce gyrogonites 745 (Soulié-Märsche 1998 and references therein). The presence of monospecific 746 assemblages of L. papulosum at Vilobí del Penedès indicates the prevalence of brackish 747 water conditions, limiting the development of other charophyte species (Fig. 10). Several 748 germinated gyrogonites indicate that seasonally the palaeosalinity could have been going 749 down to 10‰ for short periods of time. 750 751 -------------------------------------------Figure 9 near here------------------------------------- 752 753 Charophytes from permanent lakes: Based on the assemblages from el Pí Gros, two 754 charophyte species, Chara cf. hispida and Chara sp., and two ostracod taxa, Heterocypris 31 755 salina Brady 1868 (Fig. 9A‒D) and a probable leptocytherid (Fig. 9E‒J) are present. The 756 presence of articulated valves suggests in situ burial. Gyrogonites of Chara cf. hispida 757 are by far the dominant charophyte remains. Living meadows of C. hispida mainly grow 758 in alkaline permanent freshwater lakes at a depth up to 15 m, mainly between 0.5 and 7 759 m (Krause 1997). This is an eurythermal species preferring calcareous and well- 760 oxygenated waters (Barinova et al. 2014 and references therein), which can also tolerate 761 brackish water conditions, as reported from the Baltic Sea (Mannschreck 2003).. Chara 762 hispida shares similar habitat preferences with other Chara species and with Nitellopsis 763 obtusa (Barinova et al. 2014). While charophyte thalli are absent in the marls, charophyte 764 gyrogonites are very well preserved showing the endocalcine and ectocalcine layers 765 intact, without traces of corrosion or dissolution, thus evidencing that they were buried in 766 situ or near the production area. The ostracods found in these lacustrine facies provide 767 significant information about the characteristics of the palaeo-waterbody at el Pi Gros. 768 The leptocytherid species is similar to Leptocythere gr. psammophila in Gliozzi et al 769 (2005). Heterocypris salina Brady, 1868 is widely distributed across Europe, West Asia, 770 and North Africa inhabiting several types of waterbodies, from freshwater springs and 771 wells to littoral zones of seas (Meisch, 2000; Gusakov et al. 2021). This species is found 772 in slightly saline coastal and continental waters with salinities ranging from 5 to 35‰ 773 (Gusakov et al. 2021), although Meisch (2000) reports that it is more usually associated 774 with oligosaline waters, with an optimum salinity preference of around 6 ‰. Meisch 775 (2000) further maintains that the species can also live in freshwater so that its presence 776 should not be regarded as indicative of a saline waterbody in the absence of supporting 777 evidence. Although Miocene leptocytherids have been reported from both marine 778 brackish and athalassic saline environments (Gliozzi et al. 2005), the co-occurrence of 779 the specimens with H. salina, together with additional micropalaeontological evidence, 32 780 suggests that el Pí Gros corresponds to lacustrine and oligohaline waterbody (Fig. 10). 781 This interpretation is consistent with the conclusions of Gliozzi et al (2005), who have 782 previously reported the co-occurrence of leptocytherids, including Leptocythere gr. 783 psammophila with Heterocypris salina from Seravallian lacustrine or palustrine 784 sediments from the Iberian Peninsula. 785 786 -------------------------------------------Figure 10 near here------------------------------------- 787 788 Conclusions 789 Middle Miocene (Langhian–Serravallian) lacustrine deposits of the Vallès‒Penedès and 790 Vilanova i la Geltrú basins (SW of Barcelona) are here analyzed from the 791 sedimentological and palaeoenvironmental viewpoints. The three localities studied, i.e., 792 els Casots (Subirats), Vilobí del Penedès, and Mas de l´Alonso‒el Pi Gros (Vilanova i la 793 Geltrú), yielded a well-preserved charophyte assemblage compprising ten different 794 species. The recovered flora shows a characteristic palaeoenvironmental distribution and 795 sheds new light on the prevailing palaeolimnological characteristics of the wetlands in 796 the coastal basins of NE Iberia during the Middle Miocene. Facies analysis, taphonomy 797 and palaeoecology of the recovered taxa allows for the distinction of three different 798 palaeoenvironmental settings: shallow lake, coastal salina, and permanent stable lake. 799 Claystones related to shallow lakes occur in the three studied localities and have yielded 800 Sphaerochara ulmensis, Chara cf. vulgaris, Chara molassica var. notata, 801 Lychnothamnus barbatus var. antiquus, Lychnothamnus sp., Nitellopsis (Tectochara) 802 merianii, and Nitellopsis sp. This flora is unevenly distributed owing to local 803 palaeolimnological conditions. Mesotrophic to eutrophic conditions dominated the 33 804 Langhian palaeolake at els Casots, while more oligotrophic and stable conditions might 805 have prevailed at the upper part of the Vilobí del Penedès (Langhian) section and at Mas 806 de l’Alonso (Serravallian). The occurrence of abraded tests of benthic foraminifera 807 suggests that these freshwater lakes were situated close to the coastline. Facies related to 808 coastal salinas are only found at the base of the Vilobí del Penedès section (Langhian) 809 and are characterized by primary gypsum beds alternated with greenish claystones rich in 810 Lamprothamnium papulosum. Monospecific populations of this halotolerant taxon 811 indicate the prevalence of brackish waters with salinity fluctuations from low to 812 hypersaline concentrations. However salinities of around 20‰ are needed for the 813 production of gyrogonites. Finally, charophytes found in marls and limestones attributed 814 to permanent stable lakes occurs at el Pi Gros (Serravallian). The lake was inhabited by 815 Chara cf. hispida and Chara sp. as well as the ostracod species ?Leptocythere gr. 816 psammophila and Heterocypris salina, indicating the prevalence of oligohaline and well 817 oxygenated waters. 818 This is the first in-depth study of the charophytes from the Vallès-Penedès and Vilanova 819 basins. The results presented here show that charophytes are crucial for accurately 820 reconstructing palaeolimnological conditions, such as the water salinity, trophism or 821 depth. The combined study of charophytes and other microfossils, such as ostracods and 822 foraminifera, even improve such palaeoenvironmental inferences, especially regarding 823 salinity. 824 This study provides just a limited view of the total range of palaeoenvironments recorded 825 in this area. The whole Vallès sector and other associated sub-basins such as Sant Andreu 826 de la Barca, which contains mainly fluvio-lacustrine facies, have yet to be studied but 827 preliminary results indicate that they are also rich in charophyte fossils. Perhaps more 828 importantly, the combined continental record of these Catalan coastal basins covers 34 829 almost the entire Miocene, therefore allowing for the study of the evolution of terrestrial 830 wetland ecosystems along protracted time intervals. 831 832 Acknowledgments 833 This study was funded by the project IBERINSULA (PID2020-113912GB-100 / AEI / 834 10.13039/501100011033) of the Spanish Research Agency (AEI) and the European 835 Regional Development Fund (ERDF) and to project SGR2017-841 from the AGAUR 836 Research Agency (Catalan Autonomous Government). We thank Dr. Carles Ferràndez 837 (Department of Earth and Ocean Dynamics, Universitat de Barcelona) for his advice in 838 the determination of benthic foraminifera. Alejandro Gallardo (Laboratory of 839 Palaeontology, Department of Earth and Ocean Dynamics, Universitat de Barcelona) is 840 acknowledged for preparing the thin sections. We are also very thankful to David Artiaga 841 of the Centres Científics i Tecnològics, Universitat de Barcelona (CCiTUB) for his 842 technical support during the SEM sessions. Josep Rovira (Vilobí del Penedès) is 843 acknowledged for reporting us the charophyte beds from this locality. Dave Horne (Queen 844 Mary, University of London) advised on the ostracod identification. We also thank the 845 editor G. Dyke and the reviewer I. Soulié-Märsche for constructive comments during the 846 peer-review process. 847 848 849 35 850 References 851 Agardh CA. 1824. Systema Algarum. Lund, Lundae Literis Berlingianis, 312 p. 852 Agustí J, Cabrera L, Moyà-Solà S. 1985. Sinopsis estratigráfica del Neógeno de la fosa 853 del Vallés-Penedés. Paleontologia i Evolució. 18:57–81. 854 Agustí J, Cabrera L, Domènech R, Martinell J, Moyà-Solà S, Ortí F, de Porta J. 1990. 855 Neogene of the Penedès area (Prelittoral Catalan Depression, NE Spain). 856 Paleontol Evol mem espec. 2:187–207. 857 Agusti J, Cabrera L, Garces M, Pares JM. 1997. The Vallesian mammal succession in 858 the Vallès-Penedès Basin (northeast Spain): Paleomagnetic calibration and 859 correlation with global events. Palaeogeogr. Palaeoclimatol. Palaeoecol. 133 (3- 860 4): 149‒180. 861 Alonso-Zarza AM, Wright VP. 2010. Palustrine carbonates. In: Alonso-Zarza AM, 862 Tanner LH, editors. Carbonates in Continental Settings. Facies, Environments and 863 Processes: Developments in Sedimentology, 61. Elsevier: Amsterdam, p. 103– 864 126. 865 Anadón P, Julià R, De Deckker P, Rosso JC, Soulié-Märsche I. 1987. Contribución de la 866 paleolimnología del Pleistoceno inferior de la Cuenca de Baza (sector Orce-Vetna 867 Micena). Paleontol Evol. mem. espec.1:35‒72. 868 Baciu C, Feist M. 1999. Les charophytes Oligocènes du nord-ouest de la Transylvanie 869 (Roumanie). Acta Palaeontol. Roman. 2:27–29. 870 Barinova S, Romanov R, Solak CN. 2014. New record of Chara hispida (L.) Hartm. 871 (Streptophyta: Charophyceae, Charales) from the Işıklı Lake (Turkey) and Critical 872 Checklist of Turkish Charophytes. Nat Resour Conserv. 2 (3):33‒42. 36 873 Bartrina MT, Cabrera L, Jurado MJ, Guimerà J, Roca E. 1992. Evolution of the central 874 Catalan margin of the Valencia trough (western Mediterranean). Tectonophysics. 875 203(1):219–247. https://doi.org/10.1016/0040-1951(92)90225-U 876 Bathia SB, Soulié-Märsche I, Gemayel P. 1998. Late Pliocene and Early Pleistocene 877 charophyte floras of the Haripur Formation, Kerawa Group, Kashmir, India. N. 878 Jb. Geol. Paläont. Abh. 210 (2):185‒209. 879 Batllori J, Moreno J. 2003. Nota preliminar sobre la malacofauna (Gastropoda) del Miocè 880 del Pi Gros (Vilanova i La Geltrú, Garraf): Butll Inst Catalana Hist Nat. 71:117– 881 127. 882 Belaústegui Z, Domènech R, Martinell J. 2018. An ichnofossil-lagerstätte from the 883 Miocene Vilanova Basin (Ne Spain): Taphonomic and paleoecologic insights 884 related to bioerosion structures. Palaios 33:16–28. 885 Bessedik M. 1985. Reconstitution des environnements miocènes des régions nord-ouest 886 méditerranéennes à partir de la palynologie. PhD Thesis, Université des Sciences 887 et Techniques du Languedoc, Montpellier: 162 p. 888 Berger JP. 1983. Charophytes de 1’Aquitanien de Suisse occidentale. Essai de taxonomie 889 et biostratigraphie. Geobios 16 (1):5‒37. 890 Bitzer K. 2004. Estimating paleogeographic, hydrological and climatic conditions in the 891 upper Burdigalian Vallès‒Penedès Basin (Catalunya, Spain). Geol. Acta 2:321– 892 31. 893 Brady G. 1868. A monograph of the recent British Ostracoda. Transactions of the Linnean 894 Society of London 26:353‒495. 37 895 Braun A, Nordstedt O. 1882. Fragmente einer Monographie der Characeen nach den 896 hinterlassenen Manusckripte A. Braun’s, herausgegeben von Dr. O. Nordstedt. 897 Berlin, Abhandlungen der Kaiserlichen Akademie der Wisenschaften, p. 211. 898 Brzozowski M, Palomares-Cabanilles M, Kowalewski G, Pełechaty M. 2019. 899 Environmental factors responsible for the gyrogonite formation by an endangered 900 macroalga, Lychnothamnus barbatus, a fertility indicator of past and present 901 lacustrine ecosystems. Limnologica 77:125686. 902 Burne RV, Bauld JI, Deccker P. 1980. Saline lake charophytes and their geological 903 significance. J. Sediment. Petrol. 50:281‒293. 904 Cabrera L. 1979. Estudio estratigráfico y sedimentológico de los depósitos continentales 905 basales de la depresión del Vallès-Penedès [dissertation]. Barcelona: Universitat 906 de Barcelona. 907 Cabrera L. 1981a. Influencia de la tectónica en la sedimentación continental de la 908 cuenca del Vallès-Penedès (provincia de Barcelona, España) durante el Mioceno 909 inferior. Acta Geol Hisp. 16:163–169. 910 Cabrera L. 1981b. Estratigrafía y características sedimentológicas generales de las 911 formaciones continentales del Mioceno inferior de la cuenca del Vallés Penedés 912 (Barcelona, España). Estud Geol. 37:35–44. 913 Cabrera L, Calvet F. 1996. The onshore Neogene record in NE Spain: Vallès-Penedès 914 and El Camp half-grabens (NW Mediterranean). In: Friend PF, Dabrio CJ, 915 editors. Tertiary Basins of Spain: Record of Crustal Kinematics. Cambridge, 916 United Kingdom: Cambridge University Press; p. 97–105. 917 Cabrera L, Calvet F, Guimerà J, Permanyer A. 1991. El registro sedimentario miocénico 918 en los semigrabens del Vallès‒Penedès y de El Camp: organización secuencial y 38 919 relaciones tectónica-sedimentación. In: Colombo, F, editor. Libro Guía nº 4 del 920 Congreso del GET, Vic: 132 p. 921 Cabrera L, Roca E, Garcés M, de Porta J. 2004. Estratigrafía y evolución 922 tectonosedimentaria Oligocena superior-Neógena del sector central del margen 923 Catalán (Cadena Costero-Catalana). In: Vera JA. editor. Geología de España, 924 Sociedad Geológica de España, Instituto Geológico y Minero de España: 569– 925 573. 926 Casanovas-Vilar I, DeMiguel D, Galindo J, Robles JM, Garcés M, Cabrera L. 2011. The 927 continental Burdigalian (Early Miocene) of the Vallès‒Penedès Basin (Catalonia, 928 Spain). In: Pérez-García A, Gascó F, Gasulla JM, Escaso F. editors. Viajando a 929 Mundos Pretéritos. Ayuntamiento de Morella, Morella: p. 93–100. 930 Casanovas-Vilar I, Madern A, Alba DM, Cabrera L, García-Paredes I, Van den Hoek 931 Ostende LW, De Miguel D, Robles JM, Furió M, Van Dam J, Garcés M, Angelone 932 C, Moyà-Solà S. 2016a. The Miocene mammal record of the Vallès‒Penedès 933 Basin (Catalonia). C R Palevol. 15:791–812. 934 Casanovas-Vilar I, Garcés M, Van Dam J, García-Paredes I, Robles JM, Alba DM. 2016b. 935 An updated biostratigraphy for the late Aragonian and the Vallesian of the Vallès- 936 Penedès Basin (Catalonia). Geol Acta 14 (3):195‒217. 937 Casanovas-Vilar I, Garcés M, Marcuello A, Abella J, Madurell-Malapeira J, Jovells- 938 Vaqué S, Cabrera Ll, Galindo J, Beamud E, Ledo JJ, Queralt P, Martí A, Sanjuan 939 J, Martín-Closas C, Jiménez-Moreno G, Luján AH, Villa A, DeMiguel D, Sanchez 940 I, Robles JM, Furió M, Van den Hoek Ostende LW, Sanchez-Marco A, Sanisidro 941 O, Valenciano A, Garcia-Paredes I, Angelone C, Pons-Monjo G, Azanza B, 942 Delfinor M, Bolet A, Grau-Camats M, Vizcaíno-Varo V, Mormeneo D, Kimura 39 943 Y, Moyà-Solà S, Alba DM. 2022. Els Casots (Subirats, Catalonia), a key site for 944 the Miocene vertebrate record of Southwestern Europe. Hist Biol. 34 (8):1494‒ 945 1508. 946 Castel M, Grambast L. 1969. Charophytes de l´Éocène des Corbières. Bull Soc Géol Fr. 947 7 (11):936‒943. 948 Chellaï EH, Baudelot S, Crochet B, Feist M, Durand-Delga M. 1982. Preuves 949 paléontologiques de l´antériorité des Conglomérats de la Grésine (Éocène 950 supérieur) par rapport à la Molasse oligocène de l´Albigeois (Tarn). C. R. Acad. 951 Sci. Paris 295:683‒690. 952 Corillion R. 1972. Les Charophycées de France et d’Europe Occidentale. Travaux du 953 Laboratoire de Botanique de la Faculté des Sciences d’Angers, Angers: 11‒12, 954 499 p. 955 de Gibert JM, Casanovas-Vilar I. 2011. Contexto geológico del Mioceno de la cuenca del 956 Vallès-Penedès. Paleontologia i Evolució. Mem. espec. 6: 39–45. 957 Détriché S, Bréhéret JG, Soulié-Märsche I, Karrat L, Macaire JJ. 2009. Late Holocene 958 water level fluctuations of Lake Afourgagh (Middle-Atlas Mountains, Morocco) 959 inferred from charophyte remains. Palaeogeogr Palaeoclimatol Palaeoecol. 960 283:134–147. 961 Dubois, A. 1968. Observations sur la morphologie et la biologie des formes naines de 962 Lamprothamnium papulosum J. Groves (Characées). Naturalia Monspeliensia Ser 963 Bot. 19:37‒41. 964 Fan H, Gasse F, Huc A, Li Y, Sifeddine A, Soulié-Märsche I. 1996. Holocene 965 environmental changes in Bangong Co basin (Western Tibet). Part 3: Biogenic 966 remains. Palaeogeogr Palaeoclimatol Palaeoecol. 120:65‒78. 40 967 Feist-Castel M. 1977. Etude floristique et biostratigraphique des Charophytes dans les 968 series du Paleogene de Provence. Géol. Médit. 4:109‒138. 969 Feist M, Ringeade M. 1977. Étude biostratigraphique et paléobotanique (charophytes) 970 des formations continentales d’Aquitaine, de l’Éocène supérieur au Miocène 971 inférieur. Bull Soc Géol Fr. 7:341‒354. 972 Feist M, Anadón P, Cabrera L, Choi SJ, Colombo F, Saez M. 1994. Upper Eocene- 973 Lowermost Miocene charophyte in the Ebro Basin (Spain). Contribution to the 974 charophyte biozonation in Western Europe. Newsl Stratigr. 30:1‒32. 975 Feist M, Grambast-Fessard N, Guerlesquin M, Karol K, Lu H, Mccourt RM, Wang Q, 976 Shenzen Z. 2005. Treatise on Invertebrate Paleontology. Part B., Protoctista 1. 977 Charophyta, The Geological Society of America, Kansas: 170 p. 978 Fontboté JM. 1954. Las relaciones tectónicas de la depresión del Vallès‒Penedès con la 979 Cordillera Prelitoral Catalana y con la Depresión del Ebro. In: CSIC editor. Tomo 980 extraordinario de trabajos geológicos publicado con motivo del 80 aniversario del 981 nacimiento del profesor Eduardo Hernández-Pacheco. R Soc Esp Hist Nat. p. 281– 982 310. 983 Foster GL, Lear CH, Rae JWB. 2012. The evolution of pCO2, ice volume and climate 984 during the middle Miocene. Earth Planet Sci Lett. 341–344:243–254. 985 https://doi.org/10.1016/j.epsl.2012.06.007 986 García A. 1999. Quaternary charophytes from Salina del Bebedero, Argentina: Their 987 relation with extant taxa and palaeolimnological significance. J Paleolimnol. 988 21:307–323. 989 García A, Chivas AR. 2004. Quaternary and extant euryhaline Lamprothamnium Groves 990 (Charales) from Australia: Gyrogonite morphology and paleolimnological 41 991 significance. J Paleolimnol. 31(3):321‒341.Ghetti P, Anadón P, Bertini A, Esu D, 992 Gliozzi E, Rook L, Soulié-Märsche I. 2002. The Early Messinian Velona Basin 993 (Siena, Central Italy): Paleoenvironmental and paleobiogeographical 994 reconstructions. Palaeogeogr Palaeoclimatol Palaeoecol. 187:1‒33. 995 Gierlowski-Kordesch EH. 2010. Lacustrine carbonates. In: Alonso-Zarza AM, Tanner 996 LH, editors. Carbonates in Continental Settings. Facies, Environments and 997 Processes: Developments in Sedimentology, 61. Elsevier, Amsterdam: p. 1‒101. 998 Gliozzi E, Rodriguez-Lazaro J, Nachite D, Martin-Rubio M, Bekkali R. 2005. An 999 overview of Neogene brackish leptocytherids from Italy and Spain: 1000 Biochronological and palaeogeographical implications. Palaeogeogr 1001 Palaeoclimatol Palaeoecol. 225:283‒301. 1002 González-Pardos M. 2012. Carófitos del Mioceno inferior de la Formación Tudela 1003 (Cuenca del Ebro, Navarra). Master´s thesis, Universitat de Valencia: 69 p. 1004 Grambast L. 1962. Aperçu sur les Charophytes tertiaires du Languedoc et leur 1005 signification stratigraphique. C R Somm Séances Soc Géol Fr. 10:313‒314. 1006 Grambast L, Grambast-Fessard N. 1954. Sur la position systématique de quelques 1007 charophytes tertiaires. Rev Gén Bot. 61:665‒771. 1008 Grambast L, Paul P. 1965. Observations nouvelles sur la flore de charophytes du 1009 Stampien du bassin de Paris. Bull Soc Géol Fr. 7:239‒247. 1010 Grambast L, Soulié-Märsche I. 1972. Sur l’ancienneté et la diversification des Nitellopsis 1011 (Charophytes). Paléobiol cont. 3:1‒14. 1012 Groves J. 1916. On the name Lamprothamnus Braun. J Bot. 54:336–337. 42 1013 Guimerà J. 1992. Les fosses i depressions Neògenes. Història natural dels Països 1014 Catalans. Geologia 2. Enciclopedia Catalana, S.A. Barcelona: 548 p. 1015 Gusakov VA, Makhutova ON, Gladyshev MI. Golovatyuk LV, Zinchenko TD. 2021. 1016 Ecological role of Cyprideis torosa and Heterocypris salina (Crustacea, 1017 Ostracoda) in saline rivers of the Lake Elton basin: abundance, biomass, 1018 production, fatty acids. Zool Stud. 60:60‒53. 1019 Holbourn A, Kuhnt W, Kochhann KGD, Andersen N, Sebastian Meier KJ. 2015. Global 1020 perturbation of the carbon cycle at the onset of the Miocene Climatic Optimum. 1021 Geology. 43(2):123–126. https://doi.org/10.1130/G36317.1 1022 Horn af Rantzien H. 1959. Morphological types and organ-genera of Tertiary Charophyte 1023 fructifications. Stockh Contrib Geol. 4:45‒197. 1024 Huang RJ, Zhong ZQ, Nie YS. 1988. Early Tertiary Charophytes from the Dongpu 1025 region. In: Series on Stratigraphy and Paleontology of Oil and Gas bearing Areas 1026 in China. Research Institute of Exploration and Development, Zhongyuan 1027 Petroleum Exploration Bureau and Nanjing Institute of Geology and 1028 Paleontology, The Petroleum Industry Press, Beijing: 188 p. 1029 ICC (Institut Cartogràfic de Catalunya). ICGC - Vissir3 (icc.cat) (accessed 15 April 1030 2021).Janis CM. 1993. Tertiary mammal evolution in the context of changing 1031 climates, vegetation, and tectonic events. Annu Rev Ecol Syst. 24:467–500. 1032 Jovells-Vaqué S. 2020. Early Miocene cricetids from the Vallès- Penedès Basin 1033 (Catalonia): taxonomy, biostratigraphy and paleoecological implications. PhD 1034 Thesis, Universitat Autònoma de Barcelona, Bellaterra: 370 p. 1035 Jovells-Vaqué S, Casanovas-Vilar I. 2021. Dispersal and early evolution of the first 1036 modern cricetid rodents in Western Europe: new data from the Vallès-Penedès 43 1037 Basin (Catalonia). Comptes Rendus Palevol. 20(22):401–439. 1038 https://doi.org/10.5852/cr-palevol2021v20a22 1039 Julià de Agar JJ. 1991. Contribución al estudio de carófitos fósiles de la Cuenca Terciaria 1040 de Loranca (provincia de Cuenca, España). Coloquios de Paleontología 43:57‒78. 1041 Katsuhara M, Tazawa M. 1986. Salt tolerance in Nitellopsis obtusa. Protoplasma 1042 135:155‒161. 1043 Knobloch E. 1979. Fossil Charophyta from the Neogene and Holocene of Iraq. Vĕst Ústr 1044 Úst Geol. 54: 237–241. 1045 Korsch H. 2016. Chara vulgaris, In: Arbeitsgruppe Characeen Deutschlands, editors. 1046 Armleucheralgen. Die Characeen Deutschlands. Springer Verlag, Heidelberg: p. 1047 379‒387. 1048 Krause W. 1997. Charales (Charophyceae). Süsswasserflora von Mitteleuropa. Band 18. 1049 Gustav Fischer Verlag, Jena: 202 p. 1050 Leonhardi H Von. 1863. Über die böhmischen Characeen. Lothos 13:55‒62. 1051 Lindley J. 1836. A Natural System of Botany. Longman, 2nd edition, London: 526 p. 1052 Linnæus C. 1753. Species plantarum, exhibentes plantas rite cognitas, ad genera relatas, 1053 cum differentiis specificis, nominibus trivialibus, synonymis selectis, loci 1054 natalibus, secundum sexuale digestas, vol. 2. Impensis Laurentii Salvii. 1055 Stockholm: p. 561–1.200. 1056 Liu JY. 1989. Cenozoic Paleobiota of the continental shelf of the East China Sea 1057 (Donghai). Geological Publishing House, Beijing: 324 p. 1058 Lu H, Luo Q. 1990. Fossil Charophytes from the Tarim Basin. Xinjiang. Series on 1059 Stratigraphy and Palaeontology of oil and gas bearing areas in China. Science and 1060 Technical Documents Publishing House, Beijing: 261 p. 44 1061 Ludwig K. 1987. Mikropaläontologische Hinweise für die autochthone Entstehung der 1062 miozänen Braunkohle im Becken von Teruel-Ademuz (NE- Spanien) Paläontol Z. 1063 61:17–27. 1064 Mädler K. 1952. Zur Taxonomie der Tertiären Charophyten. Geol Jahrb. 70:265‒328. 1065 Mädler K. 1955. Charophyten aus dem Nordwestdeutschen Kimmeridge. Geol Jahrb. 67, 1066 1–46. 1067 Mädler K, Staesche U. 1979. Fossile Charophyten aus dem Könozoikum (Tertiär und 1068 Quartär) der Türkei. Geol Jahrb. B 33:81‒157. 1069 Mannschreck B. 2003. Chara hispida (L.) Hartm. 1820. In: Schubert I, Blindow H, 1070 editors. Charophytes of the Baltic Sea. Gantner Verlag, Ruggell, Liechtenstein: p. 1071 107–112. 1072 Martín-Closas C. 1999. Epiphytic overgrowth of charophyte thalli by stromatolite-like 1073 structures and fungi in the Lower Cretaceous of the Iberian Ranges (Spain). Aust 1074 J Bot. 47: 305‒313. 1075 Martín-Closas C, Ramos E. 2005. Palaeogene Charophytes of the Balearic Islands 1076 (Spain). Geol Acta. 3:39‒58. 1077 Meisch C. 2000. Freshwater Ostracoda of Western and Central Europe. Süßwasserfauna 1078 von Mitteleuropa 8/3. Gustav Fischer, Stuttgart, 522 p. 1079 Meyen F. 1827. Beobachtung und Bemerkungen über die Gattung Chara. Linnaea 1080 2(1):55–81. 1081 Mennad A, Adaci M, Tabuce R, Martín-Closas C, Benyoucef M, Bensalah M, Otero O, 1082 Sarr R, Zaoui D. 2021. Découverte de charophytes et ostracodes de l’Yprésien 1083 inférieur dans les Monts des Ksour (Algérie): biostratigraphie et paléoécologie. 1084 Ann Paléontol. 107:102466. 45 1085 Migula W. 1897. Die Characeen, In: Rabenhorst L, editor. Kryptogamen-Flora von 1086 Deutschland, Oesterreich und der Schweiz. E. Kummer, Leipzig: 772 p. 1087 Miller KG, Browning JV, Schmelz WJ, Kopp RE, Mountain GS, Wright JD. 2020. 1088 Cenozoic sea-level and cryospheric evolution from deep-sea geochemical and 1089 continental margin records. Science Advances. 6(20): eaaz1346. 1090 https://doi.org/10.1126/sciadv.aaz1346 1091 Ortí F. 1990. Yesos de Vilobí (Mioceno). In: Ortí F, Salvany JM, editors. Formaciones 1092 evaporíticas de la Cuenca del Ebro y cadenas periféricas, y de la zona de Levante 1093 Barcelona, ENRESA: 306 p. 1094 Ortí F, Pueyo JJ. 1976. Yeso primario y secundario de depósito de Vilobí (província de 1095 Barcelona, España). Publicaciones del Instituto de Investigaciones Geológicas, 1096 Universidad de Barcelona 31:5–34. 1097 Ortíz JE, García-Cortés E, Colliga LA, Mansilla H, Torres T. 1998. Yacimientos de 1098 carofitas en la Depresión Intermedia (Cuenca, Guadalajara, España central). 1099 Geogaceta 23:115‒118. 1100 Permanyer A. 1990. Sedimentologia i diagènesi dels esculls miocènics del Penedès. 1101 Arxius de les Seccions de Ciències, Institut d’Estudis Catalans 92: 321 p. 1102 Pukacz A, Boszke P, Pelechaty MJ, Raabe U. 2012. Comparative study of the oospore 1103 morphology of two populations of a rare species Chara baueri A. Braun in 1104 Cedynia (Poland) and Batzlow (Germany). Acta Soc Bot Pol. 81:131–136. 1105 Ramos-Guerrero E, Casas A, Pinto V, Agustí J. 1996. Estructura y relleno sedimentario 1106 de la semifosa Neógena de Vilanova (Garraf, Barcelona). Acta Geol Hisp. 29:93– 1107 106. 46 1108 Rasky K. 1945. Fossile Charophyten-Früchte aus Ungarn. Ungarischen 1109 Naturwissenschaftlichen Museums 2:1–75, 3 pl. 1110 Reichenbacher B, Schwarz J. 1997. Charophyten und Otolithen aus den Cyrenen- 1111 Schichten des nördlichen Alpenvorlandes. PalZ. 71:173–188. 1112 Roca E. 2001. The northwest Mediterranean (Valencia through, Gulf of Lyons and 1113 Liguro-Provençal basins): structure and geodynamic evolution. In: Ziegler PA, 1114 Cavazza W, Robertson AHF, Crasquin-Soleau S, editors. Peritehtyan rift/wrench 1115 basins and passive margins. Mém Mus Hist Natl Paris 186:671–706. 1116 Roca E, Guimerà J. 1992. The Neogene structure of the eastern Iberian margin: structural 1117 constraints on the crustal evolution of the Valencia through (Western 1118 Mediterranean). Tectonophysics 203:203–218. 1119 Roca E, Sans M, Cabrera L, Marzo M. 1999. Oligocene to Middle Miocene evolution of 1120 the Central Catalan margin (North-western Mediterranean). Tectonophysics 315: 1121 209‒229. 1122 Ruprecht FJ. 1845. Distributio Cryptogamarum Vascularium in Imperio Rossico. 1123 Beiträge der Pflanzenkunde des Russischen Reiches 3: 56 p. 1124 Sanjuan J, Alqudah M. 2018. Charophyte flora from Miocene deposits of Zahle (Beeka 1125 Valley, Lebanon). Biostratigraphic, Palaeoenvironmental and 1126 Palaeobiogeographical implications. Geodiversitas 40 (10):195‒209. 1127 Sanjuan J, Martín-Closas C. 2014. Taxonomy and palaeobiogeography of charophytes 1128 from the Upper Eocene-Lower Oligocene of the eastern Ebro basin (Catalonia, 1129 NE Spain). Geodiversitas 36 (3):385‒420. 1130 Sanjuan J. Martín-Closas C. 2015. Biogeographic history of two Eurasiatic Cenozoic 1131 charophyte lineages. Aquat Bot. 120 (A):18‒31. 47 1132 Sanjuan J, Vicente A, Flor-Arnau N, Monleón T, Cambra J, Martín-Closas C. 2017. 1133 Effects of light and temperature on Chara vulgaris gyrogonite productivity and 1134 polymorphism. Paleoenvironmental implications. Phycologia 56 (2):204‒212. 1135 Sanz de Siria Catalán A. 1993. Datos sobre la paleoclimatología y paleoecología del 1136 Neógeno del Vallès-Penedès según las macrofloras halladas en la cuenca y zonas 1137 próximas. Paleontologia i Evolució 26–27:281–289. 1138 Sanz de Siria Catalán A. 1994. La evolución de las paleofloras en las cuencas cenozoicas 1139 catalanas. Acta Geol Hisp. 29:169–189. 1140 Schwarz J. 1997. Charophyten aus dem Tertiär des Oberrheingrabens (Mitteleozän- 1141 Untermiozän). Palaeontographica B 243:1–84. 1142 Schwarz J. Reichenbacher B. 1989. Die Charophytenflora der Kirchberger Schichten 1143 (Unter-Miozän). Geol Bavarica. 94:179–193. 1144 Smith GM. 1938. Botany. Algae and Fungi. Charophyceae. New York, McGraw Hill, 1: 1145 127 p. 1146 Soulié-Märsche I. 1978. Contributions a la Paléontologie du Miocéne Moyen continental 1147 du Bassin du Tage. IV-Charophytes-Póvoa de Santarém, Pęro Filho et Tremęs. 1148 Cięncias da Terra. Earth Sciences Journal 4:91–102. 1149 Soulié-Märsche I. 1982. Lac pléistocène du Fezzan (Libye). In: Le Shati (editions du 1150 centre national de la recherche scientifique). Marseille, France: p. 80‒85. 1151 Soulié-Märsche I. 1989. Étude comparée de gyrogonites de charophytes actuelles et 1152 fossiles et phylogénie des genres actuels. Millau, Imprimerie des Tilleuls: 237 p. 1153 Soulié-Märsche I. 1991. Charophytes as lacustrine biomarkers during the Quaternary in 1154 North Africa. J Afr Earth Sci. 12 (1/2):341‒351. 48 1155 Soulié-Märsche I. 1998. Fossil Lamprothamnium papulosum (Charophyta), a biomarker 1156 for seasonal rainfall in northern Mauritania. In: Heine K, editor. Paleoecology of 1157 Africa 25: p. 65‒76. 1158 Soulié-Märsche I. 2007. Charophytes, indicators for low salinity phases in North sebkhet. 1159 J Afr Earth Sci. 51:69‒76. 1160 Soulié-Märsche I. Martín-Closas, C. 2003. Lychnothamnus barbatus (Charophytes) from 1161 the Upper Miocene of La Cerdanya (Catalonia, Spain): taxonomic and 1162 palaeoecological implications. Acta Micropal Sinica. 20:156–165. 1163 Soulié-Märsche I, Gemayel P, Chaimanee Y, Suteethorn V, Jaeger JJ, Ducrocq S. 1997. 1164 Nitellopsis (Charophyta) from the Miocene of northern Thailand. Alcheringa 1165 21:141‒156. 1166 Soulié-Märsche I, Benammi M, Gemayel P. 2002. Biogeography of living and fossil 1167 Nitellopsis (Charophyta) in relationship to new finds from Morocco. J Biogeogr. 1168 29:1703‒1711. 1169 Soulié-Märsche I, Bieda S, Lafond R, Maley J, Baitoudji M, Vincent PM, Faure H. 2010. 1170 Charophytes as bio-indicators for lake level high stand at “Trou au Natron”, 1171 Tibesti, Chad, during the Late Pleistocene. Glob Planet Change. 72 (4):334–340. 1172 Steinthorsdottir M, Coxall HK, Boer AM de, Huber M, Barbolini N, Bradshaw CD, Burls 1173 NJ, Feakins SJ, Gasson E, Henderiks J, Holbourn AE, Kiel S, Kohn J, Kürschner 1174 VM, Lear CH, Liebrand D, Kunt DJ, Mörs T, Pearson PN, Pound MJ, Stoll H, 1175 Strömberg E. 2020. The Miocene: the future of the past. Paleoceanogr 1176 Paleoclimatol. 36(4): e2020PA004037. https://doi.org/10.1029/2020PA004037. 1177 Straub EW. 1952. Mikropaläontologishe Untersuchungen im Tertiär zwischen Ehingen 1178 und Ulm an der Donau. Geol Jahrb. 66:433–524. 49 1179 Tang L, Di H. 1991. Fossil charophytes from Qaidam basin, Qinghai. Beijing: 220 p., 79 1180 pl. (in Chinese with abstract and table in English). 1181 Unger F. 1852. Iconographia plantarum fossilium. Denkschriften Akademie der 1182 Wissenschaften, Mathematisch-Naturwissenchaftliche Klasse, Wien 4: p. 73‒118. 1183 Van Dam JA, Krijgsman W, Abels HA, García-Paredes I, López Guerrero P, Peláez- 1184 Campomanes P, Ventra D. 2014. Updated chronology for Middle to Late Miocene 1185 mammal sites of the Daroca area (Calatayud-Montalbán Basin, Spain). Geobios. 1186 47:325–334. 1187 Van der Meulen AJ, García-Paredes I, Álvarez Sierra MÁ, Van den Hoek Ostende LW, 1188 Hordijk K, Oliver A, Pélaez-Campomanes P. 2012. Updated Aragonian 1189 biostratigraphy: small mammal distribution and its implications for the Miocene 1190 European chronology. Geol Acta. 10:159–179. 1191 https://doi.org/10.1344/105.000001710 1192 Wallroth FG. 1833. Flora cryptogamia Germaniae. Pars posterior. Continens Algas et 1193 Fungos. In: Bluff MJ, Fingerhuth CA, editors. Compendium florae Germanicae. 1194 Sectio II plantae cryptogamicae sine cellulosae. Tomus IV: 923 p. 1195 Xinlun Z. 1978. Early Tertiary Charophytes from Coastal Region of Bohai. Nanjing 1196 Institute of Geology and Paleontology. Nanjing (in Chinese): 49 p. 1197 You Y, Huber M, Müller RD, Poulsen CJ, Ribbe J. 2009. Simulation of the Middle 1198 Miocene Climate Optimum. Geophys Res Lett. 36:L04702. 1199 doi:10.1029/2008GL036571. 1200 Zachos J, Pagani M, Sloan L, Thomas E, Billups K. 2001. Trends, rhythms, and 1201 aberrations in global climate 65 Ma to present. Science. 292(5517):686–693. 1202 https://doi.org/10.1126/science.1059412. 50 1203 Zachos JC, Dickens GR, Zeebe RE. 2008. An early Cenozoic perspective on greenhouse 1204 warming and carbon-cycle dynamics. Nature. 451(7176):279–283. 1205 https://doi.org/10.1038/nature06588. 1206 51 1207 Figure captions 1208 Figure 1. Geological map of the study area. Red stars represent the location of the three 1209 studied localities in the Vallès-Penedès and Vilanova i la Geltrú basins. V=Vilobí; 1210 E.C=els Casots; M.A= Mas de l´Alonso; P.G=el Pí Gros. Modified from ICC (2021). 1211 Figure 2. Stratigraphic logs of the 4 studied localities. 1) els Casots (Subirats); 2) Vilobí 1212 del Penedès; 3) el Pí Gros (Vilanova i la Geltrú); 4) Mas de l´Alonso (Vilanova i la 1213 Geltrú). 1214 Figure 3. Facies and microfacies from els Casots (Subirats). A‒B. Field photos of the 1215 stratigraphic section. A. organic-claystones with pedogenetic features, see the location of 1216 the sample C4. B. limestone interval from where the samples C2 was extracted. C‒H. 1217 Microphotographs of the limestone sample C2. C. Packstone of cyanobacterial crusts. D. 1218 subrounded oncoid. E. fragments of mollusk shells (m.s). F. detail of a mollusk shell. G. 1219 well-developed oncoid (Onc). H. detail of cyanobacterial crusts. 1220 Figure 4. Field photos of the section at Vilobí del Penedès. A. laminated gypsarenite 1221 interval at the base of the section. B. gypsum (G) and claystone alternation at the base of 1222 the section, see the position of samples yielding charophytes V1, V3, and V4. C. Partially 1223 covered greenish claystones, see the position of the sample V5. D. Oyster bank at the top 1224 of the section. 1225 Figure 5. Facies and microfacies from the el Pi Gros locality (Vilanova i la Geltrú). A.‒ 1226 B. Field photos of the stratigraphic section. A. general overview of the upper half section. 1227 B. facies showing a shallowing upward cycle from base to top; marly limestone (M), 1228 wackestone-packstone limestone (W-P. L), stromatolitic limestone (S.L) and bioturbated 1229 claystone (B.C). C‒F. Microphotographs of the sample PG1. C. wackstone with 1230 gyrogonites (Gyr). D. Detail of a Chara gyrogonite showing the basal plate (B.P). E. 1231 wackestone‒packstone with ostracods (ostr). F. wackestone‒packstone with charophytes 52 1232 showing diagenetic dissolution. G‒H. Microphotographs of PG4 sample, wackestone‒ 1233 packstone with ostracods (ostr), gyrogonites (gyr), and thalli. 1234 Figure 6. Fossil charophytes from the Vallès-Penedès and Vilanova i la Geltrú basins. A‒ 1235 D. Sphaerochara ulmensis from sample V6/7. A. apical view, IPS122528.1; B. lateral 1236 view, IPS122528.2; C. lateral views; IPS122528.3; D. basal view, IPS122528.4. E-I. 1237 Sphaerochara ulmensis from sample MA1, Mas de l´Alonso. E. apical view, 1238 IPS127516.1; F. lateral view, IPS127516.1; G. lateral view, IPS127516.2; H. lateral view, 1239 IPS127516.3; I. basal view, IPS127516.4; J‒N. Chara cf. vulgaris from sample C4, els 1240 Casots. J. apical view, IPS127515.1; K. lateral view, IPS127515.2; L. lateral view, 1241 IPS127515.3; M. lateral view, IPS127515.4; N. basal view, IPS127515.5; O‒R. Chara 1242 molassica var. notata from sample C4, els Casots. O. apical view, IPS127515.6; P. lateral 1243 view, IPS127515.7; Q. lateral view, IPS127515.8; R. basal view, IPS IPS127515.8; S‒U. 1244 Chara molassica var. notata from sample V6/7, Vilobí del Penedès. S. apical view, 1245 IPS122528.5; T. lateral view, IPS122528.6; U. basal view, IPS122528.7 1246 Figure 7. Fossil charophyte assemblage from the Vallès-Penedès and Vilanova i la Geltrú 1247 basins (continued). A‒H. Chara cf. hispida from sample PG2. A. lateral view, 1248 IPS127517.1; B. lateral view, IPS127517.2; C. lateral view, IPS127517.3; D. lateral view, 1249 IPS127517.4; E. apical view, IPS127517.5; F. apical view, IPS127517.6; G. basal view, 1250 IPS127517.7; H. basal view, IPS127517.8; I‒M. Chara sp. from sample PG2. I. apical 1251 view, IPS127517.9; J. lateral view, IPS127517.10; K. lateral view, IPS127517.11; L. 1252 lateral view, IPS127517.12; M. basal view, IPS127517.13; N‒U. Lamprothamnium 1253 papulosum from sample V5, Vilobí del Penedès. N. lateral view, IPS127514.1; O. lateral 1254 view, IPS127514.2; P. lateral view, IPS127514.3; Q. lateral view, IPS127514.4; R. apical 1255 view, IPS127514.5; S. apical view, IPS127514.6; T. basal view, IPS127514.7; U. basal 1256 view, IPS127514.8. 53 1257 Figure 8. Fossil charophyte assemblage from the Vallès-Penedès and Vilanova i la Geltrú 1258 basins (continued). A‒C. Lychnothamnus barbatus var. antiquus from sample V6/7, 1259 Vilobí del Penedès. A. apical view, IPS122528.8; B. lateral view, IPS122528.9; C. basal 1260 view, IPS122528.10; D‒F. Lychnothamnus sp. from sample MA1, Mas de l´Alonso. D. 1261 apical view, IPS127516.5; E. lateral view, IPS127516.5; F. basal view, IPS127516.6; G‒ 1262 N. Nitellopsis (Tectochara) merianii from sample V6/7, Vilobí del Penedès. G. apical 1263 view, IPS122530.1; H. apical view, IPS122530.2; I. basal view, IPS122530.3; J. basal 1264 view, IPS122530.4; K. lateral view, IPS122530.5; L. lateral view, IPS122530.6; M. 1265 lateral view, IPS122530.7; N. lateral view, IPS122530.8; O‒Q. Nitellopsis sp. from 1266 sample V6/7, Vilobí del Penedès. O. apical view, IPS122528.11; P. lateral view, 1267 IPS122528.11; Q. basal view. IPS122528.12. 1268 Figure 9. Fossil ostracods and foraminifera from Vallès-Penedès and Vilanova i la Geltrú 1269 basins. A‒D. Heterocypris salina from sample PG2, el Pi Gros locality. A. external view 1270 of a right valve, IPS127517.14; B. internal view of a right valve, IPS127517.15; C. dorsal 1271 view of carapace, IPS127517.16; D. ventral view of carapace, IPS127517.17; E‒J. 1272 ?Leptocythere gr. L. psammophila from sample PG2, el Pi Gros locality. E. external view 1273 of a left valve, IPS127517.18; F. external view of a left valve, IPS127517.19; G. external 1274 view of a right valve, IPS127517.20; H. opened carapaces from the ventral view, 1275 IPS127517.21; I. dorsal view of carapace, IPS127517.22; J. ventral view of carapace, 1276 IPS127517.23; K‒M. Ammonia sp. from sample C4, els Casots. K. umbilical view, 1277 IPS127515.9; L. umbilical view, IPS127515.10; M. spiral view, IPS127515.11. 1278 Figure 10. Palaeoenvironmental model showing the distribution of the charophyte taxa 1279 and the distinguished depositional environments in the Vallès-Penedès and Vilanova i la 1280 Geltrú basins. Note that this is not a palaeoenvironmental reconstruction, as the Vallès- 1281 Penedès sites date back to the Langhian while those from the Vilanova i la Geltrú date 54 1282 back to the Serravallian. The figure is meant to illustrate the range of wetland 1283 environments occurring in these coastal basins during the Middle Miocene. 1284 Table 1. List of microfossils and their relative abundances based on a semi-quantitative 1285 visual estimation. 1286 Table 2 (in supplementary data). Biometric measurements of the studied charophyte 1287 samples from the Middle Miocene deposits of the E Vallès-Penedès and Vilanova i la 1288 Geltrú basins. 55 Figure 1 Sant Sadurní d´Anoia IBERIAN PENINSULA E.C N Vilobí del Penedès V Vilafranca del Penedès Garraf Massif Olivella t h ors gre tne on –M rraf Ga Sant Pere de Ribes M.A Sitges P.G Vilanova i sea El Vendrell ean La Geltrú iterran Med 4 km Legend Village Quaternary (fluvial, alluvial and beach deposits) Middle/Late Miocene (comglomerate) Section location Early Miocene (comglomerate and coarse sandstone) Middle/Late Miocene (marls and mudstone) Major normal faults Early Miocene (marls and mudstone) Jurassic and Cretaceous rocks (limestone) Minor faults Early Miocene (reefal limestone) Early/Middle Miocene (bioclastic limestone) Early Miocene (gypsum) Figure 2 1. ELS CASOTS (SUBIRATS) 2. VILOBÍ DEL PENEDÈS 3. PI GROS (VILANOVA I LA GELTRÚ) Color Lithology Color Lithology Color Lithology 700 300 PG 4 325 600 Interval 3 250 300 Lithology 500 70 200 275 60 400 V4 PG 3 50 150 250 V3 300 Interval 2 V6/7 40 V2 100 225 30 200 V1 PG 2 V5 50 20 200 100 PG 1 10 C3 0 Interval 1 m M W P G 175 C2 cm 0 C S Sf Sm Sc 0 cm m M W P G cm m M W P G C1 C S Sf Sm Sc C S Sf Sm Sc 4. MAS DE L´ALONSO (VILANOVA I LA GELTRÚ) 150 Color Lithology 125 100 MA 2 Legend 100 Limestone Ripples Gyrogonites 50 MA 1 Limonite horizon Charophyte thalli 75 Bioclastic limestone Bioturbation Ostracods 0 cm m M W P G Marly limestone C S Sf Sm Sc Plant remains Ostracod fragments 50 C4 Marlstone Carbonate nodules Freshwater gastropods Cyanobacterial crusts Potamidid gastropods 25 Claystone Root marks Mollusk´s fragments Siltstone Nodule Vertebrate remains 0 cm m M W P G Soft Samples Oysters C S Sf Sm Sc C / V/ PG / MA Gypsarenite Hard Samples Foraminifera C / PG / MA Equinoids Gypsum Rodophytes Lignites Figure 3 A B C2 C4 C D E M.S 600 µm 600 µm 200 µm F G H Onc 200 µm 600 µm 400 µm Figure 4 A B G V4 G V3 G G V1 C D V5 Figure 5 A B B.C S.L W-P. L M C D E Ostr B.P Gyr 1mm 300 µm 300 µm F G H Ostr Ostr Gyr Thalli Gyr Thalli 300 µm 300 µm 300 µm Figure 6 A B C D E F G H I 500 µm J K L M N O P Q R 400 µm S T U Figure 7 A B C D E F G H 500 µm I J K IN L M 500 µm N O P Q R S T U 500 µm Figure 8 A B C D E F 500 µm G H I J K L M N 500 µm O P Q Figure 9 A B C D 500 µm E F G H I J K L M 500 µm Figure 10 Table 1 Penedès El Pi Gros Vilobí del Sample els Casots Mas de l´Alonso Species V5 C4 V1 V4 V3 PG 3 PG 2 MA 1 V 6/7 Nitellopsis (T.) merianii Nitellopsis sp. Relative abundance Lychno. barbatus var. antiquus Lychnothamnus sp. rare Chara cf. vulgaris Chara molassica var. notata Chara cf. hispida Charophytes Chara sp. Sphaerochara ulmensis abundant Lamprothamnium papulosum ?Leptocythere gr. psammophila Ostracods Heterocypris salina rich Ammonia sp. Foraminifera indet. Fish teeth Table 2 (supplementary data). H=gyrogonite height, μm W=gyrogonite width, μm C.N=convolution number in lateral view S.C.H=spiral cell height, μm ISI=Isopolarity index. H/W x 100 ELS CASOTS Chara molassica var. notata. Sample C4 H W C.N S.C.H ISI 550 439 10 56 125 491 369 11 44 133 641 404 11 54 159 538 418 11 44 129 555 408 11 50 136 mean 491 369 10 44 125 min 641 439 11 56 159 max Chara cf. vulgaris. Sample C4 H W C.N S.C.H ISI 419 335 11 31 125 475 375 12 53 127 454 325 10 42 140 501 305 10 33 164 504 391 12 43 129 520 365 11 38 142 486 347 11 49 140 435 386 10 53 113 513 430 11 43 119 483 359 10 42 135 401 289 12 42 139 455 371 11 52 123 451 390 13 51 116 552 359 11 38 154 482 384 11 46 126 460 402 11 44 114 533 414 13 45 129 529 429 12 49 123 495 356 13 41 139 473 357 11 49 132 584 406 12 54 144 446 360 11 41 124 414 370 10 45 112 437 355 11 37 123 382 336 12 38 114 468 374 11 44 125 1 Table 2 (supplementary data). 456 293 12 40 156 465 359 10 41 130 481 375 10 54 128 442 377 10 60 117 470 299 10 48 157 444 346 11 45 128 456 333 10 39 137 481 350 10 33 137 474 301 11 42 157 378 323 11 48 117 435 345 12 43 126 487 352 12 37 138 468 323 11 48 145 543 357 10 53 152 492 330 10 57 149 426 256 10 39 166 449 325 12 40 138 428 334 11 37 128 386 319 10 39 121 470 347 10 45 135 467 361 12 46 129 501 333 12 43 150 419 325 10 42 129 421 363 12 46 116 466 352 11 44 133 mean 378 256 10 31 112 min 584 430 13 60 166 max VILOBÍ DEL PENEDÈS Chara molassica var. notata. Sample V6/7 H W C.N S.C.H ISI 488 320 11 53 153 570 402 11 56 142 542 383 10 68 142 517 331 9 60 156 498 411 10 52 121 544 389 9 71 140 531 368 10 51 144 557 384 9 72 145 547 368 11 51 149 468 333 10 58 141 523 386 10 57 135 478 398 10 64 120 509 344 9 60 148 558 367 10 50 152 451 416 8 69 108 2 Table 2 (supplementary data). 599 407 10 60 147 508 375 9 59 135 513 412 8 53 125 438 336 10 51 130 519 407 10 61 128 527 398 9 69 132 580 363 11 46 160 513 407 9 52 126 480 353 10 47 136 551 415 9 69 133 536 401 9 61 134 452 343 9 66 132 491 347 10 54 141 527 375 11 57 141 584 362 12 51 161 517 396 9 69 131 520 377 10 59 138 mean 599 416 12 72 161 max 438 320 8 46 108 min Sphaerochara ulmensis. Sample V6/7 H W C.N S.C.H ISI 590 576 10 62 102 630 619 9 73 102 652 564 9 84 116 623 569 9 74 109 596 582 9 79 102 622 567 10 73 110 614 514 10 64 119 571 514 10 63 111 607 578 10 71 105 612 565 10 71 109 mean 652 619 10 84 119 max 571 514 9 62 102 min Nitellopsis (Tectochara) merianii. Sample V6/7 H W number conv. conv H ISI 986 863 9 130 114 964 856 9 161 113 969 790 8 149 123 1035 767 9 130 135 907 832 8 145 109 781 656 8 104 119 914 774 8 170 118 1004 786 7 158 128 1120 957 8 171 117 3 Table 2 (supplementary data). 997 818 7 183 122 998 856 7 145 117 1079 914 8 141 118 947 867 8 149 109 922 886 7 179 104 933 852 7 158 110 843 786 9 107 107 894 810 7 137 110 958 866 8 130 111 1023 815 7 123 126 804 676 7 104 119 955 811 7 158 118 854 709 7 118 120 959 847 7 131 113 1036 904 8 131 115 1066 913 9 181 117 1004 865 8 152 116 899 816 7 130 110 943 813 7 164 116 988 838 7 139 118 928 697 7 131 133 951 878 9 114 108 1060 914 9 134 116 914 841 8 129 109 981 836 9 122 117 900 740 8 140 122 958 824 8 141 116 mean 1120 957 9 183 135 max 781 656 7 104 104 min Nitellopsis sp. Sample V6/7 H W C.N S.C.H ISI 1335 1053 8 164 127 1243 1048 9 146 119 1289 1051 9 155 123 mean Lychnothamnus barbatus var. antiquus. Sample V6/7 H W C.N S.C.H ISI 934 787 10 113 119 726 600 9 107 121 974 812 10 153 120 878 733 10 124 120 mean Lamprothamnium papulosum. Sample V5 H W C.N S.C.H ISI 592 464 10 88 128 4 Table 2 (supplementary data). 639 506 10 85 126 583 491 9 85 119 700 486 11 70 144 632 504 10 75 125 653 490 10 85 133 608 460 10 63 132 663 542 11 78 122 638 495 11 85 129 634 459 10 86 138 613 488 11 85 126 653 484 10 82 135 598 446 11 68 134 581 486 10 68 120 638 499 9 87 128 627 442 10 73 142 658 455 10 67 145 591 458 9 78 129 650 507 9 73 128 583 478 9 94 122 569 431 9 87 132 639 455 10 88 140 604 479 9 89 126 628 528 9 76 119 657 422 8 100 156 666 452 8 73 147 606 460 8 76 132 610 538 8 93 113 651 499 9 84 130 640 460 9 96 139 619 478 9 87 129 649 505 9 95 129 617 467 9 89 132 639 457 8 85 140 637 510 8 70 125 636 447 8 85 142 625 444 8 71 141 589 441 9 95 134 620 484 10 92 128 586 422 9 87 139 591 413 8 85 143 629 433 9 67 145 690 436 9 72 158 610 455 9 75 134 616 462 11 76 133 599 460 9 78 130 663 373 9 73 178 5 Table 2 (supplementary data). 631 511 10 90 123 661 448 8 75 148 638 501 9 84 127 601 474 8 89 127 571 391 8 95 146 649 506 10 62 128 677 459 9 73 147 622 459 9 78 136 701 528 9 81 133 682 519 8 86 131 711 452 9 86 157 682 480 9 96 142 585 348 8 75 168 631 469 9 81 135 mean 711 542 11 100 178 max 569 348 8 62 113 min MAS DE L´ALONSO‒EL PI GROS Chara cf. hispida. Sample PG2 H W C.N S.C.H ISI 783 540 10 80 145 783 540 9 88 145 783 513 10 104 153 783 540 10 88 145 783 567 9 138 567 378 8 150 810 540 10 150 621 432 10 144 621 459 9 135 810 540 10 150 864 567 8 152 594 378 8 157 783 540 10 145 756 486 9 156 810 567 10 143 810 540 11 150 648 405 10 160 540 432 12 125 594 405 147 810 567 143 675 351 192 810 540 150 756 567 133 729 540 135 756 540 140 729 513 142 6 Table 2 (supplementary data). 729 513 142 756 540 140 540 405 133 783 540 145 810 567 143 675 486 139 648 405 160 702 540 130 810 540 150 702 432 163 891 439 203 648 567 114 675 540 125 540 432 125 648 432 150 810 540 150 567 405 140 702 567 124 724 540 134 756 513 147 702 432 163 756 513 147 648 405 160 837 540 155 783 540 145 702 486 144 756 459 165 783 486 161 675 513 132 810 567 143 783 540 145 783 540 145 648 405 160 621 405 153 702 486 144 810 567 143 756 513 147 540 351 154 783 540 145 702 486 144 594 540 110 783 567 138 810 513 158 810 513 158 621 486 128 756 459 165 7 Table 2 (supplementary data). 594 432 138 756 513 147 567 432 131 594 405 147 837 405 207 810 621 130 756 486 156 783 567 138 687 540 127 756 486 156 729 486 150 729 513 142 702 540 130 810 540 150 702 540 130 783 567 138 540 378 143 675 540 125 810 540 150 810 540 150 729 540 135 621 486 128 756 540 140 810 459 176 726 467 155 774 548 141 806 572 141 854 516 166 726 500 9,6 90 145 mean 891 621 207 max 540 351 110 min Chara sp. Sample PG2 H W C.N S.C.H ISI 612 350 6 100 175 625 387 7 75 161 825 400 7 112 206 687 379 7 96 181 mean Sphaerochara ulmensis. Sample MA1 H W C.N S.C.H ISI 477 431 9 50 111 433 407 10 41 106 413 330 10 39 125 413 378 9 43 109 446 385 10 43 116 8 Table 2 (supplementary data). 398 374 9 53 106 447 379 9 44 118 447 363 9 54 123 449 409 10 61 110 446 410 10 52 109 418 375 10 43 111 447 421 10 66 106 442 404 9 45 109 426 383 9 60 111 466 402 9 53 116 460 417 9 57 110 410 388 9 62 106 437 397 10 48 110 424 402 10 57 105 436 406 9 63 107 421 375 9 62 112 453 378 10 54 120 437 390 9 51 112 437 391 9 52 112 mean 477 431 10 66 125 max 398 330 9 39 105 min Lychnothamnus sp. Sample MA1 H W C.N S.C.H ISI 1022 777 8 70 131 9