Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

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Palaeogeography, Palaeoclimatology, Palaeoecology

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Molecular and mineral biomarker record of terrestrialization in the

Rhynie Chert
T.O. Akinsanpe a, b, S.A. Bowden a, J. Parnell a, *
a
School of Geosciences, University of Aberdeen, Aberdeen AB24 3UE, United Kingdom
b
Obafemi Awolowo University, Ile Ife 220103, Osun State, Nigeria

A R T I C L E I N F O A B S T R A C T

Keywords: The colonization of the terrestrial surface by land plants involved the evolution of a complex ecosystem of plants,
Devonian animals, fungi, algae and bacteria, within a mineral framework. The record of this advance is highly fragmentary
Rhynie basin and uncertain. However, a wealth of fossil evidence is preserved in the Lower Devonian Rhynie Chert lagerstätte,
Organic geochemistry
which is consequently considered to be the world's oldest preserved terrestrial ecosystem. The physical record of
Biomarker
Fossil plants
fossils in the Rhynie Chert is accompanied by a chemical record of molecular biomarkers and mineral diagenesis.
The biomarkers include components derived from vascular and other higher plants, and decomposers fungi and
bacteria. The chert contains the earliest record of the plant diterpane ent-beyerane, identified in this study. The
biomarker profile is distinct from that in Lower Devonian organic-rich shales at Strathpeffer, 100 km to the west,
where plants are not preserved and the environment was hypersaline. In addition to organic biomarkers, the
chert contains mineralogical characters which imply biological activity, including pyrite framboids, strongly
leached monazite and garnet, and pitted micas similar to grains altered by modern fungi. The molecular and
mineral biomarkers combine with the fossil record and phylogenetic data in the toolbox available to document
plant colonization and symbiosis on the terrestrial surface.

1. Introduction growth position, dominated by forms that are considered to be vascular

plants (Cascales-Miñana et al., 2019), and laminae of organic matter
The world's oldest preserved terrestrial ecosystem is in the 407- which are composed of phytodebris (plant cuticles etc.) together with
million-year-old (Lower Devonian: Pragian-Emsian) Rhynie Chert cyanobacteria (Fig. 2; Trewin and Fayers, 2015, Strullu-Derrien et al.,
(Fig. 1), a lagerstätte of early plants and their accompanying biota of 2023). The non-photosynthesizing fraction of organic matter would
cyanobacteria, green algae, fungi, arthropods etc. (Strullu-Derrien et al., include a range of microbial matter including archaea and bacteria,
2014; Trewin and Fayers, 2015; Edwards et al., 2018; Wellman et al., which dominate many modern hot spring systems and consume or
2019). The early evolution of plants was tied to the occurrence of fungi, liberate gases including methane and hydrogen (Gomez, 2011; Escuder-
with which they developed in symbiosis (Strullu-Derrien et al., 2014; Rodríguez et al., 2022), but which are not evident in petrographic
Lutzoni et al., 2018). The chert beds at Rhynie contain plants (Figs. 2, 3) studies. Organic matter also occurs in interbedded grey shales which
with remarkably preserved cells, in which, despite silicification, primary represent an ephemeral lacustrine environment. Several types of fungal
carbon is preserved (Abbott et al., 2017; Loron et al., 2023). Combined interaction with plants have been documented in the Rhynie Chert
with only a limited burial history, and therefore minimal thermal (Powell et al., 2000; Taylor et al., 2004; Strullu-Derrien et al., 2014;
alteration (Rice et al., 2002), the cherts are a viable source of biomarkers Krings et al., 2018).
which could represent the components of the ecosystem. Further data for the Lower Devonian terrestrial ecosystem can be
Several plants are distinguished in the chert (Trewin, 1994; Trewin measured in organic-rich shales found 100 km to the west of Rhynie at
and Fayers, 2015). There are multiple interpretations of their phylog­ Strathpeffer. The organic-rich shales from Strathpeffer share a low
eny, but a broad division can be made between true vascular plants thermal maturity with Rhynie sediments and their proximity implies a
(Asteroxylon, Rhynia), and less complex pre-vascular plants (Aglaophy­ similar climate and continentality. However, deposits from Strathpeffer
ton, Nothia, Horneophyton). The chert beds include plants in upright lack megascopic plant fossils, although they contain plant spores.

* Corresponding author.
E-mail address: [email protected] (J. Parnell).

https://doi.org/10.1016/j.palaeo.2024.112101
Received 3 October 2023; Received in revised form 8 February 2024; Accepted 16 February 2024
Available online 19 February 2024
0031-0182/© 2024 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 1. Geological map of region near Rhynie village (after Rice et al., 2002).

Fig. 2. Main facies in Rhynie Chert. Contrast between plant-bearing chert

(centre) and laminae of phytodebris with intermixed sand and authigenic oxide
minerals (lower). Note phytodebris not penetrated by roots.
Fig. 3. Thin section micrographs of Rhynie Chert and associated Windyfield
Together, the Lower Devonian deposits at Rhynie and Strathpeffer allow Chert, showing brown-coloured kerogen. A, Fungal ring (dark brown) within
plant stem, Trench 03/T1Chert bed 9; B, Fungal cysts within plant stem, Trench
testing for biomarkers of the Lower Devonian terrestrial ecosystem, and
03/T1 Chert bed 9; C, Cynanobacterial crusts on plant fragments, Windyfield
particularly between a unit with a life assemblage of plants and related
Chert float sample RW20. (For interpretation of the references to colour in this
biota, and one without.
figure legend, the reader is referred to the web version of this article.)
The distribution of organic biomarkers in terrestrial sedimentary
rocks helps to delineate the evolution of land plants (Versteegh and
to synthesis from abietic acid in conifers and earlier vascular plants
Riboulleau, 2010; Böcker et al., 2013). In the pre-angiosperm evolu­
(Noble et al., 1985; Romero-Sarmiento et al., 2011).
tionary scheme, tetracyclic diterpenoids are particularly valuable. These
Only exceptionally have fossil fungi in symbiosis with plants sur­
compounds, including ent-kaurane, ent-beyerane and phyllocladane, are
vived, but where early permineralization has preserved fungi, they may
relatively stable, so have become established as biomarkers, attributed

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

yield a molecular record. Biomolecular evidence for fungi in the focussed on samples from boreholes and trenches adjacent to the fault
geological record has focussed on the polyaromatic hydrocarbon per­ bounding the Devonian basin (Rice et al., 2002), and recorded relatively
ylene (Grice et al., 2009; Marynowski et al., 2013), which is documented high thermal maturity, including blackened palynomorphs (Rice et al.,
as a fungal biomarker back to the end-Permian. Recent data, however, 1995; Wellman, 2006). This probably reflects the channelling of hot
suggests that perylene can have a more diverse origin, including plant mineralizing fluids through the fault zone (Baron et al., 2004). The
pigmentation and diagenesis (Li et al., 2022). Perylene is also found in current study is based particularly on samples from a trench excavated
modern marine/lacustrine sediments, washed off land surfaces, and by in 2003, more distant (about 300 m) from the fault (Fig. 1), in which the
analogy perylene-bearing shales back to the Middle Devonian have been organic matter is coloured brown (Fig. 3) and hence holds a higher
interpreted to signify a terrestrial hinterland (Tulipani et al., 2015; Philp potential for the determination of biomarkers. Samples containing light
and De Garmo, 2020). Notwithstanding the uncertainty in interpreta­ brown kerogen also from the nearby Windyfield Chert (Fig. 3) confirm
tion, these observations indicate that perylene survives from the Devo­ that the ambient maturity is suitable to yield biomarkers.
nian and could be found in the Rhynie Chert. The organic-rich shales at Strathpeffer are slightly younger, from
The sedimentary host/template is a fundamental part of the middle to late Emsian (Richardson and Rasul, 1978). They accumulated
ecosystem, with which biological components can interact. The Devo­ in a half-graben, as at Rhynie, but the shales passed laterally into a thick
nian Rhynie Chert is ideal for study of such interaction in the geological sequence of conglomerates banked against the boundary fault (Clarke
record. Plants, fungi and minerals are well preserved by contempora­ and Parnell, 1999). While the bounding fault at Rhynie channelled
neous silicification, hot spring siliceous fluids introduced anomalous mineralizing fluids during sedimentation, the fault at Strathpeffer did so
amounts of trace elements to the environment, and the deposit has not at a later stage after sediment lithification (Parnell et al., 2016). Also as
been overprinted by later geological events or high temperatures. Fungi at Rhynie, the Strathpeffer sediments were partly chertified pre-
particularly provide phosphorus to plants as part of their symbiotic compaction, by silicification of gypsum, and were additionally per­
relationship. Several minerals are targeted by fungi in the modern mineralized by dolomite (Parnell, 1985). Abundant sulphate evaporites
environment, to enable associated plants to obtain specific elements. associated with the organic-rich shales led to pyrite precipitation in the
Examples include the extraction of phosphorus, potassium and magne­ shales (Parnell, 1985). Long-term generation of waters rich in hydrogen
sium from monazite, feldspars and amphiboles respectively (Hoffland sulphide from the pyritic shales led to the establishment of Strathpeffer
et al., 2003; van Schöll et al., 2006; Lian et al., 2008; Kang et al., 2021). as a spa town focussed around the outcrop of shales (Manson, 1879; Fox,
Some micas localize fungal activity, especially if they are potassium- 1889).
bearing, such as muscovite (e.g. van Schöll et al., 2006; Quirk et al., The sandstones in the Rhynie Chert contain quartz, feldspar, mica
2012; Song et al., 2015). Fungal extraction of lithium from the mica and chloritoid grains, with minor amounts of heavy minerals zircon,
lepidolite is also recorded (Sedlakova-Kadukova et al., 2020). White monazite, ilmenite and garnet. The monazite and zircon are derived
(potassic) micas are abundant in beds of phytodebris in the Rhynie Chert from erosion of Caledonian granites. Many of the sandstones are highly
(Trewin and Fayers, 2015), which also contain remains of fungi (Powell micaceous, which reflects their derivation from Dalradian (Neo­
et al., 2000), so the mica was targeted for study. proterozoic) metasediments dominated by micaceous schists, and
The potential of the biomarker record in the Rhynie Chert is evalu­ deposition in low-energy fluvial settings (Trewin and Fayers, 2015).
ated here, in particular to determine: White micas are predominantly muscovite, but with variable composi­
tion. Micas in the parent Dalradian schists had been variably altered to
(i) If molecular biomarkers can be resolved to represent different phengite, with variable composition in individual crystals, during the
components of the ecosystem, especially plants and fungi. Caledonian Orogeny (Dempster, 1992; Aoki et al., 2013), and these
(ii) Identification of petrographic evidence for a biological role in variable compositions are therefore expected in the derived Devonian
mineral precipitation, in which fungal activity is likely to be sediments. Biotite micas in Caledonian granite in the region are also
important. variable altered to phengitic muscovite (Harrison, 1990). The whole
(iii) Whether the biomarker distribution in Rhynie and Strathpeffer rock at Rhynie was silicified as part of the chert formation that affected
sediments is consistent with differences in environment in these the whole deposit.
two Lower Devonian sections.
3. Methods
2. Geological setting
Sampling. Samples were sourced from archives in the School of
The Rhynie Chert occurs in the Rhynie Basin, northern Scotland, a Geosciences, University of Aberdeen. They consist of three spatially
half-graben with a basin-fill of continental Lower Devonian (Lower Old associated suites: a trench (03/T1; Fig. 1) through bedrock excavated in
Red Sandstone; LORS) sedimentary and volcanic rocks (Rice et al., 1995; 2003, loose blocks lying on the bedrock in the trench, and a borehole
Trewin and Fayers, 2015). Palynology indicates a Pragian to earliest (19C) drilled in 1988. Each suite is preserved with depth data. All
Emsian age (Wellman, 2006). Lacustrine shales and sandstones of the samples were originally collected responsibly, and according to local
Dryden Flags Formation host the Rhynie Chert, an ephemeral pool with laws. Eleven distinct beds of the Rhynie Chert from trench 03/T1, and
hot springs within a geothermal wetland (Rice et al., 2002; Trewin and four grey shales representing an ephemeral lacustrine environment from
Fayers, 2015). The chert preserved the earliest terrestrial ecosystem borehole 19C, were sampled. The trench and borehole samples are
comprising vascular plants, fungi, macro-algae, aquatic and terrestrial linked by very close proximity (Fig. 1) and the occurrence of plant-
animals, which were buried by silica-rich waters from hot-spring vent bearing cherts in both. Samples were selected from borehole 19C
pools (Trewin and Fayers, 2015). The chert consists of variable combi­ depths 12.0 m, 14.7 m, 17.0 m, 17.2 m, 17.3 m and 19.4 m because
nations of two main facies; plant-bearing silica sinter which may be in petrographic study (Powell et al., 2000) identified them as well-
growth position, and layers of compacted phytodebris mixed with colonised by fungi. A sample of andesite from borehole 97/8, archived
detrital sediment up to sand grade (Fig. 2). A separate silicified body, the with the cherts, was analysed to test that no modern fungal signal was
Windyfield Chert, lies 700 m to the north-east of the Rhynie Chert, and present. Fifty samples of Rhynie Chert dominated by either plants or
represents a distinct centre of hydrothermal activity (Fayers and Trewin, laminae of phytodebris were drilled to produce powders. Nine samples
2004). The biota of the two cherts is very similar, and they were pene­ of near-coeval shales from Strathpeffer were also sampled. Before dril­
contemporaneous. The Windyfield Chert is a few hundred metres higher ling, the rock samples were treated with dichloromethane (DCM) fol­
in the succession than the Rhynie Chert (Rice and Ashcroft, 2004). lowed by distilled water to eliminate any weathering effects on their
Previous observations of organic matter in the Rhynie Chert were surfaces or associated contamination.

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 4. Schematic environment of Rhynie Chert, distinguishing biomarkers for specific components. Chromatograms from this study highlight diagnostic peaks (red)
in numbered samples. Schematic re-designed from N.H. Trewin (unpublished) and Strullu-Derrien et al. (2021). (For interpretation of the references to colour in this
figure legend, the reader is referred to the web version of this article.)

Elemental Analysis. The total organic carbon (TOC) values were (purge 40 ml min− 1 for 2 mins). The temperature programme for the GC
determined on 200 mg of powdered decarbonated samples using the oven was 80–295 ◦ C, holding at 80 ◦ C for 2 mins, rising to 10 ◦ C min− 1
LECO CS744 Carbon Sulphur Analyser at the University of Aberdeen. for 8 mins and then 3 ◦ C min− 1, and finally holding the maximum
The decarbonisation process was done by treating the samples with 20% temperature for 10 min− 1. Quantitative biomarker data were obtained
and 10% hydrochloric acid (HCl) several times until there was no for both saturate and aromatic fractions. These include n-alkanes and
effervescence. This was used to remove the inorganic carbon present in acyclic isoprenoids, measured on the charge-to-mass ratio (m/z) 85;
the samples in the form of gases such as CO2. tetracyclic diterpenoids (m/z 109, 123, 193 and 233); β-carotane (m/z
Biomarker measurement. The biomarker extractions were achieved 125); norhopanes, hopanes and methylhopanes (m/z 177, 191 and 205);
with DCM/methanol (MeOH) (9/1, v/v) using a DIONEX Accelerated and steranes (m/z 217 and 218). The aromatic compounds were
Solvent Extractor (ASE-200) and micro-extraction method (ME) for measured on m/z 183, 202, 219 and 252, including cadalene, pyrene,
sample masses <0.3 g. The micro-extraction process involved adding 10 retene and perylene, respectively. Mass spectra for diterpenoids,
ml of DCM and MeOH mix (1/1, v/v) to the weighed sample in a vial. norhopanes, cadalene and retene are shown in Figs. S2, S3, S4 and S5.
This was mixed and placed in the ultrasonic bath for 15 min. The The different tetracyclic diterpenoids were monitored on a combined m/
resulting solvents were then transferred into clean vials with a pipette. z 109 + 123 + 193 + 233 chromatogram. The peaks were identified by
These steps were repeated with a solvent mix of DCM and MeOH (3/1, comparing their mass spectra and retention times with available pub­
v/v) and 100% DCM. The dried extracted organic matter (EOM) from lished data for well-characterized samples from Romero-Sarmiento et al.
ASE and ME was separated into aliphatic, aromatic, and polar fractions (2011; Figs. S1, S2)) and Noble et al. (1985). A sample of SKT-D from the
by silica column chromatography using hexane, hexane/DCM (3/1, v/ 2011 study was run with our samples. A standard for perylene (>99%
v), and DCM/MeOH (2/1, v/v), respectively. The glassware used was purity), was obtained from Sigma Aldrich. Thermal maturity was esti­
thoroughly cleaned with a 93:7 mixture of DCM/ MeOH. This paper only mated from the 20S/20S + 20R ratio for C29 sterane, based on the P
focuses on the saturate and aromatic hydrocarbon fractions. stereoisomeric transformation and the corresponding increase in the
Analysis used an Agilent 6890 N gas chromatograph fitted with a proportion of the S isomer with maturation (Peters et al., 2005). The
J&W DB-5 phase 50 m MSD and a quadrupole mass spectrometer gammacerane index was calculated as the ratio of gammacerane/
operating in scan and selected ion monitoring (SIM) mode (dwell time (gammacerane + C30 hopane). The concentration of all compounds is
0.1 s per ion and ionisation energy 70 eV). The samples were injected reported relative to the internal standard of 5β-cholane, with the
manually using a split/splitless injector operating in splitless mode response of 5β-cholane measured on the m/z 217 ion chromatogram.

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

4. Results

4.1. Molecular biomarker preservation

At both Rhynie and Strathpeffer, permineralization pre-compaction

preserved the organic matter and a high-resolution suite of biomarkers
(Tables S1, S2). The 20S/20S + 20R ratio for the C29 sterane is in the
range 0.45 to 0.65 which indicates the upper part of the oil window. The
plant-bearing beds and phytodebris beds (Fig. 2) have mean organic
carbon contents of 0.36% (n = 26) and 0.77% (n = 22) respectively.
Consequently, we can decipher the organic components to a degree
exceptional in rocks of Lower Devonian age. Measurements focussed on
biomarkers for specific components of the ecosystem (Fig. 4), including
higher plants (cadalene, diterpenoids), fungi (perylene??) and biodeg­
radation most likely attributable to bacteria (25-norhopanes). Quanti­
tative data are calculated based on identifications of peaks for n-
alkanes/isoprenoid hydrocarbons (Fig. 5),diterpenenoids (Fig. 6), ster­
anes and hopanes (Fig. 7). Summary plots for pristane/phytane ratios,
steranes, hopanes, perylene, and gammacerane indices (Fig. 8) show
how the compositions of plant-bearing beds and phytodebris beds
compare. A ternary plot of C27, C28 and C29 steranes (Fig. 9) shows that
the plant and phytodebris samples have a comparable sterane compo­
sition, but Rhynie shale samples (n = 4) are relatively enriched in the C27
sterane, while the Strathpeffer shale samples (n = 9) are relatively
lacking in the C27 sterane. Changes in the diterpenoid distribution
through trench 03/T1 are shown in Fig. 10.

4.2. Raman spectroscopy

Measurements on carbon in Rhynie Chert samples converted to vit­

rinite reflectance values (Ro %) using the approach of Schito and Cor­
rado (2020) yield a range of maturities from 0.63 to 1.21% (Table S3).
Omitting two outliers of data (0.63, 1.21), the data range from 0.80 to
1.10%, with a mean value (n = 20) of 0.93 ± 0.10%. The median value is
also 0.93%.
Fig. 5. Mass chromatograms m/z 85 of aliphatic fractions for plant and phy­
todebris beds. Distribution of n-alkanes and acyclic isoprenoid hydrocarbons
(pristane Pr, phytane Ph) shown from samples of Rhynie Chert, and shales at 4.3. Sediment mineralogy
Rhynie and Strathpeffer for comparison. Plant-bearing bed has higher Pr/Ph
ratio than phytodebris bed. Several minerals in the phytodebris layers exhibit alteration,
including monazite, garnet and mica. Detrital monazite grains show
The sample extracts are preserved frozen in the Organic Geochemistry dissolution pits and reprecipitation (Fig. 11) and detrital garnet also
Laboratory, School of Geosciences, University of Aberdeen. shows dissolution pits (Fig. 11). The garnet grains typically contain
Raman Spectroscopy. A Renishaw inVia reflex Raman spectrometer several percent manganese. The alteration in heavy mineral grains in the
was used for micro-Raman analyses, with a backscattering geometry in chert is not observed in sandstones in the same succession.
the range of 700–3200 cm− 1 (first- and second-order Raman spectra), The most abundant altered minerals are silicified micas. The micas
with a 2400 1 mm− 1 spectrometer grating and CCD detector under a exhibit numerous holes, typically 2 μm diameter (Fig. 12). The holes are
maximum of ×100 optical power (numerical aperture (NA) of the lens of present in both polished and unpolished samples. Some mica grains
0.90). The slit opening was 65 μm with CCD area of c. 10 pixels (80% of show layers of different composition, alternating between ideal
the total signal height hitting the CCD chip) and a confocal hole of 200 muscovite and altered muscovite. The ideal muscovite contains no or
μm. A 514.5 nm diode laser was used for excitation with an output of 50 few holes, while the altered muscovite has many holes. The margins of
mW. Optical filters (1%) were used to adjust the power of the laser to many holes are brighter in SEM images. Analyses of the host mica show
<0.5 mW. Raman backscattering was recorded after an integration time higher contents of sulphur (up to 2%) in the bright rim, in contrast to the
of 20 s for three repetitions for each measurement. The Raman system background mica (usually no sulphur, rarely up to 1%). The grains have
was calibrated against the 520.7 cm− 1 band of silica. neoformed mineral precipitates on their surfaces, including titaniu­
Electron Microscopy. Scanning electron microscopy (SEM) was con­ m‑iron oxides and monazite.
ducted in the Aberdeen Centre for Electron Microscopy, Analysis and The chert is extensively mineralized by pyrite framboids (Fig. 11),
Characterisation (ACEMAC) facility at the University of Aberdeen using and also clusters of pyrite microcrystals with a less organized
a Carl Zeiss Gemini SEM 300 VP Field Emission instrument equipped morphology. The Rhynie framboids are commonly arsenic-rich, and in
with an Oxford Instruments NanoAnalysis Xmax80 Energy Dispersive some samples they are manganese-rich.
Spectroscopy (EDS) detector, and AZtec software suite. Imaging was
done in backscattered mode. Standards were supplied by the factory. 5. Discussion
Samples were polished using corundum and alumina pastes, cleaned
with ethanol, then carbon-coated. 5.1. Thermal maturity

The thermal maturity of the Rhynie Chert samples is indicated by

several lines of evidence. The kerogen is brown (Fig. 3), distinct from the

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 6. Summed m/z 109 + 123 + 193 + 233 chromatograms showing diterpenoid distribution in selected Rhynie samples. Peak XII represents ent-beyerane. Peak
XV represents ent-16α(H)-kaurane. Peak identifications are in Table S2. The mass spectra and retention time of peaks of studied samples were compared with that of
reference sample SKT-D in Romero-Sarmiento et al. (2011).

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 7. m/z 217 and 191 ion chromatograms for samples RC28 and RC27. C21P and C22P = C21 and C22 pregnane; C27 Dia S and R = C27 13β,17α(H) 20S and 20R
diasterane; C29 ααα S = C27 5α,14α,17α(H) 20S sterane; C29 αββ R = C27 5α,14β,17β(H) 20R sterane; C23TT = C23 13β,14α(H) tricyclic terpane; C27Ts = C27 18α
(H)-22,29,30-trisnorneohopane; C27Tm = C27 17α(H)-22,29,30-trisnorhopane; C30 αβ = C30 14α,17β (H) hopane, C31 αβ R = C31 14α,17β (H) 22R hopane.

blackened samples examined by Wellman (2006) from adjacent to the and shale, with the exception that the chert may have slightly high
fault zone. proportions of n-alkanes greater than C30 (Fig. 5). The Strathpeffer shale
Carbon Preference Index (CPI) values are near-unity (Table S1), has notably greater proportions of isoprenoids compared to n-alkanes
typical of oil window conditions. The sterane isomerization values (Fig. 5). This has typically been explained as a consequence of some of
(Table S1) are near-maximum, characteristic of upper oil window con­ the continental Devonian muddy lithologies being deposited in hyper­
ditions. Raman spectroscopy of carbon in the samples, converted to saline lakes in which a significant proportion of the primary productivity
vitrinite reflectance (Table S3), yields a mean Ro value of 0.93%, also was in the form of halophilic archaea whose cell membranes comprised
characteristic of the upper oil window. tetra-ether lipids and phytol (Duncan and Hamilton, 1988; Killops and
The maturity achieved reflects hydrothermal heating, as shown by Killops, 2005).
the greater maturity increase in the fault zone. This rapid heating is Fewer/shorter n-alkanes occur in Devonian plant matter, which may
distinct from thermal maturity due to burial, and may have allowed have lacked waxy leaves (Song et al., 2017; Blumenberg et al., 2018, but
greater preservation of biomarkers for a given reflectance value (Rabi­ see Martinez et al., 2019). The paucity of n-alkanes above C25 should
nowitz et al., 2017; Savage et al., 2018). For example, perylene is pre­ not, therefore, be interpreted to indicate a lack of higher plants. How­
sent in coal tar and wood pyrolysis products (Cook et al., 1933), so is ever, the m/z 85 chromatograms for grey shales (Fig. 5) suggest that the
known to survive short term high-temperature heating of the kind found contribution from higher plants may be lower than in the cherts, and the
in hydrothermal systems. organic matter was rather dominated by algae/bacteria/cyanobacteria.
Notwithstanding the nature of the heating, previous studies show In addition to differences between the proportions of n-alkanes and
that thermal maturities up to Ro 1% can preserve abundant biomarkers isoprenoids, samples of the Rhynie Chert mostly have higher ratios of
(Dzou et al., 1995), and they imply that the Rhynie Chert has a good pristane/phytane (Pr/Ph) and C29 17α,21β(H) hopane to C30 17α,21β(H)
potential for biomarker data. hopane, than the Strathpeffer shales (Fig. 8 a,b). These ratios compare
Pyrene occurs in many Rhynie Chert samples (Figs. S6, S7), in the relative abundance of homologous compounds where the numerator
common with other sedimentary rocks containing organic matter, at a is a lower carbon number (pristane and C29 hopane), that can form by
wide range of thermal maturities (Fang et al., 2015). oxidising of a higher carbon precursor (Hughes et al., 1995; Peters et al.,
2005). Anoxic conditions in the depositional environment restrict oxi­
5.2. Molecular biomarkers dising reactions and preserve higher carbon number homologues, thus
on the basis of these biomarker proxies it might be concluded that
5.2.1. n-alkanes and isoprenoids sedimentary organic matter in the Rhynie chert experienced more
The proportions of n-alkanes are similar for samples of Rhynie Chert persistently oxic conditions than the sedimentary organic matter in the

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 8. Quantitative data for biomarkers in Rhynie Chert, and shales at Rhynie and Strathpeffer. (a) Pristane/Phytane (Pr/Ph) ratio; (b) C29/C30 hopanes; (c)
gammacerane index; (d) C29 steranes/TOC.

shales. Qualitatively, Pr/Ph values in the range 1 to 3 could be inter­ conifer plant assemblage recorded in younger rocks (Hautevelle et al.,
preted to result from dysoxic conditions, similar to that experienced by 2006).
deltaic sediments where sedimentary organic matter is at least period­ Diterpenoids were identified in samples of the Rhynie Chert,
ically subaerially exposed (Hughes et al., 1995). However, although including C18-C20 tricyclic terpanes, ent-beyerane, kaurane, and phyl­
likely uncommon in typical sedimentary settings, pristane can also locladane (Fig. 6). The distribution of diterpenoids varies through the
derive from side chain cleavage of tocopherols (Goossens et al., 1984; sample set (Fig. 10), and defines three stages. In terms of the plant as­
Maeda et al., 2006) a group of compounds found in plants. Samples of semblages (observations of Powell et al., 2000 Trewin and Fayers,
Rhynie Chert with intact plant fragments have the highest ratios of 2015), samples dominated by the vascular plants Rhynia (trench; blocks
pristane to phytane (Fig. 8a), thus the higher proportion of pristane in 4 and 5, borehole 19C) and Asteroxylon (trench beds 1 and 2) all contain
these samples may reflect production by vascular plants as well as the ent-beyerane and ent-kaurane. The occurrence of ent-beyerane in the
subaerial nature of the hydrothermal spring. cherts which contain Rhynia and Asteroxylon is consistent with earlier
suggestions that ent-beyerane is synthesized in vascular plants (Romero-
5.2.2. Higher plants Sarmiento et al., 2011; Song et al., 2017). A larger set of samples,
Aromatic compounds cadalene and retene were detected in including cherts rich in the plants Horneophyton and Nothia (trench beds
numerous samples. Cadalene was recorded in almost all Rhynie Chert 3, 5 and 14) contain ent-kaurane, but not ent-beyerane. Ancient ent-
samples and also the Rhynie shale samples, and some of the Strathpeffer kaurane is attributed to synthesis in pre-vascular plants (Romero-Sar­
samples. Retene also occurs in several, but many fewer, Rhynie Chert miento et al., 2010, 2011), and modern liverworts are a source of the
samples. The biomarkers cadalene and retene are ubiquitous signatures compound and its derivatives (Qu et al., 2008). While the kaurane
of higher plants. Cadalene is taken as an indication of higher plant skeleton was developed in compounds in the pre-vascular plants, the
contribution to organic matter, back to the Middle Devonian (Philp and beyerene skeleton may have been lacking. Ent-beyerane is a diagenetic
De Garmo, 2020), and retene in Palaeozoic rocks is probably derived degradation product of ent-beyerol, the parent hydrocarbon of a tetra­
from compounds with a kaurane-type skeleton in pre-vascular plants cyclic diterpene found in higher plants (Noble et al., 1985). Most sam­
and algae (Romero-Sarmiento et al., 2010). The uniformly low retene/ ples from the upper part of the section (trench beds 8 and 9, borehole
cadalene ratios (mostly <0.1) in Rhynie samples are typical of a pre- 19C) have not yielded diterpenoids.

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 9. Ternary plot of C27, C28 and C29 steranes in Rhynie Chert, and shales at Rhynie and Strathpeffer.

Hitherto, the earliest records of ent-beyerane were in Middle Devo­ including animal life, than the plant-rich Rhynie Chert. In contrast, in
nian coals from China (Song et al., 2017). Younger occurrences of ent- the Strathpeffer hypersaline environment, C28 and C29 sterol precursors
beyerane have been reported in a few coals of Lower Carboniferous age were produced by phytoplankton (Volkman, 2003).
(Romero-Sarmiento et al., 2011; Blumenberg et al., 2018), and then are The C29 sterol precursors can derive from both plant sterols, as well
common from the Upper Carboniferous onwards. as sterols synthesized by algae (green algae, Volkman, 2003). Although
Diterpenoids are important in plant-fungus symbiosis. By analogy the proportions of C29 and C28 steranes are similar for both the Rhynie
with the function of modern terpene derivatives, diterpenoid synthesis Chert and Rhynie shale, the Rhynie Chert contains an order of magni­
in the Devonian was likely a defence against diseases and pests, tude higher absolute concentrations of C29 sterane than the Rhynie
including fungal pathogens (Karunanithi and Zerbe, 2019). Terpenoids shale. Given the presence of fossil plants in the cherts, this feature of the
in vascular plants also deter predation by herbivorous insects and other sterane composition reflects a higher total amount of C29 sterols that can
arthropods (Mumm et al., 2008; Sharma et al., 2017). Fungi and ar­ be derived from higher plants. The very high concentrations of C29
thropods are so widespread in the Rhynie Chert (Trewin and Kerp, 2017) steranes in the Strathpeffer shale (Fig. 8d) is due to the high productivity
that diterpenoid synthesis was a likely response to protect the plants. of the environment.
Nevertheless, genes involved in diterpene synthesis in plants were in
turn acquired by fungi and became involved in fungal metabolism 5.2.4. Decomposers: fungi and bacteria
(Fischer et al., 2015). This included a positive feedback role for fungal Perylene was identified in some samples (Figs. S6, S7), including
diterpenes as hormones in plant-fungus symbiosis (Schardl et al., 2013). samples in borehole 19C which were recorded as fungi-rich by petrog­
raphy (Fig. 3) (Powell et al., 2000). Perylene has been proposed as an
5.2.3. Steranes indicator of fungal decay of woody matter (Grice et al., 2009; Mar­
A ternary diagram illustrating the proportions of 5α14α17α R ster­ ynowski et al., 2013) in younger rocks. If the identification was extended
anes (Fig. 9) shows that the samples of Rhynie Chert are intermediate to indicate degradation of plant matter in older rocks, its identification
between the Rhynie and Strathpeffer shales. The Strathpeffer shale has a would be consistent with the abundance of fungi previously observed in
higher proportion of C28 sterane, a characteristic typically associated the Rhynie Chert (Powell et al., 2000; Taylor et al., 2004; Strullu-Derrien
with sedimentary organic matter from lakes (Huang and Meinschein, et al., 2014; Krings et al., 2018). However, it is important to note that
1979; Grantham and Wakefield, 1988), whereas the Rhynie shale has a recent evaluation of perylene indicates the link to fungi is not at all
greater proportion of C27 sterane, a characteristic that in other circum­ definitive (Li et al., 2022), and the occurrence is reported here to add to
stances is typical of marine sedimentary organic matter. In this instance, the database of identifications. The occurrence at Rhynie is also in rocks
the high proportion of C27 sterane in the Rhynie shale more likely rep­ at a higher maturity than the limit of about 0.7% Ro which is hitherto
resents the production of higher proportions of precursor C27 sterols assumed for perylene. Perylene is found in moderately abundant
(such as cholesterol) by zookplankton (Huang and Meinschein, 1979; amounts at geothermal sites in sedimentary rocks in Iceland, deposited
Kodner et al., 2008). Thus, the relatively high-C27 sterane composition over a similar temperature range (largely <100 ◦ C) to the Rhynie sili­
for the lacustrine Rhynie shales suggests a broader community, ceous sediments (Geptner et al., 2005, 2006). This is an analogous

9
T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 10. Distribution of diterpenoids through Rhynie section. Most samples in lower part of combined trench 03/T1 (0 to 12 m) and borehole 19C section (21 to 36
m) contain the plant Asteroxylon, and the biomarkers ent-beyerane and ent-kaurane. Overlying section contains samples with the plants Horneophyton and Nothia, and
just ent-kaurane, followed by section with no diterpenoids.

environment which may help to explain what is otherwise an anomalous environment, rather than alteration of lithified sediments after uplift.
occurrence of perylene. Perylene was also recorded in the carbonaceous Most previous records of 25-norhopanes are signatures of biodegraded
grey shales of borehole 19C. As in other published occurrences in oils or lithified shales, but they also form where organic matter was
Devonian shales (Tulipani et al., 2015; Philp and De Garmo, 2020), the microbially altered before burial (Cao et al., 2008). As these biomarkers
perylene is implied to be washed off a terrestrial surface. No perylene are preserved within silica from hot springs, they represent degradation
was detected in any of the 9 shale samples from Strathpeffer, or from a in the surface soil zone, rather than during subsequent burial, so the
control sample of andesite at Rhynie, which excludes the possibility of a decomposers formed part of the surface biota.
contribution from modern fungi.
Similarly, the C29 25-norhopane was detected in many of the Rhynie 5.2.5. Gammacerane
Chert samples, but not in the Strathpeffer shale samples. This difference Many of the Rhynie Chert samples, but none of the Rhynie shale
suggests that the degradation was a feature of the depositional samples, contain gammacerane (Fig. 8c). All the Strathpeffer shale

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 11. Backscattered electron images showing minerals in Rhynie Chert representing biological activity. A, Pyrite framboid, typical of bacterial activity in reduced
environments. B, Detrital garnet grain exhibiting sub-micron dissolution pits. C, Irregular surface of altered monazite grain, showing stubby re-growths of monazite
on surface. D, Coatings of iron-rich oxides (bright) on and around detrital plant matter (black). Samples carbon-coated.

samples contain gammacerane. The mean gammacerane index, which Lower Devonian terrestrial plant matter in Britain (Rayner, 1984; Ken­
measures the abundance relative to a bacterially-derived hopane, is rick and Edwards, 1988), and as at Rhynie occurs directly associated
much lower in Rhynie Chert (mean 0.02) than in Strathpeffer (mean with organic matter and formed pre-compaction. Framboidal pyrite is
0.21). These values are low and high relative to a benchmark division of probably a consequence of bacterial decay (Wacey et al., 2015). Addi­
values at 0.15 (Hao et al., 2009). The high value for the Strathpeffer tionally, under anaerobic conditions, sulphate-reducing bacteria may
samples is typical of a hypersaline environment, evidenced by replaced form consortia with fungal mycelia on decomposing organic matter, and
gypsum in the shales (Parnell, 1985), and further supported by the precipitate pyrite (Borkow and Babcock, 2003), as at Rhynie. Higher
detection of β-carotane in all of the samples. Gammacerane could degrees of pyritization (DOP values) are recorded in the plant beds
conceivably indicate elevated salinity at Rhynie in waters of the hot compared to the phytodebris beds (Parnell et al., 2022).
spring system. Although gammacerane is interpreted as asalinity indi­ The study encountered grains exhibiting holes (Fig. 12) that can be
cator, it is also deposited in diverse other environments, including fossil understood by modern analogues. Mitchell et al. (2019) described grains
fungi (Drake et al., 2021), which would be possible at Rhynie. No in modern Icelandic soils which exhibit biologically mediated weath­
β-carotane was detected in Rhynie samples. ering, including holes/tunnels, authigenic mineral precipitates (Fe-rich
in this case) and fretted surfaces. Tunnelling of grains by fungi is widely
observed (Hoffland et al., 2003; Smits, 2006; van Schöll et al., 2008),
5.3. Mineral biomarkers
and the tunnels may include filamentous structures which are direct
evidence of their fungal origin (Mitchell et al., 2021). The Rhynie mica
Pyrite framboids are abundant in the plant-bearing cherts (Fig. 11),
grain surfaces similarly exhibit holes, mineral precipitates and fretted
and they were encased by the earliest silica. The pyrite must have been
surfaces. The holes are of comparable size, about 2 μm in diameter, and
precipitated almost immediately to be included in the pre-compaction
have a comparable spatial density of about thirty 2 μm holes/μm2. In
silica. This implies a waterlogged, reduced environment, which is
addition, many sub-micron holes occur in both cases. Holes in modern
consistent with the evidence of biodegradation. Framboidal-textured
grains show bright rims in SEM images due to heavier elements (Mitchell
pyrite is very commonly developed within decaying organic matter,
et al., 2021), which is replicated closely by the bright rims around holes
including in plant debris (Garcia-Guinea et al., 1998; Grimes et al.,
in Rhynie grains. In both cases the rims contain sulphur. The particular
2002). Pyrite, at least some of which is framboidal, is found in other

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 12. Backscattered electron images showing altered detrital mica grains in Rhynie Chert, attributable to biological activity. A, Rounded detrital mica grain,
exhibiting dissolution pits, and crystals of authigenic monazite (bright) on grain surface. B, Close-up of grain surface, showing pitted mica and monazite grains. C,
Further close-up of dissolution pits showing bright (sulphur-rich) rims as found around modern fungal pitting. Samples carbon-coated.

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Fig. 13. Schematic summary of mineral provenance for bioavailable elements in Devonian sediment at Rhynie. Monazite from granite, and mica and garnet from
metasediments, supply bioavailable P/REE, K and Mn respectively.

occurrence of holes in muscovite micas is consistent with the targeting of (i) The molecular and mineral data is derived from the same limited
such grains to extract potassium by fungi (e.g., van Schöll et al., 2006; suite of samples collected within an area of <100 m2.
Quirk et al., 2012; Song et al., 2015). Further, the holes occur mostly in (ii) The organic matter and minerals were both sealed at a very early
altered muscovite, which is probably phengitic, inherited from phengitic stage by hot spring silicification, so the data reflect a very limited
micas in the Dalradian Supergroup and Caledonian granites which were time window.
eroded to form the Devonian sediments. Holes are present in both pol­ (iii) There is no alternative explanation for the observed mineral
ished and unpolished samples, which excludes polishing as a cause of features.
hole formation. (iv) The plant beds and phytodebris beds show distinctions in both
In addition to the pitted micas, the detrital grain assemblage in the the molecular (Pr/Ph) and mineral (DOP) data.
Rhynie Chert includes pitted monazite and pitted garnet (Fig. 11). Both
minerals are known to be dissolved by microbiota as a strategy to release 6. Conclusions
nutrients. Monazite in particular releases phosphorus, which is essential
to plant-fungal symbiosis which supplies phosphorus to plants (Corbett The data show that it is possible to obtain good quality signatures for
et al., 2017; Castro et al., 2020). The dissolution of garnet by microbes a wide range of organic biomarkers in the Rhynie Chert. Specific bio­
would release manganese and other trace elements (Ivarsson et al., markers for several of the main components of the ecosystem can be
2018). identified readily. The components include higher plants (cadalene,
The alteration of detrital grains and the authigenic mineralogy in the diterpenoids), fungi (perylene??) and biodegradation (25-norhopanes).
chert may be linked. The garnet grains would have released manganese Each of these characteristic biomarkers is present in at least 45% of the
when altered, and the Rhynie pyrite is unusually enriched in manganese. Rhynie Chert samples. Lacustrine shales in the Rhynie Chert succession
Coatings of manganese‑iron oxides on plant debris (Parnell et al., 2022) show distinctive compositions, characterized by the lack of the C29 25-
also indicate manganese availability, and by analogy with modern soils norhopane and gammacerane. The Lower Devonian rocks of Strath­
may have been mitigated by fungi (Santelli et al., 2011). Similarly, rare peffer are also distinct, characterized by lack of perylene, and abundant
earths and phosphorus released during the alteration of detrital mona­ gammacerane. The environment at Strathpeffer was different to that at
zite could be the source of the authigenic monazite that mineralizes Rhynie, characterized by hypersalinity, high productivity and lack of
altered mica grains (Fig. 12). any evidence of decomposition. These distinctions emphasize the fi­
The authigenic mineral precipitates in Rhynie grain surfaces include delity of the data.
titanium‑iron oxides, which may be similar to the iron-rich precipitates The new data help to understand the hot spring environment, which
recorded on modern grains by Mitchell et al. (2019). However, the is important as a residence for diverse life, including during terrestri­
Rhynie grains also include widespread neoformed monazite, which is alization, and as a source of novel microbial life with potential for
exceptional in any low-temperature environment. The occurrence of technological advances (Sen et al., 2014; Jardine et al., 2018).
monazite indicates a source of both rare earth elements (REEs) and The mineral component of the ecosystem shows evidence for a bio­
phosphate, most probably from a very proximal source, given the local logical contribution to authigenesis and alteration. Pyrite framboids,
abundance required. The occurrence of REE-bearing fluorocarbonate heavily leached monazite and garnet, and dissolution pits in micas
mineralization in the shales (Parnell et al., 2023) also indicates REE comparable with pits formed by modern fungi, may all be regarded as
mobility. The most likely source would be pre-existing monazite. The mineralogical biomarkers. The geological landscape at Rhynie made
potential for localized redistribution of monazite in sedimentary rocks is several important elements bioavailable as detrital grains. Monazite
shown by monazite overgrowths on detrital monazite grains in sand­ from granite, and mica and garnet from metasediments, supplied
stone (Milodowski and Zalasiewicz, 1991; Allaz et al., 2013). Monazite phosphorus, potassium and manganese respectively (Fig. 13). Each of
overgrowths are a consequence of low-temperature non-biological pro­ these elements can be liberated by fungi.
cesses, but there is also evidence in previous studies for phosphate The data from Rhynie, supported by data from Strathpeffer, show
mineralization by fungi and bacteria, including monazite neoformation that the earliest terrestrial ecosystem is recorded by an assemblage of
(Taunton et al., 2000; Cheng et al., 2018). molecular and mineral biomarkers. Together with the physical fossil
It is difficult to prove unequivocally that the molecular and mineral record (Edwards et al., 2018; Strullu-Derrien et al., 2019) and phylo­
biomarkers are directly coupled, i.e., the molecular evidence of fungi genetic analysis (Gerrienne et al., 2016; van Straalen, 2021; Tihelka
and bacterial decomposition is related to the observed mineral diagen­ et al., 2022), the investigation of terrestrialization is open to a multi-
esis. However, a relationship is suggested by: faceted approach.

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T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

CRediT authorship contribution statement a study utilizing Western Australian monazite. Bioprocess. Bio-syst. Eng. 40,
929–942.
Dempster, T.J., 1992. Zoning and recrystallization of phengitic micas: implications for
T.O. Akinsanpe: Writing – review & editing, Validation, Supervi­ metamorphic equilibration. Contrib. Mineral. Petrol. 109, 526–537.
sion, Methodology, Investigation. S.A. Bowden: Writing – review & Drake, H., Ivarsson, M., Heim, C., Snoeyenbos-West, O., Bengston, S., Belivanova, V.,
editing, Supervision. J. Parnell: Writing – original draft, Supervision, Whitehouse, M., 2021. Fossilized anaerobic and possibly methanogenesis-fueling
fungi identified deep within the Siljan impact structure, Sweden. Communicat. Earth
Project administration, Investigation, Funding acquisition, Environ. 2, 34.
Conceptualization. Duncan, A.D., Hamilton, R.F.M., 1988. Palaeolimnology and organic geochemistry of the
Middle Devonian in the Orcadian Basin. In: Fleet, A.J., Kelts, K., Talbot, M.R. (Eds.),
Lacustrine Petroleum Source Rocks. Geological Society, vol. 40. Special Publications,
Declaration of competing interest London, pp. 173–201.
Dzou, L.I.P., Noble, R.A., Senftle, J.T., 1995. Maturation effects on absolute biomarker
The authors declare that they have no known competing financial concentrations in a suite of coals and associated vitrinite concentrates. Org.
Geochem. 23, 681–697.
interests or personal relationships that could have appeared to influence Edwards, D., Kenrick, P., Dolan, L., 2018. History and contemporary significance of the
the work reported in this paper. Rhynie cherts—our earliest preserved terrestrial ecosystem. Philos. Trans. R. Soc.
Lond. B 373, 20160489.
Escuder-Rodríguez, J.-J., DeCastro, M.-E., Saavedra-Bouza, A., Becerra, M., González-
Data availability Siso, M.-I., 2022. Insights on microbial communities inhabiting non-volcanic hot
springs. Int. J. Mol. Sci. 23, 12241. https://doi.org/10.3390/ijms232012241.
Data is supplied as Supplementary Material Fang, R., Li, M., Wang, T.-G., Zhang, L., Shi, S., 2015. Identification and distribution of
pyrene, methylpyrenes and their isomers in rock extracts and crude oils. Org.
Geochem. 83–84, 65–76.
Acknowledgement Fayers, S.R., Trewin, N.H., 2004. A review of the palaeoenvironments and biota of the
Windyfield chert. Trans. R. Soc. Edinb. Earth Sci. 94, 325–339.
Fischer, M.J., Rustenhloz, C., Leh-Louis, V., Perrière, P., 2015. Molecular and functional
The study depended on material archived by N.H. Trewin, C.M. Rice,
evolution of the fungal diterpene synthase genes. BMC Microbiol. 15, 221.
S. Fayers and L.I. Anderson. We acknowledge C. Taylor, J. Still, J. Fox, F., 1889. Strathpeffer Spa, its Climate and Waters. With Observations Historical,
Johnston, W. Ritchie and J. Bowie for skilled technical support. A. Schito Medical, and General, Descriptive of the Vicinity, Etc. H.K. Lewis, London, p. 165.
Garcia-Guinea, J., Martinez-Frías, J., Harffy, M., 1998. Cell-hosted pyrite framboids in
kindly performed the Raman spectroscopy. We are very grateful to
fossil woods. Naturwissenschaften 85, 78–81.
Professor Armelle Riboulleau, Universite de Lille, who provided a Geptner, A., Kristmannsdottir, H., Yurii Pikovskii, Y., Richter, B., 2005. Abiogenic
sample of SKT-D. The study was partly supported by the Natural Envi­ hydrocarbon’s emission in the modern rift zone, Iceland. In: Proceedings, World
ronment Research Council (grant NE/T003677/1). We are grateful for Geothermal Congress, Antalya, Turkey, 24–29 April 2005, pp. 1–7.
Geptner, A.R., Richter, B., Pikovskii, Y.I., Chernyansky, S.S., Alekseeva, T.A., 2006.
the comments of anonymous reviewers which greatly improved the Hydrothermal polycyclic aromatic hydrocarbons in marine and lagoon sediments at
manuscript. the intersection between Tjörnes Fracture Zone and recent rift zone (Skjálfandi and
Öxarfjörður bays), Iceland. Mar. Chem. 101, 153–165.
Gerrienne, P., Servais, T., Vecoli, M., 2016. Plant evolution and terrestrialization during
Appendix A. Supplementary data Palaeozoic times—the phylogenetic context. Rev. Palaeobot. Palynol. 227, 4–18.
Gomez, F., 2011. Hot Spring Microbiology. In: Gargaud, M. (Ed.), Encyclopedia of
Supplementary data to this article can be found online at https://doi. Astrobiology. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-
11274-4_74.
org/10.1016/j.palaeo.2024.112101. Goossens, H., de Leeuw, J., Schenck, P., et al., 1984. Tocopherols as likely precursors of
pristane in ancient sediments and crude oils. Nature 312, 440–442. https://doi.org/
References 10.1038/312440a0.
Grantham, P.J., Wakefield, L.L., 1988. Variations in the sterane carbon number
distributions of marine source rock derived crude oils through geological time. Org.
Abbott, G.D., Fletcher, I.W., Tardio, S., Hackz, E., 2017. Exploring the geochemical
Geochem. 12, 61–73.
distribution of organic carbon in early land plants: a novel approach. Philos. Trans.
Grice, K., Hong, L., Atahan, P., Asif, M., Hallmann, C., Greenwood, P., Maslen, E.,
R. Soc. B 373, 20160499.
Tulipani, S., Williford, K., Dodson, J., 2009. New insights into the origin of perylene
Allaz, J., Selleck, B., Williams, M.L., Jercinovic, M.J., 2013. Microprobe analysis and
in geological samples. Geochim. Cosmochim. Acta 73, 6531–6543.
dating of monazite from the Potsdam Formation, New York: a progressive record of
Grimes, S.T., Davies, K.L., Butler, I.B., Fiona Brock, F., Edwards, D., Rickard, D.,
chemical reaction and fluid interaction. Am. Mineral. 98, 1106–1119.
Briggs, D.E.G., Parkes, R.J., 2002. Fossil plants from the Eocene London Clay: the use
Aoki, K., Windley, B., Sato, K., Sawaki, Y., Kawai, T., Shibuya, T., Kumagai, H.,
of pyrite textures to determine the mechanism of pyritization. J. Geol. Soc. Lond.
Suzuki, K., Maruyama, S., 2013. Chemical composition and K–Ar age of Phengite
159, 493–501.
from Barrovian metapelites, Loch Leven, Scotland. J. Geol. Soc. Jpn. 119, 437–442.
Hao, F., Zhou, X., Zhu, Y., Bao, X., Yuanyuan, Y., 2009. Charging of the Neogene Penglai
Baron, M., Hillier, S., Rice, C.M., Czapnik, K., Parnell, J., 2004. Fluids and hydrothermal
19-3 field, Bohai Bay Basin, China: Oil accumulation in a young trap in an active
alteration assemblages in a Devonian gold-bearing hot-spring system, Rhynie,
fault zone. AAPG Bull. 93, 155–179.
Scotland. Trans. R. Soc. Edinb. Earth Sci. 94, 309–324.
Harrison, T.N., 1990. Chemical variation in micas from the Cairngorm pluton, Scotland.
Blumenberg, M., Weniger, P., Kus, J., Scheeder, G., Piepjohn, K., Zindler, M.,
Mineral. Mag. 54, 355–366.
Reinhardt, L., 2018. Geochemistry of a middle Devonian cannel coal (Munindalen)
Hautevelle, Y., Michels, R., Malartre, F., Trouiller, A., 2006. Vascular plant biomarkers as
in comparison with Carboniferous coals from Svalbard. Arktos 4, 1–8.
proxies for palaeoflora and palaeoclimatic changes at the Dogger/Malm transition of
Böcker, J., Littke, R., Hartkopf-Froder, C., Jasper, K., Schwarzbauer, J., 2013. Organic
the Paris Basin (France). Org. Geochem. 37, 610–625.
geochemistry of Duckmantian (Pennsylvanian) coals from the Ruhr Basin, western
Hoffland, E., Giesler, R., Jongmans, A.G., van Breemen, N., 2003. Feldspar tunneling by
Germany. Int. J. Coal Geol. 107, 112–126.
fungi along natural productivity gradients. Ecosystems 6, 739–746.
Borkow, P.S., Babcock, L.E., 2003. Turning pyrite concretions outside-in: Role of biofilms
Huang, W.Y., Meinschein, W.G., 1979. Sterols as ecological indicators. Geochim.
in pyritization of fossils. Sediment. Record 1, 4–7.
Cosmochim. Acta 43, 739–745.
Cao, J., Hu, K., Wang, K., Bian, L., Liu, Y., Yang, S., Wang, L., Chen, Y., 2008. Possible
Hughes, W.B., Holba, A.G., Dzou, L.I.P., 1995. The ratios of dibenzothiophene to
origin of 25-norhopanes in Jurassic organic-poor mudstones from the northern
phenanthrene and pristane to phytane as indicators of depositional environment and
Qaidam Basin (NW China). Org. Geochem. 39, 1058–1065.
lithology of petroleum source rocks. Geochim. Cosmochim. Acta 59, 3581–3598.
Cascales-Miñana, B., Steemans, P., Servais, T., Lepot, K., Gerrienne, P., 2019. An
Ivarsson, M., Skogby, H., Phichaikamjornwut, B., Bengtson, S., Siljeström, S.,
alternative model for the earliest evolution of vascular plants. Lethaia 52, 445–453.
Ounchanum, P., Boonsoong, A., Kruachanta, M., Marone, F., Belivanova, V.,
Castro, L., Blázquez, M.L., González, F., Muñoz, J.A., 2020. Bioleaching of phosphate
Holmström, S., 2018. Intricate tunnels in garnets from soils and river sediments in
minerals using Aspergillus Niger: Recovery of copper and rare earth elements. Metals
Thailand - possible endolithic microborings. PLoS One 18, e0200351. https://doi.
10, 978.
org/10.1371/journal.pone.0200351.
Cheng, Y., Zhang, L., Bian, X., Zuo, H., Dong, H., 2018. Adsorption and mineralization of
Jardine, J.L., Stoychev, S., Mavumengwana, V., Ubomba-Jaswa, E., 2018. Screening of
REE-lanthanum onto bacterial cell surface. Environ. Sci. Pollut. Res. 25,
potential bioremediation enzymes from hot spring bacteria using conventional plate
22334–22339.
assays and liquid chromatography - Tandem mass spectrometry (Lc-Ms/Ms).
Clarke, P., Parnell, J., 1999. Facies analysis of a back-tilted lacustrine basin in a strike-
J. Environ. Manag. 223, 787–796.
slip zone, lower Devonian, Scotland. Palaeogeogr. Palaeoclimatol. Palaeoecol. 151,
Kang, X., Csetenyi, L., Gadd, G.M., 2021. Colonization and bioweathering of monazite by
167–190.
Aspergillus Niger: solubilization and precipitation of rare earth elements. Environ.
Cook, J.W., Hewett, C.L., Hieger, I., 1933. The isolation of a cancer-producing
Microbiol. 23, 3970–3986.
hydrocarbon from coal tar. J. Chem. Soc. 106, 395–405.
Karunanithi, P.S., Zerbe, P., 2019. Terpene synthases as metabolic gatekeepers in the
Corbett, M.K., Eksteen, J.J., Niu, X.-Z., Croue, J.-P., Watkin, E.L., 2017. Interactions of
evolution of plant terpenoid chemical diversity. Front. Plant Sci. 10, 1166.
phosphate solubilising microorganisms with natural rare-earth phosphate minerals:

14
T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Kenrick, P., Edwards, D., 1988. The anatomy of lower Devonian Gosslingia breconensis Rice, C.M., Ashcroft, W.A., Batten, D.J., Boyce, A.J., Caulfield, J.B.D., Fallick, A.E.,
heard based on pyritized axes, with some comments on the permineralization Hole, M.J., Jones, E., Pearson, M.J., Rogers, G., Saxton, J.M., Stuart, F.M., Trewin, N.
process. Bot. J. Linn. Soc. 97, 95–123. H., Turner, G.A., 1995. Devonian auriferous hot spring system, Rhynie, Scotland.
Killops, S.D., Killops, V.J., 2005. Introduction to Organic Geochemistry, Second edition. J. Geol. Soc. Lond. 152, 229–250.
Blackwell Publishing, Oxford. Rice, C.M., Trewin, N.H., Anderson, L.I., 2002. Geological setting of the early Devonian
Kodner, R.B., Pearson, A., Summons, R.E., Knoll, A.H., 2008. Sterols in red and green Rhynie cherts, Aberdeenshire, Scotland: an early terrestrial hot spring system.
algae: quantification, phylogeny, and relevance for the interpretation of geologic J. Geol. Soc. Lond. 159, 203–214.
steranes. Geobiology 6, 411–420. Richardson, J.B., Rasul, S.M., 1978. Palynological evidence for the age and provenance
Krings, M., Harper, C.J., Taylor, E.L., 2018. Fungi and fungal interactions in the Rhynie of the lower Old Red Sandstone from the Apley Barn borehole, Witney, Oxfordshire
chert: a review of the evidence, with the description of Perexiflasca tayloriana gen. (southern England). Proc. Geol. Assoc. 90, 27–42.
Et sp. nov. Philos. Trans. R. Soc. B 373, 20160500. Romero-Sarmiento, M., Riboulleau, A., Vecoli, M., Versteegh, G.J.M., 2010. Occurrence
Li, Z., Huang, H., Yan, G., Xu, Y., George, S.C., 2022. Occurrence and origin of perylene of retene in upper Silurian–lower Devonian sediments from North Africa: Origin and
in Paleogene sediments from the Tasmanian Gateway, Australia. Org. Geochem. 168, implications. Org. Geochem. 41, 302–306.
104406. Romero-Sarmiento, M., Riboulleau, A., Vecoli, M., Laggoun-Défarge, F., Versteegh, G.J.
Lian, B., Wang, B., Pan, M., Liu, C.Q., Teng, H.H., 2008. Microbial release of potassium M., 2011. Aliphatic and aromatic biomarkers from Carboniferous coal deposits at
from K-bearing minerals by thermophilic fungus Aspergillus fumigatus. Geochim. Dunbar (East Lothian, Scotland): Palaeobotanical and palaeoenvironmental
Cosmochim. Acta 72, 87–98. significance. Palaeogeogr. Palaeoclimatol. Palaeoecol. 309, 309–326.
Loron, C.C., Dzul, E.R., Orr, P.J., Gromov, A.V., Fraser, N.C., McMahon, S., 2023. Santelli, C.M., Webb, S.M., Dohnalkova, A.C., Hansel, C.M., 2011. Diversity of Mn oxides
Molecular fingerprints resolve affinities of Rhynie chert organic fossils. Nat. produced by Mn(II)- oxidizing fungi. Geochim. et Cosmochim. 75, 2762–2776.
Commun. 14, 1387. Savage, H.M., Rabinowitz, H.S., Spagnuolo, E., Aretusini, S., Polissar, P.J., Di Toro, G.,
Lutzoni, F., et al., 2018. Contemporaneous radiations of fungi and plants linked to 2018. Biomarker thermal maturity experiments at earthquake slip rates. Earth
symbiosis. Nat. Commun. 9, 5451. Planet. Sci. Lett. 502, 253–261.
Maeda, H., Song, W., Sage, T.L., Della, Penna D., 2006. Tocopherols play a crucial role in Schardl, C.L., Young, C.A., Hesse, U., Amyotte, S.G., Andreeva, K., Calie, P.J., et al.,
low-temperature adaptation and phloem loading in Arabidopsis. Plant Cell 18, 2013. Plant-symbiotic fungi as chemical engineers: Multi-genome analysis of the
2710–2732. Clavicipitaceae reveals dynamics of alkaloid Loci. PLoS Genet. 9, e1003323.
Manson, D., 1879. On the Sulphur Waters of Strathpeffer, in the Highlands of Ross-Shire, Schito, A., Corrado, S., 2020. An automatic approach for characterization of the thermal
with District Guide. J. & A. Churchill, London. maturity of dispersed organic matter Raman spectra at low diagenetic stages. Geol.
Martinez, A., Boyer, D.L., Droser, M.L., Barrie, C., Love, G.D., 2019. A stable and Soc. Lond. Spec. Publ. 484, 107–119.
productive marine microbial community was sustained through the end-Devonian Sedlakova-Kadukova, J., Marcincakova, R., Luptakova, A., Vojtko, M., Fujda, M.,
Hangenberg Crisis within the Cleveland Shale of the Appalachian Basin, United Pristas, P., 2020. Comparison of three different bioleaching systems for Li recovery
States. Geobiology 17, 27–42. from lepidolite. Sci. Rep. 10, 14594.
Marynowski, L., Smolarek, J., Bechtel, A., Phillippe, M., Kurkiewicz, S., Simoneit, B.R.T., Sen, S.K., Raut, S., Dora, T.K., Mohapatra, P.K.D., 2014. Contribution of hot spring
2013. Perylene as an indicator of conifer fossil wood degradation by wood-degrading bacterial consortium in cadmium and lead bioremediation through quadratic
fungi. Org. Geochem. 59, 143–151. programming model. J. Hazard. Mater. 265, 47–60.
Milodowski, A.E., Zalasiewicz, J.A., 1991. Redistribution of rare earth elements during Sharma, E., Anand, G., Kapoor, R., 2017. Terpenoids in plant and arbuscular mycorrhiza-
diagenesis of turbidite/hemipelagite mudrock sequences of Llandovery age from reinforced defence against herbivorous insects. Ann. Bot. 119, 791–801.
Central Wales. Geol. Soc. Lond. Spec. Publ. 57, 101–124. https://doi.org/10.1144/ Smits, M., 2006. Mineral tunnelling by fungi. In: Gadd, G.M. (Ed.), Fungi in
GSL.SP.1991.057.01.10. Biogeochemical Cycles. Cambridge University Press, Cambridge.
Mitchell, R.L., Strullu-Derrien, C., Kenrick, P., 2019. Biologically mediated weathering in Song, M., Pedruzzi, I., Peng, Y., Li, P., Liu, J., Yu, J., 2015. K-extraction from muscovite
modern cryptogamic ground covers and the early Paleozoic fossil record. J. Geol. by the isolated fungi. Microbiol. J. 32, 771–779.
Soc. 176, 430–439. Song, D., Simoneit, B.R.T., He, D., 2017. Abundant tetracyclic terpenoids in a Middle
Mitchell, R.L., Davies, P., Kenrick, P., Volkenandt, T., Pleydell-Pearce, C., Johnston, R., Devonian foliated cuticular liptobiolite coal from northwestern China. Org.
2021. Correlative microscopy: a tool for understanding soil weathering in modern Geochem. 107, 9–20.
analogues of early terrestrial biospheres. Sci. Rep. 11, 12736. Strullu-Derrien, C., Kenrick, P., Pressel, S., Duckett, J.G., Rioult, J.-P., Strullu, D.-G.,
Mumm, R., Posthumus, M.A., Dicke, M., 2008. Significance of terpenoids in induced 2014. Fungal associations in Horneophyton ligneri from the Rhynie Chert (c. 407
indirect plant defence against herbivorous arthropods. Plant Cell Environ. 31, million year old) closely resemble those in extant lower land plants: novel insights
575–585. into ancestral plant–fungus symbioses. New Phytol. 203, 964–979.
Noble, R.A., Alexander, R., Kagi, R.I., Knox, J., 1985. Tetracyclic diterpenoid Strullu-Derrien, C., Kenrick, P., Knoll, A.H., 2019. The Rhy,nie Chert. Curr. Biol. 29,
hydrocarbons in some Australian coals, sediments and crude oils. Geochim. R1218–R1223.
Cosmochim. Acta 49, 2141–2147. Strullu-Derrien, C., Le Hérissé, A., Goral, T., Spencer, A.R.T., Kenrick, P., 2021. The
Parnell, J., 1985. Evidence for evaporites in the ORS of northern Scotland: replaced overlooked aquatic green algal component of early terrestrial environments: Triskelia
gypsum horizons in Easter Ross. Scott. J. Geol. 21, 377–380. scotlandica gen. Et sp. nov. from the Rhynie cherts. Paper. Palaeontol. 7, 709–719.
Parnell, J., Baba, M., Bowden, S., 2016. Emplacement and biodegradation of oil in Strullu-Derrien, C., Fercoq, F., Gèze, M., Kenrick, P., Martos, F., Selosse, M.-A.,
fractured basement: the ‘coal’ deposit in Moinian gneiss at Castle Leod, Ross-shire. Benzerara, K., Knoll, A.H., 2023. Hapalosiphonacean cyanobacteria (Nostocales)
Earth Environ. Sci. Trans. R. Soc. Edinb. 107, 23–32. thrived amid emerging embryophytes in an early Devonian (407-million-year-old)
Parnell, J., Akinsanpe, T.O., Armstrong, J.G.T., Boyce, A.J., Still, J.W., Bowden, S.A., landscape. iScience 26 (8), 107338.
Clases, D., de Vega, R.G., Feldmann, J., 2022. Trace element geochemistry in the Taunton, A.E., Welch, S.A., Banfield, J.F., 2000. Microbial controls on phosphate and
earliest terrestrial ecosystem, the Rhynie Chert. Geochem. Geophys. Geosyst. 23 lanthanide distributions during granite weathering and soil formation. Chem. Geol.
e2022GC010647. 169, 371–382.
Parnell, J., Akinsanpe, T.O., Still, J.W., Schito, A., Bowden, S.A., Muirhead, D.K., Taylor, T.N., Klavins, S.D., Krings, M., Taylor, E.L., Kerp, H., Hass, H., 2004. Fungi from
Armstrong, J.G.T., 2023. Low-temperature fluorocarbonate mineralization in lower the Rhynie Chert: a view from the dark side. Trans. R. Soc. Edinb. Earth Sci. 94,
Devonian Rhynie Chert. UK. Minerals 13, 595. 457–473.
Peters, K.E., Walters, C.C., Moldowan, J.M., 2005. The Biomarker Guide. Biomarkers and Tihelka, E., Howard, R.J., Cai, C., Lozano-Fernandez, J., 2022. Was there a Cambrian
Isotopes in Petroleum Exploration and Earth History. Cambridge University Press, Explosion on land? The case of arthropod terrestrialization. Biology 11, 1516.
Cambridge. https://doi.org/10.3390/biology11101516.
Philp, R.P., De Garmo, D., 2020. Geochemical characterization of the Devonian- Trewin, N.H., 1994. Depositional environment and preservation of biota in the lower
Mississippian Woodford Shale from the McAlister Cemetery Quarry, Criner Hills Devonian hot-springs of Rhynie, Aberdeenshire, Scotland. Trans. R. Soc. Edinb.
Uplift, Ardmore Basin, Oklahoma. Mar. Pet. Geol. 112, 104078. Earth Sci. 84, 433–442.
Powell, C.L., Trewin, N.H., Edwards, D., 2000. Palaeoecology and plant succession in a Trewin, N.H., Fayers, S.R., 2015. Macro to micro aspects of the plant preservation in the
borehole through the Rhynie cherts, lower Old Red Sandstone, Scotland. In: early Devonian Rhynie cherts, Aberdeenshire, Scotland. Earth Environ. Sci. Trans. R.
Friend, P.F., Williams, B.P.J. (Eds.), New Perspectives on the Old Red Sandstone, Soc. Edinb. 106, 67–80.
Geological Society, vol. 180. Special Publications, London, pp. 439–457. Trewin, N.H., Kerp, H., 2017. The Rhynie and Windyfield cherts, early Devonian, Rhynie,
Qu, J.-B., Zhu, R.-L., Zhang, Y.-L., Guo, H.-F., Wang, X.-N., Xie, C.-F., Yu, W.-T., Ji, M., Scotland. In: Fraser, N.C., Sues, H.-D. (Eds.), In: Terrestrial Conservation
Lou, H.-X., 2008. Ent-Kaurane diterpenoids from the liverwort Jungermannia Lagerstätten - Windows into the Evolution of Life on Land. Dunedin Academic Press
atrobrunnea. J. Nat. Prod. 71, 1418–1422. Ltd, Edinburgh, pp. 1–38.
Quirk, J., Beerling, D.J., Banwart, S.A., Kakonyi, G., Romero-Gonzalez, M.E., Leake, J.R., Tulipani, S., et al., 2015. Changes of palaeoenvironmental conditions recorded in late
2012. Evolution of trees and mycorrhizal fungi intensifies silicate mineral Devonian reef systems from the Canning Basin, Western Australia: a biomarker and
weathering. Biol. Lett. 8, 1006–1011. stable isotope approach. Gondwana Res. 28, 1500–1515.
Rabinowitz, H.S., Polissar, P.J., Savage, H.M., 2017. Reaction kinetics of alkenone and n- van Schöll, L., Smits, M., Hoffland, E., 2006. Ectomycorrhizal weathering of the soil
alkane thermal alteration at seismic timescales. Geochem. Geophys. Geosyst. 18, minerals muscovite and hornblende. New Phytol. 171, 805–816.
53–69. van Schöll, L., Kuyper, T.W., Smits, M.M., Landeweert, R., Hoffland, E., van Breemen, N.,
Rayner, R.J., 1984. New finds of Drepanophycus spinaeformis Göppert from the lower 2008. Rock-eating mycorrhizas: their role in plant nutrition and biogeochemical
Devonian of Scotland. Trans. R. Soc. Edinb. Earth Sci. 75, 353–363. cycles. Plant Soil 303, 35–47.
Rice, C.M., Ashcroft, W.A., 2004. The geology of the northern half of the Rhynie Basin, van Straalen, N.M., 2021. Evolutionary terrestrialization scenarios for soil invertebrates.
Aberdeenshire, Scotland. Trans. R. Soc. Edinb. Earth Sci. 94, 299–308. Pedobiologia 87–88, 150753.

15
T.O. Akinsanpe et al. Palaeogeography, Palaeoclimatology, Palaeoecology 640 (2024) 112101

Versteegh, G.J.M., Riboulleau, A., 2010. An organic geochemical perspective on Wellman, C.H., 2006. Spore assemblages from the lower Devonian ‘lower Old Red
terrestrialization. Geol. Soc. Spec. Publ. 339, 11–34. Sandstone’ deposits of the Rhynie outlier, Scotland. Trans. R. Soc. Edinb. Earth Sci.
Volkman, J.K., 2003. Sterols in microorganisms. Appl. Microbiol. Biotechnol. 60, 97, 167–211.
495–506. Wellman, C.H., Graham, L.E., Lewis, L.A., 2019. Filamentous green algae from the early
Wacey, D., Kilburn, M.R., Saunders, M., Cliff, J.B., Kong, C., Liu, A.G., et al., 2015. Devonian Rhynie chert. Paläontol. Z. 93, 387–393.
Uncovering framboidal pyrite biogenicity using nano-scale CNorg mapping. Geology
43, 27–30.

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