1 Revealing rangeomorph species characters using spatial analyses 2 Emily G. Mitchell1, Charlotte G. Kenchington2, Simon Harris3 and Philip R. Wilby3 3 4 1 Department of Earth Sciences, University of Cambridge, Downing Street, Cambridge CB2 5 3EQ, UK. 6 7 2 Department of Earth Sciences, Memorial University of Newfoundland, St. John’s, NL A1B 3X5 8 Canada. 9 10 3British Geological Survey, Nicker Hill, Keyworth, Nottingham NG12 5GG, United Kingdom. 11 12 *Corresponding author email address: [email protected]. 13 14 1 15 Abstract: Rangeomorphs dominate the Ediacaran Avalonian macrofossil assemblages of 16 Charnwood Forest, UK (~562Ma). However, their unfamiliar fractal architecture makes 17 distinguishing phylogenetically reliable characters from intraspecific features difficult. 18 Fortunately, spatial analysis of large in-situ populations offers an independent means of 19 assessing their taxonomy. Populations of a single biological species are likely to exhibit similar 20 spatial distributions due to their shared responses to the biological and ecological processes 21 acting upon them. As such, spatial analyses can be used to interrogate which are the most 22 taxonomically deductive characters in similar species. We used Random Labelling Analyses to 23 investigate the presence/absence of characters of Primocandelabrum boyntoni, P. aethelfalaedia 24 and P. aelfwynnia on the North Quarry ’B’ surface. The resultant spatial distributions were 25 compared to observed characters using goodness-of-fit tests to determine which characters were 26 associated with unique populations, and which were found across multiple populations. We 27 found that P. boyntoni and P. aelfwynnia had statistically indistinguishable character 28 distributions, suggesting that they represent a single biological species, and that they exhibited 29 significantly different distributions to P. aethelfalaedia, suggesting that there are two (rather than 30 three) Primocandelabrum species present on the B surface. Furthermore, we found that the 31 distribution of Concealed versus Unconcealed 1st order branches across all specimens exhibited 32 significantly different, density-dependant behaviour, with Unconcealed branching occurring in 33 areas of higher density populations, and Concealed branching occurring in the lower 34 Primocandelabrum density areas. We speculate that unconcealed branches may have been a 35 response to the reduced availability of resources in higher density areas, implying rangeomorphs 36 were capable of ecophenotypic responses. 37 38 Key words: Ediacaran, Rangeomorph, Spatial analyses 39 2 40 Introduction 41 The Avalonian Assemblage of Ediacaran organisms contains some of the oldest known examples 42 of macroscopic organisms in the fossil record, and is known primarily from south east 43 Newfoundland (Clapham et al. 2003, Hoffmann et al. 2008, Narbonne et al. 2009) and 44 Charnwood Forest, UK (Wilby et al. 2011, Kenchington et al. 2018). Avalonian macrofossils 45 share few similarities with living taxa, making organism biology, phylogenetic relationships and 46 ecological interactions difficult to assess (Liu et al. 2015, Erwin et al 2011, Xiao & Laflamme 47 2009 ). Their communities are dominated by rangeomorphs, a proposed clade of “fractally- 48 branching” organisms (Narbonne 2004, Hoyal Cuthill and Conway Morris 2014, Erwin et al. 49 2011). However, their unfamiliar fractal architecture makes distinguishing phylogenetically 50 reliable characters from intraspecific features difficult (Liu et al 2015b, Kenchington and Wilby 51 2017). 52 One of the largest and most diverse Avalonian communities is preserved on the so-called 53 Bed ‘B’ bedding-plane surface in Charnwood Forest (UK) which, at 115m2 surface area and with 54 at least 18 different taxa, is comparable to that on the Mistaken Point ‘E’ surface in 55 Newfoundland (Wilby et al. 2011). The multifoliate, stemmed rangeomorph Primocandelabrum 56 (Hofmann et al. 2008) comprises a significant proportion of the Bed ‘B’ community, and is 57 principally diagnosed by its coarse branching, the triangular outline of its crown, and by having 58 typically rotated, unfurled folia (first order branches) that are proximally inflated, and second 59 order branches that are typically displayed, unfurled and medially inflated (Figs. 1 and 2; 60 Hofmann et al. 2008, Kenchington and Wilby, 2017). Prior statistical analyses by Kenchington 61 and Wilby (2017) used multivariate cluster analyses to formally describe three 62 Primocandelabrum species using both continuous (e.g. length to width ratios) and discreet (e.g. 63 rotated first order branches, Fig. 2) characters, and revealed a hitherto unrecognized 64 morphological variability in Primocandelabrum. Within the Kenchington and Wilby results 65 there is overlapping variation between the individuals in the clusters (Fig. 3), with individuals in 66 different clusters sharing some of the same characters and some characters not being 100% 67 unique to a cluster. Their analyses suggested two or three Primocandelabrum species: P. 68 boyntoni, P. aelfwynnia and P. aethelflaedia. As such, there remains uncertainty regarding the 69 number of species present on Bed ‘B’. 3 70 The preservation of Avalonian macrofossil communities may be exceptional, with 71 notable examples of thousands of these organisms preserved in-situ on successive bedding planes 72 (Wood et al. 2003, Kenchington and Wilby 2014, Liu et al., 2015). Their sessile habit and their 73 preservation in life position means that their position on a bedding plane encapsulates their life- 74 history: that is, their position is a combined record of the abiological, biological and ecological 75 processes that impacted them during their life. Consequently, spatial analyses can be used to 76 investigate otherwise cryptic aspects of Ediacaran life, such as how they reproduced (Mitchell et 77 al. 2015) or how they interacted with each other (Mitchell and Kenchington 2018) and/or their 78 environment (Mitchell and Butterfield 2018) and provide a complimentary record to prior 79 ecological analyses of population size-frequency distributions (Darroch et al. 2013, Wilby et al 80 2015). 81 There are four types of processes that control population spatial distributions for extant 82 sessile organisms: 1) dispersal processes, 2) associations with local habitat heterogeneities, 3) 83 direct inter-specific interactions, and 4) density-dependent effects (Wiegand et al. 2007). The 84 first three represent inter-specific interactions and these can have either a negative impact on 85 populations, such as competition for limited resources, or a positive impact, such as facilitation, 86 whereby one species enhances the survival of another. Density-dependent effects include, for 87 example, Janzen-Connell effects, whereby a specialist consumer, such as a pathogen or a taxon- 88 specific herbivore, leads to density-dependent deaths (Velázquez et al. 2016). In non-motile 89 systems, such as the Avalonian ones, density-dependent effects are limited to pathogens because 90 macroscopic predators/herbivores are absent (e.g. Liu et al. 2015). Pathogens transfer between 91 organisms in high-density areas more easily than between those in low density areas, thus 92 creating differential mortality patterns based on relative density (Wiegand et al. 2007a). 93 Significant density-dependent mortality can be detected via the spatial distributions of 94 taphomorphs, such as ivesheadiomorphs, and/or by the incorporation of effaced characteristics 95 into analyses (cf. Mitchell and Butterfield 2018). Consequently, well preserved specimens, such 96 as those considered in the present study, can be assumed to not have been significantly impacted 97 by pathogenic mortality effects and, hence, only the first three processes need be considered. 98 Each of the three inter-specific processes have numerous factors which impact the 99 resultant spatial distributions. The spatial distributions induced by dispersal processes are 100 determined by the following factors: the type of dispersal process (e.g. propagule or 4 101 stolonifereous), the number of offspring, the height at which propagules are released, the current 102 speed (which is assumed conserved across a given community at a given time), and the relative 103 density of the settled offspring (e.g. Yamamichi et al. 2014). Habitat influences can vary within 104 any one habitat by operating at different spatial scales, having a positive or negative impact 105 and/or vayring in their relative strength (Wiegand et al. 2007b), as can interspecific interactions 106 (Lingua et al. 2008). Acting in combination, these factors produce an enormous number of 107 possible different combinations of spatial distributions: just taking each factor as discrete (which 108 would be a very simplified model, since only 2/11 of the characters are discreet) would give 109 almost 40 million different possible combinations. Consequently, if two populations exhibit the 110 same spatial distribution, it is reasonable to infer that the same abiological, biological and 111 ecological processes have impacted both populations. This situation is most likely to happen 112 when the two populations are, in fact, just a single species. 113 Spatial analyses have previously been used within the Avalon Assemblage to investigate 114 the possible taxonomic affiliations of enigmatic forms referred to as ivesheadiomorphs and 115 Lobate Discs (Mitchell and Butterfield 2018). These have variously been suggested to be either 116 the