What Lies Below
What lurks below the depths of the Jade Sea? For a misguided team of Jade Brotherhood miners, the answer may be far more sinister than anticipated. It's up to you to explore the map, investigate the mysterious circumstances, and discover the truth of what lies beneath.
What Lies Below
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Heatwave and Chief pass the image Heatwave copied on to Optimus Prime, who identifies it as energon and says he's coming to help. The next morning at breakfast, Cody notices Woodrow is missing, and takes Boulder to the mines where he finds Woodrow powering up the forger. Cody climbs aboard to try to stop him, but Woodrow accidentally fires it up. It knocks Boulder over as it turns, and begins tunneling downwards too fast for the Rescue Bot to catch. While Boulder heads back to get help, the pair experience a wild ride, including a jump across a chasm. Optimus Prime arrives at the fire station with a trailer to transport the energon moments before Boulder arrives with the news of what has happened in the mine. The rescue team is soon loaded on to Optimus's rocket-powered trailer and speeding through the tunnels in pursuit.
The crevasse that you must get to is just slightly north from Lion's Arch. It is to the east of Miraba and Leighton Cranford. Click on the crevasse and choose to enter. You will be brought to Beneath Lion's Arch. Follow the path through the small cavern and into the ancient hall, killing some undead along the way. Keep going on the singular path until you see dwarven allies. Climb up the stairs and a cinematic will begin. The Beginning of the End is the follow-up primary quest that is automatically added to your quest log.
Scientists already suspected that there were ascending motions in Venus' atmosphere all along the equator, caused by the higher levels of solar heating. This finding reveals that the amount of water and ultraviolet-dark material found in Venus' clouds is also strongly enhanced at particular places around the planet's equator. "This is caused by the mountains way down on Venus' surface, which trigger rising waves and circulating winds that dredge up material from below," says Markiewicz.
This artist's rendering shows a cross-section of the ice shell immediately beneath one of Enceladus' geyser-active fractures, illustrating the physical and thermal structure and the processes ongoing below and at the surface. Narrow cracks extend upward from the sub-surface sea all the way to the surface, through both ductile and brittle layers of the ice shell (see What Lies Beneath: Regional View). Liquid water under pressure fills the cracks, keeping them open even through the ductile layer and providing a conduit for vapor and sea water to reach the near-surface. Other processes, such as volatile exsolution of gases, can drive vapor and water droplets all the way to the surface, forming geysers and condensing close to the surface, depositing latent heat. This heat is observed by Cassini's long-wavelength infrared instruments as the small-scale hot spots (dozens of feet, or tens of meters, in size) surrounding each geyser.
Adaptive radiation of vertebrates is in part explained by genetic changes that allowed new functions to emerge [13,14,15], increasing the fitness of the organisms in new environments. One of these environments, the soil, presents several restrictive conditions, including low levels of light, high resistance to locomotion, low airborne transmission of sound and scent, and low oxygen (O2) and high carbon dioxide (CO2) levels (hypoxia and hypercapnia respectively). In addition, many microorganisms (fungi, protozoans, bacteria) and diverse invertebrates (often pathogenic) abound in especially humid and thermally stable soils [16]. Despite these challenges, several groups of vertebrates are well adapted to life in soil [17,18,19], including one of the most ancient lineages of extant terrestrial vertebrates, the caecilian amphibians that radiated in the edaphic environment during the early Mesozoic [20, 21]. Caecilians (order Gymnophiona) are limbless, elongate, mostly tropical amphibians. Adults of most species burrow in soil. Many other extant amphibians spend time in soil but feed and breed above ground [22, 23]. In contrast, many adult terrestrial caecilians are highly fossorial, dedicated burrowers that feed and breed within moist soils [24]. Terrestrial caecilians inhabit different layers of soil, from leaf litter to deeper strata, while species of one family (Typhlonectidae) are secondarily semi- or fully aquatic [22]. Caecilian evolution has clearly involved the colonisation of tropical soils. We hypothesise that, as well as providing distinctive challenges, the soil offered new ecological opportunities to caecilians with new resources and absence of, or reduction in, competitors and predators, perhaps similar to emergent islands [25,26,27,28], newly formed lakes [29, 30], and post-mass extinction environments [31, 32] for other organisms. Regions with high above-ground biodiversity, such as the tropics, exhibit low below-ground biodiversity [33] where caecilians might have encountered lower competitive pressure. In addition of the suggestive reduced competition, soil is potentially more stable and less subject to harmful fluctuations in humidity and temperature. Ancestral caecilians adapted to life in soil, developed important innovations and diversified. Given that fossoriality is a derived condition among amphibians, several morphological features of caecilians are clearly adaptations to life in soil, some of which are shared convergently with other edaphic animals. These include modified skull architecture for head-first burrowing and feeding underground [34], elongated limbless bodies with modified axial musculature [35, 36], reduced visual and hearing systems, and novel sensory tentacles [37,38,39]. The molecular changes underlying the evolutionary origin and diversification of caecilians remain unexplored thus far. In this study, we investigated molecular processes involved in the exploitation of (i) soil surface habitats, (ii) deeper soil habitats, and (iii) freshwaters and associated muds.
Recently, reference transcriptomes for several species of caecilians have been generated [21, 40], providing an opportunity to explore genomic changes in caecilian amphibians. Here, we analyse the protein-coding sequences from transcriptome data for nine different tissue types (foregut, heart, kidney, liver, lung, muscle, skin, spleen, and testis) of five species of Neotropical caecilians [40] that occur in a range of habitats (DJG, MW, DSM pers. obs.). The semi-fossorial species Rhinatrema bivittatum (Guérrin-Méneville, 1838) is encountered mostly in more superficial layers of soils as well as on the surface after heavy rain. Caecilia tentaculata Linnaeus, 1758 appears to be a much stronger burrower based on its more heavily ossified skull [34], but it is also encountered on the surface after heavy rains. Typhlonectes compressicauda (Duméril & Bibron, 1841) is a fully aquatic species that can burrow in soft substrates. Microcaecilia unicolor (Duméril, 1861) and M. dermatophaga Wilkinson, Sherratt, Starace & Gower, 2013 are more dedicated burrowers not seen on the surface and mostly found in deeper layers of the soil. The sampled caecilians include species from both sides of the basal divergence within Gymnophiona belonging to four of the ten currently described families [41, 42] of the order (Rhinatremidae, Typhlonectidae, Siphonopidae and Caeciliidae), and their phylogenetic history encompasses several major shifts in caecilian evolution. We have compared nucleotide substitution rates of candidate groups of orthologous protein-coding genes for these five caecilian species in order to identify genes that potentially have, at some time, been under positive selection. The sampled caecilians allow us to explore nine different branches of the caecilian tree of life (Fig. 1) covering the evolutionary periods in which caecilians first adapted to life in soil, and subsequently adapted to deeper soils and to aquatic environments. We identified signatures of positive selection in several protein-coding genes on all branches. Some of these candidate genes could be involved in the adaptive radiation of caecilian amphibians, plausibly in the adaptation to fossoriality, and in the evolution of their special innovative traits.
Molecular adaptive changes in caecilian amphibians are found to be associated mostly with protein-coding gene products with membrane or extracellular location. These genes present low levels of conservation and connectivity (no PPIs and only one functional network were found). The 168 genes that we infer to have been under positive selection are candidate genes with potential to further clarify adaptations of caecilians linked to their unique and variable natural histories. Several of these candidate genes are possibly causally related to differing degrees of fossoriality and hypothesized ecological shifts that might each have led to new ecological opportunities. Experiments (e.g. transfecting cell-lines with a candidate gene and in silico reconstructions of the protein structure) are required to test the function of these protein-coding genes and to identify their particular roles in important processes, such as perception, reduction-oxidation, and aging in caecilians. Functional experiments can be prompted and focused based on genome-wide studies that have narrowed down candidate genes for more thorough investigation. In this study, we identify a set of candidate genes plausibly involved in ecological and evolutionary key processes. Much biological research relies upon a small number of animal models to investigate biological processes but insights from a broader spectrum of organismal diversity, especially from neglected taxa such as caecilians, are also helpful [102].
The source data of this study were the protein-coding gene sequences (both nucleotide and amino-acid level) from reference transcriptomes of five caecilian species (R. bivittatum, C. tentaculata, T. compressicauda, M. unicolor and M. dermatophaga; assemblies are available from NCBI through BioProject ID number PRJNA387587; [40]) as well as those for the frog X. tropicalis, the only amphibian currently represented in the Ensembl database [103]. Species-specific caecilian transcriptomes were de novo assembled from paired-end RNA-seq samples of multiple tissues (kidney, liver, and skin samples for each of the five species plus a selection of other tissues for subsets of the five species: foregut, heart, lung, muscle, spleen, and testis) yielding five reference transcriptomes with a high percentage of completeness. Protein-coding sequences were identified from these assembled sequences with an open reading frame [40]. For each X. tropicalis gene, the isoform encoding the longest protein was chosen for analysis, and BLAST searches (blastp tool, version 2.2.28; E-value 041b061a72