Questioning the evidence of organic compounds called sponge biomarkers

Elevated concentrations of an organic compound, 24‐isopropylcholestane, found in the Precambrian Huqf Supergroup of Oman may provide the oldest known sponge ‘fossil’. This evidence is of critical importance for a properly balanced understanding of the origin of animals. Several different pelagophyte (Class Pelagophyceae part of the Stramenopiles within the Chromaveolata) algae are also capable of producing these exact compounds, and may similarly have done so in deep time. Modern marine algae are also reported to produce structural isomers that are compositionally identical to the sponge marker; they do this in copious quantities. Further, 24‐isopropylcholestane can be produced by diagenetic alteration of compounds produced in large quantities by algae. It is also possible that contamination by petroleum derived lubricating oil used when coring while extracting these compounds from subsurface layers, has affected the data. All extinct organisms that may have produced this compounds are unavailable for analysis by the modern organic chemist and cannot be eliminated from the list of possible producers of the sponge marker. There are also significant uncertainties regarding the dating of the strata from which these ancient compounds are found. Although the compounds are widely reported as c. 751 Ma, they are younger than 645 Ma. It seems more likely that these compounds represent algal biochemical evolution at a time when algal burial occurred in great quantity with well known coeval algal fossils but no sponge fossils. The macroalgal biomass may have declined during the agronomic revolution at the base of the Cambrian Period owing to processing by metazoans, accounting for the comparative scarcity of these sponge markers in Phanerozoic sediments, after which time sponge spicules and body fossils become evident.     

Early sponge evolution: A review and phylogenetic framework

Sponges are one of the critical groups in understanding the early evolution of animals. Traditional views of these relationships are currently being challenged by molecular data, but the debate has so far made little use of recent palaeontological advances that provide an independent perspective on deep sponge evolution. This review summarises the available information, particularly where the fossil record reveals extinct character combinations that directly impinge on our understanding of high-level relationships and evolutionary origins. An evolutionary outline is proposed that includes the major early fossil groups, combining the fossil record with molecular phylogenetics. The key points are as follows. (1) Crown-group sponge classes are difficult to recognise in the fossil record, with the exception of demosponges, the origins of which are now becoming clear. (2) Hexactine spicules were present in the stem lineages of Hexactinellida, Demospongiae, Silicea and probably also Calcarea and Porifera; this spicule type is not diagnostic of hexactinellids in the fossil record. (3) Reticulosans form the stem lineage of Silicea, and probably also Porifera. (4) At least some early-branching groups possessed biminerallic spicules of silica (with axial filament) combined with an outer layer of calcite secreted within an organic sheath. (5) Spicules are homologous within Silicea, but also between Silicea and Calcarea, and perhaps with Homoscleromorpha. (6) The last common ancestor of extant sponges was probably a thin-walled, hexactine-bearing sponge with biminerallic spicules. (7) The stem group of sponges included tetraradially-symmetric taxa that grade morphologically into Cambrian fossils described as ctenophores. (8) The protomonaxonid sponges are an early-branching group, probably derived from the poriferan stem lineage, and include the problematic chancelloriids as derived members of the piraniid lineage. (9) There are no definite records of Precambrian sponges: isolated hexactine-like spicules may instead be derived from radiolarians. Early sponges had mineralised skeletons and thus should have a good preservation potential: the lack of sponge fossils in Precambrian strata may be due to genuine absence of sponges. (10) In contrast to molecular clock and biomarker evidence, the fossil record indicates a basal Cambrian diversification of the main sponge lineages, and a clear relationship to ctenophore-like ancestors. Overall, the early sponge fossil record reveals a diverse suite of extinct and surprising character combinations that illustrate the origins of the major lineages; however, there are still unanswered questions that require further detailed studies of the morphology, mineralogy and structure of early sponges.     

Origin and early diversification of the phylum Cnidaria Verrill: major developments in the analysis of the taxon's Proterozoic–Cambrian history

Diploblastic eumetazoans of the phylum Cnidaria originated during the Neoproterozoic Era, possibly during the Cryogenian Period. The oldest known fossil cnidarians occur in strata of Ediacaran age and consist of polypoid forms that were either nonbiomineralizing or weakly so. The oldest possible anthozoans, including the genus Ramitubus, may be related to tabulate corals and occur in the Doushantuo Lagerstätte (upper Doushantuo Formation, South China), the age of which is poorly constrained (approximately 585 Ma?). Conulariid scyphozoans may first appear as early as 635–577 Ma (Lantian Formation, South China). A definite conulariid, most similar to Palaeozoic species assigned to the genus Paraconularia, occurs in association with the possible scyphozoan, Corumbella werneri, in the latest Ediacaran (c. 543 Ma) Tamengo Formation of Brazil. Basal Cambrian (c. 540 Ma) phosphorites in the upper Kuanchuanpu Formation (South China) yield solitary polyps of the oldest probable anthozoan (Eolympia pediculata), which appears to have been a stem hexacorallian. This same formation contains fossils interpreted by some authors as pentaradial cubozoan polyps; however, both the oldest known cubozoans and the oldest hydrozoans, all medusae, may actually occur in the Cambrian (Series 3, c. 505 Ma) Marjum Formation (Utah, USA). Although these recently published palaeontological data tend to corroborate the hypothesis that Cnidaria has a relatively deep Neoproterozoic history, the timing of major internal branching events remains poorly constrained, with, for example, the results of some molecular clock analyses indicating that the two cnidarian subphyla (Anthozoaria and Medusozoa) may have originated as many as one billion years ago. Further progress towards elucidating the evolution and early fossil record of cnidarians may accrue from: (1) an intensive search for phosphatized soft parts in possible anthozoans from the Ediacaran Doushantuo Formation; (2) an expanded search for Ediacaran conulariids; and (3) additional detailed analyses of the taphonomy and preservation of Ediacaran and Cambrian cnidarians, including possible pentaradial cubozoan polyps from the Fortunian upper Kuanchuanpu Formation.     

湖北宜昌乔家坪村寒武系水井沱组海绵动物化石群

海绵动物是最原始的动物门类之一,其演化史对于研究多细胞动物的起源和早期演化至关重要.现有的化石记录显示,水体较深的贫氧环境可能是寒武纪早期海绵动物演化的重要场所之一.前人在贵州、湖南、安徽等多地的斜坡至盆地相黑色页岩中已经报道了数个海绵化石群.在这样的环境中,由于水动力作用较弱,海绵骨架结构容易得到原位保存.湖北宜昌秭...     

Small shelly fossils and trace fossils near the Precambrian-Cambrian boundary in the Yukon Territory, Canada

In the past an ‘explosion’ in diversity and abundance of small shelly fossils and of trace fossils has served to mark the base of the Cambrian. However, no evidence has been presented to prove that the ‘explosions’ of the two groups were synchronous. We describe small shelly fossils and trace fossils from the same phosphatic limestone beds that indicate that the two events were separate in time. The small shelly fossils are Anabarites trisulcatus, Hyolithellus cf. H. isiticus, Microcornus? sp., Protohertzina anabarica, P. unguliformis, P. sp. A, Pseudorthotheca sp. A, Rushtonia? sp. A, four types of tuberculate plates and one type of reticulate plate. These fossils represent a restricted, ‘pre-explosion’ fauna and are assigned to the Anabarites-Circotheca-Protohertzina Assemblage Zone, an uppermost Precambrian zone in the Meishucun Stage, Yunnan Province, China. A point at the top of this zone has received strong international endorsement for future designation as the base of the Cambrian. Associated with the small shelly fossils are the trace fossils Cruziana sp. A, Cruziana? sp. B, Rusophycus sp. A, Palaeophycus rubdark and arthropod scratch marks. If found in isolation, this trace fossil assemblage would be considered as post-Precambrian because it includes large, highly organized arthropod traces that are traditionally accepted as occurring above the trace fossil ‘explosion’. We therefore conclude that the trace fossil ‘explosion’ predates the small shelly fossil ‘explosion’. If the proposed location of the base of the Cambrian in Yunnan is accepted, the small shelly fossil ‘explosion’ concept and its relationship to the boundary would not be greatly modified. The trace fossil ‘explosion’, however, would no longer indicate the base of the Cambrian and the ranges of some trace fossils would be extended into the Precambrian.     

Oldest fossil ciliates from the Cryogenian glacial interlude reinterpreted as possible red algal spores

The Cryogenian Period experienced two long lived global glaciations known as Snowball Earths. While these events were dramatic, eukaryotic life persisted through them, and fossil evidence shows that eukaryotes thrived during the c. 30-million-year interlude between the glaciations. Carbonate successions have become an important taphonomic window for this interval. One of the most notable examples is the c. 662–635 Ma Taishir Formation (Tsagaan Olom Group, Zavkhan Terrane, Mongolia) which has yielded a number of eukaryotic fossil taxa. Here, we examine more closely the morphology and taxonomic affinity of some of these Taishir fossils previously interpreted as remains of ciliate tintinnid loricae (purportedly the oldest fossil ciliates). New morphological and ultrastructural analyses indicate that these fossils are not ciliate tintinnids. Instead, we propose a new interpretation: that they are algal reproductive structures related to coeval macroscopic organic warty sheets described as putative red algae. We report the first occurrence of these fossils in the earliest Ediacaran Ol Formation, indicating that this taxon persisted through the Marinoan Snowball Earth. A new interpretation of these fossils as putative red algal spores has broad implications for our understanding of biodiversity in the Neoproterozoic Era, specifically during the Cryogenian Period, and for the antiquity of ciliates.     

贵州交榜留茶坡组化石组合特征及其生物地层学和演化意义*

埃迪卡拉纪—寒武纪之交见证了地球—生命系统不同圈层的重大变革, 是复杂多细胞生物演化的重要转折期。华南是开展相关研究的主要区域之一, 但前人工作主要集中在化石材料丰富的浅水台地相区, 深水区生物组成面貌尚未得到充分认识。本研究对斜坡相区贵州交榜剖面的留茶坡组进行了详细研究, 通过酸蚀和薄片观察发现了丰富的化石材料, 包括: 海绵骨针、软舌螺、可能的原牙形类(? Protohertzina sp.)和织金钉类(? Zhijinites sp.)、球形疑源类、Megathrix longus, 及不同类型的多细胞藻类, 亲缘关系未定的Poratusiramus sp.、球形化石和形态多样的丝状、片状化石等。该化石组合属于寒武纪纽芬兰世幸运期, 其中的骨骼化石以海绵骨针为主。化石组合中既有与浅水区共有分子, 也有深水区特有类群, 丰富了斜坡相区的化石记录, 具有潜在的地层对比和演化意义。     

Trace fossils from arenig flysch sediments of eire and their bearing on the early colonisation of the deep seas

A sequence of Lower Ordovician (Arenig) turbidites in Co. Wexford, Eire, has yielded one of the earliest diverse ichnofaunas yet recorded from deep water sediments comprising: Chondrites, Glockerichnus, Gordia, Helminthopsis, Lorenzinia, Neonereites, Palaeophycus, Paleodictyon, Planolites, Sublorenzinia, Taenidium, Taphrhelminthopsis, Teichichnus and Tomaculum. This ichnofauna is critical in any analysis of the colonisation of the deep seas by trace fossil‐producing animals.

A world‐wide review shows that the earliest trace fossils are mainly from Late Precambrian shelf sea environments, but many more evolved during very rapid diversification in the pre‐trilobite Lower Cambrian.

There was little increase in diversity in shallow water after the Lower Cambrian but a progressive colonisation of the deep ocean took place and this accelerated during the Ordovician, when the main lineages of deep sea trace fossils were established there. Rosetted, patterned, meandering and simple spiral forms evolved in shallow water in the Upper Precambrian and pre‐trilobite Lower Cambrian and only later migrated into the deep sea, whereas complex, closely programmed, spiral traces may have evolved there.     

Evidence for spicule homology in calcareous and siliceous sponges: biminerallic spicules in Lenica sp. from the Early Cambrian of South China

Botting, J.P., Muir, L.A., Xiao, S., Li, X. & Lin, J.‐P. 2012: Evidence for spicule homology in calcareous and siliceous sponges: biminerallic spicules in Lenica sp. from the Early Cambrian of South China. Lethaia, Vol. 45, pp. 463–475. The relationships of the extant sponge classes, and the nature of the last common ancestor of all sponges, are currently unclear. Early sponges preserved in the fossil record differ greatly from extant taxa, and therefore information from the fossil record is critical for testing hypotheses of sponge phylogenetic relationships that are based on modern taxa. New specimens of the enigmatic sponge Lenica sp., from the Early Cambrian Hetang Biota of South China, exhibit an unusual spicule structure. Each spicule consists of a siliceous core with an axial canal, an organic outer layer and a middle layer interpreted to have been originally calcium carbonate. This finding confirms previous work suggesting the existence of biminerallic spicules in early sponges. Combined with data from other early sponges, the new findings imply that the two fundamental spicule structures of modern sponges were derived from a compound, biminerallic precursor. Spicules are therefore homologous structures in Calcarea and Silicea, and if sponges are paraphyletic with respect to Eumetazoa, then spicules may also have been a primitive feature of Metazoa. □Calcarea, Early Cambrian, Hetang Biota, phylogeny, Silicea, taphonomy.     

寒武纪早期岩家河生物群:研究进展和展望*

埃迪卡拉纪-寒武纪转换时期动物的起源、演化和"寒武纪大爆发"一直是国际古生物学界研究的热点问题,其中寒武纪早期小壳化石群与埃迪卡拉纪化石群和寒武纪早期澄江化石库之间的内在关系是古生物学界研究的难题,其主要原因是寒武纪早期与小壳化石群伴生的宏体动、植物化石的缺乏。发现于峡东地区的寒武纪早期岩家河生物群填补了这一缺失环节,该生物群包含宏体动物、宏观藻类、小壳化石、球形化石(可能的胚胎化石)、微古植物和蓝菌类等化石,部分宏体化石显示了从埃迪卡拉纪向寒武纪过渡色彩。化石保存方式有碳质膜、黄铁矿化、磷酸盐化、硅化。因此对岩家河生物群生物多样性和埋藏学进行综合研究,将可提供纽芬兰世(梅树村期)碳酸盐台地—碳酸盐台地内部的局部凹陷盆地相的一个较完整的生物景观图,对探索"寒武纪大爆发主幕"前夕生物的辐射、演化模式及保存机制具有重要的科学意义。     

Teeth,fossil record and evolutionary history of the cowtail stingray Pastinachus Rüppell, 1829

Abstract

Hypolophin ‘dasyatids’ are a common group of large stingrays today frequenting the Indo-Pacific inshores. Being often harvested in their restricted area, few are known about their biology and their evolutionary history despite a very peculiar dental pattern making it easy to track their fossil record. An abundant material consisting of isolated teeth from Late Bartonian (38–40 Ma) lagoonal deposits of Djebel el Kébar, Tunisia, allows to describe a new stingray, Pastinachus kebarensis nov. sp. This taxon represents the oldest occurrence for this genus but also the oldest fossil record for hypolophins. A dental comparison of these fossils with 3D rendered models of fresh specimens testifies that early hypolophin representatives had already a strongly arcuate and bulbous upper jaw, interlocking with a broad and elongated tooth plate on the lower jaw. This new fossil and its fossil relatives (here updated), indicate a pre-Bartonian origination for hypolophins in western Neotethys, and reveal a rapid and widespread colonization of the proto-Mediterranean Sea, western Atlantic and Indo-Pacific coasts during the late Paleogene–early Neogene. Finally, it is worth noting that early hypolophin representatives seemingly entered freshwater habitats occasionally as modern cowtail stingrays do.     

The first named Ediacaran body fossil, Aspidella Terranovica

Aspidella terranovica Billings, 1872 was first described from the late Neoproterozoic Fermeuse Formation (St. John's Group) on the Avalon Peninsula of eastern Newfoundland, approximately 1km stratigraphically above the famous Ediacaran biota at Mistaken Point, and several kilometres below the base of the Cambrian. Aspidella has been reinterpreted perhaps more than any other Precambrian taxon, and has variously been regarded as a fossil mollusc or ‘medusoid’, a gas escape structure, a concretion, or a mechanical suction mark. Our studies indicate that Aspidella includes a wide variety of preservational morphs varying from negative hyporeliefs with a raised rim and ridges radiating from a slit (Aspidella‐type preservation), to flat discs with a central boss and sharp outer ring (Spriggia preservation), to positive hyporeliefs with concentric ornamentation (Ediacaria preservation). Specimens occur in a continuum of sizes, with preservational styles dependent on the size of the specimen and the grain size of the host lithology; the elongation of specimens is tectonic. Aspidella is confirmed as a body fossil from observations of complex radial and concentric ornamentation, mutually deformed borders in clusters of specimens, and occurrence on the same bedding planes as certain distinctive Ediacaran taxa. Aspidella is indistinguishable from, and has priority over, several of the most common genera of late Neoproterozoic discoidal body fossils worldwide. Similar fossils from Australia are interpreted as holdfasts of frond‐like organisms. The density of specimens in the Aspidella beds suggests levels of benthic biomass in the Neoproterozoic that could rival those of modern marine communities. The serial growth forms, PalaeopascichnusIntrites, Neonereites renariusYelovichnus, associated with Aspidella, are interpreted as body fossils of unknown affinities rather than trace fossils. A new, trilobed, Ediacaran body fossil, Triforillonia costellae gen. et sp. nov., is described from the Aspidella beds of the Fermeuse Formation.     

埃迪卡拉纪至寒武纪苗岭世刺细胞动物化石研究现状与展望

刺细胞动物是一类具有刺细胞的水生无脊椎动物,分布在世界各地的海洋和淡水中.作为后生动物最早分化出的一支,刺细胞动物对研究后生动物的起源和早期演化具有极其重要的意义,也为研究后生动物系统发育、地层对比和古地理恢复等方面提供了重要的科研线索.本文简要介绍了刺细胞动物早期(埃迪卡拉纪至寒武纪苗岭世)的化石记录和研究现状,将刺...     

The proterozoic and earliest cambrian trace fossil record; patterns, problems and perspectives

The increase in trace fossil diversity across the Neoproterozoic-Cambrianboundary often is presented in terms of tabulations of ichnogenera.However, a clearer picture of the increase in diversity andcomplexity can be reached by combining trace fossils into broadgroups defined both on morphology and interpretation. This alsofocuses attention on looking for similarites between Neoproterozoicand Cambrian trace fossils. Siliciclastic sediments of the Neoproterozoicpreserve elongate tubular organisms and structures of probablealgal origin, many of which are very similar to trace fossils.Such enigmatic structures include Palaeopascichnus and Yelovichnus,previously thought to be trace fossils in the form of tightmeanders. A preliminary two or tripartite terminal Neoproterozoic tracefossil zonation can be be recognized. Possibly the earliesttrace fossils are short unbranched forms, probably younger thanabout 560 Ma. Typical Neoproterozoic trace fossils are unbranchedand essentially horizontal forms found associated with diverseassemblages of Ediacaran organisms. In sections younger thanabout 550 Ma a modest increase in trace fossil diversity occurs,including the appearance of rare three-dimensional burrow systems(treptichnids), and traces with a three-lobed lower surfaces.     

贵州新元古代到寒武纪早期特异埋藏后生生物群及其研究意义

贵州化石丰富,素有古生物王国之称,其中早期特异埋藏后生生物十分发育,由老至新有埃迪卡拉系的瓮安生物群、江口庙河生物群、寒武系第1统的小壳动物群、第2统下部的牛蹄塘生物群及中上部的杷榔动物群、第3统底部的凯里生物群。这些特异埋藏生物群与云南寒武系的澄江动物群、关山生物群及陕南埃迪卡拉系高家山生物群构成了我国早期后生生物的演化链,反映了从动物起源（瓮安生物群）、可疑多门类动物出现（江口庙河生物群）、动物骨骼化（高家山生物群）、生物矿化质和量的变化或骨骼动物爆发性辐射（小壳动物群）及海绵动物辐射（牛蹄塘生物群）再到动物主要门类出现的寒武纪大爆发（澄江动物群）、海洋生物多样化及生态复杂化（凯里生物群）的演化过程。     

Contribution of sponge genes to unravel the genome of the hypothetical ancestor of Metazoa (Urmetazoa)

Recently the term Urmetazoa, as the hypothetical metazoan ancestor, was introduced to highlight the finding that all metazoan phyla including the Porifera (sponges) are derived from one common ancestor. Sponges as the evolutionarily oldest, still extant phylum, are provided with a complex network of structural and functional molecules. Analyses of sponge genomes from Demospongiae (Suberites domuncula and Geodia cydonium), Calcarea (Sycon raphanus) and Hexactinellida (Aphrocallistes vastus) have contributed also to the reconstruction of the evolutionary position of Metazoa with respect to Fungi. Furthermore, these analyses have provided evidence that the characteristic evolutionary novelties of Metazoa, such as the extracellular matrix molecules, the cell surface receptors, the nervous signal transduction molecules as well as the immune molecule existing in Porifera, share high sequence and in some aspects also functional similarities to related polypeptides found in other metazoan phyla. During the transition to Metazoa new domains occurred; as one example, the formation of the death domain from the ankyrin is outlined. In parallel, domanial proteins have been formed, such as the receptor tyrosine kinases. The metazoan essentials have been defined by analyzing and comparing the sponge sequences with the related sequences from the metazoans Homo sapiens, Caenorhabditis elegans and Drosophila melanogaster, the fungus Saccharomyces cerevisiae and the plant Arabidopsis thaliana. The data revealed that those sponge molecules grouped to cell adhesion cell recognition proteins are predominantly found in Protostomia and Deuterostomia while they are missing in Fungi and Viridiplantae. Moreover, evidence is presented allowing the conclusion that the sponge molecules are more closely related to the corresponding molecules from H. sapiens than to those of C. elegans or D. melanogaster. Especially surprising was the finding that the Demospongiae are provided with elements of adaptive immunity.     

On the origin of Metazoan adhesion receptors: cloning of integrin alpha subunit from the sponge Geodia cydonium

Integrins are prominent receptors known from vertebrates and the higher phyla of invertebrates. Until now, no evidence has been provided for the existence of integrins in the lowest Metazoa, the sponges (Porifera). We have isolated and characterized a cDNA clone encoding the alpha subunit of integrin from the marine sponge Geodia cydonium (GCINTEG). The open reading frame encodes a polypeptide of 1,086 residues (118 kDa). The intracellular domain features the sequence Tyr- Phe-x-Gly-Phe-Phe-x-Arg, which is different in one residue from the characteristic consensus pattern for integrin alpha subunits. We conclude that sponges, the oldest multicellular animal phylum, already utilize the structural elements which are required for a tuned and controlled interaction among cells, and between cells and the extracellular matrix.      

Origin and the Early Evolution of the Phylum Mollusca

The paper gives an overview of the modern hypotheses on the origin of the phylum Mollusca and the formation of its main classes. The Cambrian stage of molluscan evolution is characterized based on the paleontological material. The doubtfulness of assignment of the Precambrian (Vendian) soft-bodied fossil Kimberella to mollusks is argued. Judging from the interpretation of the morphologically diverse Cambrian fossils, it is suggested that the classes Polyplacophora, Monoplacophora, Gastropoda and Bivalvia formed already near the Precambrian–Cambrian boundary, i.e., from the beginning of the paleontologically documented evolutionary history of the phylum, whereas the assumption of the later origin of these taxa is unconvincing. The remaining classes of mollusks arose later, i.e., Cephalopoda, in the Late Cambrian; Scaphopoda, in the Ordovician; and Aplacophora, in the Silurian.     

Isolation and characterization of two T-box genes from sponges,the phylogenetically oldest metazoan taxon

It is now well established that all metazoan phyla derived from one common ancestor, the hypothetical Urmetazoa. Due to the basal position of Porifera (Demospongiae) in the phylogenetic tree of Metazoa, studies on the mechanisms controlling the development of these animals can provide clues on the understanding of the origin of multicellular animals and on how the first organization of the body plan evolved. In this report we describe the isolation and genomic characterization of two T-box genes from the siliceous sponge Suberites domuncula. The phylogenetic analysis classifies one into the subfamily of Brachyury, Sd-Bra, and the second into the Tbx2 subfamily, Sd-Tbx2. Analyses of the Sd-Bra and Sd-Tbx2 sequences and their intron-exon structures demonstrate their basal position in the phylogeny of the T-box family, and allows us to hypothesize a model of the phylogenetic evolution of all T-box genes. Furthermore, we report the presence of two different products of alternative splicing of Sd-Bra, and demonstrate that they exist in different phosphorylation and glycosylation states in the sponge tissue. Sd-Bra expression in tissue and 3D-cell aggregates (primmorphs) is analyzed, suggesting that Sd-Bra might also have a role in Porifera morphogenesis.Edited by N. SatohThe sequences from Suberites domuncula reported here are deposited in the EMBL/GenBank data base: the cDNA of Brachyury (Sd-Bra; accession number AJ544242) as well as the second T-box gene Sd-Tbx2 (AJ544241).