Leaf evolution in early-diverging ferns: insights from a new fern-like plant from the Late Devonian of China

Background and Aims With the exception of angiosperms, the main euphyllophyte lineages (i.e. ferns sensu lato, progymnosperms and gymnosperms) had evolved laminate leaves by the Late Devonian. The evolution of laminate leaves, however, remains unclear for early-diverging ferns, largely represented by fern-like plants. This study presents a novel fern-like taxon with pinnules, which provides new insights into the early evolution of laminate leaves in early-diverging ferns.Methods Macrofossil specimens were collected from the Upper Devonian (Famennian) Wutong Formation of Anhui and Jiangsu Provinces, South China. A standard degagement technique was employed to uncover compressed plant portions within the rock matrix.Key Results A new fern-like taxon, Shougangia bella gen. et sp. nov., is described and represents an early-diverging fern with highly derived features. It has a partially creeping stem with adventitious roots only on one side, upright primary and secondary branches arranged in helices, tertiary branches borne alternately or (sub)oppositely, laminate and usually lobed leaves with divergent veins, and complex fertile organs terminating tertiary branches and possessing multiple divisions and numerous terminal sporangia.Conclusions Shougangia bella provides unequivocal fossil evidence for laminate leaves in early-diverging ferns. It suggests that fern-like plants, along with other euphyllophyte lineages, had independently evolved megaphylls by the Late Devonian, possibly in response to a significant decline in atmospheric CO₂ concentration. Among fern-like plants, planate ultimate appendages are homologous with laminate pinnules, and in the evolution of megaphylls, fertile organs tend to become complex.     

BOTRYOPTERIS FORENSIS (BOTRYOPTERIDACEAE), A TRUNK EPIPHYTE OF THE TREE FERN PSARONIUS

Basal parts of Botryopteris forensis have been discovered rooted within the mantle of the tree fern Psaronius. Specimens occur in Upper Pennsylvanian coal balls from near Steubenville, Ohio, USA. The Botryopteris stems branch profusely, and these shoots are intertwined with the Psaronius roots near the surface of the mantle. They also produce adventitious roots that extend among the Psaronius roots. This material demonstrates that B. forensis was a trunk epiphyte, rather than a rhizomatous terrestrial fern. The B. forensis plant is interpreted to have branched continuously, to ramify, and to maintain itself at the periphery of the growing mantle of Psaronius roots. A new reconstruction of B. forensis is offered showing the large, globose fructifications hanging pendulously from horizontal fronds on emergent shoots. Epiphytes and lianas are common on the trunks of Psaronius, indicating that some Marattiales did not produce leaf skirts.     

Oligocene ferns from the Rancahué Formation (Aluminé, Neuquén, Argentina): Cuyenopteris patagoniensis nov. gen., nov. sp. (Polypodiales: Blechnaceae/Dryopteridaceae) and Alsophilocaulis calveloi Menéndez emend. Vera (Cyatheales: Cyatheaceae)

The study of abundant permineralized fern remains collected in the Oligocene Rancahué Formation (Neuquén Province, Argentina) allowed the identification of two different taxa. The name Cuyenopteris patagoniensis nov. gen., nov. sp. is proposed for several stems and fronds characterized by the presence of caulinar dictyosteles with root gaps, producing C-shaped leaf traces composed by two hippocampiform bundles flanked by eight to ten small round bundles, half of them diverging from cauline meristeles and the rest from the hippocampiform bundles. The presence of these features, along with frond bases with adaxial grooves and a pair of pneumatophores, suggest affinities with the Blechnaceae or Dryopteridaceae. The second identified taxon is Alsophilocaulis calveloi Menéndez. Based on features observed in these new materials, including characteristics of the mantle of adventitious roots and anatomy of the petiole bases, an emended diagnosis for this latter species is proposed.     

THE LEPIDOPHYTIC AFFINITIES OF THE GENUS CHINLEA AND OSMUNDITES WALKERI

Chinlea campii Daugherty and Osmundites walkeri Daugherty are species of petrified stems from the Upper Triassic Chinle Formation of Arizona that were described as members of the fern family Osmundaceae. Investigation of additional material indicates that the two species are conspecific and belong to the Lepidophyta. The stems are radially symmetric and have an ectophloic siphonostele in which the xylem cylinder is thick and deeply furrowed. Internal pressure against the xylem cylinder caused by the lateral expansion of the pith in some stems produces what appears in transverse section to be a ring of up to 60 separate xylem strands. Leaf traces are small, terete, collateral and have exarch xylem. They are arranged in a tight spiral. Adventitious roots, secondary xylem, and secondary cortex are lacking. The stems are classified under the binomial Chinlea campii, and other axes that have similar cortical anatomy but in which all vascular tissues have decayed are treated as Chinlea sp. Both types of stems are interpreted as ephemeral aerial shoots of an herbaceous plant. Of the known fossil Lepidophyta, Chinlea is most similar to Pleuromeia and Nathorstiana, but it differs from each of these genera in a number of respects and is therefore included in Lepidophyta incertae sedis.     

STRUCTURALLY PRESERVED PLANTS FROM SOUTHEASTERN KENTUCKY: STAUROPTERIS BISERIATA SP. NOV.

Several axes of the coenopterid fern Stauropteris are described from permineralized peat associated with Lower-Middle Pennsylvanian coal deposits of southeastern Kentucky. This represents the first documented report of the genus in North America. The specimens are regarded as representatives of a new species—S. biseriata—based on the distinctive branching habit. Three branch orders are described, and in each case, branches are singular and distichous, arranged in a two-ranked pattern. This is in contrast to other species of Stauropteris in which the branches are paired and form a quadriseriate pattern. A pair of vascularized aphlebiae subtend each branch through all branching orders. Aphlebiae associated with first- and second-order branches are three-parted at the point of insertion; those that subtend third-order branches are singular. Stauropteris and a number of Devonian fern-like plants are compared on the basis of certain morphological similarities.     

VASCULAR CONNECTION BETWEEN LATERAL ROOTS AND STEM IN THE OPHIOGLOSSACEAE

The vascular connection between lateral roots and stem in the Ophioglossaceae and in two leptosporangiate fern species was examined. Two types of connections were found: “gradual” connections, which resemble leaf traces in ontogeny and morphology, and “abrupt” connections, which resemble the connections between lateral roots and their parent roots. Gradual root-stem connections occur in the genera Ophioglossum and Helminthostachys and in Woodwardia virginica. They are initiated in shoot apices distal to the level where cauline xylem elements mature. They resemble leaf traces in being provascular (procambial) strands that connect the cauline stele with the future vasculature of lateral appendages. As with leaf traces, gradual connections are part of the provascular and, later, protoxylem continuity between stems and lateral appendages. Gradual connections have many features in common with leaf traces, and the term root trace is applicable to them. The order of radial maturation of the primary xylem in gradual connections varies in different parts of the connections. It is endarch near the intersection with the cauline stele and exarch where the connections intersect root steles. Gradual connections resemble the transition regions of certain seed plants where protoxylem is also continuous from stem to root and the order of maturation is found to change continuously from stem to root. Abrupt connections occur in Botrychium and Osmunda cinnamomea. They develop in shoot apices at levels where cauline xylem is mature or maturing. The mature xylem does not dedifferentiate, so provascular and protoxylem continuity of the kind found in root traces does not occur. Also, reorientation of the order of maturation does not occur in abrupt connections. Xylem connectors are found in the region where radially oriented elements of the connections abut the longitudinally oriented cauline elements. Abrupt connections resemble the connection of secondary roots with their parent root systems since xylem connectors and the lack of continuity are also features found in these vascular systems. The resemblance of the vascular pattern of the fern root trace to the transition region of seed plants suggests that the radicle is more closely comparable to the cladogenous roots of pteridophytes than hitherto supposed.     

ANACHOROPTÉRIS INVOLUTA AND ITS ATTACHMENT TO A TUBICAULIS TYPE OF STEM FROM THE PENNSYLVANIAN OF IOWA

Hall , John W. (U. Minnesota, Minneapolis.) Anachoropteris involuta and its attachment to a Tubicaulis type of stem from the Pennsylvanian of Iowa. Amer. Jour. Bot. 48(8): 731–737. Illus. 1961.—Petioles referable to Anachoropteris involuta are described, attached to a stem which most nearly corresponds to a member of the genus Tubicaulis. These petioles are attached in a 2/5 phyllotaxy. At their points of departure petiole traces are massive and C-shaped but become involute in regions away from their attachment. Adventitious roots were borne on the stem in partial whorls. It is suggested that Anachoropteris petioles were extremely long and that they bore adventitious stems at intervals. These, in turn, bore adventitious roots, perhaps to serve for uptake of minerals or support in regions removed from the true stem. Such a “petiole unit” may also have functioned as a vegetative propagule. This would account for the apparent rarity of true stems and the abundance of petioles in coal balls.     

Conantiopteris schuchmanii, gen. et sp. nov., and the Role of Fossils in Resolving the Phylogeny of Cyatheaceae s.l.

Conantiopteris schuchmanii gen. et sp. nov. The specimen, 23.2 cm long and 11.7 cm wide, shows helically arranged persistent frond bases embedded in adventitious roots, and is clothed by multicellular trichomes. A parenchymatous pith with mucilaginous cells and sclerotic nests is surrounded by an amphiphloic distyostele, parenchymatous inner cortex, and outer sclerenchymatous cortex. Sclerenchyma also surrounds the cauline vasculature and leaf traces. Medullary and cortical bundles are absent. Phloem contains both axially elongated and tangential sieve elements. Frond bases are oval in outline with three vascular bundles, including an undulating abaxial arc and an adaxial pair. Protoxylem of the stipe is endarch and is associated with cavity parenchyma. These characters are indicative of tree fern affinities. A cladistic analysis using trunk characters of both living and fossil tree ferns was conducted to help establish relationships of the new species and other fossil ferns, and to test hypotheses of general tree fern relationships. Additional analyses of living taxa only were also performed. Results from the analysis using both living and fossil taxa compare favorably with those that included only living species when either morphological characters or molecular sequences of the chloroplast gene rbcL are utilized. Although there are variations in the topologies of the various trees, results indicate that the new genus is nested among a paraphyletic assemblage of dicksoniaceous, lophosoriaceous, and metaxyaceous species that subtend a monophyletic Cyatheaceae s.s. Received 26 April 1999/ Accepted in revised form 15 July 1999     

A comparison of mineral uptake and translocation by above-ground and below-ground root systems ofSalix syringiana

Nutrient uptake and translocation by above-ground adventitious roots and below-ground roots of woodySalix syringiana saplings were studied with gamma spectrometry. Each of four radionuclides (⁷⁵Se,¹³⁸Cs,⁵⁴Mn, and⁶⁵Zn) administered to adventitious and belowground roots were detected in stems and leaves within one month. Nuclides tended to be immobilized in the leaves and branches closest to the adventitious roots that absorbed them, while nuclides absorbed from below-ground sources were distributed more evenly throughout the plant. The capacity of adventitious roots to acquire nutrients from above-ground sources suggests they function as a potential auxiliary pathway of nutrient uptake and might enhance plant nutrient status where below-ground root uptake it hindered by adverse soil conditions.     

A novel life cycle arising from leaf segments in plants regenerated from horseradish hairy roots

Summary Horseradish (Armoracia rusticana) hairy root clones were established from hairy roots which were transformed with the Ri plasmid in Agrobacterium rhizogenes 15834. The transformed plants, which were regenerated from hairy root clones, had thicker roots with extensive lateral branches and thicker stems, and grew faster compared with non-transformed horseradish plants. Small sections of leaves of the transformed plants generated adventitious roots in phytohormone-free G (modified Gamborg's) medium. Root proliferation was followed by adventitious shoot formation and plant regeneration. Approximately twenty plants were regenerated per square centimeter of leaf. The transformed plants were easily transferable from sterile conditions to soil. When leaf segments of the transformed plants were cultured in a liquid fertilizer under non-sterile conditions, adventitious roots were generated at the cut ends of the leaves. Adventitious shoots were generated at the boundary between the leaf and the adventitious roots and developed into complete plants. This novel life cycle arising from leaf segments is a unique property of the transformed plants derived from hairy root clones.     

Growth responses to waterlogging and drainage of woody Hakea (Proteaceae) seedlings,originating from contrasting habitats in south-western Australia

We investigated the responses of seven woody Hakea (Proteaceae) species (two populations of each), to two months of waterlogging and subsequent drainage, in a controlled glasshouse experiment. The species originated from contrasting environments (winter-wetland versus non-wetland habitats), and differed in abundance (endangered ironstone species versus common species). Waterlogging arrested growth of the main root system, and stimulated the formation of superficial adventitious roots just below the root/shoot junction in all species. Wetland species produced at least twice the amount of adventitious root dry mass of that of non-wetland species, due to differences in number, length or degree of branching. Their adventitious roots also tended to have higher porosities (7–10% versus 5–6% gas spaces). The relative amount of adventitious roots formed was strongly, positively correlated with the maintenance of shoot growth, and only the non-wetland species showed significant shoot growth reductions (19%) upon waterlogging. Dry mass percentage of stems and leaves, and leaf dry mass per area (LMA) increased considerably during waterlogging in all species (averages of 15, 29 and 27%, respectively), returning to the values of continuously drained control plants after drainage. Similarly, upon drainage, a suppression of shoot growth (average 35%) and a stimulation of root growth (average 50%) restored the root mass ratios to those of control plants. We found a negative correlation between the maintenance of growth during waterlogging versus that after waterlogging, suggesting a trade-off in functioning of the superficial adventitious roots between waterlogged and drained conditions. The rare winter-wet ironstone endemics resembled the common winter-wet species in most of their responses to waterlogging and drainage. Therefore, the results presented here cannot offer an explanation for their different distribution patterns. Our results suggest that non-wetland species may be disadvantaged in a wetland environment, due to their lower capacity to form adventitious roots resulting in stronger growth reductions.     

An auxin-inducible gene from loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis> L.) is differentially expressed in mature and juvenile-phase shoots and encodes a putative transmembrane protein

We have isolated a gene from loblolly pine, 5NG4, that is highly and specifically induced by auxin in juvenile loblolly pine shoots prior to adventitious root formation, but substantially down-regulated in physiologically mature shoots that are adventitious rooting incompetent. 5NG4 was highly auxin-induced in roots, stems and hypocotyls, organs that can form either lateral or adventitious roots following an auxin treatment, but was not induced to the same level in needles and cotyledons, organs that do not form roots. The deduced amino acid sequence shows homology to the MtN21 nodulin gene from Medicago truncatula. The expression pattern of 5NG4 and its homology to a protein from Medicago involved in a root-related process suggest a possible role for this gene in adventitious root formation. Homology searches also identified similar proteins in Arabidopsis thaliana and Oryza sativa. High conservation across these evolutionarily distant species suggests essential functions in plant growth and development. A 38-member family of genes homologous to 5NG4 was identified in the A. thaliana genome. The physiological significance of this redundancy is most likely associated with functional divergence and/or expression specificity of the different family members. The exact biochemical function of the gene is still unknown, but sequence and structure predictions and 5NG4::GFP fusion protein localizations indicate it is a transmembrane protein with a possible transport function.Electronic Supplementary Material Supplementary material is available in the online version of this article at Abbreviations ABA Abscisic acid - BA Benzylaminopurine - EST Expressed sequence tag - NAA 1-Naphthaleneacetic acid - GFP Green fluorescent protein - ORF Open reading frame     

Ellesmeris sphenopteroides,gen. ET SP. NOV., a new zygopterid fern from the Upper Devonian (Frasnian) of Ellesmere,N.W.T., Arctic Canada

A new fern-like fossil plant is described from the lower Upper Devonian of southern Ellesmere Island, Canadian Arctic Archipelago. The plant occurs in an Archaeopteris-dominated flora preserved in the Nordstrand Point Formation (Mid-Late Frasnian) near Bird Fiord. The plant has a pinnate vegetative system with three branch orders and laminate sphenopteroid pinnules. Primary pinnae usually diverge from the main axis in distichous pairs (quadriseriate), but can depart singly (biseriate). Each primary pinna bears a basal catadromic aphlebia. Anatomically, the plant exhibits a mesarch, bipolar protostele that is ribbon- to clepsydropsoid-shaped in the main axis. Primary pinna traces are also initially bipolar and crescent-shaped, but may become four-ribbed before dividing into a pair of bipolar traces. The morphology and anatomy of this plant are nongymnospermous and are most similar to Zygopteridales (particularly Rhacophytaceae and Zygopteridaceae). The Frasnian age of Ellesmeris shows that laminated foliage had evolved in some zygopterid ferns much earlier than previously recognized. The Sphenopteris-like pinnules of Ellesmeris indicate the need for caution when attributing such a convergent foliar design to other plant groups, such as the Devonian gymnosperms.     

A new tree fern stem,Heilongjiangcaulis keshanensis gen. et sp. nov., from the Cretaceous of the Songliao Basin,Northeast China: a representative of early Cyatheaceae

A new genus and species of Cretaceous Cyatheacean tree fern, Heilongjiangcaulis keshanensis gen. et sp. nov., is erected for several permineralized stems collected at the Keshan County in Songliao Basin, Heilongjiang Province, Northeast China. The new taxon is characterized by a dictyostelic, erect stem with dense multicellular scales and surrounded by persistent petiole bases and adventitious roots. The stem contains a central pith lacking medullary bundles, which is surrounded by a dictyostele, and the cortex externally. Each meristele of the dictyostelic ring is enclosed by a sclerenchyma sheath. The pith and cortex are parenchymatous. The proximal petiole bases present a frond trace composed of numerous meristeles, arranged in 1 abaxial and 2 adaxial arcs, with internally projecting bundles on the upper and lateral sides. The feature combination of the new genus is nearly identical to the anatomical structures of modern scaly genera of the Cyatheaceae apart from the absence of medullary bundles. It is interpreted as a primitive representative of early Cyatheaceae, that closely resembles the modern scaly genera, which suggests that in the Cretaceous, the tree ferns in this family were already in possession of most of the anatomical characteristics observed in extant taxa. The fossil records of the stems, petioles, and spores indicate that during the Jurassic and Cretaceous, the Northeastern region of Asia may have been one of the distribution centers of early Cyatheaceae.     

AUREALCAULIS CROSSII GEN. ET SP. NOV., AN ARBORESCENT,OSMUNDACEOUS TRUNK FROM THE FORT UNION FORMATION (PALEOCENE), WYOMING

Aurealcaulis crossii gen. et sp. nov., is based on permineralized trunks of an osmundaceous tree fern from the Paleocene Fort Union Formation from near Bitter Creek Station of southwestern Wyoming. This new species is characterized by centripetal (exarch) development of its xylem strands which form part of the leaf traces. Most of the leaf traces depart the stele as two segments that fuse into a single C-shaped petiole vascular strand outside of the outer cortex. Stipular expansions of the petiole bases of this species lack sclerenchyma, and roots arise from the lateral edges of leaf traces in the inner cortex. The family Osmundaceae and subfamily Osmundoideae are slightly emended to accept genera assignable to this family and subfamily with exarch protoxylem in their steles. Foliage similar to Osmunda greenlandica (Heer) Brown, which is possibly the leaf form of A. crossii, occurred next to an axis of this species which was in growth position. This axis was anchored in a lignite suggesting that this species grew under swampy conditions. Aurealcaulis crossii is the first arborescent member of the Osmundaceae of Tertiary age and the second arborescent form in this family reported from the Northern Hemisphere.     

Effects of explant types and media salt strength on growth and secondary metabolite accumulation in adventitious roots of <Emphasis Type="Italic">Periploca sepium</Emphasis> Bunge

In order to study the capabilities of Periploca sepium adventitious root induction in different types of explants, we selected leaves, roots and stems with or without buds. The growth of adventitious roots and periplocin content in these roots were determined. In order to investigate the suitable media salt strength, we cultured the adventitious roots in different salt strength (0.25, 0.50, 1.0, 1.5, 2.0) of Murashige and Skoog (MS) media supplemented with 1 mg/l indole butyric acid (IBA) and 30 g/l sucrose. The results showed that both leaf and root explants were proven suitable for the adventitious root induction; however, the stems could hardly induce adventitious roots no matter whether the stems had buds or not. Further studies reported that adventitious root proliferation and periplocin production derived from root explants were higher than those derived from leaf explants. So the root explants were the optimum explants for adventitious root induction, growth and periplocin production. The salt strength experiment showed that with the increasing salt strength (1.0–2.0 MS), adventitious root growth decreased significantly, as well as periplocin content in comparison with lower (0.25–0.5 MS) salt strength media.     

ITOPSIDEMA,A NEW GENUS OF THE OSMUNDACEAE FROM THE TRIASSIC OF ARIZONA

Daugherty , Lyman H. (San Jose State College, San Jose, Calif.) Itopsidema, a new genus of the Osmundaceae from the Triassic of Arizona. Amer. Jour. Bot 47(9): 771–777. Illus. 1960.—Itopsidema vancleavei, a new genus and species of the family Osmundaceae, is described. The specimen consists of several segments of an arborescent stem obtained from the Upper Triassic of the Petrified Forest National Monument near Holbrook, Arizona. The surfaces on 2 of the segments are covered by adventitious roots and the remaining segments are covered by leaf bases. The fronds are spirally arranged and have an 8/21 phyllotaxy. The cortex of the stem, which contains numerous leaf traces and adventitious roots, consists of parenchyma with cell walls of medium thickness. The leaf traces are oblong to crescent-shaped in the inner cortex and horseshoe-shaped in the outer cortex. The base of the petiole contains a single, large vascular bundle and is covered by multicelled, glandular spines. The adventitious roots originate on the abaxial side of the leaf traces in the region of the inner cortex. The center of the stem is occupied by an ectophloic, mesarch siphonostele without leaf gaps. The pith is composed of firm-walled parenchyma cells which have isolated tracheids with reticulate pitting scattered among them. These tracheids are so rare the pith cannot be considered a “mixed pith.” The relationship and morphological significance of Itopsidema with respect to other members of the Osmundaceae are briefly discussed.     

Adventitious roots inGlycine subg.Glycine (Leguminosae): Morphological and taxonomic indicators of the B genome

Adventitious roots were observed on 3 wild perennialGlycine species, in short-poddedG. clandestina, G. latifolia and mostG. tabacina (2x, 4x, 6x), while other species lacked adventitious roots. This suggests that the adventitious roots trait is associated with B genome species. Intra- and interspecific F₁ hybrids reveal that adventitious roots apparently are inherited recessively. The presence of adventitious roots or short-poddedG. clandestina, coupled with infertility with longer-poddedG. clandestina, and enzymatic and leaflet morphology differences between the twoG. clandestina subgroups, supports segregation of the short-podded form as a separate taxon.     

THE VEGETATIVE ANATOMY OF SCHOPFIASTRUM DECUSSATUM FROM THE MIDDLE PENNSYLVANIAN OF THE ILLINOIS BASIN

Schopfiastrum decussatum, a monostelic pteridosperm, has been recovered from Middle Pennsylvanian age coal balls from six localities within the Illinois Basin. Additional features of stem anatomy include the presence of horizontal sclerotic plates in the inner cortex, secretory ducts in the inner and outer cortex, and adventitious roots. The primary xylem is interpreted as bilobed, emitting massive leaf traces in a distichous manner. Five orders of frond members are described, including pinnules. All orders of the frond contain secretory ducts and tissues characteristic of Schopfiastrum stems. The rectangular, bilobed protostele, and method of leaf trace origin indicate that Schopfiastrum is more closely related to certain lower Carboniferous seed ferns than to contemporary Pennsylvanian pteridosperms.