Marbleseeds are gromwells – Systematics and evolution of Lithospermum and allies (Boraginaceae tribe Lithospermeae) based on molecular and morphological data

Phylogenetic relationships are complex within the Lithospermeae, a large subgroup of the Boraginaceae s.str. The relationships of New World Lasiarrhenum, Macromeria, Nomosa, Onosmodium, Perittostoma, and Psilolaemus to subcosmopolitan and much larger Lithospermum have not been critically investigated in the recent past. No molecular data on the phylogeny of these genera and Lithospermum have so far been published. We investigated the relationships within Lithospermeae using three loci (nuclear ITS plus 5.8S rRNA, chloroplast trnL-F-spacer, and trnS-G-spacer) and micromorphological character traits (pollen, nutlets). Lithospermum s.l. constitutes the sistergroup of Asian Ulugbekia and is monophyletic only when its American segregates “Macromeria”, monotypic Nomosa, and Onosmodium are included. Both the African and the South American species groups of Lithospermum are monophyletic, but North American representatives are not resolved in a single clade. Morphological characters that have been considered as important for generic delimitation in the past (such as large, yellow corollas without faucal scales, particular pollen types, coarsely veined leaves, shrubby habit) have evolved in at least two only distantly related lineages within Lithospermum s.l. The reduction of American “Macromeria”, Nomosa, and Onosmodium as well as Asian Ulugbekia under Lithospermum is proposed to render the latter monophyletic. This redefined Lithospermum s.l. appears to have undergone a type of recent “island radiation” in the Americas, reflected in a morphological diversity far exceeding that found in the Old World.     

Nomenclatural changes in <Emphasis Type="Italic">Lithospermum</Emphasis> (Boraginaceae) and related taxa following a reassessment of phylogenetic relationships

Lithospermum (Boraginaceae) comprises approximately 40 species in both the Old and New Worlds, with a center of diversity in the southwestern United States and Mexico. Using ten cpDNA regions, a phylogeny of Lithospermum and related taxa was reconstructed. Lithospermum (including New World and Old World species) and related New World members of Lithospermeae form a monophyletic group, with Macromeria, Onosmodium, Nomosa, Lasiarrhenum, and Psilolaemus nested among species of Lithospermum. New World Lithospermeae also is a monophyletic group, with Eurasian species of Lithospermum sister to this group. Because Lithospermum is not monophyletic without the inclusion of the other New World genera, species from these genera are transferred to Lithospermum, and appropriate nomenclatural changes are made. New combinations are Lithospermum album, Lithospermum barbigerum, Lithospermum dodrantale, Lithospermum exsertum, Lithospermum helleri, Lithospemum leonotis, Lithospermum notatum, Lithospermum oaxacanum, Lithospermum pinetorum, Lithospermum rosei, Lithospermum trinverium, and Lithospermum unicum; new names are Lithospermum chiapense, Lithospermum johnstonii, Lithospermum macromeria, Lithospermum onosmodium, Lithospermum rzedowskii, and Lithospermum turneri.     

Trigonotis jiaochengensis sp. nov. (Boraginaceae) from Shanxi,China

Trigonotis jiaochengensis Q. R. Liu & R. Y. Yan, a new species from Shanxi, China, is described and illustrated here. It is similar to T. peduncularis in having multiple diffuse stems, ovate to elliptic leaf blades, scorpioid cyme with only basal bracts and trigonous‐tetrahedral nutlets, but differs in having slender pedicels that are 5.2–18.4 mm long (vs clavate pedicels 1.5–4.8 mm in T. peduncularis), smooth nutlets with a short carpophore and a flat bottom (vs verrucose nutlets with a long carpophore and convex bottom in T. peduncularis). Morphological characters, including pollen and nutlets, with illustrations, diagnostic comparisons and habitat, are presented.     

A phylogenetic analysis of morphological and molecular characters of Lithospermum L. (Boraginaceae) and related taxa: evolutionary relationships and character evolution

Lithospermum (Boraginaceae) includes ca. 60 species and exhibits a wide range of floral, palynological, and vegetative diversity. Phylogenetic analyses based on 10 chloroplast DNA regions and 22 morphological characters were conducted in order to (i) examine evolutionary relationships within Lithospermum and among related genera of Boraginaceae, and (ii) investigate patterns of morphological evolution. Several morphological features, such as long‐funnelform corollas, faucal appendages, reciprocal herkogamy, and evident secondary leaf venation, have evolved multiple times within the genus. In contrast, other morphological features, including the presence of glands and the position and number of pollen pores, are less plastic and tend to characterize larger clades. Some features, including the presence of glands, are interpreted as symplesiomorphic for Lithospermum, while others, such as evident secondary leaf venation, appear to have originated repeatedly. The range of structural diversity that occurs among the species of Lithospermum suggests the potential utility of this genus as a model for integrative studies of evolution, development, and molecular biology.© The Willi Hennig Society 2011.     

Nutlet sculpturing of selectedTeucrium species (Lamiaceae): A character of taxonomic significance

Nutlet sculpturing of 23Teucrium species belonging to six sections was studied. The nutlets of analysed species were characterized by the presence of the oil glands of various density. SectionTeucrium is well defined by very densely distributed trichomes and oil glands on nutlets. Trichomes on nutlets were absent in all other species, except inT. marum from sect.Chamaedrys. Sect.Polium is a homogeneous group, distinguished from the other sections. There are some similarities of this sect. with sect.Scordium. Species of sect.Chamaedrys are heterogeneous and overlap according to the surface sculpturing characters with the other sections. The significance of nutlet microcharacters as additional taxonomical markers in the infrageneric classification of the genusTeucrium is discussed.     

Floral evolution in Lithospermum (Boraginaceae): independent origins of similar flower types

In Lithospermum (Boraginaceae), floral diversity is quite large, with variation in individual quantitative and qualitative traits as well as suites of floral characteristics. The present study utilizes phylogenetic, morphometric and developmental methodologies to investigate patterns of floral evolution, of individual and suites of traits, in the genus and among related genera. The evolutionary patterns of eight quantitative and five qualitative traits were reconstructed, and morphometrics and the evolution of floral morphospace were examined in the genus. Floral developmental patterns were established with light and scanning electron microscopy. Phylogenetic analyses provided evidence that derived flower types have each evolved multiple times, with differences in the manner in which these flower types arose. Floral morphospace has increased throughout the evolution of Lithospermum. Floral developmental patterns and cell lengths of floral organs at anthesis provided evidence that, in Lithospermum, an increase in the length of the sexual organs of flowers involves both longer cells and a greater number of mitotic divisions, but an increase in the length of the corolla is primarily a result of a larger number of cell divisions. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2016, 180 , 213–228.     

Stable transformation of Lithospermum erythrorhizon by Agrobacterium rhizogenes and shikonin production of the transformants

Seedling hypocotyls of Lithospermum erythrorhizon were infected with Agrobacterium rhizogenes (strain 15834) harboring a binary vector with an intron-bearing the β-glucuronidase (GUS) gene driven by cauliflower mosaic virus (CaMV) 35S promoter as well as the hygromycin phosphotransferase (HPT) gene as the selection marker. About 20% of the hairy roots isolated were hygromycin resistant and had co-integrated GUS and HPT genes in their Lithospermum genomic DNA. Because GUS activity was detected in almost all the hygromycin-resistant root tissues, the CaMV 35S promoter seems to be ubiquitously active in L. erythrorhizon hairy roots. In pigment production medium M9, the hairy root cultures had shikonin productivity similar to that of cell suspension cultures of Lithospermum. They also showed light-dependent inhibition of shikonin biosynthesis similar to that of Lithospermum cell cultures. These findings suggest that this hairy root system transformable with A. rhizogenes is a suitable model system for molecular characterization of shikonin biosynthesis via reverse genetics. Received: 2 March 1998 / Revision received: 25 May 1998 / Accepted: 8 July 1998     

Micromorphological and anatomical characteristics of Salvia amplexicaulis Lam., S. jurisicii Košanin and S. ringens Sibth. & Sm. (Lamiaceae)

Abstract

In this study, we examined Salvia amplexicaulis Lam., S. jurisicii Ko?anin and S. ringens Sibth. & Sm. collected in the Republic of North Macedonia, which are for the first time subjected to detailed micromorphological and structural analysis using light and scanning electron microscopy. The nutlets and mucilage were additionally subjected to spectroscopic analysis using Raman and Attenuated Total Reflectance Fourier Transform Infrared spectroscopy. The anatomical structure of stems and leaves is described and compared. The stems, leaves and calyces bear numerous one- and multi-cellular nonglandular trichomes, and various peltate, capitate and digitiform glandular trichomes. The nutlets differ in size and shape, as well as in myxocarpy. The nutlets predominantly contained α-linolenic and linoleic acid, whereas the mucilages are primarily formed of polysaccharides. The results obtained in this study confirmed the importance of micromorphological and anatomical analysis of Salvia L. spp. plant parts, particularly trichomes and nutlets, and hence contributing to the knowledge about the variety of micromorphological characteristics within the genus Salvia L.     

Nutlet micromorphology of Iranian Nepeta (Lamiaceae) species

To evaluate nutlets characteristics for systematic relationships, a comprehensive morphological and micro‐morphological study of the nutlets of 16 Iranian taxa of Nepeta (Lamiaceae) was conducted using scanning electron microscopy (SEM). Differences in surface ornamentation, size, shape and color were observed between the species. The studied taxa were categorized in two basic types based on surface ornamentation: non‐sculptured (smooth‐type) and sculptured nutlets. The non‐sculptured nutlets type could be further divided into four sub‐types, including smooth, granulate, undulate‐cellular and reticulate. In the sculptured nutlets, three subtypes of tuberculate, tuberculate‐cellular and verrucate were recognized. The shape of nutlets were described as oblong, ovoid‐oblong, oblong‐linear, to elliptic‐oblong and their size range are 1.2–2.0 mm in length and 0.5–2.0 mm in width. Based on micrographs, the areole form and location were defined as bi‐lobed, straight, basal, sub‐basal or lateral. Nutlet micromorphological characteristics such as surface ornamentation can be useful for classification and identification of e.g. medicinal species of Nepeta in Iran.     

Wachstum und Ausbildungsformen des Gynoeceums vonRochelia (Boraginaceae)

The ontogeny of the gynoecium ofRochelia disperma has been investigated by LM and SEM. From the floral apex only one carpel primordium arises abaxially and eventually shifts into a subterminal position. Neither an initial stadium of a second carpel nor an adaxial vascular strand in gynobase and style could be observed. InR. stylaris two vascular strands run through the style and two undifferentiated lobes in adaxial position may be regarded as rudimentary mericarps. Only from comparison with related taxa the conclusion can be drawn thatRochelia is really pseudomonomerous, more so inR. disperma than inR. stylaris. The primary gynoecial bulge splits up into three parts inR. disperma: style/stigma, nutlets with gynobase, and disc. While the mericarps originate in mostBoraginoideae from the symplicate region and the ascidiate one is restricted to the very basal parts, inR. disperma the ascidiate part extends and forms the nutlets. The hood-shaped mouth of the carpel (the plicate zone) is closed to a triangular slit in lateral position, the stigma.The nutlets are triangular with broad base and do not surround the adaxial part of the gynobase inR. disperma, R. persica, R. bungei, R. stylaris, andR. macrocalyx. In contrast,R. peduncularis, R. cardiosepala, andR. cancellata have nutlets clasping the gynobase; they may be more closely related than was assumed up to now. The glochids ofRochelia are fascicled unicellular hairs (with different shapes) and not emergences as in theCynoglosseae. There is an evolutionary trend towards fruit formation with only one mericarp, especially inR. disperma.

     

Anatomical and nutlet differentiation between <Emphasis Type="Italic">Teucrium montanum</Emphasis> and <Emphasis Type="Italic">T. polium</Emphasis> from Turkey

Teucrium montanum L. and T. polium L. are the two closest Teucrium L. species from sect. Polium (Mill.) Schreb in Turkey. In addition, they are sympatric for some part of their range in Turkey. In this study, comparative anatomical and micromorphological studies of the two species are carried out. They have been investigated by their leaf and stem anatomical features, as well as nutlet micromorphological characteristics. The results of anatomical studies show that the anatomical characters of both taxa are observed to be similar to the general features of the family Lamiaceae anatomy, except for lacking rich collenchyma at the corners. Both taxa are similar in stem anatomy and their leaves exhibit xeromorphy. However, trichome morphology on the stems and the leaves appear to have a taxonomic value in segregation of the two taxa. Light and scanning electron microscope studies on the nutlets also show that nutlet shapes, measures and surface micromorphologies are different in the two species. Whereas nutlet surfaces are bireticulate in both species, the nutlets are larger and primary sculpturing is more distinct in T. polium than in T. montanum. Moreover, the nutlets are oblong to oblong-ovoid and larger in T. polium, but ovoid and smaller in T. montanum.     

Nutlets micromorphology of some <Emphasis Type="Italic">Onosoma</Emphasis> L. (Boraginaceae) species from Turkey

Morphological and micromorphological characters of the nutlets of ten Onosma species in Turkey were reported. Studied fruit surfaces show some variations. Based on the structure and patterns of epidermis cells of the nutlets, two principal types can be distinguished. In Type 1, the epidermal cells of the nutlet surface are formed in a reticulate pattern with varied sizes and shapes of the meshes. This type is the most common type in the studied species. Seven of the species observed belong to the reticulate type, i.e. O. bracteosum, O. mutabile, O. aleppicum, O. bornmuelleri, O. aucheranum, O. araraticum and O. tauricum. In Type 2, the epidermal cells of the nutlet surface are formed in an elongated type with varied sizes and shapes. Three of the observed species belong to this type, i.e. O. roussaei, O. isauricum and O. stenolobum. Significant taxonomic differences between species were determined on the surface of the nutlets.     

Taxonomic revision of the genus Heliotropium (Boraginaceae s.l.) in south Yemen

The flora and fauna of Arabia, particularly southern Yemen, has recently attracted the interest of many authors. In this study, the genus Heliotropium L. (Boraginaceae) is taxonomically revised in southern Yemen. Ten species are recognized. Nomenclature, typification, representative specimens and a diagnostic key to all species are provided, along with their distribution in southern Yemen. The taxonomically most valuable characters in the genus are those of flowers (calyx, corolla, androecium and gynoecium) and nutlets, and those characters are thoroughly investigated and discussed. Both light and scanning microscopy are used in the investigation. Papillate anther apices (brush‐like) were recognized in some species. Connate anthers and deep or shallow ventral circular depressions on the nutlets were found to be useful in distinguishing members of H. sect. Orthostachys (viz. H. strigosum, H. rariflorum and H. ovalifolium), while winged nutlets were found only in H. pterocarpum. The length and hairiness of the style and stigma also proved to be taxonomically useful.     

The first subfossil records of Urtica kioviensis Rogow. and their consequences for palaeoecological interpretations

Among plant remains from Mesolithic layers dating from 9249 to 7779 B.C. at the excavation site of Friesack IV in north-eastern Germany, nutlets of Urtica kioviensis were identified. Morphological studies have shown that they clearly differed from all other European Urtica species investigated. In contrast, pollen morphological investigations revealed only slight differences between the central European Urtica species, which could hardly have been noticed during routine or normal pollen analyses. The records of U. kioviensis nutlets are the first subfossil finds reported and prove the indigenous status of this taxon in north-eastern Germany. The records are discussed in the context of the overall species spectrum of the Mesolithic layers and consequences for the interpretation of pollen analytical studies concerning human impact are pointed out. This paper is dedicated to the memory of the botanist and palaeoecologist Klaus Kloss (1934–2004).     

Onosma lhokaensis (Boraginaceae), a new species from Xizang,China

A new species of Boraginaceae, Onosma lhokaensis, is described and illustrated from Xizang, China. The new species closely resembles O. waddellii and O. waltonii in having azure corollas, coherent anthers and glabrous filaments, but differs in having particularly bristly indumentum, observably narrow corolla, longer calyxes and anthers, and remarkable nutlets with stipitate cicatrix. An updated key to the species of Onosma known from Xizang is also provided.     

Cynoglossopsis somaliensis,sp. nov., und die GattungCynoglossopsis (Boraginaceae)

Cynoglossopsis somaliensis H. Riedl, sp. nov., is described as a new species of the hitherto monotypic genusCynoglossopsis Brand which is closest related toCynoglossum L. but has to be included inBoraginoideae-Eritrichieae from the way the nutlets are attached to the gynobasis.

     

Die Perikarpentwicklung geflügelter Klausen aus demParacaryum-Verwandtschaftskreis (Cynoglosseae,Boraginaceae)

The anatomy and development of mericarps in four representatives ofBoraginaceae-Cynoglosseae has been investigated. In each case, the nutlets grow from a six-layered carpel. While the mesocarp becomes multilayered and thick in correlation with epizoochory inCynoglossum, it remains six-layered inMattiastrum, Paracaryum, andRindera. The wings are formed by both exo- and mesocarp and later on the inflated rim becomes hollow. Development of wings and reduction of glochids are discussed as adaptations to anemochory.

     

Nutlet morphology in Hemigenia R.Br. and Microcorys R.Br. (Lamiaceae)

Nutlets of Hemigenia R.Br. and Microcorys R.Br. were examined using SEM. Significant variation, mainly useful at the infrageneric level, was found in nutlet shape, nature of the attachment scar, nature of surface sculpturing, exocarp cell shape and sculpturing, and nature of the indumentum. Typical nutlets are ovoidal, strongly reticulate or rugose. The exocarp cells are isodiametric and convex to papillate. Also common are cylindrical nutlets, often with longitudinal ridging and papillate exocarp cells. Surface pitting and concave exocarp cells are rare. A cladistic analysis of nutlet characters suggests both Hemigenia and Microcorys are polyphyletic, and Microcorys paraphyletic with respect to Westringia Sm. Notwithstanding that, the infrageneric classification of Hemigenia was largely supported, while in Microcorys, there was support for sect. Hemigenioides, but sects Anisandra and Microcorys were not resolved as distinct.     

Pogostemon nudus sp. nov. (Lamiaceae) from Thailand

A new species of Pogostemon Desf., P. nudus Bongcheewin & Pramali, is described, illustrated and compared with the similar P. menthoides from Thailand. Morphological characters, especially the micromorphology of nutlets and leaf epidermis, and conservation status in accordance with the IUCN red list categories and criteria are described and discussed. In addition, a key to all species of P. sect. Pogostemon known from Thailand is presented.     

Maternal provisioning and possible joint breeding in the burrower bug Adomerus triguttulus (Heteroptera: Cydnidae)

Subsocial burrower bugs (Heteroptera: Cydnidae) provide unique opportunities to investigate evolutionary ecological questions regarding parental provisioning and family dynamics. Observations and marked nutlet‐setting experiments in the field showed that Adomerus triguttulus females progressively delivered mint nutlets into nests harbouring nymphs under the litter. More than one female often attended nymphs, but not eggs, in a nest in the field. The number of nymphs aggregating in a nest with a single female was usually smaller than that in a nest with two females, suggesting the joining of different families and facultative joint parental care. There was a positive correlation between the number of nutlets delivered and the number of nymphs in a nest. The number of attendant females also affected the amount of provisioning; more nutlets were found for second‐instar broods with more females. The effect of brood size on provisioning was confirmed for families under laboratory rearing. Maternal provisioning also varied with the developmental stage of offspring; second‐instar broods received more nutlets than first‐instar broods, with a temporal decrease in provisioning during the moulting of nymphs. Considering the growing evidence of food solicitation signals of young in subsocial insects, the observed finely tuned supply of food by the female could be induced by begging signals from the nymphs.