Five vicariant genera from Gondwana: the Velloziaceae as shown by molecules and morphology [corrected

Background and Aims

The amount of data collected previously for Velloziaceae neither clarified relationships within the family nor helped determine an appropriate classification, which has led to huge discordance among treatment by different authors. To achieve an acceptable phylogenetic result and understand the evolution and roles of characters in supporting groups, a total evidence analysis was developed which included approx. 20 % of the species and all recognized genera and sections of Velloziaceae, plus outgroups representatives of related families within Pandanales.

Methods

Analyses were undertaken with 48 species of Velloziaceae, representing all ten genera, with DNA sequences from the atpB-rbcL spacer, trnL-trnF spacer, trnL intron, trnH-psbA spacer, ITS ribosomal DNA spacers and morphology.

Key Results

Four groups consistently emerge from the analyses. Persistent leaves, two phloem strands, stem cortex divided in three regions and violet tepals support Acanthochlamys as sister to Velloziaceae s.s., which are supported mainly by leaves with marginal bundles, transfusion tracheids and inflorescence without axis. Within Velloziaceae s.s., an African Xerophyta + Talbotia clade is uniquely supported by basal loculicidal capsules; an American clade, Barbacenia s.l. + Barbaceniopsis + Nanuza + Vellozia, is supported by only homoplastic characters. Barbacenia s.l. (= Aylthonia + Barbacenia + Burlemarxia + Pleurostima) is supported by a double sheath in leaf vascular bundles and a corona; Barbaceniopsis + Nanuza + Vellozia is not supported by an unambiguous character, but Barbaceniopsis is supported by five characters, including diclinous flowers, Nanuza + Vellozia is supported mainly by horizontal stigma lobes and stem inner cortex cells with secondary walls, and Vellozia alone is supported mainly by pollen in tetrads.

Conclusions

The results imply recognition of five genera (Acanthochlamys (Xerophyta (Barbacenia (Barbaceniopsis, Vellozia)))), solving the long-standing controversies among recent classifications of the family. They also suggest a Gondwanan origin for Velloziaceae, with a vicariant pattern of distribution.     

Hummingbird Pollination in Two Species of Vellozia (Liliiflorae: Velloziaceae) in Southeastern Brazil*

Pollination by hummingbirds was observed in two species of Vellozia (Velloziaceae) in southeastern Brazil. Both species have purplish-pink to dark purple tepals, tube-like arrangement of stamens, large and protected nectar chamber, and no odour. Tepals are reflexed in V. leptopetala and narrow in V. declinans, further enhancing the tube-like effect given by the long yellow anthers. Nectar is very dilute (ca. 10%). Three species of hummingbirds were observed pollinating these Vellozia species: Colibri serrirostris, Augastes scutatus, and Chlorostilbon aureoventris. The first species established feeding territories in the vicinity of V. leptopetala flowers. A bee-pollinated Vellozia flower-type may give rise to a hummingbird-flower by relatively simple adjustments, consisting mainly of the reflexion of tepals, increase in stamen length, enhancement of reddish pigments, enlargement of nectar chamber and dilution of nectar. Vellozia declinans may be likened to one of the hypothetical stages of this trend towards ornithophily, whereas V. leptopetala is possibly one of the extremes within the genus.     

THE PHYLOGENETIC POSITION OF ANEMIA COLIMENSIS

The unique leaf structure of the genus Anemia with its upright fertile basal pinnae has been interpreted as derived from the typical fern leaf with unmodified basal pinnae. Reported herein for the first time is the leaf morphology of a species, Anemia colimensis Mickel, which shows the most primitive condition known in the genus and clearly confirms the course of evolution. The fertile pinnae are but little modified in size, lamination, and posture. Although A. colimensis has characters of two subgenera, it is shown to be a primitive element in subgen. Anemiorrhiza and not a member of subgen. Coptophyllum as previously reported.     

MODIFIED REMNANTS OF CENTRIPETAL XYLEM IN THE LEAF TRACE OF CEPHALOTAXUS

A study of the leaf traces at the nodes in various species and varietal forms of Taxus, Torreya, Amentotaxus and Cephalotaxus reveals, only in Cephalotaxus, an unusual type of parenchymatous tissue associated with the xylem of the leaf trace. The cells of this tissue occur in one to three layers, have abundant cytoplasm and conspicuous nuclei. The thin walls of these cells are devoid of pits and show spiral or spiral-reticulate thickenings. These thickenings, although readily taking the counterstain, reveal the presence of lignin as determined by the phloroglucin test and by fluorescence microscopy. This tissue is always internal—that is, adaxial—to the protoxylem of the trace. From the node it accompanies the trace for a short distance into the leaf base, where it gives way centrally to the typical leaf parenchyma and laterally to the transfusion tissue on the flanks of the bundle. In the basipetal direction it is in contact with the inner face of the annular protoxylem of the stele, eventually disappearing in the typical pith parenchyma. This tissue occasionally reveals instances of well-lignified tracheid-like centripetal elements. On the basis of the characteristics of this tissue, it is suggested that its origin lies in former centripetal xylem. The significance of this tissue to the evolution of the stele and the systematic position of Cephalotaxus is discussed.     

Cytogenetics and cytotaxonomy ofVelloziaceae

Chromosome number and other cytological features are reported from 35 species ofVelloziaceae, including several African and Brazilian populations. All analyzed species show areticulate interphase nuclei and prophase/prometaphase chromosomes with proximal early condensation. Most heteropycnotic blocks do not seem to correspond to heterochromatin since, at least inVellozia patens, they do not stain differentially after C-banding procedures. Regarding the chromosome number, three main groups could be identified. The first comprised diploid species of the generaNanuza, Vellozia and the Brazilian species ofXerophyta with 2n = 14 or 16; the second comprised tetraploid species with 2n = 34, and included all Brazilian species of subfam.Barbacenioideae; the third group, of hexaploid species, comprised the African representatives of the genusXerophyta. A single population ofVellozia, possibly of hybrid origin, had 2n 32. A basic number of x = 8 is proposed for the family. The karyological information supports the hypothesis that theVelloziaceae originated on the South American, rather than on the African continent.     

Effects of ultraviolet-B radiation on leaf elongation, production and phenylpropanoid concentrations of Deschampsia antarctica and Colobanthus quitensis in Antarctica

Stratospheric ozone depletion by anthropogenic chlorofluorocarbons has lead to increases in ultraviolet‐B radiation (UV‐B; 280–320 nm) along the Antarctic Peninsula during the austral spring. We manipulated UV‐B levels around plants of Antarctic hair grass (Deschampsia antarctica; Poaceae) and Antarctic pearlwort (Colobanthus quitensis; Caryophyllaceae) for one field season near Palmer Station along the west coast of the Antarctic Peninsula. Treatments involved placing frames over naturally growing plants that either (1) held filters that absorbed most biologically effective radiation (UV‐B_BE; ‘reduced UV‐B’, 22% of ambient UV‐B_BE levels), (2) held filters that transmitted most UV‐B_BE (‘near‐ambient UV‐B’, 87% of ambient UV‐B_BE levels), or (3) lacked filters (‘ambient UV‐B’). Leaves on D. antarctica exposed to near‐ambient and ambient UV‐B were 16–17% shorter than those exposed to reduced UV‐B, and this was associated with shorter epidermal cells at the leaf base and tip. Leaves on C. quitensis exposed to near‐ambient and ambient UV‐B tended to be shorter (P=0.18) and epidermal cells at the leaf base tended to be smaller than those under reduced UV‐B (P<0.10). In order to further explain reductions in leaf length, we examined leaf concentrations of insoluble (cell‐wall bound) phenylpropanoids, since it has been proposed that wall‐bound phenylpropanoids such as ferulic acid may constrain cell expansion and leaf elongation. In both species, HPLC analysis revealed that ferulic and p‐coumaric acid were major components of both insoluble and soluble phenylpropanoids. Although there were no significant differences in concentrations between UV‐B treatments, concentrations of insoluble ferulic acid in D. antarctica tended to be higher under ambient and near‐ambient UV‐B than under reduced UV‐B (P=0.17). We also examined bulk‐leaf concentrations of soluble (methanol extractable) UV‐B‐absorbing compounds and found that concentrations were higher in plants exposed to near‐ambient and ambient UV‐B than in plants exposed to reduced UV‐B. We also assessed the UV‐B‐screening effectiveness of leaves that had developed on plants at the field site with a fiber‐optic microprobe. Leaf epidermal transmittance of 300‐nm UV‐B was 4.0 and 0.6% for D. antarctica and C. quitensis, respectively, which is low compared to grasses and herbaceous dicotyledonous plants found in more temperate climates. While the leaves of Antarctic vascular plants are relatively effective at screening UV‐B, levels of UV‐B in Antarctica are sufficient to reduce leaf epidermal cell size and leaf elongation in these species, although the mechanisms for these reductions remain unclear.     

THE ROLES OF HETEROCHRONY AND HETEROBLASTY IN THE DIVERSIFICATION OF LEAF SHAPES IN BEGONIA DREGEI (BEGONIACEAE)

The relationship between shape variation in the transitional series of leaves and in adult leaves was examined in seedlings of seven morphs of Begonia dregei using several quantitative methods of shape analysis. There is variation in the shape of adult leaves among individuals as well as in juvenile leaves within individuals in B. dregei. As an individual grows, there is a gradual transition in leaf shape from the symmetrical, oval, smooth-margined leaves through a series of more than ten transitional leaves to a stable adult leaf shape. There appear to be two basic patterns to the acquisition of adult traits. Traits that differ among morphs are acquired gradually throughout the entire transitional series while those that are similar among morphs are acquired by about leaf 5 and remain stable through the later juvenile leaves. There is no identity of leaf shape between the earlier leaves of some morphs and the later leaves of others. Evolutionary diversification in adult leaf morphology in this species is not related to simple changes in ontogeny of the whole plant.     

A NUMERICAL–TAXONOMIC STUDY OF THE GENUS BULNESIA (ZYGOPHYLLACEAE): CLUSTER ANALYSIS,ORDINATION AND SIMULATION OF EVOLUTIONARY TREES

Cluster analysis by four methods, ordination by principal component analysis (PCA) and simulation of evolutionary trees (Wagner Trees) were performed on morphological data from 43 characters of eight species of the South American genus Bulnesia (Zygophyllaceae). The results of cluster analysis and principal component analysis in general agree and show that there are three pairs of taxa that appear, or obviously are, closely related. These are the pairs B. arborea–B. carrapo, B. foliosa–B. schickendantzii and B. retama–B. chilensis. These methods also indicate that the southern species B. bonariensis occupies an intermediate position between the pair of northern tropical species (B. arborea, B. carrapo) and the remaining southern species. From the beginning it was assumed that these three multifoliolate species with large flowers may be rather primitive. The Prim network indicates that these three species are closely related among themselves. Also in two of the three Wagner Trees they are placed in a group. In all cases B. sarmientoi is shown as the more remote and isolated of all species. It is regarded as a unique, specialized arboreal species showing extreme reduction in number of leaflets and carpels, leaf and flower size, etc. All graphic representations (Fig. 1–3) show the phenetic similarity or the close phylogenetic relationships of the pairs B. foliosa–B. schickendantzii and B. retama–B. chilensis to each other. These four species would represent a rather advanced group. The most xerophytic species B. retama and B. chilensis are regarded as the most advanced taxa and the most specialized histophysiologically. These occupy extreme and distant positions in PCA diagrams and Prim network, and top positions in the Wagner Trees.     

ROLE OF THE NITROGEN-FIXING BACTERIAL MICROFLORA IN THE EPIPHYTISM OF TILLANDSIA (BROMELIACEAE)

Nitrogen-fixing activity in the phyllosphere of 12 species of Tillandsia from different Mexican habitats was evaluated by the acetylene reduction assay, and nitrogen-fixing microorganisms were isolated and characterized. The leaves from eight of the 12 Tillandsia species examined exhibited nitrogenase activity in enrichment cultures. Among the microorganisms implicated—Agrobacterium, Bacillus, Erwinia, Pseudomonas, Rahnella, Vibrio, and Xanthomonas—only Bacillus megatherium reduced acetylene in pure culture. Our findings suggest that nitrogen fixation in the phyllosphere of the sampled epiphytes occurs under suitable conditions and that most of the bacteria involved are primarily soil and water inhabitants. The results also suggest a relationship between the composition of the nitrogen-fixing microbial communities grown on the leaf and the different development of the leaf area in Tillandsia due to the aerial components (wings) of the trichomes.     

THE ALLIUM INFLORESCENCE: SOME SPECIES OF THE SECTION MOLIUM

Mann , Louis K. (U. California, Davis.) The Allium inflorescence: some species of the section Molium. Amer. Jour. Bot. 46(10): 730–739. Illus. 1959.—The inflorescence is described for Allium neapolitanum, A. roseum, A. hirsutum, A. subhirsutum, A. zebdanense and A. erdelii, species of Molium, a Mediterranean section of the genus. Inflorescence structure is similar among these 6 species. The single spathe appears to consist of 4 coalesced bracts, each of which bears in its axil a bostryx (helicoid cyme) of 3–7 flowers. Central to these 4 peripheral bostryxes are several smaller ones which differentiate later and decrease in flower number toward the inflorescence center. Bracts and bracteoles (prophylls) are generally absent within the spathe. The inflorescence is not radially symmetrical and the bostryx on the side opposite the uppermost foliage leaf differentiates and develops first. Among the peripheral bostryxes there is a definite sequence of development and certain regularities of coiling (clockwise vs. counterclockwise). Within each bostryx the flowers open in a strict sequence from oldest to youngest; among the bostryxes, the 4 peripheral ones flower first, usually starting with the first bostryx to be differentiated. The central bostryxes flower last.     

Circadian Rhythms of Leaf and Stomatal Movements in Gymnosperm Species

It is generally accepted that various physiological, morphological and gene expression phenomena are under the control of a circadian clock, and that this time keeping mechanism is universally present. Although such endogenously regulated phenomena have first been documented in plants more than 250 years ago and much work has been accumulated particularly in the past 70 years, it was not obvious from the literature whether such time keeping mechanisms exist in gymnosperms. Two prominent parameters were investigated in several gymnosperm species which have been demonstrated to be under the control of a circadian clock in many plants: (i) leaf movement and (ii) stomata movement. In young plants of Pinus sylvestris, Picea abies, Taxus baccata, Araucaria angustifolia, Araucaria heterophylla and Ginkgo biloba leaf oscillations could be recorded for about 5 days. However, compared to angiosperm plants, the amplitude was small. The period length under free running conditions (constant temperature and continuous light) was characteristic for the species. Stomatal movement was observed in Ginkgo biloba leaves by electron microscopy. Stomata were open at noon and closed at midnight under normal day/night conditions (LD) as well as under constant light conditions (LL), indicating that stomatal aperture is under circadian control in the gymnosperm Ginkgo biloba. Online recordings of stomata conductance however, exhibited diurnal but not circadian oscillations of net CO₂-exchange in G. biloba leaves. Our results show that a circadian clock controls leaf and stomatal movements in gymnosperm species indicating that endogenous time keeping mechanisms are present.     

Drought responses,phenotypic plasticity and survival of Mediterranean species in two different microclimatic sites

• Climate models predict a further drying of the Mediterranean summer. One way for plant species to persist during such climate changes is through acclimation. Here, we determine the extent to which trait plasticity in response to drought differs between species and between sites, and address the question whether there is a trade‐off between drought survival and phenotypic plasticity.
• Throughout the summer we measured physiological traits (photosynthesis – A_max, stomatal conductance – g_s, transpiration – E, leaf water potential – ψl) and structural traits (specific leaf area – SLA, leaf density – LD, leaf dry matter content – LDMC, leaf relative water content – LRWC) of leaves of eight woody species in two sites with slightly different microclimate (north‐ versus south‐facing slopes) in southern Spain. Plant recovery and survival was estimated after the summer drought period.
• We found high trait variability between species. In most variables, phenotypic plasticity was lower in the drier site. Phenotypic plasticity of SLA and LDMC correlated negatively with drought survival, which suggests a trade‐off between them. On the other hand, high phenotypic plasticity of SLA and LDMC was positively related to traits associated with rapid recovery and growth after the drought period.
• Although phenotypic plasticity is generally seen as favourable during stress conditions, here it seemed beneficial for favourable conditions. We propose that in environments with fluctuating drought periods there can be a trade‐off between drought survival and growth during favourable conditions. When climate become drier, species with high drought survival but low phenotypic plasticity might be selected for.

     

A CORRELATION BETWEEN THE APICAL CELL AND THE HETEROBLASTIC LEAF SERIES IN MARSILEA

Plants of three species of Marsilea (M. vestita, M. villosa, M. drummondii) were grown in sterile culture under controlled conditions, and stem apices were sampled at one of the three heteroblastic leaf forms typical of this plant: spatulate, bifid, or quadrifid leaves. Statistical analyses were made of the relationship between the area of the apical cell and the leaf form which the plant produces under varied growth conditions. For all three species there is a statistically significant correlation (1% level) between apical cell area and leaf form. The analysis indicates that 83% of the variation in apical cell area in Marsilea vestita, 52% in M. villosa, and 54% in M. drummondii can be related to the change in leaf form. An increase in the glucose concentration of the culture medium increases the average apical cell area, and the addition of the protein synthesis inhibitor 2-thiouracil at concentrations of 10 mg/liter and 25 mg/liter decreases the average apical cell area. A certain average apical cell area is necessary for the production of a particular leaf form in the series. The area is a relative rather than an absolute size under any one particular growth condition. Any growth condition which inhibits or reverses the increase in apical cell size (which is typical of the normal growth pattern) will inhibit or reverse the normal heteroblastic series. Under constant conditions, the average size at which particular leaf forms develop appears to be species specific. These results confirm a generally held idea that the apex size is related to the heteroblastic leaf series, and they indicate that the area of the apical cell is the key factor rather than the volume or the number of cells of the apex. Undoubtedly the area of the apical cell is only a reflection of the physiological or morphological characteristics of the apical meristem that underlie the heteroblastic leaf series and which currently do not appear to lend themselves to more direct quantitative analysis.     

Coccinella septempunctata (Coleoptera,Coccinellidae): a species complex?

Marin, J., Crouau‐Roy, B., Hemptinne, J.‐L., Lecompte, E. & Magro, A. (2010). Coccinella septempunctata (Coleoptera, Coccinellidae): a species complex? —Zoologica Scripta, 39, 591–602. Coccinella septempunctata L., the seven spot ladybird is widely distributed across the Paleartic region. Based on a few morphological characteristics and geographical origin, some populations are recognized as distinct species, e.g., the North African Coccinella algerica Ková? and Japanese Coccinella brucki Mulsant later considered to be a subspecies –C. septempunctata brucki. The objective of this study is to discuss the taxonomic status of the seven spot ladybird, in particular whether it is a complex of species or the same species throughout the Paleartic region. The relationship between populations was clarified by using a combination of molecular and limited morphometric data and assessing potential reproductive barriers by means of cross breeding. Although there is considerable variation in the size of the spots on the elytra, the results of this study confirm that all studied populations belong to the same species.     

AN EXPANDED SURVEY OF THE ULTRASTRUCTURE OF RED ALGAL PIT PLUGS1

The fine structure of pit plugs in 90 species of red algae was examined, bringing the total number of species in the continuing survey to 153. The organization of plug caps was confirmed to be a stable, predictable trait within thalli, between generations in heteromorphic life histories, and within the presently recognized orders, with one exception—the Acrochaetiales. Two forms of the outer cap were found in this group, a thin plate, as in the Nemaliales and Palmariales, and a dome, as in Batrachospermales and Corallinales. Variation of pit plug structure indicates that the Acrochaetiales are a heterogeneous assemblage and that pit plugs will be useful in reappraising their systematics. The systematic affinities of several species of uncertain affinities are clarified. Schmitziella endophloea Bornet et Batters is excluded from both orders, Corallinales and Acrochaetiales, with which it previously was allied. Although other ordinal attributions are not precluded by pit plug structure alone, pit plug structure is consistent with placement of Apophlaea sinclairii Harvey and Hildenbrandia rivularis (Liebman) J. Agardh in the Hildenbrandiales, Plagiospora gracilis Kuckuck, Schmitziella endophloea, and Wurdemannia miniata (Duby) J. Feldmann et Hamel in the Gigartinales, and Pseudorhododiscus nipponicus Masuda in the Palmariales.     

THE ORGANIZATION OF THE VASCULAR SYSTEM IN THE STEMS OF THE NYMPHAEACEAE. I. NYMPHAEA SUBGENERA CASTALIA AND HYDROCALLIS

The vascular system in the stems of Nymphaea odorata and N. mexicana subgenus Castalia, and N. blanda subgenus Hydrocallis consists of continuing axial stem bundles with eight being the usual number. The stem bundles are concentric and xylem maturation is mesarch. Xylem elements consist of tracheids with spirally or weakly reticulated secondary wall thickenings. The phloem is made up of companion cells and short sieve tube members with simple sieve plates that are nearly transverse. At the node each leaf is supplied with two lateral leaf traces and a median leaf trace. A root trace is also present and supplies a series of adventitious roots borne on the leaf base. Flowers and vegetative buds develop directly from the apical meristem and occupy leaf sites in a single genetic spiral. Each flower or vegetative bud is related to a leaf through specific spatial and vascular association. The related leaf is separated from the related flower by three members of the genetic spiral and occupies an adjacent orthostichy. Vascular tissue for the related flower arises from the inner surfaces of the four stem bundles supplying leaf traces to the related leaf and extends through the pith to the flower or vegetative bud via a peduncle fusion bundle. The vascular system organization in the investigated species of Castalia and Hydrocallis is not typically monocotyledonous or dicotyledonous, nor can it be considered transitional between them. The ontogeny of the vascular system is similar to typical dicotyledons and the investigated species of Nymphaea can, therefore, be considered to represent highly specialized and modified dicotyledons.     

A CONTRIBUTION TO THE EARLY TERTIARY FOSSIL RECORD OF THE SALVINIACEAE

Megaspores and massulae of Azolla, Azollopsis, and Salvinia are recorded from the Paleocene and Eocene of Montana and the Dakotas. Three new species of Azolla are described and two species of Azolla are redescribed. The columella of the floating apparatus of the megaspore apparatuses of Azolla appears to be a phylogenetic localization of the perispore around the megaspore. A new section of Azolla (section Kremastospora) is created for species whose megaspore apparatuses have many floats and whose massulae have hooked (anchor-shaped) glochidia. Salvinia preauriculata is based only on leaf remains from the Eocene of North America. Megaspores and massulae which probably are conspecific with the leaf remains are described. The megaspores and massulae of the fossil species are much smaller than those of the living species, S. auriculata, with which S. preauriculata has been compared. The similar size of the megaspores and massulae of the fossil species suggests a less specialized condition than in living species, where these two structures differ greatly in size. Azollopsis tomentosa, previously known from the late Cretaceous, has been found in the Eocene.     

The molecular phylogenetics of Trachymyrmex Forel ants and their fungal cultivars provide insights into the origin and coevolutionary history of ‘higher‐attine’ ant agriculture

The fungus‐growing ants and their fungal cultivars constitute a classic example of a mutualism that has led to complex coevolutionary dynamics spanning c. 55–65 Ma. Of the five agricultural systems practised by fungus‐growing ants, higher‐attine agriculture, of which leaf‐cutter agriculture is a derived subset, remains poorly understood despite its relevance to ecosystem function and human agriculture across the Neotropics and parts of North America. Among the ants practising higher‐attine agriculture, the genus Trachymyrmex Forel, as currently defined, shares most‐recent common ancestors with both the leaf‐cutter ants and the higher‐attine genera Sericomyrmex Mayr and Xerolitor Sosa‐Calvo et al. Although previous molecular‐phylogenetic studies have suggested that Trachymyrmex is a paraphyletic grade, until now insufficient taxon sampling has prevented a full investigation of the evolutionary history of this group and limited the possibility of resolving its taxonomy. Here we describe the results of phylogenetic analyses of 38 Trachymyrmex species, including 27 of the 49 described species and at least 11 new species, using four nuclear markers, as well as phylogenetic analyses of the fungi cultivated by 23 species of Trachymyrmex using two markers. We generated new genetic data for 112 ants (402 new gene sequences) and 95 fungi (153 new gene sequences). Our results corroborate previous findings that Trachymyrmex, as currently defined, is paraphyletic. We propose recognizing two new genera, Mycetomoellerius gen.n. and Paratrachymyrmex gen.n. , and restricting the continued use of Trachymyrmex to the clade of nine largely North American species that contains the type species [Trachymyrmex septentrionalis (McCook)] and that is the sister group of the leaf‐cutting ants. Our fungal cultivar phylogeny generally corroborates previously observed broad patterns of ant–fungus association, but it also reveals further violations of those patterns. Higher‐attine fungi are divided into two groups: (i) the single species Leucoagaricus gongylophorus (Möller); and (ii) its sister clade, consisting of multiple species, recently referred to as Leucoagaricus Singer ‘clade B’. Our phylogeny indicates that, although most non‐leaf‐cutting higher‐attine ants typically cultivate species in clade B, some species cultivate L. gongylophorus, whereas still others cultivate fungi typically associated with lower‐attine agriculture. This indicates that the attine agricultural systems, which are currently defined by associations between ants and fungi, are not entirely congruent with ant and fungal phylogenies. They may, however, be correlated with as yet poorly understood biological traits of the ants and/or of their microbiomes.     

AN EXPERIMENTAL STUDY OF VARIATION IN THE PHACELIA SERICEA COMPLEX

Gillett , Georce W. (Michigan State U., East Lansing.) An experimental study of variation in the Phacelia sericea complex. Amer. Jour. Bot. 48(1): 1–7. Illus. 1961.—The Phacelia sericea complex consists of 2 diploid (n = 11), intergrading species, P. idahoensis and P. sericea. The experimental culture of several races of this complex demonstrated that differences in pubescence, leaf shape, and flower shape persist in plants grown in a common environment. Experimental interspecific F₁ hybrids demonstrated high fertility; portrayed intermediate expressions of pubescence, leaf shape, and flower shape; and were found to be, in many cases, indistinguishable from many wild intermediates. A study of herbarium specimens revealed numerous intergrades in which pubescence, leaf shape, and flower shape are highly variable, though loosely correlated. The evidence obtained from herbarium specimens, greenhouse cultures, field investigations, chromosome studies, and experimental hybridizations suggests a hybrid origin for the wild intermediates, recognized as Phacelia sericea (Graham) A. Gray subsp. ciliosa (Rydb.) Gillett.