Comparative leaf anatomy of wild Manihot Mill. species (Euphorbiaceae) from Chapada Diamantina,Bahia, Brazil

The leaf anatomy of nine species of Manihot Mill. has been studied with the objective of finding anatomical features that contribute to our understanding of the taxonomy of the genus. Leaf samples were collected in the field and from herbarium specimens, and standardized methods were used for both light and scanning electron microscopy (SEM). The following species were studied: M. bellidifolia P. Carvalho & M. Martins, M. brachyandra Pax & K. Hoffm., M. caerulescens Pohl, M. diamantinensis Allem, M. elongata sp. nov. ined., M. jacobinensis Müll. Arg. (I), M. jacobinensis Müll. Arg. (II), M. longiracemosa P. Carvalho & M. Martins, M. maracasensis Ule, M. reniformis Pohl, M. tripartita (Spreng) Müll. Arg. and Manihot sp. Anatomically, the analysis revealed differences that indicate the importance of leaf anatomy in the species‐level taxonomy. Presence of fibers in the bundle sheath, and presence or absence of papillae and fibers in the midrib are diagnostic characters. Although these characteristics were not enough to separate all of the species, or to group them into a section, they did show taxonomic importance at the species level. In addition, morphoanatomical differences and geographical distribution support Manihot sp. as a distinct species.     

Manihot allemii sp. nov. (Euphorbiaceae s.s.) with entire and unlobed leaves from northern Brazil,with notes about foliar anatomy

Manihot allemii M. J. Silva is described and illustrated as a new species, and its morphological affinities and conservation status is discussed. It is most similar to M. salicifolia Pohl in having a subshrubby and erect habit and in the general aspect of its unlobed leaves, but differs from it in having leaves that are conspicuously petiolate, an inflorescence that is a congested spike‐like thyrse, entire and diminute bracts and bracteoles subtending flowers of both sexes, pistillate calyx that is deeply lobed with oblong lobes, and staminate flowers with 8 or 10 stamens. Both species belong to a group of 14 species recognized by their possession of entire and unlobed leaves, which are the subject of taxonomic studies by the second author. Some notes on the leaf anatomy of species of Manihot that have entire and unlobed leaves are also presented.     

An Assessment of Genetic Diversity Within a Collection of Cassava (Manihot esculenta Crantz) Germplasm Using Molecular Markers

Cassava (Manihot esculenta Crantz) cDNA clones were used todetect restriction fragment length polymorphisms in a collectionof Manihot germplasm maintained as in vitro plants at ORSTOM,Montpellier. The collection consisted of mostly African cultivarsof M. esculenta, together with a few M. glaziovii Mueller vonArgau and M. caerulescens Pohl, and some interspecific hybridsbetween M. esculenta and M. glaziovii. The clones revealed significantlevels of polymorphism both within and between the species;sufficient to construct dendrograms indicating the genetic diversitywithin the collection.Copyright 1993, 1999 Academic Press Cassava, Manihot esculenta (Crantz), restriction fragment length polymorphisms, RFLPs, genetic diversity     

Manihot gratiosa and M. lourdesii spp. nov. (Manihoteae,Euphorbiaceae) from the Brazilian Cerrado

During a botanic expedition in 2013 to study the taxonomy of the genus Manihot in the Cerrado biome, numerous specimens were collected in the Chapada dos Veadeiros region, in central Brazil. After careful analyses, two new species of this genus were discovered. Manihot gratiosa and M. lourdesii are herein described and illustrated. Both species can be distinguished from morphologically similar species by a combination of characters such as shape and overlapping of leaf lobes, indumentum, type and aspect of inflorescence, integrity of pistillate sepals, as well as shape, integrity, and size of bracts of both male and female flowers.     

Evolutionary biogeography of Manihot (Euphorbiaceae), a rapidly radiating Neotropical genus restricted to dry environments

Aim The aims of this study were to reconstruct the phylogeny of Manihot, a Neotropical genus restricted to seasonally dry areas, to yield insight into its biogeographic history, and to identify the closest wild relatives of a widely grown, yet poorly known, crop: cassava (Manihot esculenta). Location Dry and seasonally dry regions of Meso‐ and South America. Methods We collected 101 samples of Manihot, representing 52 species, mostly from herbaria, and two outgroups (Jatropha gossypiifolia and Cnidoscolus urens). More than half of the currently accepted Manihot species were included in our study; our sampling covered the whole native range of the genus, and most of its phenotypic and ecological variation. We reconstructed phylogenetic relationships among Manihot species using sequences for two nuclear genes and a non‐coding chloroplast region. We then reconstructed the history of traits related to growth form, dispersal ecology and regeneration ability. Results Manihot species from Mesoamerica form a grade basal to South American species. The latter species show a strong biogeographic clustering: species from the cerrado form well defined clades, species from the caatinga of north‐eastern Brazil form another, and so do species restricted to forest gaps along the rim of the Amazon basin. Vine and tree growth habits evolved repeatedly in the genus, as did fruit indehiscence and loss of ant‐mediated seed dispersal. Main conclusions The genus Manihot probably originated in Mesoamerica, where it diversified prior to colonizing South America. Within South America, several groups then radiated southwards and eastwards within all kinds of seasonally dry lowland habitats. Some species also adapted to more humid environments, such as forest gaps, around the rim of the Amazon basin. Given the limited dispersal abilities of Manihot species, we propose that this radiation is most likely to have occurred only after, or shortly before, the completion of the Isthmus of Panama, around 3.5 Ma. Our results are in agreement with the past existence of a corridor of dry vegetation through Amazonia or along the eastern South American coast. In addition, our phylogeny allows identification of cassava’s closest wild relatives.     

Attempts to hybridize wildManihot species with cassava

Hybridization between the wildManihot species,anomala Pohl,M. oligantha Pax emend. Nassar subsp.nesteli, M. gracilis Pax andM. zehntneri Ule, and cassava have been tried. Seeds were obtained from all hybrids. Moreover, crosses ofM. oligantha subsp.nesteli and cassava were highly fertile.     

SNPs,SSRs and inferences on cassava’s origin

Despite its importance as a staple food throughout the tropics, the root crop cassava (it Manihot esculenta ssp. esculenta) has traditionally not been a major focus of research. One basic question about cassava that remained unresolved until recently concerns the crop’s origin. This paper describes analyses of SNPs (single nucleotide polymorphisms) and SSR (simple sequence repeat) variation as a means of tracing cassava’s evolutionary and geographical origins. Genetic diversity was examined in a sample of 20 cassava varieties that are representative of germplasm diversity within the crop, and in 212 individuals collected from wild populations of two closely related Manihot species. SNP and indel variation was examined in portions of two low copy nuclear genes, BglA and Hnl. Inferences from these genes were compared to those from previously examined loci, including the low copy nuclear gene G3pdh and 5 SSR loci. For all genes examined, SNPs and SSR alleles are shared between domesticated cassava and a specific geographical subset of wild Manihot populations, which suggests the following: (1) Cassava was likely domesticated from a single wild Manihot species, M. esculenta ssp. flabellifolia, rather than from multiple hybridizing species, as traditionally believed; and (2) the crop most likely originated in the southern Amazon basin.     

Energetics of free existence in swallows and martins (hirundinidae) during breeding: a comparative study using doubly labeled water

Energy metabolism of three sympatric swallows (Hirundinidae) was investigated during the breeding season using doubly labeled water (²H₂ ¹⁸O). Interspecific and intraspecific differences in energy metabolism were examined in relation to the habits, size and environment of the birds. To facilitate comparisons we expressed energy metabolism (M) as the ratio of average daily metabolic rate (ADMR, cm³CO_2g ^-1h^-1) to basal metabolic rate (BMR). We observed adults during incubation and when feeding nestlings. Then, both sexes of Sand Martins Riparia riparia and House Martins Delinchon urbica were either at the nest or on the wing. Incubation reduced activity levels during the day resulting in M (incubation) being 17–26% lower than during rearing. Differences in energy costs for rearing chicks depended mainly on flight behaviour, the smaller Sand Martin doing nearly twice as much flapping during flight as the House Martin, giving higher values for M. In Swallows Hirundo rustica the female incubates alone, alternating between short feeding trips and incubating in daytime. This pattern was linked with a relatively high value for M in the only individual behaving like our controls. Both sexes of Swallows feed the chicks, and they showed similar values of M. They also closely resembled House Martins, despite contrasts in the time spent flying and their behaviour during flight. Feeding conditions affected activity, and thereby M, in a species specific way. The House Martin did more gliding in poor weather, taking less mobile prey, reducing M. Swallows reduced foraging costs further by using body reserves, as in the House Martin. The smaller Sand Martin, in contrast, showed a high expenditure in poor weather. Over two breeding seasons ADMR reached values around 5 BMR for all three species.     

Microsatellites in Cassava (Manihot esculenta Crantz): discovery, inheritance and variability

 Fourteen microsatellites containing GA-repeats were isolated and characterized in cassava (Manihot esculenta Crantz, Euphorbiaceae). Microsatellite heterozygosity (h) was estimated in 48 accessions using (³²P)-end-labeled primers and in more than 500 accessions using fluorescence-based genotyping. Heterozygosity values ranged from 0.00 to 0.88 and the number of alleles detected varied from 1 to 15. The reproducibility of allele sizing was also assessed using fluorescence-based genotyping. The average inter-gel size difference was 1.03 nucleotides. Chi-square tests (χ²) were performed to analyse segregation distortion and the linkage between alleles segregating from either or both parents in an F₁ mapping population. Most microsatellite loci segregated in the expected 1 : 1, 1 : 2 : 1 or 1 : 1 : 1 : 1 ratio. Linkage was detected between loci segregating from either parent, and segregation distortion from the male parent was detected for locus GA-131. Approximately 80% of the microsatellites detected one or two alleles per accession, suggesting a low degree of microsatellite locus duplication, an unexpected finding for a putative allopolyploid, highly heterozygous species. The high h values of most microsatellites, their amplification in other Manihot taxa and their suitability for high-throughput, fluorescence-based genotyping, make microsatellites the marker of choice for germplasm characterization and saturation of the cassava map. Received: 4 September 1997 / Accepted 16 March 1998     

Manihot pachycaulis sp. nov. (Euphorbiaceae) from the Brazilian Cerrado

A new species of Manihot was discovered in the course of revising the taxonomy of some species of the genus occurring in the Midwest Region of Brazil. Its morphology resembles species which belong to different sections of the genus. However, a set of characters related to stem, leaves, inflorescences and flowers serves to differentiate it from the closest species, especially from M. falcata, to which it is most similar. The geographical distribution and conservation status of the new species as well as its flowering and fruiting period are also provided. Additionally, anatomical information about the aerial parts of the stem, a novelty regarding the wild species of the genus, is provided.     

Pollination and breeding system of Melochia tomentosa L. (Malvaceae), a keystone floral resource in the Brazilian Caatinga

The main goals of the present paper were to investigate the floral biology and the breeding system of Melochia tomentosa in a semi-arid region in Brazil, comparing the role of Apis mellifera with other native pollinators, and to discuss the importance of this plant species as a floral resource for the local fauna in maintaining different guilds of specialized pollinators in the Caatinga. M. tomentosa is very common in Caatinga areas and blooms year-round with two flowering peaks, one in the wet and another in the dry period. The pink, bright-colored flowers are distylous and both morphs are homogamous. The trichomatic nectary is located on the inner surface of the connate sepals, and the nectar (ca. 7 μl) is accumulated in the space between the corolla and the calyx. Nectar sugar concentration reaches an average of 28%. The results of controlled pollination experiments show that M. tomentosa is self-incompatible. Pollen viability varies from 94% to 98%. In spite of being visited by several pollen vectors, flower attributes of M. tomentosa point to melittophily, and A. mellifera was the most frequent visitor and the principal pollinator. Although honeybees are exotic, severely competing with native pollinators, they are important together with other native bees, like Centris and Xylocopa species, for the fruit set of M. tomentosa. This species represents a keystone floral resource favoring the maintenance of many species of bees, butterflies and hummingbirds through the year in this deciduous tropical dry forest.     

Variability of chloroplast DNA and nuclear ribosomal DNA in cassava (Manihot esculenta Crantz) and its wild relatives

Chloroplast DNA (cp) and nuclear ribosomal DNA (rDNA) variation was investigated in 45 accessions of cultivated and wild Manihot species. Ten independent mutations, 8 point mutations and 2 length mutations were identified, using eight restriction enzymes and 12 heterologous cpDNA probes from mungbean. Restriction fragment length polymorphism analysis defined nine distinct chloroplast types, three of which were found among the cultivated accessions and six among the wild species. Cladistic analysis of the cpDNA data using parsimony yielded a hypothetical phylogeny of lineages among the cpDNAs of cassava and its wild relatives that is congruent with morphological evolutionary differentiation in the genus. The results of our survey of cpDNA, together with rDNA restriction site change at the intergenic spacer region and rDNA repeat unit length variation (using rDNA cloned fragments from taro as probe), suggest that cassava might have arisen from the domestication of wild tuberous accessions of some Manihot species, followed by intensive selection. M. esculenta subspp flabellifolia is probably a wild progenitor. Introgressive hybridization with wild forms and pressures to adapt to the widely varying climates and topography in which cassava is found might have enhanced the crop's present day variability.     

Microstructural morphology in early dermal denticles of hammerhead sharks (Elasmobranchii: Sphyrnidae) and related taxa

Mello, W.C., de Carvalho, J.J., Brito, P.M.M. 2011. Microstructural morphology in early dermal denticles of hammerhead sharks (Elasmobranchii: Sphyrnidae) and related taxa. —Acta Zoologica (Stockholm) 00 : 1–7. This study uses scanning electron microscopies to investigate and describe the microstructural diversity of dermal denticles in the family Sphyrnidae, which comprises all living hammerhead shark species, comparing them to other related taxa (i.e. Carcharhinus dussumieri, Carcharhinus plumbeus, Carcharhinus acronotus, Rhizoprionodon acutus, Negaprion brevirostris and Hemigaleus microstoma). The results reveal that sphyrnids present noticeable microstructures in the dermal denticles, distinguishing them from the other related species investigated. Additionally, scale patterns are the same in three distinct body regions (i.e. cephalic, branchial and dorsal fin). Species of Sphyrnidae that reach bigger total lengths and that are widely distributed (i.e. Sphyrna lewini and Sphyrna mokarran) presented more, smaller and nearly hexagonal microstructures that do not cover the entire scale surface, unlike species reaching smaller sizes and restricted to coastal habits (i.e. Sphyrna tiburo, Sphyrna tudes, Sphyrna media and Eusphyra blochii). The sphyrnid scales are similar to R. acutus and C. dussumieri rather than to the other species, but it is not possible to identify the sphyrnid species only by scale features. It is clear that a similar morphology of scales is not necessarily related to similar life habits, and that they are candidates to provide new characters in phylogenetical studies among sphyrnids.     

Germination of Prosopis juliflora (Sw) DC seeds after scarification treatments

Invasive plant species are the second most important threat to global biodiversity loss after land‐use change. Invasive species can modify native community composition, deplete species diversity and affect ecosystem processes. The Caatinga is one of the most human‐affected Brazilian ecosystems owing to non‐sustainable use of its natural resources. Prosopis juliflora is an important invasive plant species in the Caatinga ecosystem. Seed germination is a critical stage in plant life cycles and is a major factor in the establishment and success of invasive plant species. Among the factors that affect seed germination and dormancy, coat‐imposed seems to be the most important for P. juliflora. In Prosopis species, the ingestion of fruits by wild and domestic animals may promote and accelerate germination, enhancing the dispersal of seeds and fruits of these species. We investigated the germination capacity of P. juliflora seeds after artificial mechanical and chemical scarification and analyzed the changes in seedling vigor caused by the scarification treatments. Germination rate, germination time (TMG) and germination synchrony (E) differed significantly with the length of the scarification treatments in H₂SO₄ for both seeds with endocarps and seeds without endocarps (non‐endocarp seeds). Sulfuric acid affected plant survival more strongly than germination rate, particularly in non‐endocarp seeds.     

MANTONIELLA IN ANTARCTIC WATERS INCLUDING THE DESCRIPTION OF M. ANTARCTICA SP. NOV. (PRASINOPHYCEAE)1

We report the finding of two species of Mantoniella from the Weddell Sen and Prydz Bay, Antarctica. As well as M. squamata, a cosmopolitan species, we describe M. antarctica sp. nov., the second species in this genus. M. antarctica is covered with two distinctive types of discoid scales. The flagella bear only the smaller of the scale types whereas the cell body has both types, the larger overlying the smaller.     

Pollination and seed dispersal of Melocactus ernestii Vaupel subsp. ernestii (Cactaceae) by lizards: an example of double mutualism

Recent studies show that the mutualistic role of lizards as pollinators and seed dispersers has been underestimated, with several ecological factors promoting such plant–animal interactions, especially on oceanic islands. Our aim is to provide a quantitative assessment of pollination and seed dispersal mutualisms with lizards in continental xeric habitats. We carried out focal observations of natural populations of Melocactus ernestii (Cactaceae) in the Caatinga, a Brazilian semiarid ecosystem, in order to record the frequency of visits, kind of resource searched and behaviour of visiting animals towards flowers and/or fruits. We made a new record of the lizard Tropidurus semitaeniatus foraging on flowers and fruits of M. ernestii. During the search for nectar, T. semitaeniatus contacted the reproductive structures of the flowers and transported pollen attached to its snout. Nectar production started at 14:00 h, with an average volume of 24.4 μl and an average concentration of solutes of 33%. Approximately 80% of the seeds of M. ernestii found in the faeces of T. semitaeniatus germinated under natural conditions. The roles of T. semitaeniatus as pollinator and seed disperser for M. ernestii show a clear relationship of double mutualism between two endemic species, which may result from the environmental conditions to which both species are subject. Seasonality, low water availability and arthropod supply in the environment, high local lizard densities, continuous nectar production by the flower and fruits with juicy pulp may be influencing the visits and, consequently, pollination and seed dispersal by lizards in this cactus.     

Speciation and gene flow in two sympatric small mammals from Madagascar,Microgale fotsifotsy and M. soricoides (Mammalia: Tenrecidae)

Madagascar's shrew tenrecs (Mammalia: Tenrecidae; Microgale, Nesogale) represent an excellent system for studying speciation. Most species are endemic to the island's eastern humid forests, a region renowned for high levels of biodiversity and a high rate of in situ diversification. We set out to understand the speciation dynamics in a clade of recently described taxa: Microgale fotsifotsy and M. soricoides, which have nearly identical distributions in the moist evergreen forest, and M. nasoloi, which occurs in the western dry deciduous forest. A phylogenetic analysis using mitochondrial DNA data recovered two distinct clades of M. fotsifotsy: a south clade that is sister to, and broadly sympatric with, M. soricoides, and a north clade that is sister to the dry‐forest and distantly allopatric species M. nasoloi. To better understand this result, we analysed cranioskeletal measurements and performed demographic analyses using nuclear sequence data from ultraconserved elements. Nuclear data did not support a sister relationship between M. soricoides and the south clade of M. fotsifotsy but did demonstrate introgression between these clades, which probably explains the discordance between nuclear and mitochondrial phylogenies. Demographic analyses also revealed the absence of gene flow between the north and south clades of M. fotsifotsy. Morphometric data revealed several major differences between M. soricoides and M. fotsifotsy, as well as more subtle differences between the two clades of M. fotsifotsy. In light of these results, we treat the south clade of M. fotsifotsy as a new candidate species. Our findings demonstrate the utility of integrating multiple data types to understand complex speciation histories, and contribute to a growing body of evidence that species diversity on Madagascar is underestimated.     

Impact of orphaning on field colonies of Southeast Asian Macrotermes gilvus (Hagen) and M. carbonarius (Hagen) (Termitidae, Macrotermitinae)

Field colonies of Macrotermes gilvus (Hagen) and M. carbonarius (Hagen) were experimentally orphaned to examine their potential for producing replacement reproductives. Orphaned colonies were investigated only once for caste composition at selected time intervals at 3, 6, 9 or 12 months after orphaning. Of the 38 orphaned colonies of M. gilvus, 15 colonies (39.5%) re-established. In M. carbonarius, three colonies out of 20 (15%) re-established. Re-established colonies were headed by adultoids which were morphologically indistinguishable from primary reproductives. In naturally orphaned colonies of M. gilvus, we often found multiple adultoids with normal pigmentation but torn wings, i.e. the colonies retained alates as replacement reproductives. The number of reproductives probably declines over time. It may take alates of M. gilvus 6 months to develop into functional adultoids, and up to 12 months for alates of M. carbonarius. Our results also demonstrate that the presence of sexual castes (nymphs or alates) at the time of orphaning does not necessarily guarantee the success of colonies in re-establishing themselves as breeding colonies. We also found a high prevalence of occupation of the mounds by other termite species, after the death of M. gilvus (18.4%) or M. carbonarius (30.0%) colonies, probably using them as foraging sites.