Stomatal size,ploidy and polyembryony in Eriotheca Stellate Trichome Species Complex (Bombacoideae – Malvaceae) in the Cerrados of Brazil

• Geographic parthenogenesis, range expansion of apomictic plants after climate changes, has been described for Northern Hemisphere gametophytic apomicts. But similar trends have been observed for sporophytic apomicts of Cerrado, the savannas in Brazil. Eriotheca pubescens is a common Cerrado tree, an agamic complex of either hexaploid/polyembryonic apomicts or tetraploid/monoembryonic sexual individuals. Some populations have been described as a new species, Eriotheca estevesiae, all included in the Eriotheca Stellate Trichome Species Complex (ESTSC). Since breeding systems and ploidy are clearly associated with polyembryony and stomatal size, we used these ancillary features to map the reproductive and ploidy level traits of E. pubescens and E. estevesiae.
• Leaves and seeds were collected from individuals of 19 populations. Seeds were evaluated for the presence of polyembryony and leaves for stomatal measurements.
• Eight populations were monoembryonic while another eight were polyembryonic and for other three, the embryonic pattern was not readily verified. E. pubescens polyembryonic and hexaploid populations formed a homogeneous group, but monoembryonic plants were more variable. E. estevesiae populations were monoembryonic with smaller stomata. In contrast, some E. pubescens monoembryonic populations further south presented larger stomata. Despite these outliers, possibly mixed populations, stomatal size and embryonic pattern differed from northern to southern populations.
• Embryonic pattern and stomatal size indicated that northernmost populations of Eriotheca STSC (E. estevesiae) are diploid and sexual. Southernmost populations, mostly polyembryonic and with large stomata, are hexaploid and apomictic. This is in agreement with geographic parthenogenesis and range expansion of apomictic lineages to southern habitats available after the last glacial maximum.

     

Pollen and stomata morphometrics and polyploidy in Eriotheca (Malvaceae‐Bombacoideae)

Approximately 70% of the angiosperm species are polyploid, an important phenomenon in the evolution of those plants. But ploidy estimates have often been hindered because of the small size and large number of chromosomes in many tropical groups. Since polyploidy affects cell size, morphometric analyses of pollen grains and stomata have been used to infer ploidy level. Polyploidy is present in many species of the Cerrado, the Neotropical savanna region in Central Brazil, and has been linked to apomixis in some taxa. Eriotheca gracilipes and Eriotheca pubescens are common tree species in this region, and present cytotypes that form reproductive mosaics. Hexaploid individuals (2n = 6x = 276) are polyembryonic and apomictic, while tetraploid and diploid individuals (2n = 2x = 92, 2n = 4x = 184) are sexual and monoembryonic. We tested whether morphometric analysis can be used to estimate ploidy levels in E. gracilipes and E. pubescens individuals. Pollen material from diploid and hexaploid individuals of E. gracilipes, and tetraploid and hexaploid individuals of E. pubescens, were fixed in 50% FAA, and expanded leaves were dried in silica gel. Pollen grains and stomata of at least five individuals from each population were measured. The results demonstrate that all measures were significantly different among cytotypes. Individuals with higher levels of ploidy (hexaploid) all presented measurements that were higher than those with lower levels (diploid and tetraploid). There was no overlap between ploidy levels in each species at 95% confidence interval. Thus, the size of the pollen grains and stomata are effective parameters for analysis of ploidy levels in E. gracilipes and E. pubescens.     

RAPD Evidence for Apomixis and Clonal Populations in Eriotheca (Bombacaceae)

Abstract: Two woody species of Eriotheca (Bombacaceae) of the Central Brazilian Cerrados were submitted to RAPD analyses. Both species are bee pollinated and have a similar flower structure, but E. pubescens presents adventitious embryony and apomixis, while E. gracilipes is self-incompatible. The RAPD screening reflects these differences in breeding systems, with very low genetic variation in the apomictic species, while the sexual species presented much higher variability with no similar genotypes among the sampled trees. The results suggest that adventitious embryony in E. pubescens effectively results in clonal populations or population mosaics of clonal individuals. Since recent studies have indicated poly-embryony and possibly apomixis in a number of Cerrado woody species otherwise considered obligatorily allogamous, the RAPD results presented here indicate the technique will be a useful tool to detect clonal populations of apomictic origin among Cerrado woody species with mixed mating systems and will help to assess the importance of apomixis as a breeding system for the Cerrado flora.     

无融合生殖与柑橘多胚现象的研究进展

植物无融合生殖是指植物的胚珠组织不经历正常的减数分裂和受精作用,而直接进行胚发育形成种子的无性生殖方式。无融合生殖植物完全继承了母体的全部基因型,因而具有独特研究与育种意义。芸香科柑橘属植物具有独特的多胚现象,其珠心组织能够发育成不定胚(称为珠心胚)进行无融合生殖。文中介绍了柑橘类植物的珠心胚生殖现象、细胞学和遗传学研究进展。珠心胚现象虽然对柑橘杂交后代获得有较大影响,但在生产上可产生性状整齐一致的后代,可以培育无病毒苗木。     

Nuclear microsatellite variation in Malagasy baobabs (Adansonia,Bombacoideae, Malvaceae) reveals past hybridization and introgression

Background and Aims Adansonia

comprises nine species, six of which are endemic to Madagascar. Genetic relationships between the Malagasy species remain unresolved due to conflicting results between nuclear and plastid DNA variation. Morphologically intermediate individuals between distinct species have been identified, indicative of interspecific hybridization. In this paper, microsatellite data are used to identify potential cases of hybridization and to provide insights into the evolutionary history of the genus on Madagascar.

Methods

Eleven microsatellites amplified with new primers developed for Adansonia rubrostipa were used to analyse 672 individuals collected at 27 sites for the six Malagasy species and morphologically intermediate individuals. Rates of individual admixture were examined using three Bayesian clustering programs, STRUCTURE, BAPS and NewHybrids, with no a priori species assignment.

Key Results

Population differentiation was coherent, with recognized species boundaries. In the four Malagasy species of section Longitubae, 8·0, 9·0 and 9·5 % of individuals with mixed genotypes were identified by BAPS, NewHybrids and STRUCTURE, respectively. At sites with sympatric populations of A. rubrostipa and A. za, NewHybrids indicated these individuals to be F₂ and, predominantly, backcrosses with both parental species. In northern Madagascar, two populations of trees combining A. za and A. perrieri morphology and microsatellite alleles were identified in the current absence of the parental species.

Conclusions

The clear genetic differentiation observed between the six species may reflect their adaptation to different assortments of climate regimes and habitats during the colonization of the island. Microsatellite variation reveals that hybridization probably occurred in secondary contact between species of section Longitubae. This type of hybridization may also have been involved in the differentiation of a local new stabilized entity showing specific microsatellite alleles and morphological characters, suggesting a potential role of hybridization in the recent history of diversification on Madagascar.     

State-of-knowledge on Astronium fraxinifolium Schott (Anacardiaceae) for genetic conservation in Brazil

This study explores the basis for conservation action on Astronium fraxinifolium Schott (Anacardiaceae). This is a wide-ranging forest species occurring in Brazil and other South American countries, and typical of gallery forest along watercourses in the Cerrado region. Information about this species is scanty and scattered. This paper aims to provide a structured review of available knowledge of its biology, ecology, silviculture and management. Widely-scattered published reports have been critically considered and efforts made to highlight and resolve contradictions and inconsistencies. Because little effort has been applied to its domestication and improvement, knowledge of silviculture and management of Astronium fraxinifolium is particularly meagre. Gaps in current knowledge relevant for conservation are identified and steps to fill them proposed. Where in situ conservation was considered appropriate, recommendations are made for the location of additional protected areas. Complementary ex situ and enrichment conservation action is suggested for specific parts of the range where resource losses are already so extensive that in situ measures alone are insufficient. Provision for refining the limited management and conservation knowledge is made through highlighting priorities for study of the taxon. Finally, future action is discussed in the context of the infrastructure of the national conservation sector of Brazil.     

Comparative cytomolecular analyses reveal karyotype variability related to biogeographic and species richness patterns in Bombacoideae (Malvaceae)

Bombacoideae is one out of nine subfamilies of Malvaceae and encompasses 160 tree species. The subfamily is karyotypically characterized by small and numerous chromosomes and is traditionally known by a remarkable inter- and intraspecific chromosome number variation. We conducted a comparative cytogenetic analysis to investigate karyotype diversity and chromosome evolution within Bombacoideae. To achieve this, we performed new chromosome counts, CMA/DAPI double staining, genome size estimations, and localization of 5S and 45S rDNA by fluorescence in situ hybridization for 21 species distributed across the Bombacoideae phylogeny. We performed ancestral states reconstruction analyses to elucidate chromosome evolution and provide insights into the systematics and evolution of Bombacoideae in comparison with other Malvaceae species. Newly generated data on chromosome number on Bombacoideae revealed diploids (Ochroma (2n = 84), Cavanillesia, Pochota, Pseudobombax (2n = 88), and Pachira (2n = 92)) and polyploids (Adansonia digitata (2n = 160) and Eriotheca species (2n = ca. 194 and 2n = 276)). For most species, in situ hybridization revealed karyotype, with two pairs of 45S rDNA sites co-located with CMA⁺ bands, and 5S rDNA sites in only one chromosome pair. Taken together, our results provide support to the hypothesis of karyotypic stability in Bombacoideae. Only the Pachira s.l. clade displayed some variability in ploidy level, number of CMA⁺ bands and 45S rDNA sites, and genome size compared to other Bombacoideae clades. The Striated bark clade was characterized by comparatively small genomes and low cytomolecular variability. Karyotypic data were related to biogeographic and species richness patterns of Bombacoideae.     

Preliminary Information on the Home Range and Movement Patterns of Sturnira lilium (Phyllostomidae) in a Naturally Fragmented Landscape in Bolivia

Here, we used radiotelemetry to determine home range and movement patterns of the frugivorous bat Sturnira lilium in a naturally fragmented landscape in Beni, Bolivia. Four females were monitored for up to 7 d. Based on 575 radiotelemetry positions, kernel home ranges were between 36.5 and 90.7 ha. During each night, bats visited most of their home range, which included forest islands and continuous forest; however activity was concentrated in core-use areas representing 6–12 percent of the home ranges. There was moderate overlap in home range and core-use areas among the four females. The mean long axis across the kernel home ranges was 1324 m. The results reported here show that S. lilium is able to move freely among forest elements in the landscape while crossing a savanna matrix. Hence, S. lilium may contribute to maintain connectivity and ecological processes in a fragmented landscape.     

Sex and Season Affect Individual-Level Diet Variation in the Neotropical Marsupial Gracilinanus microtarsus (Didelphidae)

Individual-level diet variation was investigated in Gracilinanus microtarsus , an insectivorous marsupial whose diet is affected by sex and season in the highly seasonal Brazilian Cerrado. To measure individual-level diet variation, the diets of individual males and females were compared to that of their population in the warm-wet and cool-dry seasons using the proportional similarity index (PS _i ). This index varies from 1 (complete overlap between the individual i  's and population's diets) toward 0 (decreasing overlap). Mean PS _i values were computed as a measure of the degree of interindividual diet variation (the larger the mean PS _i value, the smaller the variation among individuals' diets). Interindividual diet variation among females was similar between seasons, whereas among males it decreased from the warm-wet to the cool-dry season. Diet variation among males is probably reduced in the cool-dry season because of constraints on food consumption generated by interactions between endogenous (physiological needs associated with high rates of body mass growth) and exogenous (food limitation) factors.     

Circumscription of Malvaceae (Malvales) as determined by a preliminary cladistic analysis of morphological, anatomical, palynological, and chemical characters

We report a phylogenetic analysis of “core” Malvales (Tiliaceae, Sterculiaceae, Bombacaceae, and Malvaceae) based on morphological, anatomical, palynological, and chemical features. The results of the analyses lead to the conclusion that Tiliaceae, Sterculiaceae, and Bombacaceae, as variously delimited, are paraphyletic; only the Malvaceae are likely monophyletic. The genera of “core” Malvales form a well-defined clade. Genera of “Tiliaceae” constitute the basal complex within “core” Malvales. The “Sterculiaceae” (most genera)+ “Bombacaceae” + Malvaceae form a clade on the basis of a monadelphous androecium; “Bombacaceae”+ Malvaceae also form a clade, which is diagnosable on the basis of monoloculate anthers. It is clear that the traditional classification, with its arbitrarily delimited evolutionary grades, is unsatisfactory, especially if one seeks to reflect phylogeny accurately. Thus, Malvaceae is redefined to refer to the most recent common ancestor of plants previously considered to be “Tiliaceae,” “Sterculiaceae,” “Bombacaceae,” and Malvaceae, and all of the descendants of that ancestor. This broadly circumscribed Malvaceae can be diagnosed by several presumed synapomorphies, but we draw special attention to the unusual floral nectaries that are composed of densely packed, multicellular, glandular hairs on the sepals (or less commonly on the petals or androgynophore).     

State-of-knowledge on Myracrodruon urundeuva Fr. Allemão (Anacardiaceae) for genetic conservation in Brazil

This study explores the basis for conservation action on Myracrodruon urundeuva Fr. Allemão (Anacardiaceae). This is a wide-ranging forest species occurring in Brazil and other South American countries, notably in the Cerrado region. This paper aims to provide a structured review of available knowledge of its biology, ecology, silviculture and management. Widely-scattered published reports have been critically considered and efforts made to highlight and resolve contradictions and inconsistencies. Information about this species is scanty and scattered, particularly in respect to its biology and ecology. Some effort has been applied to its domestication and improvement, but opinion on its silviculture and management is still controversial. The taxon is typical of open and deciduous forest in the Cerrado. Gaps in current knowledge relevant for conservation are identified and steps to fill them proposed. Where in situ conservation proved to be more appropriate, recommendations are made for the location of additional protected areas. Complementary ex situ and enrichment conservation actions are suggested for specific parts of the range where resource losses are already so extensive that in situ measures alone are inadvisable. Opportunities for refining the limited management and conservation knowledge are proposed by highlighting priorities for study of the taxon. Finally, future action is discussed in the context of the infrastructure of the national conservation sector.     

Sacred Giants: Depiction of Bombacoideae on Maya Ceramics in Mexico,Guatemala, and Belize

Sacred Giants: Depiction of the Malvaceae Subfamily Bombacoideae on Maya Ceramics in Mexico, Guatemala, and Belize This study categorized and identified plants depicted on Maya ceramics from the Classic Period (250 a.d.–900 a.d.). We chose art objects with a predominance of iconographic images of Malvaceae subfamily Bombacoideae, which are easily identified morphologically and have culinary, medicinal, ceremonial, economic, and cosmological significance to the Maya. Among ten species of Bombacoideae native to the Southern Lowlands region of Central America (Belize, parts of Guatemala, and Mexico), the Maya utilized at least six, which also have Maya names. We observed four or five bombacoid species depicted on Maya ceramics; most images were identifiable to genus. Burial urns and incensarios (incense burners) commonly had images of trunk spines of Ceiba pentandra, the Maya “World Tree.” Flowers of Pseudobombax ellipticum, a plant used to make ceremonial beverages, were most similar to floral images portrayed on vessels, bowls, and plates, although the morphologically similar flowers of Pachira aquatica may also be depicted. Plants representing Quararibea funebris or Q. guatemalteca, which were used during preparation of cacao beverages, were discernable on drinking vessels.     

Phylogenetic relationships of Malvatheca (Bombacoideae and Malvoideae; Malvaceae sensu lato) as inferred from plastid DNA sequences

Previous molecular phylogenetic analyses have revealed that elements of the former families Malvaceae sensu stricto and Bombacaceae together form a well-supported clade that has been named Malvatheca. Within Malvatheca, two major lineages have been observed; one, Bombacoideae, corresponds approximately to the palmate-leaved Bombacaceae, and the other, Malvoideae, includes the traditional Malvaceae (the mallows or Eumalvoideae). However, the composition of these two groups and their relationships to other elements of Malvatheca remain a source of uncertainty. Sequence data from two plastid regions, ndhF and trnK/matK, from 34 exemplars of Malvatheca and six outgroups were analyzed. Parsimony, likelihood, and Bayesian analyses of the sequence data provided a well-resolved phylogeny except that relationships among five lineages at the base of Malvatheca are poorly resolved. Nonetheless, a 6-bp insertion in matK suggests that Fremontodendreae is sister to the remainder of Malvatheca. Our results suggest that the Malvoideae originated in the Neotropics and that a mangrove taxon dispersed across the Pacific from South America to Australasia and later radiated out of Australasia to give rise to the ca. 1700 living species of Eumalvoideae. Local clock analyses imply that the plastid genome underwent accelerated molecular evolution coincident with the dispersal out of the Americas and again with the radiation into the three major clades of Eumalvoideae.     

State-of-knowledge on Amburana cearensis (Fr. Allem.) A.C. Smith (Leguminosae: Papilionoideae) for genetic conservation in Brazil

This study explores the basis for conservation action on Amburana cearensis (Fr. Allem.) A.C. Smith (Leguminosae: Papilionoideae), a wide-ranging forest species occurring in Brazil, notably in the Caatinga and Cerrado regions, and other South American countries. It offers insight into the taxon through structured reviews of knowledge of its biology, ecology, silviculture and management. Widely scattered published reports are critically considered and efforts made to highlight and resolve contradictions and inconsistencies therein. Information about this species is scanty and scattered, particularly in respect to its biology and ecology. Because little effort has been applied to its domestication and improvement, knowledge of its silviculture and management is meagre. The taxon is typical of open and deciduous forest in the Caatinga and Cerrado. Gaps in current knowledge relevant for conservation are identified and steps to fill them proposed. Where in situ conservation proved to be more appropriate, recommendations are made for the location of additional protected areas. Ex situ and enrichment conservation action is highly recommended for specific parts of the range where resource losses are already so extensive that in situ measures alone are inadvisable. Provision for refining the limited management and conservation knowledge is made through highlighting priorities for study of the taxon. Finally, future action is discussed in the context of the infrastructure of the national conservation sector.     

Genetic analysis of apomixis in Citrus and Poncirus by molecular markers

Propagation of citrus rootstocks depends upon the production of clonal plants from nucellar seedlings. This makes apomixis one of the host important traits in breeding programs for citrus rootstocks. The genetic control of apomixis was studied in a 50-tree progeny derived from the cross C. volkameriana×P. trifoliata using 69 molecular markers and bulked segregant analysis. The proportion of nucellar seedlings was estimated by isoenzymatic analysis of 25 seedlings per tree for 2 consecutive years. The type of embryony (polyembryonic versus monoembryonic seeds) was also determined for fruit-yielding trees. Separate genetic maps for each parental species were developed. The integration and comparison of these maps could be accomplished using common multiallelic segregant loci. Differences in gene synteny between the two species-specific genetic maps were shown. Important distortions in the segregation of markers at several genomic regions, some of them also involving differences in the C-methylation pattern, have been observed, especially for the pollen parent. Analysis of quantitative trait loci (QTLs) revealed the presence of six genomic positions (two in P. trifoliata and four in C. volkameriana) contributing individually up to 24% of the total variation for apomixis. Within the same species, QTLs with positive and negative allele effects were present, even in the same linkage group. One of the markers associated to apomixis (Apo2) is also associated to embryony type. Therefore, the genetic control of apomictic reproduction found in citrus (nucellar embryony) is quite complex compared to what has been reported for gametophytic apomixis. Molecular markers linked to QTLs governing apomixis will be useful to assist selection of future apomictic rootstocks for citrus varieties. Received: 30 November 1998 / Accepted: 28 December 1998     

The best of both worlds: Apomixis and sexuality co-occur in species of Microlicia,Melastomataceae

Apomixis, the asexual formation of embryos and seeds, occurs in approximately 18% of angiosperm families. Melastomataceae exhibits a remarkable number of apomictic species, distributed among different tribes. This mode of reproduction has been elucidated in Miconieae, but remains unclarified for other groups, such as Microlicieae. Although apomixis has been previously described for Microlicieae species, the cytological basis for this phenomenon is entirely unknown in this group. Thus, populations of Microlicia fasciculata and M. polystemma were used in order to (a) investigate the presence of autonomous apomixis; (b) verify if this mode of reproduction leads to polyembryony; and (c) investigate whether apomixis may occur in parallel with the sexual process. We tested these species for autonomous fruit set and polyembryony, and pollen viability, and analyzed pollen tube growth. Anatomical techniques were used to elucidate the micro- and megasporogenesis and gametogenesis. The species showed autonomous fruit and seed formation and exhibited polyembryony. Apospory and adventitious embryony were the developmental mechanisms of apomixis in M. fasciculata and M. polystemma, respectively. Both species exhibited low pollen viability. However, some viable pollen, reduced embryo sac formation, natural pollination and pollen tube growth enable sexual reproduction and characterize these species as facultative apomicts. The independence of pollinators for fruit set, uniparental reproduction and the possibility of sexual reproduction, confer reproductive assurance and flexibility, bringing together advantages of sexual and asexual reproduction. In this sense, apomixis may have played an important role in the evolution and diversification of Microlicia, a widely distributed genus in the Brazilian Cerrado.     

A hawkmoth crossroads? Species richness,seasonality and biogeographical affinities of Sphingidae in a Brazilian Cerrado

Aim The aim of the study was (1) to describe the biodiversity of the sphingid assemblage in a Cerrado area in the Triângulo Mineiro region, south‐east of Brazil; (2) to evaluate the seasonal variations in species composition; (3) to compare the faunistic relationships between the Cerrado biome and adjacent ecosystems; and (4) to analyse the biogeographical pattern of species distribution in the Neotropical region in a historical context. Location Panga Ecological Station (PES), 30 km south of the city of Uberlândia, and other areas of the Triângulo Mineiro region, Minas Gerais state, south‐eastern Brazil. Methods Moth richness and abundance were monitored monthly at the PES from August 2003 to July 2004, with additional collections at this locality in 2001/2002, 2005 and 2006. Complementary moth richness and abundance data were also collected in other areas of the Triângulo Mineiro region. All collections were made using light traps, and the hawkmoths were mounted and identified. Cluster analysis, rarefaction curves and estimators of total species richness were used to compare the Cerrado hawkmoth assemblage with assemblages derived from other surveys in the Neotropics. Results In total, 61 hawkmoth species were recorded for the study region and their occurrence was markedly seasonal. The hawkmoth assemblage in the study area presented the closest similarity with rain forest areas and with a tropical dry forest area in Central America. The area shared species with both rain forest and seasonally dry tropical forest (SDTF) ecosystems, including supposedly endemic species previously recorded only in the latter areas. Rarefaction curves and estimators of the total number of species showed species richness to be comparable with other highly diverse forest areas in the Neotropics, such as the Brazilian Amazon and Costa Rica. Main conclusions This short‐term study is the first systematic survey of hawkmoths in the Cerrado. It has recorded around 22% of the South American fauna and highlighted the high species richness of the region, which compares favourably with that in other rain forest ecosystems. The survey indicates high regional diversity, and has shown that the Cerrado harbours a hawkmoth fauna comprising both rain forest elements, probably distributed along humid gallery forest corridors, and SDTF elements, supporting the idea of a historical Pleistocene arc connecting the Caatinga domain and other seasonal dry forest areas across the Cerrado region.     

Structure, development and evolution of the androecium in Adansonieae (core Bombacoideae, Malvaceae s.l.)

Androecium development and vasculature were studied in nine species of the Adansonieae clade (core Bombacoideae, Malvaceae s.l.). In early androecium development either distinct pentagonal androecial ring walls or five common petal/androecium primordia are present. Ring walls give rise to five antepetalous and five alternipetalous primary androecial primordia. Common primordia divide into peripheral petal primordia and antepetalous primary androecial primordia. Antepetalous primary androecial primordia split anticlinally into ten primordia-halves, on which secondary androecial primordia are initiated in a centrifugal succession. Androecial lobes are formed by fusion of an alternipetalous primary androecial primordium and its two neighbouring antepetalous primary primordia-halves, a pattern that also occurs in other Malvatheca. Later, tertiary androecial primordia are formed by the subdivision of secondary androecial primordia (except in Adansonia and Ceiba). Each tertiary primordium differentiates into a two-locular androecial unit. At anthesis these two-locular androecial units are often present in pairs, corresponding to the two halves of the same secondary androecial primordium. Androecium development and vasculature imply that the alternipetalous androecial sectors have been reduced in Bombacoideae, a tendency that is shared with other subfamilies of Malvaceae.