Taxonomic treatments of Camellia (Theaceae) species with secretory structures based on integrated leaf characters

Multivariate analysis of leaf shape, anatomy, and Fourier-transform infrared (FTIR) data of 27 Camellia species with secretory structures (sects. Archecamellia, Stereocarpus, Furfuracea, Chrysantha), together with three species from related genera, Gordonia and Tutcheria (Theacea), was conducted to clarify some taxonomic problems. Our results show that crystals occurring in adaxial epidermal cells are firstly observed in Chrysantha species, and the secretory structures described are in fact cork warts. Furthermore, we introduce a form coefficient (F _c) to assess the shape of epidermal cells, since they are usually irregular and difficult to describe. Pearson correlation analysis indicates that F _c is useful to assess epidermal cell shape. Principal component analysis (PCA) of leaf shape indicates that two species from section Archecamellia and two species from section Stereocarpus are significantly different from those in section Furfuracea. Cluster analysis of FTIR data visualizes the degree of affinity among the 30 species examined here, which is consistent with the cluster analysis (CA) of anatomical data, as illustrated in the dendrogram. Therefore, our study indicates that integrated leaf characters based on leaf shape, anatomy, and FTIR data are useful in the taxonomic treatment of Camellia species with secretory structures. Taxonomic controversies among the Camellia species with secretory structures could thus be successfully addressed using only a few intact or small portions of leaves. Moreover, our results tend to support that Chrysantha species should not be merged into section Archecamellia, and that section Heterogenea should not be recognized in taxonomic treatments of Camellia species with secretory structures.     

Leaf architecture of the genus Didymaea Hook. f. (Rubiaceae)

We examined 532 specimens representing eight species of Didymaea (Rubiaceae) to describe their leaf architecture. Characters observed in the herbarium specimens and cleared leaves were leaf base, apex, and margin; trichome type and distribution; vein orders from primary to higher order veins, veinlets, and ultimate marginal vein. Six morphometric variables were analyzed using variance and principal component analysis to identify which characters are useful to distinguish species. Most of the species present brochidodromous venation pattern, except D. hispidula, which has eucamptodromous venation pattern; Didymaea floribunda is unique in having cylindrical and bicellular trichomes. Principal component analysis revealed that 78% of the total variation is mainly related to leaf maximum width. Variance analysis showed significant differences between D. microflosculosa and the other species. Architectural features were useful for differentiating all the species, and an identification key for the species of Didymaea based only on leaf characters is presented.     

Leaf surfaces as a taxonomic tool: the case of Carex section Phacocystis (Cyperaceae) in the British Isles

An investigation into the potential taxonomic value of leaf surface characteristics using large sample size and multivariate analysis was undertaken in Carex section Phacocystis. A case study examined micromorphological characters of the leaf surface for six members of the section in Britain. The six split into three groups based on distribution of stomata: Carex recta amphistomous, C. aquatilis/C. nigra epistomous, and C. bigelowii/C. elata/C. acuta hypostomous. There was considerable variation within samples in stomatal density and cell dimensions. Discriminant Analysis of nine quantitative characters clearly separated four species and a cluster of two species, C. elata and C. acuta. Multivariate analysis indicated that the intercostal cell lengths from both leaf surfaces, abaxial costal cell lengths, and widths of the adaxial costal and abaxial intercostal cells were taxonomically useful in this group and potentially of use within the genus.     

Phylogenetic relationships and infrageneric classification of Epidendrum subgenus Amphiglottium (Laeliinae, Orchidaceae)

Epidendrum L. is the largest genus of Orchidaceae in the Neotropical region; it has an impressive morphological diversification, which imposes difficulties in delimitation of both infrageneric and interspecific boundaries. In this study, we review infrageneric boundaries within the subgenus Amphiglottium and try to contribute to the understanding of morphological diversification and taxa delimitation within this group. We tested the monophyly of the subgenus Amphiglottium sect. Amphiglottium, expanding previous phylogenetic investigations and reevaluated previous infrageneric classifications proposed. Sequence data from the trnL-trnF region were analyzed with both parsimony and maximum likelihood criteria. AFLP markers were also obtained and analyzed with phylogenetic and principal coordinate analyses. Additionally, we obtained chromosome numbers for representative species within the group. The results strengthen the monophyly of the subgenus Amphiglottium but do not support the current classification system proposed by previous authors. Only section Tuberculata comprises a well-supported monophyletic group, with sections Carinata and Integra not supported. Instead of morphology, biogeographical and ecological patterns are reflected in the phylogenetic signal in this group. This study also confirms the large variability of chromosome numbers for the subgenus Amphiglottium (numbers ranging from 2n = 24 to 2n = 240), suggesting that polyploidy and hybridization are probably important mechanisms of speciation within the group.     

Factors Underlying the Narrow Distribution of the Mediterranean Annual Plant Arenaria provincialis (Caryophyllaceae)

The factors responsible for the patterns of niche differentiation and narrow endemism have rarely been investigated in annual Mediterranean plants. This topic was addressed here by performing comparative studies on realized niches, regional occupancy, global biogeographical distribution and seed traits of a set of Arenaria (Caryophyllaceae) species inhabiting southern Provence (SE France), focusing in particular on Arenaria provincialis, a narrow endemic restricted to the hills and modest calcareous mountains around the city of Marseille. Field studies were carried out from 2008 to 2009 at 624 sites in the limestone mountains where Arenaria species are likely to occur. The Arenaria species occurring in southern Provence vary greatly in their patterns of regional occupancy and their biogeographical distribution. Multivariate analysis of the realized niche showed that they have some similar limiting environmental factors in common, but they do not all occupy the same habitats. Studies on the fruits and seeds showed that the endemic A. provincialis has a larger seed than its congeners and a capsule that remains closed and falls from the plant at maturity. This well-differentiated diaspore specific to A. provincialis may be responsible for the limited dispersal capacity of this species. Phylogenetic analyses showed that A. provincialis is not closely related to the other taxa inhabiting southern Provence, but rather to Arenaria occurring in highland areas in the south of France. The restricted pattern of distribution of A. provincialis is probably due to the past isolation event from which it originated and to its limited dispersal capacity, rather than resulting from its ecological niche range.     

Branching, nitrogen distribution, and carbon gain in solitary plants of an annual herb, Xanthium canadense

The theory of optimal nitrogen (N) distribution predicts that the carbon gain of plants will be maximised when leaves of higher irradiance have higher N content per area (N _area). Most previous studies have examined optimal N distribution without explicitly considering the branching status of plants. I investigated light environment, N distribution and photosynthetic traits of individual leaves of an herbaceous species, Xanthium canadense. X. canadense was grown solitary under high (HN) and low nutrients (LN). Light availability, leaf mass per unit area and N _area were measured for all leaves within plants. Daily photosynthesis of the plants of actual N distribution was compared with those of optimal and constant N distribution. Branch production was facilitated in HN but not in LN plants. N _area was correlated more with leaf order than with leaf light environment. Although N was more limited and the light environment was less heterogeneous within crowns in LN than in HN plants, leaf N distribution was closer to optimal in the latter. These results suggest that leaf N distribution was not optimised in solitary plants of X. canadense. Because this species often regenerates in a dense stand, leaf N distribution might be selected to maximise carbon gain only in such a stand. Leaf N distribution might thus be constrained by the regeneration strategy of the species.     

Morphological Variation in Leaf Dissection of Rheum palmatum Complex (Polygonaceae)

Aims

Rheum palmatum complex comprises all taxa within section Palmata in the genus Rheum, including R. officinale, R. palmatum, R. tanguticum, R. tanguticum var. liupanshanense and R. laciniatum. The identification of the taxa in section Palmata is based primarily on the degree of leaf blade dissection and the shape of the lobes; however, difficulties in species identification may arise from their significant variation. The aim of this study is to analyze the patterns of variation in leaf blade characteristics within and among populations through population-based sampling covering the entire distribution range of R. palmatum complex.

Methods

Samples were taken from 2340 leaves from 780 individuals and 44 populations representing the four species, and the degree of leaf blade dissection and the shape of the lobe were measured to yield a set of quantitative data. Furthermore, those data were statistically analyzed.

Important Findings

The statistical analysis showed that the degree of leaf blade dissection is continuous from lobed to parted, and the shape of the lobe is also continuous from broadly triangular to lanceolate both within and between populations. We suggested that taxa in section Palmata should be considered as one species. Based on the research on the R. palmatum complex, we considered that the quantitative characteristics were greatly influenced by the environment. Therefore, it is not reliable to delimitate the species according to the continuously quantitative vegetative characteristics.     

Comparative morphological studies on spikes of Indian Selaginella Beauv.

The present study provides a comparative morphological analysis of spikes in different Indian species of Selaginella. The salient features of spikes including their types and also the shape and margin of sporophylls of the closely related taxa are characteristically useful in taxonomic delineation. We have used the term Leaf-flap, for the first time, and the Sporophyll-pteryx to denote the occurrence of vertical/oblique projection on the dorsal surface of leaf and sporophyll, respectively. The shape and colour of both mega- and micro-sporangia are also helpful in taxonomy; however, their size is not much significant as these are variable within the same spike of the species. A new pattern of sporangial distribution is being reported for the first time in the spike of S. plana (Desv. ex Poir.) Hieron.     

A Role for Auxin in Triggering Lamina Outgrowth of Unifacial Leaves

A common morphological feature of typical angiosperms is the patterning of lateral organs along primary axes of asymmetry—a proximodistal, a mediolateral, and an adaxial–abaxial axis. Angiosperm leaves usually have distinct adaxial–abaxial identity, which is required for the development of a flat shape. By contrast, many unifacial leaves, consisting of only the abaxial side, show a flattened morphology. This implicates a unique mechanism that allows leaf flattening independent of adaxial–abaxial identity. In this study, we report a role for auxin in outgrowth of unifacial leaves. In two closely related unifacial-leaved species of Juncaceae, Juncus prismatocarpus with flattened leaves, and Juncus wallichianus with transversally radialized leaves, the auxin-responsive gene GLYCOSIDE HYDROLASE3 displayed spatially different expression patterns within leaf primordia. Treatment of J. prismatocarpus seedlings with exogenous auxin or auxin transport inhibitors, which disturb endogenous auxin distribution, eliminated leaf flatness, resulting in a transversally radialized morphology. These treatments did not affect the radialized morphology of leaves of J. wallichianus. Moreover, elimination of leaf flatness by these treatments accompanied dysregulated expression of genetic factors needed to specify the leaf central-marginal polarity in J. prismatocarpus. The findings imply that lamina outgrowth of unifacial leaves relies on proper placement of auxin, which might induce initial leaf flattening and subsequently act to specify leaf polarity, promoting further flattening growth of leaves.

Lamina outgrowth of unifacial leaves, which lack adaxial identity, relies on proper localization of auxin, which might induce initial leaf flattening and subsequently act to specify leaf polarity, promoting further flattening growth of leaves.     

Variation of leaf shape of Salix herbacea in Europe

The morphology of Salix herbacea leaves was examined to find possible reasons for the observed intra-specific morphological variation. Seventeen samples were collected from the following regions: the Southern and Eastern Carpathians, the Northern Carpathians, the Alps: Eastern, Central and Western, the Pyrenees, Western Scandinavia, and Northern Scandinavia. The leaves were dried in an herbarium, scanned and measured. A total of 3,890 leaves from 503 individuals were statistically analyzed. The analyses were based on the shape-describing characters. A notable variation of shape characters of leaves of S. herbacea was found on different levels: intra- and inter-individual, between samples and between regions. The local environmental conditions did not affect the leaf shape much, but the regional climatic factors influenced them statistically significantly. The most important factor—the summer precipitation—was positively correlated with leaf elongation, while the mean wind velocity was negatively correlated with leaf elongation. The latter was also positively correlated with the tooth number. The hypothesis that glacial migrations and isolations of S. herbacea populations were responsible for the contemporary morphological differentiation of the species was not confirmed; however, some similarities between the biogeographical structures revealed in the previously published results of molecular analyses and in the present morphological study were also noticed.     

Leaf nitrogen distribution in relation to crown architecture in the tall canopy species,Fagus crenata

The theory of optimal leaf N distribution predicts that the C gain of plants is maximized when the N content per unit area (N _area) scales with light availability, but most previous studies have demonstrated that the N distribution is not proportional to light availability. In tall trees, the leaves are often clustered on twigs (leaf cluster) and not evenly distributed within the crowns. Thus, we hypothesized that the suboptimal N distribution is partly caused by the limited capacity to translocate N between leaf clusters, and consequently, the relationship between light and N _area differs for leaves in different clusters. We investigated the light availability and N content of all individual leaves within several leaf clusters on tall trees of a deciduous canopy species Fagus crenata in Japan. We observed that the within-cluster leaf N distribution patterns differed from the between-cluster patterns and the slopes of the relationships between light and N _area were lower within clusters than between clusters. According to the detailed analysis of the N distribution within leaf clusters, N _area was greater for current-year shoots with greater light availability or a larger total leaf area. The latter pattern was probably caused by the greater sink strength of the current-year shoots with a larger leaf area. These N distribution patterns suggest that leaf clusters are fairly independent with respect to their N use, and the productivity of real F. crenata crowns may be less than optimal.     

Diversity in forms of C4 in the genus Cleome (Cleomaceae)

Background and Aims

Cleomaceae is one of 19 angiosperm families in which C₄ photosynthesis has been reported. The aim of the study was to determine the type, and diversity, of structural and functional forms of C₄ in genus Cleome.

Methods

Plants of Cleome species were grown from seeds, and leaves were subjected to carbon isotope analysis, light and scanning electron microscopy, western blot analysis of proteins, and in situ immunolocalization for ribulose bisphosphate carboxylase oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPC).

Key Results

Three species with C₄-type carbon isotope values occurring in separate lineages in the genus (Cleome angustifolia, C. gynandra and C. oxalidea) were shown to have features of C₄ photosynthesis in leaves and cotyledons. Immunolocalization studies show that PEPC is localized in mesophyll (M) cells and Rubisco is selectively localized in bundle sheath (BS) cells in leaves and cotyledons, characteristic of species with Kranz anatomy. Analyses of leaves for key photosynthetic enzymes show they have high expression of markers for the C₄ cycle (compared with the C₃–C₄ intermediate C. paradoxa and the C₃ species C. africana). All three are biochemically NAD-malic enzyme sub-type, with higher granal development in BS than in M chloroplasts, characteristic of this biochemical sub-type. Cleome gynandra and C. oxalidea have atriplicoid-type Kranz anatomy with multiple simple Kranz units around individual veins. However, C. angustifolia anatomy is represented by a double layer of concentric chlorenchyma forming a single compound Kranz unit by surrounding all the vascular bundles and water storage cells.

Conclusions

NAD-malic enzyme-type C₄ photosynthesis evolved multiple times in the family Cleomaceae, twice with atriplicoid-type anatomy in compound leaves having flat, broad leaflets in the pantropical species C. gynandra and the Australian species C. oxalidea, and once by forming a single Kranz unit in compound leaves with semi-terete leaflets in the African species C. angustifolia. The leaf morphology of C. angustifolia, which is similar to that of the sister, C₃–C₄ intermediate African species C. paradoxa, suggests adaptation of this lineage to arid environments, which is supported by biogeographical information.     

中国北方鸢尾属植物叶片解剖结构特征及分类学价值研究

以中国北方生长的鸢尾属9种植物为研究对象,利用光学显微镜对叶表皮细胞形状及大小、气孔大小、气孔分布及气孔密度、气孔指数等叶表皮构造特征进行观察,并探讨了叶表皮特征在鸢尾属内各个亚属、亚组间的分类学价值,结果表明：表皮细胞的形状和大小显示了同亚属或同亚组植物之间较为明显的亲缘关系,在区分鸢尾属下不同亚属或亚组植物上也有一定的分类学价值。气孔密度和气孔指数在鸢尾属内分类学价值不大,但却能很好地反映不同鸢尾属植物与生境之间的密切关系。并从表皮构造特征上证实了野鸢尾与射干有很强的亲缘关系,且近于鸢尾属内的其他植物的结论;此外,通过对鸢尾属植物叶横切解剖结构的观察：从维管束、横切面形状、表皮细胞与角质层、乳突、气孔、气室大小、叶肉细胞的分化这7个较为明显的特征上的相关性和差异性,探讨了由于生境的不同而产生的植物形态结构的差异。     

20种山茶科植物的叶表皮微形态研究

为解决一些山茶科(Theaceae)的分类学问题,该研究在光学显微镜下对山茶科2亚科6属共20种植物的13种叶表皮微形态特征进行了观察,并利用聚类分析和主成分分析对这些特征进行分析,为山茶科的科学分类提供参考资料。结果表明:(1)山茶科20种植物的13种叶表皮微形态特征能够区分种间差异和分类等级,显示了比较全面的分类价值。(2)基于微形态特征的聚类分析和主成分分析结果显示,20个种分为2个大类群以及10个分支,很好地区分了山茶亚科(Theoideae)与厚皮香亚科(Ternstroemoideae),并将大部分分类地位相近的物种归类到一起,较好地反映了物种之间的亲缘关系。(3)该研究结果支持将小黄花茶组(Sect.Luteoflora)作为独立的一个组;支持将重庆山茶(Camellia chungkingensis Chang)从瘤果茶组(Sect.Tuberculata Chang Tax.)中分离;支持将山茶亚科与厚皮香亚科划分为两个较大类群,认为将厚皮香亚科并入五列木科(Pentaphylacaceae)的观点还有待更多实验证据的支持。     

Mangrove sesarmid crab feeding experiments in Peninsular Malaysia

The feeding ecology of mangrove sesarmid crabs in Peninsular Malaysia was investigated by field and laboratory experiments using four mangrove leaf species (Avicennia officinalis, Bruguiera gymnorrhiza, B. parviflora and Rhizophora apiculata) and leaves of different condition (fresh and senescent). Leaves tethered on strings at high (Bruguiera zone) and low (Rhizophora zone) intertidal positions, both upstream (Sungai Pasir) and downstream (Lower Merbok) showed significant amounts of leaf litter removal in 24 h (mean 79±3% initial dry mass). Significantly more B. gymnorrhiza was consumed in Bruguiera zones and significantly less senescent A. officinalis in the upstream Rhizophora zone. In Bruguiera zones, significant numbers of leaves were taken down burrows but there were no preferences for leaf species or condition of leaf taken down burrows at all sites. In 24 h, under laboratory conditions, the sesarmid crabs Sesarma (Perisesarma) eumolpe and S. (Perisesarma) onychophorum were offered with a mangrove species choice of either fresh or senescent leaves. There was no difference in mangrove species taken when the leaves were senescent for both crab species, but when the leaves were fresh, significantly more A. officinalis leaves were consumed by both sesarmid crab species. S. onychophorum ate significantly more B. parviflora than did S. eumolpe. The crab distribution in the field was related to the preferred tree species dominance, indicating that tree species may be important for crab species distribution, or vice versa. The mean rate of leaf consumption was not significantly different between the crab species; S. eumolpe was 29.9±5.9 and S. onychophorum was 35.3±7.2 mg dry mass per wet mass gram of crab in 24 h. Rhizophora spp. were the least preferred species in all feeding experiments, a finding which may have implications for ecosystem functioning in monoculture rehabilitation projects.     

风毛菊属3种植物叶的解剖结构比较

采用石蜡切片法对分布于祁连山海拔5 000 m左右流石滩上菊科风毛菊属水母雪兔子（Saussurea medusa Maxim）、鼠曲雪兔子（Saussurea gnaphalodes (Royle) Sch.）、红叶雪兔子（Saussurea paxiana Diels.）3种植物叶片的解剖结构进行了比较研究,结果表明：叶片表皮细胞均为单层,上下表皮都有气孔分布,气孔不下陷;角质层较厚,叶表面均被单列细胞的表皮毛。3种植物均为异面叶;叶肉栅栏组织较发达,通常由2～3层细胞组成,但栅栏组织细胞排列较疏松;海绵组织存在大量的细胞间隙;叶肉中通气组织发达,且均有不规则裂生性气腔。叶脉维管束中韧皮部都具有异细胞存在。这些共同特征是3种植物对高山地区缺氧、低温、强辐射等自然条件长期适应的结果。但是,3种植物在叶片的外部形态特征、叶肉栅栏组织细胞的特点、维管束发育程度、内分泌结构、不规则裂生气腔等方面又存在明显的不同,表现出3种植物对环境的适应也是存在差异的。     

A Cost-Benefit Analysis of Leaves of Eight Australian Savanna Tree Species of Differing Leaf Life-Span

Cost-benefit analysis of foliar construction and maintenance costs and of carbon assimilation of leaves of differing life-span were conducted using two evergreen, three semi-deciduous, and three deciduous tree species of savannas of north Australia. Rates of radiant-energy-saturated CO₂ assimilation (P _max) and dark respiration were measured and leaves were analysed for total nitrogen, fat, and ash concentrations, and for heat of combustion. Specific leaf area, and leaf N and ash contents were significantly lower in longer-lived leaves (evergreen) than shorter-lived leaves (deciduous) species. Leaves of evergreen species also had significantly higher heat of combustion and lower crude fat content than leaves of deciduous species. On a leaf area basis, P _max was highest in leaves of evergreen species, but on a leaf dry mass basis it was highest in leaves of deciduous species. P _max and total Kieldahl N content were linearly correlated across all eight species, and foliar N content was higher in leaves of deciduous than evergreen species. Leaf construction cost was significantly higher and maintenance costs were lower for leaves of evergreen than deciduous species. Maintenance and construction costs were linearly related to each other across all species. Leaves of evergreen species had a higher cost-benefit ratio compared to leaves of deciduous species but with longer lived leaves, the payback interval was longer in evergreen than deciduous species. These results support the hypotheses that longer lived leaves are more expensive to construct than short-lived leaves, and that a higher investment of N into short-lived leaves occurs which supports a higher P _max over a shorter payback interval.