Large-scale histological analysis of leaf mutants using two simple leaf observation methods: identification of novel genetic pathways governing the size and shape of leaves

Observations of cellular organization are essential in understanding the mechanisms underlying leaf morphogenesis. These observations require several preparative steps, such as fixation and clearing of organs, and such procedures are time-consuming and labor-intensive for large-scale analyses. Thus, we have developed simple methods for the observation of leaf epidermal and mesophyll cells. To visualize the epidermis, a gel cast was made of the leaf surface, which was then observed under a light microscope. To visualize the leaf mesophyll cells, leaves were immersed in a solution containing Triton X-100, briefly centrifuged, and then viewed under a light microscope. These methods allowed us to conduct a histological phenome analysis for a large number of known and newly isolated leaf-shape/size mutants of Arabidopsis thaliana by measuring various parameters, including cell number, size, and distribution of cells within a leaf blade. Mutants showed changes in leaf size caused by specific increases or decreases in the number and/or size of cells. In addition, altered cell distributions in the leaf blade were observed, resulting from increases or decreases in the number of cells along the proximo-distal or medio-lateral axis, or recruitment of cells along a particular axis at the expense of other leaf parts. These results provide a phenomic view of the cellular behavior involved in organ size control and leaf-shape patterning.     

雌雄异株的大戟科植物中叶片大小和形状的性别差异

雌雄异株的大戟科植物中叶片大小和形状的性别差异 作为自然选择的对象,叶片大小和形状能够发挥适应性作用,并且随叶龄而改变。在雌雄异株的植物中,叶片大小和形状因性别而不同,在大多数情况下,雌性的叶片较大。以往研究表明,Adriana tomentosa在叶裂方面存在性别差异。在本研究中,我们探讨了在叶片大小、形状和生理生态等方面是否存在其他性别差异,以及这些差异是否与A. tomentosa的性别适应性和繁殖作用有关。我们测定了生 长在澳大利亚东部的两个不连续种群的雌性和雄性植物的幼叶和老叶的物理化学特征,主要包括：叶面积、周长、锯齿、圆形度、长宽比、圆度以及生态生理指标,包括SLA、干物质质量、叶片水分、RWC、δ ¹³C、δ ¹⁵N同位素比、碳氮含量和碳氮比。同时还测定了叶裂、叶片损伤程度和光合色素含量。在这两个种群中,植物性别显著影响几乎所有与叶片形态相关的参数,如面积、周长、圆形度、长宽比和圆度。与预期相反,我们发现两个种群的雄性具有较大的叶面积且与叶龄无关。雄叶裂片较多,周长较长,但它们较少伸长且锯齿较少。雌性和雄性叶片的生理生态指标差异不大。叶片损伤程度因性别而异,但也因种群而异。叶面积和叶形在性别间的差异不能被生理生态因素所补偿。然而,叶面积可能由其他与叶片形态相关的生理生态机制补偿,因为与雄性相比,雌性的叶面积较小,但叶片锯齿较大。     

Hydraulics link leaf shape and environmental niche in terrestrial bromeliads

Terrestrial species of the megadiverse bromeliad family display a wide variety of leaf shapes, many of which have evolved convergently in different lineages. Here, I examine the links between leaf shape, venation architecture, hydraulic function, and bioclimatic relations in two bromeliad groups displaying diverse leaf shapes, the genus Pitcairnia (Pitcairnioideae) and the terrestrial grade of the Bromelioideae subfamily. Leaf shapes with broader leaf blades, notably petiolate and lanceolate morphologies, tend to show wider vein spacing, which is associated with reduced hydraulic capacity and higher hydraulic vulnerability. In turn, these leaf shapes tend to occur in species restricted to moist, aseasonal environments, suggesting that hydraulic function is an important mediator of the relationship between leaf shape and bromeliad environmental niches. This network of trait–trait and trait–environment relationships may have been of profound important in the ecological and evolutionary diversification of the bromeliads. Similar structure–function principles are likely to apply in other tropical herbaceous monocots, which are of great ecological importance but generally neglected in plant hydraulic research.     

Canine teeth of bats (Microchiroptera): size, shape and role in crack propagation

Upper canines in microchiropteran bats show a variety of cross-sectional shapes. A consistent feature of all species studied here is that the tooth is edged and not simply round or oval. Prominent sharp edges are positioned in several directions but particularly antero-medially toward the incisors and posteriorly toward the premolars. These edges appear to direct the cracks made in food items to the incisors or to the premolars. A continuous cutting edge is apparent in the occlusal view of the palate running from tip of canine to the ectoloph of the molars. Size and shape analysis indicates that larger bats have slender, rather than stouter, canines for their height, a condition that may be attributable to the nature of the prey. Most bats take prey that have little hard substance imbedded within. The compromises in tooth shape may vary between that of a terrestrial predator with short, conical canines for processing endoskeletal prey to that of a small flying predator with long, slender, edged canines for capturing and processing exoskeletal prey. Unicuspid teeth and how they might function in food break-up have been overlooked to the literature; such a study could lead to an understanding of how more complex teeth function.     

Hydraulic limitations imposed by crown placement determine final size and shape of Quercus rubra L. leaves

The canopies of large broad‐leaf trees exhibit significant heterogeneity in both micro‐environmental conditions and leaf morphology. Whether the visible differences in the size and shape of leaves from the top and bottom of the crown are determined prior to bud break or result from different patterns of leaf expansion is not known. Analysis of ontogenetic changes of both the degree of lobing and vein density in Quercus rubra demonstrates that leaves throughout the crown are identical in size and shape at the time of bud break. Morphological adaptation to the local micro‐environment takes place during the expansion phase and starts after the determination of the vascular architecture has been completed. Leaves from the bottom of the crown undergo greater expansion in the tissue close to the main veins than occurs either in the more peripheral tissue of the same leaf or anywhere in leaves from the top of the crown. This results in a water transport system that is well suited to the low evaporative rates near the bottom of the crown, but inadequate for the conditions found at the top of the tree. Acclimation of leaf form and function based upon differential expansion may be entirely driven by the local hydraulic demand during the expansion phase, resulting in leaf size and vein density being determined during development by the same hydraulic properties which will constrain the size of leaf that can be functionally supported at maturity.     

Application of leaf size and leafing intensity scaling across subtropical trees

Understanding the scaling between leaf size and leafing intensity (leaf number per stem size) is crucial for comprehending theories about the leaf costs and benefits in the leaf size–twig size spectrum. However, the scaling scope of leaf size versus leafing intensity changes along the twig leaf size variation in different leaf habit species remains elusive. Here, we hypothesize that the numerical value of scaling exponent for leaf mass versus leafing intensity in twig is governed by the minimum leaf mass versus maximum leaf mass (M _min versus M _max) and constrained to be ≤−1.0. We tested this hypothesis by analyzing the twigs of 123 species datasets compiled in the subtropical mountain forest. The standardized major axis regression (SMA) analyses showed the M _min scaled as the 1.19 power of M _max and the ‐α (−1.19) were not statistically different from the exponents of M _min versus leafing intensity in whole data. Across leaf habit groups, the M _max scaled negatively and isometrically with respect to leafing intensity. The pooled data''s scaling exponents ranged from −1.14 to −0.96 for M _min and M _max versus the leafing intensity based on stem volume (LIV). In the case of M _min and M _max versus the leafing intensity based on stem mass (LIM), the scaling exponents ranged from −1.24 to −1.04. Our hypothesis successfully predicts that the scaling relationship between leaf mass and leafing intensity is constrained to be ≤−1.0. More importantly, the lower limit to scaling of leaf mass and leafing intensity may be closely correlated with M _min versus M _max. Besides, constrained by the maximum leaf mass expansion, the broad scope range between leaf size and number may be insensitive to leaf habit groups in subtropical mountain forest.     

鼎湖山季风常绿阔叶林两种优势树种的生境适应研究:叶片功能性状和水力结构特征

植物在长期的进化和发展过程中,通过与环境相互作用形成了适应环境的形态结构及生理特征,反映了植物适应环境的生态策略;在森林群落中,地形和土壤等的变异常导致生境的异质性,从而直接或者间接影响物种的分布格局。因此,在生境异质性较强的森林群落中,植物物种分布格局与其生态适应策略有何关系,是值得关注的问题。该文以鼎湖山南亚热带季风常绿阔叶林20 hm2监测样地为平台,针对两种常绿优势树种罗伞树(Ardisia quinquegona)和光叶山黄皮(Aidia canthioides),对比研究了两种植物的叶片功能性状和水力结构特征在山脊、山坡、山谷三种不同生境中的生态适应策略,以阐明物种分布格局与其生态适应策略的关系。结果表明:罗伞树主要是通过调整叶面积(LA)、木材密度(WD)及渗透调节来适应不同生境;光叶山黄皮主要通过调整比叶面积(SLA)、WD及渗透调节,采取养分有效保存(低SLA,高干物质含量)及慢生长高存活的策略以适应不同生境,适应环境能力更强,尤其是在山脊和山坡生境;而且影响两个树种叶片功能性状和水力结构的主导土壤因子有所不同。研究结果说明罗伞树和光叶山黄皮对山脊和山坡生境比山谷更为适应,但在叶片功能性状和水力结构特征方面的生境适应策略不同。     

Functional and historical drivers of leaf shape evolution in palms (Arecaceae)

Aim

Leaves display a remarkable variety of shapes, each with potential ecological advantages in specific climates. While the relations between leaf shape and either climate or height have been relatively well studied in eudicots, the macroecological drivers of shape remain poorly known in monocots. Here, we investigated the associations between climate and plant height with the evolution of leaf shape in a clade with high species and morphological diversity.

Location

Global.

Time period

Cretaceous to contemporary.

Major taxa studied

Palms (Arecaceae).

Methods

We apply a Bayesian phylogenetic mixed model to test for associations between climate and leaf shape (all – entire-leaved, pinnate-dissected, palmate-dissected and costapalmate). We further reconstruct the ancestral leaf shape using multistate speciation and extinction models and compare the frequency of shapes with global temperatures through time.

Results

We find that plant height associates with dissected leaves and that annual precipitation associates with pinnate shapes. The ancestral leaf shape is unclear, but early diversification was dominated by pinnate-dissected palms, which has remained the most species-rich form of leaves throughout palm history.

Main Conclusions

Palms that are tall and live in humid regions are more likely to have pinnate leaves. Through geological time scales, temperature did not play an obvious role in determining leaf shapes. This study contributes to our understanding of how the diversity of leaf shapes is linked to biological and climatic factors.     

Morphometrics of the callitrichid forelimb: A case study in size and shape

Mosimann and colleagues formulated a technique that distinguishes between size and shape, based on the concept of geometric similarity and the distinction between log size-and-shape and log shape variables. We extend these formulations in an examination of the forelimb of three callitrichid species (adultSaguinus oedipus, Saguinus fuscicollis, andCallithrix jacchus). We employ principal components analysis to explore the relationship between variance explained by size-and-shape versus shape alone. Independence of shape vectors is examined via correlation analysis. Then we use log shape data to construct intersample (species means) and total sample (between all paris of individuals) matrices of average taxonomic distances. These distance matrices are subjected to cluster analysis and principal coordinate ordinations. Results of principal components analysis suggest that after isometric size is removed, there remains sufficient shape information to discriminate among the three taxa. Careful examination and quantification of the relationships between shape and size suggest that size information (e.g., geometric mean) is fundamental for understanding shape differences within and among callitrichid species; in other words, most aspects of forelimb shape are significantly correlated with size. Contrary to conventional wisdom, we also demonstrate that such correlations are not spurious. Ordinations and clustering of log shape distance matrices (based on means and individuals) support the notion that, despite differences in size, the two tamarins are more similar in shape than either is toC. jacchus (despite size similarity betweenS. fuscicollis andC. jacchus). Although shape variation in the forelimb of calliirichids may have a functional component, the phylogenetic signal remains strong and serves to group individuals accordingly.     

蚕用桑不同品种叶表皮形态和结构的比较研究Ⅰ

运用光学显微镜和扫描电镜对11种蚕用桑品种的叶表皮形态和结构进行观察研究。结果表明,品种间在表皮、叶肉和叶脉上的构造基本相似;主要差别在于气孔密度、上表皮细胞中的钟乳体细胞数目及叶肉中晶体的分布。基于表皮形态学特征的数量分析,结果表明,11个品种可分为6大类,这与传统分类结果基本一致。     

Global evaluation of growth in a rural population: comparative analysis of shape and size

In this work the analogies and differences in shape and size between a rural school population (6–14 aged) of the Lozoya-Somosierra region (Madrid) and several recent and past Spanish populations from different environments have been studied. The results show the growth trend as well as the influences of ecological and socioeconomical factors. Paper presented at the 4^th congress of the European Anthropology Association (Florence, Sept. 1984).     

Determination of molecular size and shape of hyperbranched polysaccharide in solution

The chemical structure of a water-soluble polysaccharide, coded as TM3b, extracted from sclerotia of Pleurotus tuber-rigium was analyzed to be a hyperbranched beta-D-glucan with beta-(1-->6), beta-(1-->4), and beta-(1-->3)-linked residues, with degree of branching (DB) of 57.6%. The results from size-exclusion chromatography combined with laser light scattering (SEC-LLS) revealed that the hyperbranched polysaccharide easily aggregated in 0.15 M aqueous NaCl, whereas it dispersed as individual chains in DMSO. The weight-average molecular weight (M(w)), radius of gyration, intrinsic viscosity, and chain density of TM3b in DMSO and in 0.15 M aqueous NaCl were measured with SEC-LLS, LLS, and viscometry. The results indicated that single chains and aggregates with aggregation number of 12 coexisted in the aqueous solution, whereas individual molecules of TM3b occurred in DMSO. In view of the molecular parameters, the aggregates in aqueous solution exhibited more compact chain structure than the individual molecules in DMSO. Furthermore, transmission electron microscopy and atomic force microscopy showed that all of the aggregates and individual molecules exhibited spherical particles in the solutions. This work provided the valuable information of chain conformation and molecular morphology of the hyperbranched polysaccharide in different solvents.     

The effect of leaf size on mutual shading and cultivar differences in soybean leaf photosynthetic capacity

This study investigated the basis of the negative relationship between leaf size and photosynthetic rate per unit of area among five cultivars of soybeans. Exposure of developing mainstem leaves to light, and sizes and light saturated photosynthesis rates of those leaves at maturity were compared in cultivars grown in field plots for two years at Beltsville, Maryland, USA. Plants were grown both in stands at 2.5 cm by 1 m spacing and as isolated plants. While cultivar differences in leaf size were large and consistent in both planting arrangements, significant cultivar differences in light saturated photosynthetic rates were found only in plants grown in stands. Similarly, leaf size was significantly correlated with specific leaf weight only for plants grown in stands. The mainstem apex and developing mainstem leaves experienced more severe shading in large-leaved cultivars than in small-leaved cultivars when plants were grown in stands. Thus, cultivar differences in photosynthetic capacity were probably a consequence of differences in the exposure of developing leaves to light.     

Frugivore gape size and the evolution of fruit size and shape in southern hemisphere floras

Abstract The present study uses differences among frugivore faunas of the southern hemisphere landmasses to test whether frugivore characteristics have influenced the evolution of fruit traits. Strong floristic similarities exist among southern landmasses; for example, 75% of New Zealand vascular plant genera also have species in Australia. However, plants in Australia and South America have evolved in the presence of a range of mammalian frugivores, whereas those in New Zealand, New Caledonia and the Pacific Islands have not. In addition, the avian frugivores in New Zealand and New Caledonia are generally smaller than those of Australia. If frugivore characteristics have influenced the evolution of fruit traits, predictable differences should exist between southern hemisphere fruits, particularly fruit size and shape. Fruit dimensions were measured for 77 New Zealand species and 31 Australian species in trans‐Tasman genera. New Zealand fruits became significantly more ellipsoid in shape with increasing size. This is consistent with frugivore gape size imposing a selective pressure on fruit ingestability. This result is not a product of phylogenetic correlates, as fruit length and width scaled isometrically for Australian species in genera shared with New Zealand. Within‐genus contrasts between New Zealand and Australian species in 20 trans‐Tasman genera showed that New Zealand species have significantly smaller fruits than their Australian counterparts. Within‐genus contrasts between New Zealand and South American species in nine genera gave the same result; New Zealand species had significantly smaller fruits than their South American counterparts. No difference was found in fruit size or shape between New Zealand and New Caledonia congeneric species from 12 genera. These results are consistent with the broad characteristics of the frugivore assemblage influencing the evolution of fruit size and shape in related species. The smaller‐sized New Zealand frugivore assemblage has apparently influenced the evolution of fruit size of colonizing taxa sometimes within a relatively short evolutionary timeframe.     

Myxosporida (Protozoa): The determination and maintenance of spore size and shape

Data are presented which suggest that the size of fish myxosporidan spores is primarily determined by the parasite's development in the tissues or organ's cavities and the spore's shape is determined by the presence of physiologically and behaviorally suitable fish.     

Comparison of allometric relationships and morphological differences in Tilia cordata Mill. outer leaves exposed to different environmental conditions

The study is based on four leaf parameters: leaf width (LW), lobe length (LL), leaf size (LS) and leaf shape which is calculated as LW to leaf length (LW/LL) ratio. Under different environmental conditions, LL is an isometric character, LW shows positive allometry, whereas LW/LL shows negative allometry. Regression analysis results indicated that there is no significant difference either in slopes or in regression coefficients between investigated sites. Thus, in this study, we found that allometric relationships between leaf parameters and LS are character specific and that they tended not to differ significantly between Tilia cordata Mill. outer leaves exposed to different environmental conditions. Also, there are no significant interpopulation differences for both principal component PC1 and PC2 scores. The stepwise discriminant functional analysis results allowed us to identify a set of two leaf parameters (LS and LL) with a moderate discriminating ability (59.8%). T. cordata outer leaves are significantly larger and broader in the reference area (R-leaves) than leaves from polluted (P-leaves) site. The data also indicated that there is a relatively larger petiole size in R-leaves than in P-leaves. We found that in P-leaves, LW increased faster with increasing LS than in R-leaves.