Effects of Supplementary Blue and UV-A LED Lights on Morphology and Phytochemicals of Brassicaceae Baby-Leaves.

Brassicaceae baby-leaves are good source of functional phytochemicals. To investigate how Chinese kale and pak-choi baby-leaves in response to different wavebands of blue (430 nm and 465 nm) and UV-A (380 nm and 400 nm) LED, the plant growth, glucosinolates, antioxidants, and minerals were determined. Both agronomy traits and phytochemical contents were significantly affected. Blue and UV-A light played a predominant role in increasing the plant biomass and morphology, as well as the contents of antioxidant compounds (vitamin C, vitamin E, phenolics, and individual flavonols), the antioxidant activity (DPPH and FRAP), and the total glucosinolates accumulation. In particular, four light wavebands significantly decreased the content of progoitrin, while 400 nm UV-A light and 430 nm blue light were efficient in elevating the contents of sinigrin and glucobrassicin in Chinese kale. Meanwhile, 400 nm UV-A light was able to increase the contents of glucoraphanin, sinigrin, and glucobrassicin in pak-choi. From the global view of heatmap, blue lights were more efficient in increasing the yield and phytochemical levels of two baby-leaves.

Microscopic investigation of pollen morphology of Brassicaceae from Central Punjab-Pakistan.

The intention of the current study is to provide an account on the palynological features of Brassicaceae from Central Punjab-Pakistan as a basis for future studies. Different morpho-palynological characteristics both qualitative and quantitative were analyzed during this research which includes shape of pollen, diameter of pollen, P/E ratio, exine sculpturing, thickness of exine, type of pollen, shape and size of lumens, and thickness of murus. Taxonomic keys were also constructed based on pollen morphological characters for correct identification of species. This study aims to provide detailed information of pollen diversity and their exine structure based on both qualitative and quantitative characters by using Light microscopy and Scanning electron microscopy. Shape of pollen is mostly prolate, but some species also have sub-prolate to spheroidal prolate types. Exine ornamentation in most species was reticulate, whereas micro reticulate (one species) and coarsely reticulate (one species) exine also observed in some pollen. All the pollen mentioned in this study have tricolpate apertures. Variation found in thickness of exine and other characters proved to be helpful at generic and specific level. The results reinforced the significance of pollen morphological features of family Brassicaceae and aid for valuable taxonomic tool in plant systematics.

Diversification of Biscutella ser. Biscutella (Brassicaceae) followed post-Miocene geologic and climatic changes in the Mediterranean basin.

Biscutella ser. Biscutella (= ser. Lyratae Malin.) is a group of mostly annual or short-lived perennial plants, with petals gradually tapering at the base and lateral intrastaminal nectaries, endemic to the Mediterranean basin and the Middle East. Recent taxonomic work has revealed that a relative morphological homogeneity occurs in Europe and Asia, but a high plasticity is found in N Africa for most of the characters traditionally used for taxonomic arrangements. This fact had generally led to overestimation of the number of taxa, which currently is reduced to ten (namely 7 species and 3 additional varieties), some of them being narrow endemics. In the present contribution, on the basis of a previous detailed morphological study carried out by the authors, the first comprehensive phylogeny based on 47 DNA sequence data including concatenation of two plastid (rpl32-trnL and trnV) and one nuclear (ITS) regions, together with the first time-calibrated phylogenetic tree, allows reappraisal of evolutionary and biogeographic relationships among the accepted taxa in the series. According to all evidence gathered in the present study, the current distribution of B. ser. Biscutella, mostly centred in the southern parts of the Mediterranean basin and the Middle East, suggests that it evolved in relation with the major geological and climatic events occurred in the Mediterranean basin and Eurasia within the last 20 million years. The origin of Biscutella is dated ca 18.75 Mya, and the radiation of the series triggered ca 5.87 Mya with the Messinian Salinity Crisis. Rapid diversification occurred coetaneously to the Intensification of Northern Hemisphere Glaciation (ca 2.86 Mya) onwards, with parallel large-amplitude aridity cycles in Africa and southwestern Asia. In recent times, the divergence of lineages became faster in the W Mediterranean (ca 1.54 to 0.43 Mya), mostly related to geographical and ecological patterns of specialisation. In many cases, the distribution of the current species is apparently linked to ancient glacial refuges in S Mediterranean basin.

Natural Variation in Flower Color and Scent in Populations of Eruca sativa (Brassicaceae) Affects Pollination Behavior of Honey Bees.

Plants of Eruca sativa Mill. (Brassicaceae) from desert and Mediterranean populations in Israel differ in flower color and size. In the desert habitat, the population has higher abundance of flowers with cream color and longer petals, whereas in the Mediterranean habitat, the population has higher abundance of flowers with yellow and shorter petals. Choice experiments with honey bee foragers (Apis mellifera Linn., Apidae, Hymenoptera), the main pollinator in the natural habitat in Israel, confirmed that they are more attracted to the yellow flower morph than to the cream one. A proboscis extension response test indicated that honey bees are able to discriminate between flower scents of the desert and Mediterranean populations. Considering the advantage of plants of the yellow morph in attracting pollinators, we further tested in a common garden experiment whether these possess higher fitness than plants of the desert population. Indeed, a significant association was found between flower color and fruit set, and seed mass. In general, our results provide evidence for ecotypic differentiation between populations imposed by pollinators. The advantage of the yellow color morph in attracting pollinators may explain its dominance among plants of the Mediterranean population. We discuss why the cream color morph may be dominant in the desert habitat, considering the possibility of different pollinators, tradeoffs between traits, or pleiotropy.

Brassicaceae flowers: diversity amid uniformity.

The mustard family Brassicaceae, which includes the model plant Arabidopsis thaliana, exhibits morphological stasis and significant uniformity of floral plan. Nonetheless, there is untapped diversity in almost every aspect of floral morphology in the family that lends itself to comparative study, including organ number, shape, form, and color. Studies on the genetic basis of morphological diversity, enabled by extensive genetic tools and genomic resources and the close phylogenetic distance among mustards, have revealed a mosaic of conservation and divergence in numerous floral traits. Here I review the morphological diversity of the flowers of Brassicaceae and discuss studies addressing the underlying genetic and developmental mechanisms shaping floral diversity. To put flowers in the context of the floral display, I describe diversity in inflorescence morphology and the variation that exists in the structures preceding the floral organs. Reconstructing the floral morphospace in Brassicaceae coupled with next-generation sequencing data and unbiased approaches to interrogate gene function in species throughout the mustard phylogeny offers promising ways to understand how developmental mechanisms originate and diversify.

Kin discrimination allows plants to modify investment towards pollinator attraction.

Pollinators tend to be preferentially attracted to large floral displays that may comprise more than one plant in a patch. Attracting pollinators thus not only benefits individuals investing in advertising, but also other plants in a patch through a 'magnet' effect. Accordingly, there could be an indirect fitness advantage to greater investment in costly floral displays by plants in kin-structured groups than when in groups of unrelated individuals. Here, we seek evidence for this strategy by manipulating relatedness in groups of the plant Moricandia moricandioides, an insect-pollinated herb that typically grows in patches. As predicted, individuals growing with kin, particularly at high density, produced larger floral displays than those growing with non-kin. Investment in attracting pollinators was thus moulded by the presence and relatedness of neighbours, exemplifying the importance of kin recognition in the evolution of plant reproductive strategies.

Successive duplication-divergence mechanisms at the RCO locus contributed to leaf shape diversity in the Brassicaceae.

Gene duplication is a major driver for the increase of biological complexity. The divergence of newly duplicated paralogs may allow novel functions to evolve, while maintaining the ancestral one. Alternatively, partitioning the ancestral function among paralogs may allow parts of that role to follow independent evolutionary trajectories. We studied the REDUCED COMPLEXITY (RCO) locus, which contains three paralogs that have evolved through two independent events of gene duplication, and which underlies repeated events of leaf shape evolution within the Brassicaceae. In particular, we took advantage of the presence of three potentially functional paralogs in Capsella to investigate the extent of functional divergence among them. We demonstrate that the RCO copies control growth in different areas of the leaf. Consequently, the copies that are retained active in the different Brassicaceae lineages contribute to define the leaf dissection pattern. Our results further illustrate how successive gene duplication events and subsequent functional divergence can increase trait evolvability by providing independent evolutionary trajectories to specialized functions that have an additive effect on a given trait.

When the BRANCHED network bears fruit: how carpic dominance causes fruit dimorphism in Aethionema.

Life in unpredictably changing habitats is a great challenge, especially for sessile organisms like plants. Fruit and seed heteromorphism is one way to cope with such variable environmental conditions. It denotes the production of distinct types of fruits and seeds that often mediate distinct life-history strategies in terms of dispersal, germination and seedling establishment. But although the phenomenon can be found in numerous species and apparently evolved several times independently, its developmental time course or molecular regulation remains largely unknown. Here, we studied fruit development in Aethionema arabicum, a dimorphic member of the Brassicaceae family. We characterized fruit morph differentiation by comparatively analyzing discriminating characters like fruit growth, seed abortion and dehiscence zone development. Our data demonstrate that fruit morph determination is a 'last-minute' decision happening in flowers after anthesis directly before the first morphotypical differences start to occur. Several growth experiments in combination with hormone and gene expression analyses further indicate that an accumulation balance of the plant hormones auxin and cytokinin in open flowers together with the transcript abundance of the Ae. arabicum ortholog of BRANCHED1, encoding a transcription factor known for its conserved function as a branching repressor, may guide fruit morph determination. Thus, we hypothesize that the plasticity of the fruit morph ratio in Ae. arabicum may have evolved through the modification of a preexisting network known to govern correlative dominance between shoot organs.

Molecular characterization of a double-flower mutation in Matthiola incana.

The double flower is one of the most important traits in the floricultural plant Matthiola incana. Although a "doubleness" locus (S/s) was defined by genetic analysis a century ago, the gene responsible for double flowers has not been identified in M. incana. We isolated MiAG from M. incana cultivars, and its sequence and genomic structure were found to be highly similar to the AGAMOUS gene in Arabidopsis. Two independent mutated alleles miag1 and miag2 were identified from the double-flowered individuals of M. incana cultivars. Deletions of 135 bp (from the 2nd exon to the 2nd intron) and 89 bp (from the 7th intron to the 8th exon) were detected in miag1 and miag2, respectively. No transcript was detected in flower buds from miag1 alleles in corresponding cultivars, whereas three mRNA variants with frameshifts were transcribed from the miag2 allele in other cultivars. Thus, two mutated alleles corresponding to the s locus contributed to the 'eversporting' type double-flower cultivars in M. incana. Moreover, we also developed co-dominant molecular markers to describe the genotypes of the three alleles of MiAG. Using these DNA markers allows for selection of single- or double-flowered individuals among seedlings that do not display phenotypic differences.

Silique valves as sails in anemochory of Lunaria (Brassicaceae).

The generally held opinion that seeds of Lunaria remain at the replum after detachment of the two valves and then wind causes a shaking or rattling of the replum with its diaphragm, thus launching the seeds, is challenged. In a sparse forest in the Swabian Alb, the first author noticed flying valves of Lunaria rediviva to which the narrow-winged flat seeds are attached. Investigations with SEM and histology have shown that the valves secrete a glue only at those sites where the seeds rest on the valves before valve tissues die. Further analysis has shown (using the periodic acid-Schiff reaction) that the glue consists of polysaccharides. After detachment and dispersal of the valves, the adhesive strength continuously decreases. This is the first report for a sticky valve exudate in the Brassicaceae. Because of the adhesion of Lunaria seeds to their valves for some time, the 1st order diaspore is a mericarp, in a broad sense, and can be interpreted as an adaptation to long-distance dispersal by stronger winds. In this context, the 'flying carpets' of Lunaria are more effective and transport more than one seed. Molecular studies assigned Lunaria to the tribe Biscutelleae, which now contains the angustiseptate genera Biscutella and Megadenia as well as the latiseptate genera Lunaria and Ricotia. The valves in Ricotia can easily be detached (studied in herbarium material and a living plant), but, in contrast to Lunaria, the ripe seeds remain at the replum and its diaphragm, respectively.

Is floral structure a reliable indicator of breeding system in the Brassicaceae?

This study investigated the usefulness of floral characters as a potential indicator of breeding system in the Brassicaceae. Initially, pod set, seed set and pollen tube growth experiments were carried out to confirm the breeding systems of 53 lines representing 25 different cultivated and weedy species from the Brassicaceae. The results of the pod set tests clearly differentiated between self-compatible and self-incompatible species. Floral characters were then evaluated on one or more lines of each of the 25 species. Fourteen floral characters were evaluated including, flower diameter, Cruden's outcrossing index, timing and direction of dehiscence and pollen-ovule ratio. Significant differences between species were evident in all of the floral characteristics evaluated. Flower diameter was generally larger in self-incompatible species than self-compatible species and pollen/ovule ratio was generally higher in self-incompatible species than self-compatible species. However, none of the floral characteristics was able to clearly differentiate the self-compatible and self-incompatible species and allow prediction of the breeding system with absolute confidence. The floral characteristic which was most effective at differentiating the two groups was anther direction at dehiscence.

Photosynthesis in C3-C4 intermediate Moricandia species.

Evolution of C4 photosynthesis is not distributed evenly in the plant kingdom. Particularly interesting is the situation in the Brassicaceae, because the family contains no C4 species, but several C3-C4 intermediates, mainly in the genus Moricandia Investigation of leaf anatomy, gas exchange parameters, the metabolome, and the transcriptome of two C3-C4 intermediate Moricandia species, M. arvensis and M. suffruticosa, and their close C3 relative M. moricandioides enabled us to unravel the specific C3-C4 characteristics in these Moricandia lines. Reduced CO2 compensation points in these lines were accompanied by anatomical adjustments, such as centripetal concentration of organelles in the bundle sheath, and metabolic adjustments, such as the balancing of C and N metabolism between mesophyll and bundle sheath cells by multiple pathways. Evolution from C3 to C3-C4 intermediacy was probably facilitated first by loss of one copy of the glycine decarboxylase P-protein, followed by dominant activity of a bundle sheath-specific element in its promoter. In contrast to recent models, installation of the C3-C4 pathway was not accompanied by enhanced activity of the C4 cycle. Our results indicate that metabolic limitations connected to N metabolism or anatomical limitations connected to vein density could have constrained evolution of C4 in Moricandia.

A comparative study of the seed structure between resynthesized allotetraploid and their diploid parents.

Brassicaceae is at the forefront of evolution because of its frequent hybridization. Hybridization is responsible for the induction of widespread genetic and phenotype changes, making it important in agricultural production. In this study, we obtained resynthesized allotetraploid Brassica napus by performing interspecific crossing of B. rapa × B. oleracea combined with embryo rescue. We applied light microscopy and electronic microscopy to analyze the microstructure and ultrastructure of seeds of diploid parents and their allotetraploid progeny. Results showed that pigments in the seed coat were mainly distributed in the palisade layer. B. rapa presented the highest amount of pigment followed by B. napus and B. oleracea. B. napus had the thickest palisade layer followed by B. rapa and B. oleracea. The seed coat microsculpturing in B. rapa and B. napus was characterized as reticulate or reticulate-foveate, whereas that in B. oleracea was observed to be rugose and sulcate. The area index of the protein body was higher in central meristematic cells than in parenchyma cells. By contrast, the area index of the oil body was the lowest in central meristematic cells. Protein bodies were found to be heterogeneous with crystal globoids in two diploid parents and resynthesized allotetraploid progenies. Oil bodies consisted of large and small oil bodies, the sizes of which differed between two parents and allotetraploid progenies. Small oil bodies were spheroid, whereas large oil bodies were ovoid in shape. The quantity of oil bodies indicated that oil bodies were spheroid in two parents, ranging in size from 0.12 to 1.18 µm. In comparison, the size of large oil bodies in allotetraploid progenies exceeds 2.0 µm. These findings suggest that the anatomy of resynthesized allotetraploid seeds remarkably differs from that of two diploid parents, and these differences definitely affect the nutritional components of rapeseeds.

Developmental Control and Plasticity of Fruit and Seed Dimorphism in Aethionema arabicum.

Understanding how plants cope with changing habitats is a timely and important topic in plant research. Phenotypic plasticity describes the capability of a genotype to produce different phenotypes when exposed to different environmental conditions. In contrast, the constant production of a set of distinct phenotypes by one genotype mediates bet hedging, a strategy that reduces the temporal variance in fitness at the expense of a lowered arithmetic mean fitness. Both phenomena are thought to represent important adaptation strategies to unstable environments. However, little is known about the underlying mechanisms of these phenomena, partly due to the lack of suitable model systems. We used phylogenetic and comparative analyses of fruit and seed anatomy, biomechanics, physiology, and environmental responses to study fruit and seed heteromorphism, a typical morphological basis of a bet-hedging strategy of plants, in the annual Brassicaceae species Aethionema arabicum Our results indicate that heteromorphism evolved twice within the Aethionemeae, including once for the monophyletic annual Aethionema clade. The dimorphism of Ae. arabicum is associated with several anatomic, biomechanical, gene expression, and physiological differences between the fruit and seed morphs. However, fruit ratios and numbers change in response to different environmental conditions. Therefore, the life-history strategy of Ae. arabicum appears to be a blend of bet hedging and plasticity. Together with the available genomic resources, our results pave the way to use this species in future studies intended to unravel the molecular control of heteromorphism and plasticity.

Fruit shape diversity in the Brassicaceae is generated by varying patterns of anisotropy.

Fruits exhibit a vast array of different 3D shapes, from simple spheres and cylinders to more complex curved forms; however, the mechanism by which growth is oriented and coordinated to generate this diversity of forms is unclear. Here, we compare the growth patterns and orientations for two very different fruit shapes in the Brassicaceae: the heart-shaped Capsella rubella silicle and the near-cylindrical Arabidopsis thaliana silique. We show, through a combination of clonal and morphological analyses, that the different shapes involve different patterns of anisotropic growth during three phases. These experimental data can be accounted for by a tissue-level model in which specified growth rates vary in space and time and are oriented by a proximodistal polarity field. The resulting tissue conflicts lead to deformation of the tissue as it grows. The model allows us to identify tissue-specific and temporally specific activities required to obtain the individual shapes. One such activity may be provided by the valve-identity gene FRUITFULL, which we show through comparative mutant analysis to modulate fruit shape during post-fertilisation growth of both species. Simple modulations of the model presented here can also broadly account for the variety of shapes in other Brassicaceae species, thus providing a simplified framework for fruit development and shape diversity.

Networks of plants: how to measure similarity in vegetable species.

Despite the common misconception of nearly static organisms, plants do interact continuously with the environment and with each other. It is fair to assume that during their evolution they developed particular features to overcome similar problems and to exploit possibilities from environment. In this paper we introduce various quantitative measures based on recent advancements in complex network theory that allow to measure the effective similarities of various species. By using this approach on the similarity in fruit-typology ecological traits we obtain a clear plant classification in a way similar to traditional taxonomic classification. This result is not trivial, since a similar analysis done on the basis of diaspore morphological properties do not provide any clear parameter to classify plants species. Complex network theory can then be used in order to determine which feature amongst many can be used to distinguish scope and possibly evolution of plants. Future uses of this approach range from functional classification to quantitative determination of plant communities in nature.

Anatomía e histoquímica de Tarenaya hassleriana (cleomaceae), especie de interés medicinal / Anatomy and histochemistry of Tarenaya hassleriana (cleomaceae), an interesting medicinal species

La decocción de la planta de Tarenaya hassleriana es utilizada en la región del Río de La Plata por sus propiedades como rubefaciente, digestiva, y antiescorbútica. El objetivo del estudio fue investigar caracteres de la hoja y tallo para identificar esta especie a partir de muestras enteras o fragmentadas. Se emplearon técnicas habituales de microscopia óptica y análisis histoquímico para identificar almidón, sustancias lipofílicas, fenólicas y mirosina. Los caracteres diagnósticos fueron: en la hoja, folíolos con venación eucamptódroma, con estomas anomocíticos presentes en ambas caras; tricomas glandulares uniseriados y pluriseriados, con cabezas unicelulares y pluricelulares conteniendo sustancias lipofílicas; mesofilo dorsiventral; en el tallo, la corteza primaria formada por varias capas de colénquima tangencial seguido de parénquima; el cilindro vascular secundario rodea la médula sólida, en cuyo parénquima se halló almidón, cristales poliédricos y esferocristales. Se detectaron idioblastos de mirosina en la epidermis y parénquimas de la hoja y el tallo. Los parámetros micrográficos descriptos garantizan una correcta identificación de T. hassleriana.

Plant decoction of Tarenaya hassleriana is used as a traditional medicine in the Río de La Plata area. It has rubefacient, digestive, and antiscorbutic properties. The aim of this study was to investigate leaf and stem microcharacters to identify this species from whole or fragmented samples. The usual techniques of optical microscopy were employed. Histochemical tests for starch, lipophilic substances, phenolic substances, and mirosina were used. The main differential traits were: leaflets with eucamptodromous venation, amphistomatic with anomocytic stomata type; uniseriate and pluriseriate glandular trichomes, with unicellular and pluricellular heads containing lipophilic substances; dorsiventral mesophyll; stem with a primary cortex formed by tangential collenchyma followed by parenchyma; the secondary vascular cylinder surrounding the pith with starch, polyhedral microcrystals and spherocrystals. Idioblast of myrosin were detected in the epidermis and parenchyma of leaves and stems. The micrographic parameters described ensure a correct identification of T. hassleriana.

Clade-specific positive selection on a developmental gene: BRANCHLESS TRICHOME and the evolution of stellate trichomes in Physaria (Brassicaceae).

Positive selection is known to drive the evolution of genes involved in evolutionary arms races, but what role does it play in the evolution of genes involved in developmental processes? We used the single-celled epidermal trichomes of Brassicaceae as a model to uncover the molecular evolutionary processes that contributed to the transition from dendritic trichomes, as seen in most species of Brassicaceae, to the distinctive stellate trichomes of the genus Physaria. We explored the role of positive selection on the evolution of BRANCHLESS TRICHOME (BLT), a candidate gene for changes in trichome branching pattern. Maximum likelihood models of codon evolution point to a shift in selective pressure affecting the evolution of BLT across the entire Physaria clade, and we found strong evidence that positive selection has acted on a subset of Physaria BLT codons. Almost all of the 10 codon sites with the highest probability of having evolved under positive selection are clustered in a predicted coiled-coil domain, pointing to changes in protein-protein interactions. Thus, our findings suggest that selection acted on BLT to modify its interactions with other proteins. The fact that positive selection occurred throughout the radiation of Physaria could reflect selection to stabilize development in response to an abrupt switch from the dendritic form to the stellate form, divergent selection for diversification of the stellate form, or both. These results point to the need for evolutionary developmental studies of BLT and its interacting proteins in Physaria.

Anatomical abnormalities of the intertribal cybrid between Brassica napus and Lesquerella fendleri.

The anatomical research of the vegetative organs of the cytoplasmatic hybrid in vitro plants containing the Brassica napus nucleus and the Lesquerella fendleri chloroplasts had been conducted in comparison to the parental forms. It was found, that the anatomical structure of the cybrid had been similar to rapeseed. Anomalous changes in the epithelial, parenchymal and connective tissue of the leaf, stalk, stem, and root of the cybrid were detected. The appearance of the anatomical defects can be explained by nuclear-cytoplasmatic incompatibility which is the cause of low adaptability of the cybrid to in vivo conditions and takes place due to alien chloroplast gene expression in the remote species.

Feeding damage to plants increases with plant size across 21 Brassicaceae species.

Plant size is a major predictor of ecological functioning. We tested the hypothesis that feeding damage to plants increases with plant size, as the conspicuousness of large plants makes resource finding and colonisation easier. Further, large plants can be attractive to herbivores, as they offer greater amounts and ranges of resources and niches, but direct evidence from experiments testing size effects on feeding damage and consequently on plant fitness is so far missing. We established a common garden experiment with a plant size gradient (10-130 cm height) using 21 annual Brassicaceae species, and quantified plant size, biomass and number of all aboveground components (flowers, fruits, leaves, stems) and their proportional feeding damage. Plant reproductive fitness was measured using seed number, 1000 seed weight and total seed weight. Feeding damage to the different plant components increased with plant size or component biomass, with mean damage levels being approximately 30 % for flowers, 5 % for fruits and 1 % for leaves and stems. Feeding damage affected plant reproductive fitness depending on feeding damage type, with flower damage having the strongest effect, shown by greatly reduced seed number, 1000 seed weight and total seed weight. Finally, we found an overall negative effect of plant size on 1000 seed weight, but not on seed number and total seed weight. In conclusion, being conspicuous and attractive to herbivores causes greater flower damage leading to higher fitness costs for large plants, which might be partly counterbalanced by benefits such as enhanced competitive/compensatory abilities or more mutualistic pollinator visits.