Revisiting an ecophysiological oddity: Hydathode-mediated foliar water uptake in Crassula species from southern Africa.

Hydathodes are usually associated with water exudation in plants. However, foliar water uptake (FWU) through the hydathodes has long been suspected in the leaf-succulent genus Crassula (Crassulaceae), a highly diverse group in southern Africa, and, to our knowledge, no empirical observations exist in the literature that unequivocally link FWU to hydathodes in this genus. FWU is expected to be particularly beneficial on the arid western side of southern Africa, where up to 50% of Crassula species occur and where periodically high air humidity leads to fog and/or dew formation. To investigate if hydathode-mediated FWU is operational in different Crassula species, we used the apoplastic fluorescent tracer Lucifer Yellow in combination with different imaging techniques. Our images of dye-treated leaves confirm that hydathode-mediated FWU does indeed occur in Crassula and that it might be widespread across the genus. Hydathodes in Crassula serve as moisture-harvesting structures, besides their more common purpose of guttation, an adaptation that has likely played an important role in the evolutionary history of the genus. Our observations suggest that ability for FWU is independent of geographical distribution and not restricted to arid environments under fog influence, as FWU is also operational in Crassula species from the rather humid eastern side of southern Africa. Our observations point towards no apparent link between FWU ability and overall leaf surface wettability in Crassula. Instead, the hierarchically sculptured leaf surfaces of several Crassula species may facilitate FWU due to hydrophilic leaf surface microdomains, even in seemingly hydrophobic species. Overall, these results confirm the ecophysiological relevance of hydathode-mediated FWU in Crassula and reassert the importance of atmospheric humidity for some arid-adapted plant groups.

Callose deposition analysis with special emphasis on plasmodesmata ultrastructure during megasporogenesis in Sedum (Crassulaceae).

In this study, the results of the first detection of callose within the ovules of the representatives of the family Crassulaceae are presented. This study was carried out on three species of the genus Sedum. Data analysis showed differences in the callose deposition pattern between Sedum hispanicum and Sedum ser. Rupestria species during megasporogenesis. Callose was present mostly in the transversal walls of dyads and tetrads in S. hispanicum. Furthermore, a complete loss of callose from the cell walls of the linear tetrad and a gradual and simultaneous deposition of callose within the nucellus of S. hispanicum were observed. The findings of this study showed the presence of hypostase with callose in the ovules of S. hispanicum, which is not common in other angiosperms. The remaining species tested in this study-Sedum sediforme and Sedum rupestre-showed a typical, well-known callose deposition pattern for plants with the monospore type of megasporogenesis and the Polygonum type of embryo sac. The functional megaspore (FM) in all studied species was located most chalazally. FM is a mononuclear cell, which wall is callose-free in the chalazal pole. The study presents the causes of different patterns of callose deposition within Sedum and their relationship with the systematic position of the study species. Moreover, embryological studies present an argument for excluding callose as a substance that forms an electron-dense material near the plasmodesmata in megaspores of S. hispanicum. This research expands the knowledge about the embryological processes of succulent plants from the family Crassulaceae.

Morphometric Characteristics and Genetic Issr Marker Variability in Rhodiola rosea L. (Crassulaceae) in Different Ecological and Geographic Conditions in the Altai Republic.

Rhodiola rosea L. is a vulnerable species in the Altai Republic (AR) and Russia in general. For the first time on the territory of AR, studies of the adaptive capabilities of the species and genetic differentiation using ISSR markers were carried out in seven cenopopulations (CP) of R. rosea in 2018 and 2020. The research was founded on the notion of conducting a comparative analysis of the morphogenetic structure of Rhodiola rosea populations in various ecological and geographical conditions of AR. The aim of this work is to evaluate the variability of morphometric traits of sexually mature living female R. rosea plants and to conduct a comparative analysis of genetic variability in cenopopulations (CP) both under undisturbed conditions and under stressful conditions of anthropogenic impact (grazing). Of the 8 primers used, HB12 turned out to be the most informative. The percentage of polymorphic loci in the populations between 0 and 88%. Two populations, located in favorable conditions at relatively low absolute altitudes (2000 m above sea level) (masl) in the undisturbed habitats of the Katun and Altai reserves of AR, were characterized by higher polymorphism. The share of polymorphic loci reached 80%. According to the analysis of statistical data, the highest values of morphometric parameters of the aerial parts of R. rosea plants and the highest potential seed productivity were also recorded in these habitats. Representatives of two high-mountain CPs (2400-2500 masl) in the Sailyugemsky National Park (SNP) were characterized by the lowest genetic polymorphism. Their genetic structure is the most homogeneous, since we have not found polymorphic loci. Due to spatial isolation, these individuals are reliably genetically differentiated. In addition, individuals of one type were subjected to stressful anthropogenic impact (grazing). Therefore, the smallest sizes and lowest potential seed productivity were recorded. Our research shows that alpine populations of R. rosea in AR, under conditions of anthropogenic stress, need protection for their gene pool.

Ultrasonographic assessment of rectus abdominis muscle adaptation after deep inferior epigastric artery perforator (DIEP) flap surgery: Single institution retrospective study.

The impact of deep inferior epigastric artery perforator (DIEP) flap on abdominal wall integrity has been the topic of an ongoing debate with previous studies having reported conflicting results using various imaging modalities. Ultrasonography is a noninvasive, cost-effective, and readily available method for evaluating the changes to the rectus muscle after DIEP flap surgery. In the present study, we aimed to compare rectus abdominis muscle thickness between the operated and non-operated sides using ultrasound imaging. The muscle thickness was measured at the cross point of the midclavicular line and the level of the umbilicus and anterior superior iliac spine using real-time B-mode ultrasonography. The muscle anteroposterior diameters of the pedicle-dissected side and the control side were compared using paired t test. In total 31 patients with a mean follow-up of 70.18 weeks were included. The mean diameters at the level of the umbilicus of the operated and non-operated sides were 8.16 ±â€…1.83 and 8.14 ±â€…1.43 mm, respectively (P = .94). The mean thicknesses at the anterior superior iliac spine level were 7.74 ±â€…1.85 on the flap harvested side and 8.04 ±â€…1.84 mm on the control side (P = .35). There was no statistically significant difference between the 2 groups. Ultrasonography can be a reliable, inexpensive, and easily usable modality for evaluating donor site complication following DIEP flap. DIEP flap seems to have minimal impact on the abdominal donor site, and it may be safe and versatile to reconstruct the breast after mastectomy.

Improved efficiency of Sedum lineare (Crassulaceae) in remediation of arsenic-contaminated soil by phosphate-dissolving strain P-1 in association with phosphate rock.

The selection of appropriate plants and growth strategies is a key factor in improving the efficiency and universal applicability of phytoremediation. Sedum lineare grows rapidly and tolerates multiple adversities. The effects of inoculation of Acinetobacter sp. phosphate solubilizing bacteria P-1 and application of phosphate rock (PR) as additives on the remediation efficiency of As-contaminated soil by S. lineare were investigated. Compared with the control, both the single treatment and the combination of inoculation with strain P-1 and application of PR improved the biomass by 30.7-395.5%, chlorophyll content by 48.1-134.8%, total protein content by 12.5-92.4% and total As accumulation by 45.1-177.5%, and reduced the As-induced oxidative damage. Inoculation with strain P-1 increased the activities of superoxide dismutases and catalases of S. lineare under As stress, decreased the accumulation of reactive oxygen species in plant tissues and promoted the accumulation of As in roots. In contrast, simultaneous application of PR decreased As concentration in S. lineare tissues, attenuated As-induced lipid peroxidation and improved As transport to shoots. In addition, the combined application showed the best performance in improving resistance and biomass, which significantly increased root length by 149.1%, shoot length by 33%, fresh weight by 395.5% and total arsenic accumulation by 159.2%, but decreased the malondialdehyde content by 89.1%. Our results indicate that the combined application of strain P-1 and PR with S. lineare is a promising bioremediation strategy to accelerate phytoremediation of As-contaminated soils.

A Review of the Popular Uses, Anatomical, Chemical, and Biological Aspects of Kalanchoe (Crassulaceae): A Genus of Plants Known as "Miracle Leaf".

Species of the genus Kalanchoe have a long history of therapeutic use in ethnomedicine linked to their remarkable healing properties. Several species have chemical and anatomical similarities, often leading to confusion when they are used in folk medicine. This review aims to provide an overview and discussion of the reported traditional uses, botanical aspects, chemical constituents, and pharmacological potential of the Kalanchoe species. Published scientific materials were collected from the PubMed and SciFinder databases without restriction regarding the year of publication through April 2023. Ethnopharmacological knowledge suggests that these species have been used to treat infections, inflammation, injuries, and other disorders. Typically, all parts of the plant are used for medicinal purposes either as crude extract or juice. Botanical evaluation can clarify species differentiation and can enable correct identification and validation of the scientific data. Flavonoids are the most common classes of secondary metabolites identified from Kalanchoe species and can be correlated with some biological studies (antioxidant, anti-inflammatory, and antimicrobial potential). This review summarizes several topics related to the Kalanchoe genus, supporting future studies regarding other unexplored research areas. The need to conduct further studies to confirm the popular uses and biological activities of bioactive compounds is also highlighted.

Assessing radical scavenging capacity of Sempervivum tectorum L. leaf extracts: An integrated high-performance thin-layer chromatography/in silico/chemometrics approach.

High-Performance Thin-Layer Chromatography (HPTLC)-radical scavenging capacity (RSC) assays are standard techniques for the separation and identification of antioxidants from complex mixtures. HPTLC coupled with DPPH· visualization of chromatograms allows for the detection of individual antioxidants. However, other HPTLC-RSC assays that recognize compounds exhibiting different mechanisms of radical-scavenging activity are rarely reported. In this study, we developed an integrated approach that combines five HPTLC-RSC assays, principal component analysis (PCA) and quantum chemical calculations to assess the antioxidant capacity of Sempervivum tectorum L. leaf extracts. Two HPTLC assays - potassium hexacyanoferrate(III) total reducing power assay (TRP) and total antioxidant capacity by phosphomolybdenum method (TAC) - were developed for the first time. The method supports a more in-depth study of the RSC of natural products, as it compares the radical scavenging fingerprints of S. tectorum leaf extracts and recognizes differences in their individual bioactive constituents. Kaempferol, kaempferol 3-O-glucoside, quercetin 3-O-glucoside, caffeic acid, and gallic acid were identified as the compounds that discriminate HPTLC-RSC assays according to their mechanism of action and capture the similarities between 20 S. tectorum samples. Additionally, DFT calculations on M06-2X/6-31+G(d,p) level were applied to map thermodynamic feasibility of hydrogen atom transfer (HAT) and single electron transfer (SET) mechanisms of the identified compounds. Based on experimental and theoretical results, a combination of HPTLC-ABTS and HPTLC-TAC assays were proposed as the optimal method for mapping the antioxidants from S. tectorum. This study represents a step forward in identifying and quantifying individual antioxidants from complex food and natural product matrices in a more rational manner.

Genomic data provide a robust phylogeny backbone for Rhodiola L. (Crassulaceae) and reveal extensive reticulate evolution during its rapid radiation.

The Tibetan Plateau and adjacent mountain regions (TP; including the Tibetan Plateau, Himalaya, Hengduan Mountains and Mountains of Central Asia) harbor great biodiversity, some lineages on which may have undergone rapid radiations. However, only a few studies have investigated the evolutionary pattern of such diversification in depth using genomic data. In this study, we reconstructed a robust phylogeny backbone of Rhodiola, a lineage that may have undergone rapid radiation in the TP, using Genotyping-by-sequencing data, and conducted a series of gene flow and diversification analyses. The concatenation and coalescent-based methods yield similar tree topologies, and five well-supported clades were revealed. Potential gene flow and introgression events were detected, both between species from different major clades and closely related species, suggesting pervasive hybridization and introgression. An initial rapid and later slowdown of the diversification rate was revealed, indicating niche filling. Molecular dating and correlation analyses showed that the uplift of TP and global cooling in the mid-Miocene might have played an important role in promoting the rapid radiation of Rhodiola. Our work demonstrates that gene flow and introgression might be an important contributor to rapid radiation possibly by quickly reassembling old genetic variation into new combinations.

Genetic Diversity in Natural Populations of Rhodiola Species of Different Adaptation Strategies.

Representatives of the Crassulaceae family's genus Rhodiola are succulents, making them distinctive in a changing environment. One of the most significant tools for analyzing plant resources, including numerous genetic processes in wild populations, is the analysis of molecular genetic polymorphism. This work aimed to look at the polymorphisms of allelic variations of the superoxide dismutase (SOD) and auxin response factor (ARF) gene families, as well as the genetic diversity of five Rhodiola species, using the retrotransposons-based fingerprinting approach. The multi-locus exon-primed intron-crossing (EPIC-PCR) profiling approach was used to examine allelic variations in the SOD and ARF gene families. We implemented the inter-primer binding site (iPBS) PCR amplification technique for genome profiling, which demonstrated a significant level of polymorphism in the Rhodiola samples studied. Natural populations of Rhodiola species have a great capacity for adaptation to unfavorable environmental influences. The genetic variety of wild populations of Rhodiola species leads to their improved tolerance of opposing environmental circumstances and species evolutionary divergence based on the diversity of reproductive systems.

A revision of the Oriental antlion genera Indoleon Banks, 1913, Indophanes Banks, 1940 and Yunleon Yang, 1986 (Neuroptera: Myrmeleontidae: Nemoleontinae).

This paper deals with three Oriental genera of the antlion subfamily Nemoleontinae. The validity of two junior synonyms of Indoleon Banks, 1913, i.e., Indophanes Banks, 1940 and Yunleon Yang, 1986, are formally restored. Indoleon is treated as a monotypic genus represented by Indol. tacitus (Walker, 1860). Indophanes comprises eight species, including five known species: Indop. audax (Walker, 1853), Indop. barbarus (Walker, 1853), Indop. infestus (Walker, 1853), Indop. sinensis Banks, 1940, and Indop. vartianorum Hölzel, 1972; two new species: Indop. amicus sp. nov. and Indop. nanliae sp. nov.; and a new combination: Indop. fuscilomus (Yang, 1986) comb. nov. Yunleon comprises two species: Y. fluctosus Yang, 1988 and Y. longicorpus Yang, 1986. We also restore the validity of a junior synonym of Indop. audax (Walker, 1853), i.e., Myrmeleon gravis Walker, 1853, and transfer it to Distoleon Banks, 1910, i.e., D. gravis stat. rev. et comb. nov. Besides, we describe the larval morphology and biology of Indop. nanliae sp. nov. and Indop. sinensis Banks, 1940. It is the first record of the immature stage of Indophanes. A key to species of these three genera is provided. Meanwhile, we discuss the taxonomic position of the aforementioned three genera and their related genera, i.e., Paraglenurus van der Weele, 1909 and Thaumatoleon Esben-Petersen, 1923, and we consider that they should be placed into Megistopini rather than Nemoleontini.

Inter- and intra-island speciation and their morphological and ecological correlates in Aeonium (Crassulaceae), a species-rich Macaronesian radiation.

BACKGROUND AND AIMS: The most species-rich and ecologically diverse plant radiation on the Canary Islands is the Aeonium alliance (Crassulaceae). In island radiations like this, speciation can take place either within islands or following dispersal between islands. Aiming at quantifying intra- and inter-island speciation events in the evolution of Aeonium, and exploring their consequences, we hypothesized that (1) intra-island diversification resulted in stronger ecological divergence of sister lineages, and that (2) taxa on islands with a longer history of habitation by Aeonium show stronger ecological differentiation and produce fewer natural hybrids. METHODS: We studied the biogeographical and ecological setting of diversification processes in Aeonium with a fully sampled and dated phylogeny inferred using a ddRADseq approach. Ancestral areas and biogeographical events were reconstructed in BioGeoBEARS. Eleven morphological characters and three habitat characteristics were taken into account to quantify the morphological and ecological divergence between sister lineages. A co-occurrence matrix of all Aeonium taxa is presented to assess the spatial separation of taxa on each island. KEY RESULTS: We found intra- and inter-island diversification events in almost equal numbers. In lineages that diversified within single islands, morphological and ecological divergence was more pronounced than in lineages derived from inter-island diversification, but only the difference in morphological divergence was significant. Those islands with the longest history of habitation by Aeonium had the lowest percentages of co-occurring and hybridizing taxon pairs compared with islands where Aeonium arrived later. CONCLUSIONS: Our findings illustrate the importance of both inter- and intra-island speciation, the latter of which is potentially sympatric speciation. Speciation on the same island entailed significantly higher levels of morphological divergence compared with inter-island speciation, but ecological divergence was not significantly different. Longer periods of shared island habitation resulted in the evolution of a higher degree of spatial separation and stronger reproductive barriers.

Echeveria Pallida: Inhibiting Adhesion of Fibroblasts From Pterygium and Neutrophil Extracellular Traps Production.

Context: Pterygium, meaty eyes, is a disease that produces a triangular, conjunctival-epithelial, neovascularized overgrowth covering the cornea, which can cause vision loss. Histological characterization of Pterygium reveals the presence of proliferating fibroblasts (FBs) that remodel the extracellular matrix, with infiltration of immune cells, causing chronic inflammation. The fresh juice of Echeveria pallida E. Walther (Crassulaceae), mechanically extracted from the leaves, can be used to lubricate the eyes and remove Pterygium, even in advanced, degenerative ocular disease. Objective: This study aimed to explore the healing mechanisms of an ethanolic extract of E. pallida on pterygium-derived FBs, lymphocytes, and neutrophils. Design: The research team designed an in-vitro study. Primary cultures of FBs were obtained from fresh, surgical pterygium tissues, and neutrophils and mononuclear cells were purified from the peripheral blood of healthy donors. Intervention: An ethanolic extract of E. pallida was evaluated at 30, 50, 80, 100, 200, and 300 µg/mL-the intervention groups-for viability and proliferation of FBs and lymphocytes. The study included a negative control with no extract, and a positive control, Mitomycin C (MMC), used as a FB proliferation inhibitor and anti-inflammatory. Because some reports have suggested that DMSO at low concentrations can stimulate or inhibit lymphocyte proliferation depending on the cell type, the study also included a DMSO control. Outcome Measures: The measures included an analysis of E. pallida's effects on the proliferation and viability of FBs, the proliferation of human lymphocytes, and human neutrophil extracellular traps (NETs) production. NETs were induced using biochemical and microbiological stimuli-phorbol myristate acetate (PMA), hypochlorous acid (HOCl), Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans-through fluorescence microscopy. Results: The ethanolic extract didn't affect the viability or proliferation of pterygium-derived FBs and human blood lymphocytes, but it showed significant inhibitory activity, from 100 µg/mL, on FB adhesion and the production of NETs. Conclusion: The study found scientific evidence that supports the effects of an extract of the medicinal plant E. pallida in inhibiting the adhesion of FBs derived from human pterygium and NET production.

Flavonoids from Sedum japonicum subsp. oryzifolium (Crassulaceae).

Twenty-two flavonoids were isolated from the leaves and stems of Sedum japonicum subsp. oryzifolium (Crassulaceae). Of these compounds, five flavonoids were reported in nature for the first time, and identified as herbacetin 3-O-xyloside-8-O-glucoside, herbacetin 3-O-glucoside-8-O-(2'''-acetylxyloside), gossypetin 3-O-glucoside-8-O-arabinoside, gossypetin 3-O-glucoside-8-O-(2'''-acetylxyloside) and hibiscetin 3-O-glucoside-8-O-arabinoside via UV, HR-MS, LC-MS, acid hydrolysis and NMR. Other seventeen known flavonoids were identified as herbacetin 3-O-glucoside-8-O-arabinoside, herbacetin 3-O-glucoside-8-O-xyloside, gossypetin 3-O-glucoside-8-O-xyloside, quercetin, quercetin 3-O-glucoside, quercetin 3-O-xylosyl-(1→2)-rhamnoside-7-O-rhamnoside, quercetin 3-O-rhamnoside-7-O-glucoside, kaempferol, kaempferol 3-O-glucoside, kaempferol 7-O-rhamnoside, kaempferol 3,7-di-O-rhamnoside, kaempferol 3-O-glucoside-7-O-rhamnoside, kaempferol 3-O-glucosyl-(1→2)-rhamnoside-7-O-rhamnoside, kaempferol 3-O-xylosyl-(1→2)-rhamnoside, kaempferol 3-O-xylosyl-(1→2)-rhamnoside-7-O-rhamnoside, myricetin 3-O-glucoside and cyanidin 3-O-glucoside. Some flavonol 3,8-di-O-glycosides were found in Sedum japonicum subsp. oryzifolium as major flavonoids in this survey. They were presumed to be the diagnostic flavonoids in the species. Flavonoids were reported from S. japonicum for the first time.

Anther Wall Formation in Aeonium balsamiferum and A. ciliatum (Crassulaceae).

Similarities (features of the family Crassulaceae) and differences in anther development and structure were observed between Aeonium balsamiferum and A. ciliatum. Microsporangia are rounded in A. ciliatum and elongate oval in A. balsamiferum. The septum between microsporangia is consequently longer in A. ciliatum and extremely short in A. balsamiferum. In the latter, pollen can spill out through both the stomium and the apical pore (the phenomenon first discovered in the family Crassulaceae), and both modes of theca opening occur simultaneously, suggesting a greater specialization for pollination for the A. balsamiferum anther. The outer tapetum is single-layered in both species, but its cells are mononuclear in A. ciliatum and binuclear in A. balsamiferum. The inner tapetum is usually single-layered and irregularly two-layered in rare cases; its cells increase in size by a factor of 2 in A. ciliatum and 3-4 in A. balsamiferum (resembling papilloid cells in structure in the latter). Similarities in anther structure were observed between the genera Aeonium and Sedum. In both genera, the anther was isobilateral (on a transverse section) and had an epiconnective, a four-beamed connective, and an annular fibrous layer on the inner side of the connective in the lower region, where there is no fusion of the anther and filament; pollen grains were of the same structural type: tricolporate with a striated surface. The findings were consistent with the cladistic reconstructions that put species of the polyphyletic genus Sedum in the tribes Aeonieae, Semperviveae, and Sedeae. Aeonium balsamiferum and A. ciliatum fall into the Aeonium clade and occupy an intermediate position between the Telephium clade (Sedum kamtschaticum) and the Acre clade (S. palmeri).

Plastome evolution of Aeonium and Monanthes (Crassulaceae): insights into the variation of plastomic tRNAs, and the patterns of codon usage and aversion.

MAIN CONCLUSION: This study reported 13 new plastomes from Aeonium and Monanthes, and observed new markers for phylogeny and DNA barcoding, such as novel tRNA structures and codon usage bias and aversion. The Macaronesian clade of Crassulaceae consists of three genera: Aichryson, with about 15 species; Monanthes, with about 10 species; Aeonium, with about 40 species. Within this clade, Aeonium, known as "the botanical equivalent of Darwin's finches", is regarded as an excellent model plant for researching adaptive evolution. Differing from the well-resolved relationships among three genera of the Macaronesian clade, the internal branching patterns within the genus Aeonium are largely unclear. In this study, we first reported 13 new plastomes from genus Aeonium and the closely related genus Monanthes. We further performed comprehensive analyses of the plastomes, with focuses on the secondary structures of pttRNAs and the patterns of codon usage and aversion. With a typical circular and quadripartite structure, the 13 plastomes ranged from 149,900 to 151,030 bp in size, and the unique pattern in IR junctions might become a family-specific marker for Crassulaceae species. Surprisingly, the π values of plastomes from Monanthes were almost twice those from Aeonium. Most importantly, we strongly recommend that highly polymorphic regions, novel putative pttRNA structures, patterns of codon usage bias and aversion derived from plastomes might have phylogenetic implications, and could act as new markers for DNA barcoding of plants. The results of phylogenetic analyses strongly supported a clear internal branching pattern in Macaronesian clade (represented by Aeonium and Monanthes), with higher nodal support values. The findings reported here will provide new insights into the variation of pttRNAs, and the patterns of codon usage and aversion of the family Crassulaceae.

Embryogenesis in Crassulaceae: structural aspect of suspensor development / Embriogeneza u Crassulaceae: strukturalny aspekt rozwoju wieszadelka.

The suspensor in the majority of angiosperms is an evolutionally conserved embryonic organ functioning as a conduit that connects ovule tissues with the embryo proper for nutrients and growth regulators flux. In this article the present knowledge on the embryo-suspensor ultrastructure and function in representatives of Crassulaceae genera: Sedum, Jovibarba, Sempervivum, Aeonium, Monanthes, Aichryson and Echeveria. The role of the suspensor in the transport of nutrients from the tissues of the ovule to the proper embryo is confirmed by the structure of the basal cell, especially the nature of the micropylar part of its wall, the "transfer wall". The basal suspensor cell is a site of intense metabolic activity. The special attention is paid to the plasmodesmata. The correlation between types of suspensors and structure of plasmodesmata was investigated. Final conclusions are given and the presented data summarized.

How challenging RADseq data turned out to favor coalescent-based species tree inference. A case study in Aichryson (Crassulaceae).

Analysing multiple genomic regions while incorporating detection and qualification of discordance among regions has become standard for understanding phylogenetic relationships. In plants, which usually have comparatively large genomes, this is feasible by the combination of reduced-representation library (RRL) methods and high-throughput sequencing enabling the cost effective acquisition of genomic data for thousands of loci from hundreds of samples. One popular RRL method is RADseq. A major disadvantage of established RADseq approaches is the rather short fragment and sequencing range, leading to loci of little individual phylogenetic information. This issue hampers the application of coalescent-based species tree inference. The modified RADseq protocol presented here targets ca. 5,000 loci of 300-600nt length, sequenced with the latest short-read-sequencing (SRS) technology, has the potential to overcome this drawback. To illustrate the advantages of this approach we use the study group Aichryson Webb & Berthelott (Crassulaceae), a plant genus that diversified on the Canary Islands. The data analysis approach used here aims at a careful quality control of the long loci dataset. It involves an informed selection of thresholds for accurate clustering, a thorough exploration of locus properties, such as locus length, coverage and variability, to identify potential biased data and a comparative phylogenetic inference of filtered datasets, accompanied by an evaluation of resulting BS support, gene and site concordance factor values, to improve overall resolution of the resulting phylogenetic trees. The final dataset contains variable loci with an average length of 373nt and facilitates species tree estimation using a coalescent-based summary approach. Additional improvements brought by the approach are critically discussed.

Evidence linking life-form to a major shift in diversification rate in Crassula.

PREMISE: Plants have evolved different ecological strategies in response to environmental challenges, and a higher lability of such strategies is more common in plant groups that adapt to various niches. Crassula (Crassulaceae), occurring in varied mesic to xeric habitats, exhibits a remarkable diversity of life-forms. However, whether any particular life-form trait has shaped species diversification in Crassula has remained unexplored. This study aims to investigate diversification patterns within Crassula and identify potential links to its life-form evolution. METHODS: A phylogenetic tree of 140 Crassula taxa was reconstructed using plastid and nuclear loci and dated based on the nuclear DNA information only. We reconstructed ancestral life-form characters to estimate the evolutionary trends of ecophysiological change, and subsequently estimated net diversification rates. Multiple diversification models were applied to examine the association between certain life-forms and net diversification rates. RESULTS: Our findings confirm a radiation within Crassula in the last 10 million years. A configuration of net diversification rate shifts was detected, which coincides with the emergence of a speciose lineage during the late Miocene. The results of ancestral state reconstruction demonstrate a high lability of life-forms in Crassula, and the trait-dependent diversification analyses revealed that the increased diversification is strongly associated with a compact growth form. CONCLUSIONS: Transitions between life-forms in Crassula seem to have driven adaptation and shaped diversification of this genus across various habitats. The diversification patterns we inferred are similar to those observed in other major succulent lineages, with the most-speciose clades originating in the late Miocene.

Purification, Structural Characterization, and Anti-Inflammatory Effects of a Novel Polysaccharide Isolated from Orostachys fimbriata.

The present study elucidated the structural characteristics and anti-inflammatory activity of a novel polysaccharide isolated from Orostachys fimbriata, which is a traditional Chinese medicinal plant. O. fimbriata polysaccharide (OFP) was extracted and subsequently purified by chromatography using a DEAE cellulose-52 and Sephadex G-75 column. The molecular weight was determined as 6.2 kDa. HPGPC and monosaccharide composition analysis revealed a homogeneous polysaccharide containing only Glc. Chromatography and spectral analysis showed that the possible chemical structure consisted of →4)-α-Glcp-(1→ and a small quantity of →4,6)-ß-Glcp-(1→ in the main chain and →6)-ß-Glcp-(1→, α-Glcp-(1→, and ß-Glcp-(1→ in the side chain. Morphological analysis using scanning electron microscopy (SEM) and atomic force microscopy (AFM) indicated that OFP had a multi-branched structure, and the sugar chain molecules of polysaccharide appeared aggregated. OFP was found to exhibit anti-inflammatory activity by reducing the secretion of inflammatory factors in RAW264.7 cells and by decreasing the extent of xylene-induced ear swelling in mice.

Graptopetalum paraguayense Extract Ameliorates Proteotoxicity in Aging and Age-Related Diseases in Model Systems.

Declines in physiological functions are the predominant risk factors for age-related diseases, such as cancers and neurodegenerative diseases. Therefore, delaying the aging process is believed to be beneficial in preventing the onset of age-related diseases. Previous studies have demonstrated that Graptopetalum paraguayense (GP) extract inhibits liver cancer cell growth and reduces the pathological phenotypes of Alzheimer's disease (AD) in patient IPS-derived neurons. Here, we show that GP extract suppresses ß-amyloid pathology in SH-SYS5Y-APP695 cells and APP/PS1 mice. Moreover, AMP-activated protein kinase (AMPK) activity is enhanced by GP extract in U87 cells and APP/PS1 mice. Intriguingly, GP extract enhances autophagy in SH-SYS5Y-APP695 cells, U87 cells, and the nematode Caenorhabditis elegans, suggesting a conserved molecular mechanism by which GP extract might regulate autophagy. In agreement with its role as an autophagy activator, GP extract markedly diminishes mobility decline in polyglutamine Q35 mutants and aged wild-type N2 animals in C. elegans. Furthermore, GP extract significantly extends lifespan in C. elegans.