Chloroplast isolation protocols have been extensively developed for various species of plants, particularly model organisms with easily manipulable physical characteristics. However, succulent plants, such as Agave angustifolia Haw., which possess adaptations for arid environments like the Crassulacean acid metabolism (CAM) and a thicker cuticle, have received less attention, resulting in a potential knowledge gap. This chapter presents a specialized protocol focusing on isolating chloroplast from A. angustifolia, a species exhibiting adaptations to arid conditions and holding ecological and economic significance due to its role in producing bacanora and mezcal beverages. By successfully isolating chloroplast from A. angustifolia plant growth in ex vitro and in vitro conditions, this protocol enables comprehensive future analyses to elucidate metabolic processes and explore potential applications in related species. Consequently, this research aims to bridge this knowledge gap in chloroplast isolation for succulent plants, providing new insights for future investigations in the field.

Chile\'s distinctive flora, geographical isolation, and complex topography collectively contribute to a notable endemic species diversity, particularly within central regions identified as critical areas for biodiversity conservation. The cactus genus Eriosyce, as currently circumscribed, encompasses seven sections, with Eriosycesect.Horridocatus presenting a notably complex species group. This study investigates the E.curvispina complex, a member of the Notocacteae tribe common in central Chile, by incorporating new populations and examining phylogenetic relationships using four plastid and one nuclear molecular marker. The phylogenetic analysis of sampled individuals identified nine independent lineages, each warranting recognition at the species rank. Despite minimal morphological differences among taxa, morphological characters were utilized to support and stabilize the DNA-based phylogenetic hypothesis. The results highlight the high taxonomic diversity in these cactus lineages and have implications for the classification of the E.curvispina complex, including new combinations and proposals of conservation status.
ResumenLa flora distintiva de Chile, su aislamiento geográfico y topografía compleja contribuyen colectivamente a una notable diversidad de especies endémicas, particularmente dentro de las regiones centrales identificadas como áreas críticas para la conservación de la biodiversidad. El género de cactus Eriosyce, tal como está circunscrito actualmente, abarca siete secciones, presentando Eriosycesect.Horridocatus un grupo de especies notablemente complejo. Este estudio investiga el complejo E.curvispina, un miembro de la tribu Notocacteae común en Chile central, incorporando nuevas poblaciones y examinando las relaciones filogenéticas utilizando cuatro marcadores moleculares del cloroplasto y uno nuclear. El análisis filogenético de las poblaciones muestreadas identificó nueve linajes independientes, cada uno mereciendo reconocimiento a nivel de especie. A pesar de las mínimas diferencias morfológicas entre los taxones, se utilizaron caracteres morfológicos para apoyar y estabilizar la filogenia basada en ADN. Los resultados resaltan la alta diversidad taxonómica en estos linajes de cactus y tienen implicaciones para la clasificación del complejo E.curvispina, incluyendo nuevas combinaciones y propuestas de estado de conservación.

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.

Studies of litter decomposition in salt marshes have been mainly focused on the measurement of decomposition rates, being litter quality, the type of microbial decomposers and their extracellular enzyme activity, rarely considered. Moreover, most of these studies have been conducted in Poaceae and Cyperaceae species, being scarce the literature on Chenopodiaceae species, which are abundant in Mediterranean salt marshes. Here we analyse the litter decomposition process of two Chenopodiaceae (Sarcocornia fruticosa and Halimione portulacoides) and one Poaceae (Elytrigia atherica) species, belonging S. fruticosa to a halophilous scrub habitat and the other two to a salt meadow habitat of a Mediterranean salt marsh. For each species, we analysed litter decomposition rates, litter quality, fungal and bacterial biomass and potential extracellular enzymes activities. In order to embrace the spatial heterogeneity, two zones were considered within each habitat. Litter of E. atherica decomposed 7- and 13-fold slower than those of S. fruticosa and H. portulacoides, respectively, suggesting that this species is the one that would favour most the carbon sequestration into the soil. The different decomposition rates would be explained by the higher initial lignin and cellulose content of E. atherica rather than by the initial carbon and nitrogen content and C/N ratio. Moreover, enzyme efficiency, compared to enzyme activity, better contributes to explain the different decomposition rates observed. Bacteria dominated throughout the litter decomposition process regardless the species, but fungi increased their relevance in the later stages, when the relative lignin litter content increased. Litter decomposition was affected by microhabitat spatial differences, although the responses depended on the species. Hence, flooding (in the habitat of S. fruticosa) or soil texture (in the habitat of E. atherica and H. portulacoides) might have modulated the decomposition process, being H. portulacoides the most sensitive species to the spatial differences of the salt meadow habitat.

The leaf calorific value (LCV) is an important trait that indicates how efficiently a plant utilizes natural resources to capture energy. However, little is known about the LCV characteristics of plants in arid and hyper-arid environments. To investigate the spatial patterns and variations in LCV of desert plants and their possible causes, we collected 343 leaf samples of 52 species along a 1000-km transect in the desert area of northwestern China. We analyzed the gross calorific value (GCV), ash-free calorific value (AFCV), carbon content (CC), nitrogen content (NC), and ash content (AC) of the leaves. The mean leaf GCV and AC were 16.2 kJ g-1 (range from 8.9 to 20.1 kJ g-1), and 189.8 mg g-1 (range from 61.5 to 495.1 mg g-1) respectively, which differ significantly from the values for plants growing in more humid areas of China. Succulence was the dominant trait that drove the differences in leaf GCV and AFCV among plant functional groups. Succulent plants had significantly lower leaf GCV and AFCV, and significantly higher AC, than non-succulent plants, indicating that the investment of energy for succulent plants in response to drought stress may be lower than that for non-succulent plants. Among the biological factors that affected LCV, the CC and AC were the main determinants of leaf GCV, whereas CC and NC were the main determinants of leaf AFCV. Drought stress is an environmental constraint that has a direct negative effect on both leaf GCV and AFCV, but its contribution may be weaker than phylogenetic effects. Our results suggest that LCV is a useful leaf trait that can be used to evaluate plant-environment interactions from an energy perspective.

Succulent plants represent a large functional group of drought-resistant plants that store water in specialized tissues. Several co-adaptive traits accompany this water-storage capacity to constitute the succulent syndrome. A widely reported anatomical adaptation of cell walls in succulent tissues allows them to fold in a regular fashion during extended drought, thus preventing irreversible damage and permitting reversible volume changes. Although ongoing research on crop and model species continuously reports the importance of cell walls and their dynamics in drought resistance, the cell walls of succulent plants have received relatively little attention to date, despite the potential of succulents as natural capital to mitigate the effects of climate change. In this review, we summarize current knowledge of cell walls in drought-avoiding succulents and their effects on tissue biomechanics, water relations, and photosynthesis. We also highlight the existing knowledge gaps and propose a hypothetical model for regulated cell wall folding in succulent tissues upon dehydration. Future perspectives of methodological development in succulent cell wall characterization, including the latest technological advances in molecular and imaging techniques, are also presented.

Best known as low maintenance houseplants, sansevierias are a diverse group of flowering plants native to Africa, Madagascar, the Arabian Peninsula, and the Indian subcontinent. Traditionally recognised as a distinct genus, Sansevieria was recently merged with the larger genus Dracaena based on molecular phylogenetic data. Within the Sansevieria Clade of Dracaena, taxonomic uncertainties remain despite attempts to unravel the relationships between the species. To investigate the evolutionary relationships, morphological evolution and biogeographical history in the group, we aim to reconstruct a robust dated phylogenetic hypothesis. Using genome skimming, a chloroplast genome (cpDNA) dataset and a nuclear ribosomal (nrDNA) dataset were generated. The sampling included representatives of all sections and informal groups previously described in Sansevieria based on morphology. Analysis of the cpDNA dataset using a maximum likelihood approach resulted in a well-supported phylogeny. The time-calibrated phylogeny indicated a recent radiation with five main clades emerging in the Pliocene. Two strongly supported clades align with previously defined groups, i.e., Sansevieria section Dracomima, characterised by the Dracomima-type inflorescence, and the Zeylanica informal group, native to the Indian subcontinent. Other previously defined groups were shown to be polyphyletic; a result of convergent evolution of the identifying characters. Switches between flat and cylindrical leaves occurred multiple times in the evolution of the Sansevieria Clade. Similarly, the Cephalantha-type inflorescence has originated multiple times from an ancestor with a Sansevieria-type inflorescence. Analysis of the nrDNA dataset resulted in a phylogenetic hypothesis with low resolution, yet it supported the same two groups confirmed by the cpDNA dataset. This study furthers our understanding of the evolution of the Sansevieria Clade, which will benefit taxonomic and applied research, and aid conservation efforts.

UNASSIGNED: Plants have evolved different ecological strategies in response to environmental challenges, and a higher lability of such strategies is more common in plant groups adapting 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 reveal 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. This article is protected by copyright. All rights reserved.

The genus Mammillaria occupies diverse habitats and exhibits diverse growth patterns and a large range of morphologies. Most of the species of this genus are used as ornamental plants and are subject to mass habitat loss. Due to these factors, they are being submitted to selective pressure that might affect conservational efforts and management plans. We obtained the 133 gene chloroplast genome as part of the project of sequencing the complete genome of pincushion cactus, including 88 protein-coding genes, 8 rRNA genes, and 37 tRNA genes. The phylogenetic tree indicates the pincushion cactus is a sister species of M. supertexta and M. huitzilopochtli.

We re-examined the taxonomic status of plants treated as Sedum formosanum (Crassulaceae) from Miyako-jima Island of the Ryukyu Islands, Japan, using morphological comparison and molecular phylogenetic analyses with related species. In morphology, plants from Miyako-jima Island bore a close resemblance to the other plants of S. formosanum, but differed in being perennial, polycarpic, and having lateral axillary branches. Molecular analyses based on ITS of nrDNA and six regions of cpDNA sequencing indicated that the Miyako-jima plants formed a distinct subclade. This subclade was part of a polytomy with three other subclades comprising nine taxa endemic to Taiwan and S. formosanum from other areas, including the type locality. Therefore, we propose and describe the Miyako-jima plants as a new subspecies, Sedum formosanum subsp. miyakojimense.