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Clethra barbinervis Sieb. et Zucc. is a tree species that grows naturally at several mine sites and seems to be tolerant of high concentrations of heavy metals, such as Cu, Zn, and Pb. The purpose of this study is to clarify the mechanism(s) underlying this species' ability to tolerate the sites' severe heavy-metal pollution by considering C. barbinervis interaction with root fungal endophytes. We measured the heavy metal concentrations of root-zone soil, leaves, branches, and fine roots collected from mature C. barbinervis at Hitachi mine. We isolated fungal endophytes from surface-sterilized root segments, and we examined the growth, and heavy metal and nutrient absorption of C. barbinervis seedlings growing in sterilized mine soil with or without root fungal endophytes. Field analyses showed that C. barbinervis contained considerably high amounts of Cu, Zn, and Pb in fine roots and Zn in leaves. The fungi, Phialocephala fortinii, Rhizodermea veluwensis, and Rhizoscyphus sp. were frequently isolated as dominant fungal endophyte species. Inoculation of these root fungal endophytes to C. barbinervis seedlings growing in sterilized mine soil indicated that these fungi significantly enhanced the growth of C. barbinervis seedlings, increased K uptake in shoots and reduced the concentrations of Cu, Ni, Zn, Cd, and Pb in roots. Without root fungal endophytes, C. barbinervis could hardly grow under the heavy-metal contaminated condition, showing chlorosis, a symptom of heavy-metal toxicity. Our results indicate that the tree C. barbinervis can tolerate high heavy-metal concentrations due to the support of root fungal endophytes including P. fortinii, R. veluwensis, and Rhizoscyphus sp. via growth enhancement, K uptake promotion and decrease of heavy metal concentrations.

Clethra fargesii, an essential ecological and endemic woody plant of the genus Clethra in Clethraceae, is widely distributed in Central China. So far, there have been a paucity of studies on its chloroplast genome. In the present study, we sequenced and assembled the complete chloroplast genome of C. fargesii. We also analyzed the chloroplast genome features and compared them to Clethra delavayi and other closely related species in Ericales. The complete chloroplast genome is 157,486 bp in length, including a large single-copy (LSC) region of 87,034 bp and a small single-copy (SSC) region of 18,492 bp, separated by a pair of inverted repeat (IR) regions of 25,980 bp. The GC content of the whole genome is 37.3%, while those in LSC, SSC, and IR regions are 35.4%, 30.7%, and 43.0%, respectively. The chloroplast genome of C. fargesii encodes 132 genes in total, including 87 protein-coding genes (PCGs), 37 tRNA genes, and eight rRNA genes. A total of 26,407 codons and 73 SSRs were identified in C. fargesii chloroplast genome. Additionally, we postulated and demonstrated that the structure of the chloroplast genome in Clethra species may present evolutionary conservation based on the comparative analysis of genome features and genome alignment among eight Ericales species. The low Pi values revealed evolutionary conservation based on the nucleotide diversity analysis of chloroplast genome in two Clethra species. The low selection pressure was shown by a few positively selected genes by adaptive evolution analysis using 80 coding sequences (CDSs) of the chloroplast genomes of two Clethra species. The phylogenetic tree showed that Clethraceae and Ericaceae are sister clades, which reconfirm the previous hypothesis that Clethra is highly conserved in the chloroplast genome using 75 CDSs of chloroplast genome among 40 species. The genome information and analysis results presented in this study are valuable for further study on the intraspecies identification, biogeographic analysis, and phylogenetic relationship in Clethraceae.

Aims This study reveals the accumulation properties of Co from viewpoints of the analogy with Ni and also the variation of the other essential micro elements in the Co accumulator tree, Clethra barbinervis. Methods Seedlings of C. barbinervis were grown in single and mixed treatments in the rhizosphere containing Co, Ni, or both at three different concentrations (5, 50, and 500 μM). In roots, stems, and leaves, the concentrations of Cu, Fe, Mn, and Zn, together with Co and Ni, were determined. Results Biomass was reduced in plants treated with 500 μM Ni, but not when treated with Co. The Co concentration in leaves was higher than those in other tissues, and the maximum Co accumulation in leaves was 2012 μg g−1. Accumulation of Co competed with Zn transport and stimulated Cu transport. Conclusion Our results indicate that C. barbinervis is a Co hyperaccumulator and is more tolerant to Co than to Ni. The Co hyperaccumulation capacity is likely achieved by Zn transport systems involved in root to shoot translocation, and Co accumulation has some competitive and facilitative interactions with the other heavy metals.

Objective Plant carnivory is distributed across the tree of life and has evolved at least six times independently, but sequenced and annotated nuclear genomes of carnivorous plants are currently lacking. We have sequenced and structurally annotated the nuclear genome of the carnivorous Roridula gorgonias and that of a non-carnivorous relative, Madeira’s lily-of-the-valley-tree, Clethra arborea , both within the Ericales. This data adds an important resource to study the evolutionary genetics of plant carnivory across angiosperm lineages and also for functional and systematic aspects of plants within the Ericales. Results Our assemblies have total lengths of 284 Mbp ( R. gorgonias ) and 511 Mbp ( C. arborea ) and show high BUSCO scores of 84.2% and 89.5%, respectively. We used their predicted genes together with publicly available data from other Ericales’ genomes and transcriptomes to assemble a phylogenomic data set for the inference of a species tree. However, groups of orthologs showed a marked absence of species represented by a transcriptome. We discuss possible reasons and caution against combining predicted genes from genome- and transriptome-based assemblies.

Key messageThe accumulation and tolerance mechanisms for Co are clearly different from those for Ni in the leaves of C. barbinervis in terms of both the distribution and speciation.Clethra barbinervis is a Co-hyperaccumulating tree and also accumulates Ni at high concentrations. The mechanism and role of accumulation in tree physiology remains unclear. The aim of this study was to determine the localization and speciation of Co and Ni in the leaves of C. barbinervis to reveal the mechanisms behind its tolerance to high concentrations of these elements. C. barbinervis seedlings were grown for 3 years under treatments with Co or Ni in the rhizosphere. X-ray fluorescence (XRF) and X-ray absorption near edge structure (XANES) analyses were then used to evaluate the distribution and chemical states of Co, Ni, and S in the adaxial leaf epidermis. In addition, the treated leaves were cut into several parts according to the XRF imaging results on Co or Ni, and the concentrations of elements, sulfate, and organic acids were determined in each part by chemical analyses. XRF images showed that Co was present at the tip of the leaf at a high concentration, whereas Ni was mainly distributed around the leaf edge. Results of chemical analyses on leaf parts containing Co or Ni indicated that sulfate acts as a counter ion for Co and that Ni combined with succinic and/or oxalic acid. In addition, XANES analysis showed that sulfate tended to be reduced and glutathione was generated in the tip of the leaf. Our results indicate that C. barbinervis distinguishes Co and Ni and translocates them to different parts of the leaf.

Honey produced by native stingless bees in the Araucaria Forest region of Rio Grande do Sul, southern Brazil, specifically the municipality of Cambará do Sul, is highly valued for its characteristic white color, floral odor and taste. In this study, we investigated the botanical origin of white honey stored in colonies of five Meliponini species of the genera Melipona (n = 3) and Plebeia (n = 2). During the production period of white honey, from January to March, flowers were sampled fortnightly along pre-established trails to identify plants used by bees. For all sampled plant species, exsiccates and pollen reference slides were prepared. Honey samples from stingless bees were processed for extraction and preparation of pollen grains for identification of pollen types. In all analyzed honey samples, pollen grains of Clethra scabra (Clethraceae) were predominant (between 46-94%). Pollen grains from other botanical families, including Myrtaceae, Fabaceae and Melastomataceae were frequently identified in honey samples of the Melipona species, while Cunoniaceae was also found in samples of Plebeia species. In this study, we concluded that Clethra scabra is predominantly used by Meliponini bees in the production of white honey in the municipality of Cambará do Sul. Resumo: Na região da Floresta de Araucária do Rio Grande do Sul, especificamente no município de Cambará do Sul, as abelhas nativas sem ferrão produzem mel característico devido à sua cor branca e gosto apreciado. Neste estudo, investigamos a origem botânica do mel branco armazenado em colônias de cinco espécies de Meliponini dos gêneros Melipona (n = 3) e Plebeia (n = 2). Durante o período de produção do mel branco, de janeiro a março, as flores foram amostradas quinzenalmente ao longo de trilhas pré-estabelecidas, a fim de identificar as espécies utilizadas pelas abelhas. A partir de amostras de plantas foram preparadas exsicatas e lâminas de referência de pólen. As amostras de mel de abelhas sem ferrão foram processadas para extração e preparo dos grãos de pólen para a determinação dos tipos polínicos presentes. Em todas as amostras de méis branco analisadas os grãos de pólen de Clethra scabra (Clethraceae) predominaram (entre 46-94%). Grãos de pólen de outras espécies das famílias botânicas Myrtaceae, Fabaceae e Melastomataceae foram frequentemente identificados nas amostras de méis das espécies de Melipona, enquanto Cunoniaceae também o foi nas amostras das espécies de Plebeia. Neste estudo, nós concluímos que Clethra scabra é predominantemente utilizada pelas abelhas Meliponini na produção de méis branco no município de Cambará do Sul.

Mature leaves of tree seedlings were exposed to high light in four experimental gaps in the Jamaican upper montane rainforest (UMRF). Two of the six species studied were light-demanders: Alchornea latifolia and Clethra occidentalis. Two were gap-favoured: Pittosporum undulatum (an invasive) and Palicourea alpina (a subcanopy shrub). One was intermediate: Hedyosmum arborescens, and one was shade-tolerant: Guarea glabra. After five months, the following significant changes occurred in shade leaves that were exposed to gaps (‘shade-to-gap’ leaves; values as % of those in the pre-gap shade): maximum rate of photosynthesis + 40% (Alchornea), +35% (Clethra), −34% (Pittosporum), +72% (Palicourea); dark respiration +120% (Alchornea), +140% (Clethra), +60% (Pittosporum), +233% (Palicourea), +175% (Hedyosmum), +100% (Guarea); leaf thickness +18% (Alchornea), +18% (Clethra), +14% (Palicourea); leaf mass per unit area +18% (Alchornea), +15% (Pittosporum). Leaves produced in the gaps were (as a percentage of total live leaf number) 74% (Alchornea), 71% (Clethra), 50% (Pittosporum), 71% (Palicourea), 62% (Hedyosmum) and 50% (Guarea). Photosynthetic rates of leaves produced in the gaps were 53–120% higher than ‘shade-to-gap’ leaves. Overall, shade leaves on the three native, more light-demanding species (Alchornea, Clethra and Palicourea) showed photosynthetic acclimation, while the more shade-tolerant species (Hedyosmum and Guarea and Pittosporum undulatum) showed little acclimation in shade-to-gap leaves.

The elevation of altitudinal treelines is generally believed to occur where low mean temperatures during the growing season limit growth and prevent trees from establishing at higher elevations. Accordingly, treelines should move upslope with increasing global temperatures. Contrary to this prediction, tropical treelines have remained stable over the past several decades despite increasing mean temperatures. The observed stability of tropical treelines, coupled with the drastically different temperature profiles between temperate and tropical treelines, suggests that using mean measures of temperature to predict tropical treeline movements during climate change may be overly simplistic. We hypothesize that frost events at tropical treelines may slow climate driven treeline movement by preventing tree recruitment beyond the established forest canopy. To assess this hypothesis, we measured freezing resistance of four canopy-forming treeline species ( Weinmannia fagaroides , Polylepis pauta , Clethra cuneata , and Gynoxys nitida ) at two life stages (juvenile and adult) and during two seasons (warm-wet and cold-dry). Freezing resistances were then compared to microclimatic data to determine if freezing events in the grassland matrix above treeline are too harsh for these forest species. Freezing resistance varied among species and life stages from −5.7°C for juveniles of P. pauta to −11.1°C for juveniles of W. fagaroides . Over a four-year period, the lowest temperatures recorded at 10 cm above ground level in the grasslands above treeline and at treeline itself were −8.9°C and −6.8°C, respectively. Juveniles maintained freezing resistances similar to adults during the coldest parts of the year and ontogenetic differences in freezing resistance were only present during the warm season when temperatures did not represent a significant threat to active plant tissue. These findings support the hypothesis that rare extreme freezing events at and above tropical treelines can prevent recruitment outside of closed canopy forest for some tree species and may significantly slow treeline advancement despite warming mean temperatures. Predictions of treeline shifts under climate change should be reevaluated to include species-specific climatic tolerances and measures of climatic variability.

We analysed the dead-wood characteristics that determine the presence of saproxylic pseudoscorpion species in remnants of riparian cloud forest. We examined 98 dead-wood pieces (70 logs and 28 stumps), and recorded tree species, decaying wood stages, presence of the pseudoscorpion Lustrochernes grossus (Banks, 1893) (Chernetidae) and the Bess beetle Helicus tropicus. In these wood samples we found 24 L. grossus and one chela. We recorded the highest number of individuals in Clethra mexicana (11), followed by Quercus corrugata (6) and Liquidambar styraciflua (6). In Annona cherimola and Trema micrantha, one chela and one female were recorded, respectively. The presence of this pseudoscorpion is likely due to its relationship with the Bess beetle, which coexists in decaying wood. The distribution of L. grossus in dead wood may also be influenced by tree species and stage of decay. Forest fragmentation and the extraction of firewood from the remnant riparian fragments of cloud forest are factors that could jeopardize the saproxylic pseudoscorpion species and other arthropod diversity associated with decaying wood in this threatened ecosystem.

Shredders play a crucial role in litter decomposition in streams. However, in oceanic islands, many streams have low shredder density and richness, and microbes seem to be the main litter decomposers. Here, we evaluate the effects of shredders and aquatic hyphomycetes on litter decomposition in insular streams. Three leaf species differing in physical and chemical characteristics, Alnus glutinosa, Clethra arborea, and Cryptomeria japonica, were enclosed in bags of coarse and fine mesh to allow and avoid macroinvertebrate access to the litter, respectively, and incubated in six streams along a gradient of Limnephilus atlanticus (Trichoptera) density in São Miguel Island. In streams with higher L. atlanticus density, leaf mass loss was higher in coarse than fine mesh bags. However, no difference in litter mass loss was found between bag types in streams with no L. atlanticus, despite the presence of other shredder taxa. These results suggest that when L. atlanticus are present at relatively high densities, they significantly contribute to litter decomposition, while litter decomposition is mainly driven by microbes when L. atlanticus density is low, or they are absent. Moreover, litter decomposition depends on litter quality, with leaves with high nutrient concentration and low concentration of secondary compounds being preferred by shredders.