Impact of trace elements on invasive plants: Attenuated competitiveness yet sustained dominance over native counterparts.

Trace element pollution has emerged as an increasingly severe environmental challenge owing to human activities, particularly in urban ecosystems. In farmlands, invasive species commonly outcompete native species when subjected to trace element treatments, as demonstrated in experiments with individual invader-native pairs. However, it is uncertain if these findings apply to a wider range of species in urban soils with trace elements. Thus, we designed a greenhouse experiment to simulate the current copper and zinc levels in urban soils (102.29 mg kg-1 and 148.32 mg kg-1, respectively). The experiment involved four pairs of invasive alien species and their natural co-existing native species to investigate the effects of essential trace elements in urban soil on the growth and functional traits of invasive and native species, as well as their interspecific relationship. The results showed that adding trace elements weakened the competitiveness of invasive species. Nonetheless, trace element additions did not change the outcome of competition, consistently favoring invasion successfully. Under trace element addition treatments, invasive species and native species still maintained functional differentiation trend. Furthermore, the crown area, average leaf area and leaf area per plant of invasive species were higher than those of native species by 157 %, 177 % and 178 % under copper treatment, and 194 %, 169 % and 188 % under zinc treatment, respectively. Additionally, interspecific competition enhanced the root growth of invasive species by 21 % with copper treatment and 14 % with zinc treatment. The ability of invasive species to obtain light energy and absorb water and nutrients might be the key to their successful invasion.

Genomics insights into flowering and floral pattern formation: regional duplication and seasonal pattern of gene expression in Camellia.

BACKGROUND: The formation and domestication of ornamental traits are influenced by various aspects, such as the recognition of esthetic values and cultural traditions. Camellia japonica is widely appreciated and domesticated around the world mainly due to its rich variations in ornamental traits. Ornamental camellias have a diverse range of resources, including different bud variations from Camellia spp. as well as inter- and intra- specific hybridization. Despite research on the formation of ornamental traits, a basic understanding of their genetics and genomics is still lacking. RESULTS: Here, we report the chromosomal-level reference genome of C. japonica through combining multiple DNA-sequencing technologies and obtain a high-density genetic linkage map of 4255 markers by sequencing 98 interspecific F1 hybrids between C. japonica and C. chekiangoleosa. We identify two whole-genome duplication events in C. japonica: one is a shared ancient γ event, and the other is revealed to be specific to genus Camellia. Based on the micro-collinearity analysis, we find large-scale segmental duplication of chromosome 8, resulting to two copies of the AGAMOUS loci, which may play a key role in the domestication of floral shapes. To explore the regulatory mechanisms of seasonal flowering, we have analyzed year-round gene expression patterns of C. japonica and C. azalea-a sister plant of continuous flowering that has been widely used for cross breeding. Through comparative analyses of gene co-expression networks and annual gene expression patterns, we show that annual expression rhythms of some important regulators of seasonal growth and development, including GIGANTEA and CONSTANS of the photoperiod pathway, have been disrupted in C. azalea. Furthermore, we reveal that the distinctive expression patterns of FLOWERING LOCUS T can be correlated with the seasonal activities of flowering and flushing. We demonstrate that the regulatory module involved in GIGANTEA, CONSTANS, and FLOWERING LOCUS T is central to achieve seasonality. CONCLUSIONS: Through the genomic and comparative genomics characterizations of ornamental Camellia spp., we propose that duplication of chromosomal segments as well as the establishment of gene expression patterns has played a key role in the formation of ornamental traits (e.g., flower shape, flowering time). This work provides a valuable genomic platform for understanding the molecular basis of ornamental traits.

Nitrogen Deposition Effects on Invasive and Native Plant Competition: Implications for Future Invasions.

Nitrogen (N) deposition has increased dramatically in recent decades, which is significantly affecting the invasion and growth of exotic plants. Whether N deposition leads to invasive alien species becoming competitively superior to native species remains to be investigated. In the present study, an invasive species (Oenothera biennis L.) and three co-occurring native species (Artemisia argyi Lévl. et Vant., Inula japonica Thunb., and Chenopodium album L.) were grown in a monoculture (two seedlings of the same species) or mixed culture (one seedling of O. biennis and one seedling of a native species) under three levels of N deposition (0, 6, and 12 g∙m-2∙year-1). Nitrogen deposition had no effect on soil N and P content. Nitrogen deposition enhanced the crown area, total biomass, leaf chlorophyll content, and leaf N to phosphorus ratio in both invasive and native plants. Oenothera biennis dominated competition with C. album and I. japonica due to its high resource acquisition and absorption capacity (greater height, canopy, leaf chlorophyll a to chlorophyll b ratio, leaf chlorophyll content, leaf N content, leaf mass fraction, and lower root-to-shoot ratio). However, the native species A. argyi exhibited competitive ability similar to O. biennis. Thus, invasive species are not always superior competitors of native species; this depends on the identities of the native species. High N deposition enhanced the competitive dominance of O. biennis over I. japonica by 15.45% but did not alter the competitive dominance of O. biennis over C. album. Furthermore, N deposition did not affect the dominance of O. biennis or A. argyi. Therefore, the species composition of the native community must be considered when preparing to resist future biological invasions. Our study contributes to a better understanding of the invasion mechanisms of alien species under N-loading conditions.

Effects of salt stress on interspecific competition between an invasive alien plant Oenothera biennis and three native species.

Biological invasions and soil salinization have become increasingly severe environmental problems under global change due to sea-level rise and poor soil management. Invasive species can often outcompete native species, but few studies focus on whether invasive alien species are always superior competitors under increasing stressors. We grew an invasive grass species, Oenothera biennis L., and three native grass species (Artemisia argyi Lévl. et Vant., Chenopodium album L., and Inula japonica Thunb.) as a monoculture (two seedlings of each species) or mixture (one seedling of O. biennis and one native species seedling) under three levels of salt treatments (0, 1, and 2 g/kg NaCl) in a greenhouse. We found that invasive O. biennis exhibited greater performance over native C. album and I. japonica, but lower performance compared to A. argyi, regardless of the soil salinity. However, salinity did not significantly affect the relative dominance of O. biennis. Interspecific competition enhanced the growth of O. biennis and inhibited the growth of I. japonica. Although O. biennis seedlings always had growth dominance over C. album seedlings, C. album was not affected by O. biennis at any salt level. At high salt levels, O. biennis inhibited the growth of A. argyi, while A. argyi did not affect the growth of O. biennis. Salt alleviated the competitive effect of O. biennis on I. japonica but did not mitigate the competition between O. biennis and the other two native species. Therefore, our study provides evidence for a better understanding of the invasive mechanisms of alien species under various salinity conditions.

Temporal transcriptome profiling reveals candidate genes involved in cold acclimation of Camellia japonica (Naidong).

Cold is a common problem that limits the distribution of Camellia. Camellia japonica (Naidong) is the northernmost species of camellia in China, which is a Tertiary remnant species that can adapt to large changes in temperature. An analysis of the transcriptional response of C. japonica (Naidong) to cold is very important for the planting and distribution of camellia. In this study, the rate of H2O2 levels, electrolyte leakage, chlorophyll and sugar content had a higher degree of cold response during 12-72 h period, than other periods (0-12h, 72h-120h) in C. japonica (Naidong) response to cold treatment. We constructed the first full-length C. japonica (Naidong) transcriptome and identified 4544 significantly differentially expressed genes (DEGs). A weighted gene coexpression network analysis showed that carbon metabolism, lipid metabolism, and transcription factors played important roles in the resistance of C. japonica (Naidong) to cold stress, and three hub transcription factor regulatory networks were constructed. In addition, overexpressing CjRAV1 led to cold sensitivity in Arabidopsis thaliana, thus CjRAV1 likely plays a negative regulatory role during cold stress in Camellia japonica. This study deepens our understanding of the regulatory mechanism of C. japonica (Naidong) under cold stress and will benefit genetic improvement of camellia.

CBF transcription factors involved in the cold response of Camellia japonica (Naidong).

CBFs belong to the ERF subfamily of the AP2 supergene family and often play an important role in the cold acclimation of temperate plants. However, the role of CBFs in Camellia japonica (Naidong), the only Camellia japonica population found in the temperate zones of China, remains unclear. It is very important to study the genetic composition of C. japonica (Naidong) to adapt to low temperature for Camellia species. Using full-length transcriptome data, we identified four CjCBF genes that respond to cold stress and analyzed their evolutionary relationships, domains, and expression patterns. The phylogeny of CBFs of 19 angiosperms divided the genes into three categories, and the four CjCBFs belong to a small subcluster. The strong response of CjCBF1 to cold treatment and its sustained high level of expression indicated that it plays an important role in the process of cold acclimation. A yeast two-hybrid assay revealed an interaction between CjCBF1, CjCBF2, and CjCBF5, and subcellular localization confirmed this finding. The expression of CjCBFs was tissue-specific: CBF1 was mainly expressed in leaves, and CBF3 was mainly expressed in stem. The responses of the four CjCBFs to drought and high temperature and the effect of light were also characterized. Our study provides new insight into the role of CBFs in the cold response in C. japonica (Naidong).

The complete chloroplast genome of the subtropical species Camellia japonica 'Huaheling'.

Camellia japonica 'Huaheling' is a rare subtropical Camellia species in China with high ornamental and medicinal value. The complete chloroplast genome of C. japonica 'Huaheling' is a 157,001-bp circular DNA molecule containing a large single-copy region (LSC, 86,704 bp), a small single-copy region (SSC, 18,393 bp), and two inverted repeat sequences (IR). Of the 131 genes identified, 86 are protein-coding genes, 8 are rRNA genes, and 37 are tRNA genes. A total of 54 simple sequence repeats (SSRs) were identified in the chloroplast genome. The phylogenetic analysis showed that C. japonica 'Huaheling' is clustered with C. japonica. This work provides valuable information for future study of the evolution and genetic diversity of C. japonica 'Huaheling.'

The complete chloroplast genome of Camellia grijsii 'zhenzhucha', a variant cultivar with floral aroma.

Camellia grijsii 'zhenzhucha' is a variant cultivar from Camellia grijsii, which is also called Camellia grijsii 'juhuacha'.C. grijsii 'zhenzhucha' is an ornamental shrub with a floral aroma, and is oftenused in landscape. To provide genetic information for genetic research, we have sequenced and assembled the complete chloroplast (cp) genome of C. grijsii 'zhenzhucha' based on the Illumina Hiseq platform. The assembled complete cp genome of C. grijsii 'zhenzhucha' was 161,478 bp in length with 37.24% GC, including a large single-copy (LSC) region of 59,942 bp, a small single-copy (SSC) region of 17,294 bp, and a pair of inverted repeats (IRs) of 20,293 bp. The cp genome was annotated with 130 functional genes, consisting of 81 protein-coding genes, 45 transporter RNAs, and 4 ribosomal RNAs. To obtain the phylogeny relationship, the cp genome of C. grijsii 'zhenzhucha'has been compared with other Camellia species, and the results indicate that C. grijsii 'zhenzhucha' is closely related to C. grijsii. This study provides fundamental information of C. grijsii 'zhenzhucha' cp genome, and has an important reference value for the evolutionary analysis.

The complete chloroplast genome of Camellia grijsii, an ornamental shrub with floral aroma.

Camellia grijsii is an ornamental shrub with a floral aroma, which is widely cultivated and used for landscaping in China. To obtain the genetic information of C. grijsii, we have sequenced and assembled the complete chloroplast (cp) genome based on the Illumina Hiseq platform. The total genome size is 161,078 bp in length with 37.18% GC, which contains a large single copy (LSC, 84,645 bp) region, a small single copy (SSC, 15,772 bp) region, and a pair of inverted repeat (IRs, 30,330 bp) regions. It is composed of 81 protein-coding genes, 4 ribosomal RNAs, and 43 transfer RNAs. The cp genome of C. grijsii has also been compared with other species of Camellia, and the results showed that the C. grijsii and the C. grandbibracteata are closely related. This study provides the complete cp genome of C. grijsii and has an important reference value for the evolutionary analysis.

Nitrogen deposition does not alleviate the adverse effects of shade on Camellia japonica (Naidong) seedlings.

Camellia japonica (Naidong), a Tertiary relict species with a unique biological and cultural characteristic, is a special ecotype of C. japonica and is the northernmost distributed populations of C. japonica in the world. This study investigated the interactive responses of C. japonica (Naidong) to shade and nitrogen deposition focusing on seedling growth, leaf morphology and leaf physiology under two light regimes (15% and 65% of full sunlight to represent deep shade and slight shade respectively) and three nitrogen deposition regimes (0, 6 and 12 g N m-2 year-1) in a greenhouse. After 123 d of treatment, the results showed that the deep shade reduced the growth of seedlings significantly compared to slight shade, but improved the specific leaf area, leaf water content, chlorophyll content and Fv/Fm of plants. Moderate nitrogen (6 g N m-2 year-1) supply increased the crown area, specific leaf area, leaf water content, chlorophyll content and water use efficiency of seedlings. However, high nitrogen (12 g N m-2 year-1) supply reduced the basal diameter, crown area, specific leaf area and leaf water content. No significant interaction of shade and nitrogen deposition on C. japonica (Naidong) was found. There is a threshold of nitrogen deposition for the growth of C. japonica (Naidong). Camellia japonica (Naidong) populations should be protected by collecting of germplasm resources and carrying out the ex situ conservation.

Capacity of six shrub species to retain atmospheric particulates with different diameters.

Garden plants can absorb and retain atmospheric particles and are important for remediating environmental pollution. In this study, the dust retention characteristics of six typical shrub species were measured in the greenbelt of a road in the Chengyang District of Qingdao, China, and the maximum capacity for dust retention of each species was determined. The different diameters and areas occupied by particulate matter (PM) were analyzed on the leaf surfaces of the plants. Based on the results for the six shrub species, the rank order of average content of dust retention per unit leaf area was Euonymus japonicus > Pyracantha fortuneana > Ligustrum vicaryi > Amygdalus triloba > Ligustrum sinense > Forsythia suspensa, whereas the rank order of average content per unit volume was E. japonicus > A. triloba > P. fortuneana > L. vicaryi > L. sinense > F. suspensa. The maximum content of dust retention per unit leaf area was reached in approximately 24 days. Plants retained atmospheric PM primarily on the upper leaf surfaces. The primary portion of particles on the leaves was PM10 (over 80%), and PM2.5 was the principal component of PM10. Leaf surface structure significantly affected the abilities of plants to retain PM, and the plants with a thick wax layer or large and dense stomata adsorbed more PM, such as E. japonicus. This study provides a scientific basis for the capacity of landscape plants to retain different diameter particulates.

Distinct double flower varieties in Camellia japonica exhibit both expansion and contraction of C-class gene expression.

BACKGROUND: Double flower domestication is of great value in ornamental plants and presents an excellent system to study the mechanism of morphological alterations by human selection. The classic ABC model provides a genetic framework underlying the control of floral organ identity and organogenesis from which key regulators have been identified and evaluated in many plant species. Recent molecular studies have underscored the importance of C-class homeotic genes, whose functional attenuation contributed to the floral diversity in various species. Cultivated Camellia japonica L. possesses several types of double flowers, however the molecular mechanism underlying their floral morphological diversification remains unclear. RESULTS: In this study, we cloned the C-class orthologous gene CjAG in C. japonica. We analyzed the expression patterns of CjAG in wild C. japonica, and performed ectopic expression in Arabidopsis. These results revealed that CjAG shared conserved C-class function that controls stamen and carpel development. Further we analyzed the expression pattern of CjAG in two different C. japonica double-flower varieties, 'Shibaxueshi' and 'Jinpanlizhi', and showed that expression of CjAG was highly contracted in 'Shibaxueshi' but expanded in inner petals of 'Jinpanlizhi'. Moreover, detailed expression analyses of B- and C-class genes have uncovered differential patterns of B-class genes in the inner organs of 'Jinpanlizhi'. CONCLUSIONS: These results demonstrated that the contraction and expansion of CjAG expression were associated with the formation of different types of double flowers. Our studies have manifested two different trajectories of double flower domestication regarding the C-class gene expression in C. japonica.

Up-regulated HMGB1 in EAM directly led to collagen deposition by a PKCß/Erk1/2-dependent pathway: cardiac fibroblast/myofibroblast might be another source of HMGB1.

High mobility group box 1 (HMGB1), an important inflammatory mediator, is actively secreted by immune cells and some non-immune cells or passively released by necrotic cells. HMGB1 has been implicated in many inflammatory diseases. Our previous published data demonstrated that HMGB1 was up-regulated in heart tissue or serum in experimental autoimmune myocarditis (EAM); HMGB1 blockade could ameliorate cardiac fibrosis at the last stage of EAM. And yet, until now, no data directly showed that HMGB1 was associated with cardiac fibrosis. Therefore, the aims of the present work were to assess whether (1) up-regulated HMGB1 could directly lead to cardiac fibrosis in EAM; (2) cardiac fibroblast/myofibroblasts could secrete HMGB1 as another source of high-level HMGB1 in EAM; and (3) HMGB1 blockade could effectively prevent cardiac fibrosis at the last stage of EAM. Our results clearly demonstrated that HMGB1 could directly lead to cardiac collagen deposition, which was associated with PKCß/Erk1/2 signalling pathway; furthermore, cardiac fibroblast/myofibroblasts could actively secrete HMGB1 under external stress; and HMGB1 secreted by cardiac fibroblasts/myofibroblasts led to cardiac fibrosis via PKCß activation by autocrine means; HMGB1 blockade could efficiently ameliorate cardiac fibrosis in EAM mice.

The checkpoint 1 kinase inhibitor LY2603618 induces cell cycle arrest, DNA damage response and autophagy in cancer cells.

Chemotherapy- or radiotherapy-induced DNA damage activates the Chk1-dependent DNA damage response (DDR) and cell cycle checkpoints to facilitate cell survival. Numerous attempts have been made to identify specific Chk1 inhibitors to enhance the efficiency of chemotherapy or radiotherapy. In this study, we investigated the molecular mechanisms underlying the antitumor activity of LY2603618, a potent and selective small molecule inhibitor of Chk1 protein kinase, in human lung cancer cells. Treatment of cancer cells with LY2603618 caused cell cycle arrest in the G2/M phase. A marked induction of DDR, including the phosphorylation of ATM, Chk2, p53 and histone H2AX, was observed after LY2603618 treatment. LY2603618 inhibited Chk1 autophosphorylation (S296 Chk1) and increased DNA damage-mediated Chk1 phosphorylation (S345 Chk1). In addition, LY2603618-treated lung cancer cells transitioned from LC3-I to LC3-II, a hallmark of autophagy. Blocking autophagy with chloroquine (CQ) further enhanced LY2603618's inhibitory effect on cell viability/proliferation. LY2603618 also significantly increased p38 and c-Jun N-terminal kinase (JNK) phosphorylation. Pretreatment with the JNK inhibitor reduced cleavage of caspase-3 and PARP levels in LY2603618-treated cells. These results suggest the following: (i) the biological consequences of LY2603618 in lung cancer cells is associated with both inhibition of Chk1 phosphorylation on S296 and activation of the DNA damage response network; and (ii) the anticancer property of LY2603618 might be increased by inhibiting autophagy.

Th17 cell expansion in gastric cancer may contribute to cancer development and metastasis.

Th0 cells differentiate into Th1 or Th2 depending on multiple transcription factors acting on specific time points to regulate gene expression. Th17 cells, a subset of IL-17-producing T cells distinct from Th1 or Th2 cells has been described as key players in inflammation and autoimmune diseases as well as cancer development. In the present study, 66 patients with gastric cancer were included; the expression level of Th1- and Th17-related IFN-γ, IL-17, T-bet, RORγt in gastric cancer tissues and peripheral blood mononuclear cell (PBMC) were detected, analyzed the relationship between Th17 or Th1 infiltration and metastasis and explored the possible mechanism. Our results showed that IL-17 and RORγt expression were significantly increased in gastric cancer tissues and PBMC, especially, in metastasis patients; plasma IL-17 also increased; furthermore, the mRNA and protein levels of IL-1ß, IL-21 and TGF-ß were up-regulated. All the data indicated that Th17 was infiltrated the cancer tissue; IL-1ß, IL-21 and TGF-ß were also involved in gastric cancer development by promoting Th17 cell generation. From the above data, we speculated that Th17 cell expansion in gastric cancer may contribute to cancer development and metastasis.