Complete Chloroplast Genomes and Phylogenetic Relationships of Bougainvillea spectabilis and Bougainvillea glabra (Nyctaginaceae).

Bougainvillea L. (Nyctaginaceae) is a South American native woody flowering shrub of high ornamental, economic, and medicinal value which is susceptible to cold damage. We sequenced the complete chloroplast (cp) genome of B. glabra and B. spectabilis, two morphologically similar Bougainvillea species differing in cold resistance. Both genomes showed a typical quadripartite structure consisting of one large single-copy region, one small single-copy region, and two inverted repeat regions. The cp genome size of B. glabra and B. spectabilis was 154,520 and 154,542 bp, respectively, with 131 genes, including 86 protein-coding, 37 transfer RNA, and 8 ribosomal RNA genes. In addition, the genomes contained 270 and 271 simple sequence repeats, respectively, with mononucleotide repeats being the most abundant. Eight highly variable sites (psbN, psbJ, rpoA, rpl22, psaI, trnG-UCC, ndhF, and ycf1) with high nucleotide diversity were identified as potential molecular markers. Phylogenetic analysis revealed a close relationship between B. glabra and B. spectabilis. These findings not only contribute to understanding the mechanism by which the cp genome responds to low-temperature stress in Bougainvillea and elucidating the evolutionary characteristics and phylogenetic relationships among Bougainvillea species, but also provide important evidence for the accurate identification and breeding of superior cold-tolerant Bougainvillea cultivars.

Elevated NO2 induces leaf defensive mechanisms in Bougainvillea spectabilis seedlings.

With a growing economy, the living standard of people has improved which has led to increased use of urban motor vehicles globally. Consequently, the concentration of nitrogen dioxide (NO2) has increased in the ambient air, becoming a major pollutant in urban areas. Plant leaves can absorb, adsorb and fix nitrogen oxides to some extent. Interestingly, NO2 has been recognized as a positive/negative regulator of plant growth. To comprehensively understand the effect of NO2-induced pollution on plants, Bougainvillea spectabilis seedlings were fumigated with different concentrations of nitrogen dioxide (NO2) for a short period in the current study. Further, the induced morphological, physiological, and biochemical changes were measured in the treated as well as untreated seedlings. NO2 exposure caused yellow-brown spotting on the leaf blades in B. spectabilis, which could be the symptoms of oxidative damage. Our findings also reflected the changes in antioxidant enzyme activity and peroxidation of membrane lipids. In addition, the levels of osmotic regulatory substances were also found to be altered to different degrees. In addition, the activities of nitrogen metabolism-related enzymes varied, mainly affecting amino acid metabolism. Overall, the current study would provide a theoretical and scientific basis for selecting and allocating plants in NO2-contaminated areas to manage the pollutants level.

Comparative Study of the Petal Structure and Fragrance Components of the Nymphaea hybrid, a Precious Water Lily.

Nymphaea hybrid, a precious water lily, is a widely-cultivated aquatic flower with high ornamental, economic, medicinal, and ecological value; it blooms recurrently and emits a strong fragrance. In the present study, in order to understand the volatile components of N. hybrid and its relationship with petals structure characteristics, the morphologies and anatomical structures of the flower petals of N. hybrid were investigated, and volatile compounds emitted from the petals were identified. Scanning and transmission electron microscopy were used to describe petal structures, and the volatile constituents were collected using headspace solid-phase microextraction (HS-SPME) fibers and analyzed using gas chromatography coupled with mass spectrometry (GC-MS). The results indicated that the density and degree of protrusion and the number of plastids and osmiophilic matrix granules in the petals play key roles in emitting the fragrance. There were distinct differences in the components and relative contents of volatile compounds among the different strains of N. hybrid. In total, 29, 34, 39, and 43 volatile compounds were detected in the cut flower petals of the blue-purple type (Nh-1), pink type (Nh-2), yellow type (Nh-3) and white type (Nh-4) of N. hybrid at the flowering stage, with total relative contents of 96.78%, 97.64%, 98.56%, and 96.15%, respectively. Analyses of these volatile components indicated that alkenes, alcohols, and alkanes were the three major types of volatile components in the flower petals of N. hybrid. The predominant volatile compounds were benzyl alcohol, pentadecane, trans-α-bergamotene, (E)-ß-farnesene, and (6E,9E)-6,9-heptadecadiene, and some of these volatile compounds were terpenes, which varied among the different strains. Moreover, on the basis of hierarchical cluster analysis (HCA) and principal component analysis (PCA), the N. hybrid samples were divided into four groups: alcohols were the most important volatile compounds for Nh-4 samples; esters and aldehydes were the predominant volatiles in Nh-3 samples; and ketones and alkenes were important for Nh-2 samples. These compounds contribute to the unique flavors and aromas of the four strains of N. hybrid.

Effects of dietary fish meal replacement by fermented moringa (Moringa oleifera Lam.) leaves on growth performance, nonspecific immunity and disease resistance against Aeromonas hydrophila in juvenile gibel carp (Carassius auratus gibelio var. CAS III).

This study was aimed to evaluate the effects of partial replacement of fish meal by fermented moringa leaves (FMLs) on growth performance, serum biochemistry, antioxidant status, nonspecific immunity, and resistance against Aeromonas hydrophila in juvenile gibel carp (Carassius auratus gibelio var. CAS III). Four isonitrogenous and isoenergetic balanced diets, including three FML diets (substituting 20%, 40%, 60% of the fish meal in basal diet, F20, F40 and F60, respectively) and a basal diet (a diet containing 10% fish meal) were used. Each diet was randomly allocated to four fish groups (F20, F40, F60 and control) reared in a recirculating system. After 50 days of the feeding trial, fish were challenged by A. hydrophila. The result revealed that final mean body weight (FBW), weight gain rate (WGR), specific growth rate (SGR), feed efficiency (FE) and survival rate (SR) were significantly increased (P < 0.05) in F20 and F40 groups compared with the control group. Decreased hepatosomatic index (HSI), body crude lipid, serum aspartate transaminase (AST) and serum alanine aminotransferase (ALT) activities, and increased serum alkaline phosphatase (AKP) and serum glutathione peroxidase (GPx) activities were observed in F40 and F60 groups compared with the control and F20 groups. All FMLs-supplemented groups increased (P < 0.05) serum superoxide dismutase (SOD), catalase (CAT) and lysozyme activities, complement component 3 (C3) and serum immunoglobulin M (IgM) concentration, or decreased serum malondialdehyde (MDA) and protein carbonyl (PCC) contents (P < 0.05). After the challenge test, the significant downregulation of toll-like receptors2 (TLR2), tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß and IL-8 mRNA transcription levels was observed in spleens of FMLs supplemented groups. Dietary F40 and F60 showed higher (P < 0.05) relative percent survival (RPS) (48.72% and 43.59%, respectively) against A. hydrophila infection than control. These results indicate that, as a dietary fish meal substitute, FMLs enhance the growth, and antioxidant and immune response, and regulate the expression of immune-related genes and increase disease resistance against A. hydrophila via TLR2 pathway in gibel carp, with greatest effects of 40% fish meal substitution.

Impacts of Climate Change on the Biogeography and Ecological Structure of Zelkova schneideriana Hand.-Mazz. in China.

This study utilized the platform for ensemble forecasting of species distributions, biomod2, to predict and quantitatively analyze the distribution changes of Zelkova schneideriana Hand.-Mazz. under different climate scenarios (SSP1-2.6 and SSP5-8.5) based on climate and land-use data. This study evaluated the geographic range changes in future distribution areas and the results indicated that, under both SSP1-2.6 and SSP5-8.5 scenarios, the distribution area of Zelkova schneideriana would be reduced, showing a trend towards migration to higher latitudes and elevations. Particularly, in the more extreme SSP5-8.5 scenario, the contraction of the distribution area was more pronounced, accompanied by more significant migration characteristics. Furthermore, the ecological structure within the distribution area of Zelkova schneideriana also experienced significant changes, with an increasing degree of fragmentation. The variables of Bio6 (minimum temperature of the coldest month), Bio2 (mean diurnal temperature range), Bio15 (precipitation seasonality), and elevation exhibited important influences on the distribution of Zelkova schneideriana, with temperature being particularly significant. Changes in land use, especially the conversion of cropland, had a significant impact on the species' habitat. These research findings highlight the distributional pressures faced by Zelkova schneideriana in the future, emphasizing the crucial need for targeted conservation measures to protect this species and similar organisms.

Regional environmental differences significantly affect the genetic structure and genetic differentiation of Carpinus tientaiensis Cheng, an endemic and extremely endangered species from China.

Differences in topography and environment greatly affect the genetic structure and genetic differentiation of species, and endemic or endangered species with limited geographic ranges seem to be more sensitive to changes in climate and other environmental factors. The complex topography of eastern China is likely to affect genetic differentiation of plants there. Carpinus tientaiensis Cheng is a native and endangered plants from China, and exploring its genetic diversity has profound significance for protection and the collection of germplasm resources. Based on AFLP markers, this study found that C. tientaiensis has low genetic diversity, which mainly came from within populations, while Shangshantou and Tiantai Mountain populations have relatively high genetic diversity. The Nei genetic distance was closely related to geographical distance, and temperature and precipitation notablely affected the genetic variation and genetic differentiation of C. tientaiensis. Based on cpDNA, this study indicated that C. tientaiensis exhibits a moderate level of genetic diversity, and which mainly came from among populations, while Tiantai Mountain population have the highest genetic diversity. It demonstrated that there was genetic differentiation between populations, which can be divided into two independent geographical groups, but there was no significant phylogeographic structure between them. The MaxEnt model showed that climate change significantly affects its distribution, and the suitable distribution areas in Zhejiang were primarily divided into two regions, eastern Zhejiang and southern Zhejiang, and there was niche differentiation in its suitable distribution areas. Therefore, this study speculated that the climate and the terrain of mountains and hills in East China jointly shape the genetic structure of C. tientaiensis, which gived rise to an obvious north-south differentiation trend of these species, and the populations located in the hilly areas of eastern Zhejiang and the mountainous areas of southern Zhejiang formed two genetic branches respectively.

Variation in the Floral Scent Chemistry of Nymphaea 'Eldorado', a Valuable Water Lily, with Different Flowering Stages and Flower Parts.

Nymphaea 'Eldorado', a valuable water lily, is a well-known fragrant plant in China. Studying the temporal and spatial characteristics of the floral components of this plant can provide a reference for the further development and utilization of water lily germplasm resources. In this study, headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS) was used to explore the types and relative contents of floral components at different flowering stages (S1: bud stage; S2: initial-flowering stage; S3: full-flowering stage; S4: end-flowering stage) and in different floral organs of N. 'Elidorado', combined with the observation of the microscopic structure of petals. A total of 60 volatile organic compounds (VOCs) were detected at different flowering stages, and there were significant differences in floral VOCs at different flowering stages and in different flower organs. The volatile compounds of N. 'Eldorado' can be divided into seven chemical classes,, namely, alkenes, alcohols, esters, aldehydes, ketones, alkanes, and others; the most common were alkenes and alkanes. A total of 39, 44, 47, and 42 volatile compounds were detected at S1, S2, S3, and S4. The VOCs present in high concentrations include benzaldehyde, benzyl alcohol, benzyl acetate, trans-α-bergamotene, α-curcumene, cis-α-farnesene, and so on. The types and total contents of volatiles at the full-flowering stage were higher than at other flowering stages. Comparing the VOCs in different parts of flower organs, it was found that the contents of alcohols, esters, and aldehydes were greatest in the petals, the alkenes in stamens were abundant with a relative content of up to 54.93%, and alkanes in the pistil were higher than in other parts. The types and total contents of volatiles in the stamens of N. 'Eldorado' were higher than those in other flower organs; they were the main part releasing fragrance. The observation of petal microstructure revealed that the size and quantity of the papillae on the epidermises of petals, the number of intracellular plastids, and the aggregates of floral components (osmophilic matrix granules) were significantly higher at the full-flowering stage than at the other flowering stages. This study suggested the main flowering stage and location at which the floral VOCs are released by N. 'Eldorado' and provided a reference for guiding the breeding of this water lily, exploring genetic patterns and developing related products.

Physiological Responses and Salt Tolerance Evaluation of Different Varieties of Bougainvillea under Salt Stress.

Soil salinization significantly impacts the ecological environment and agricultural production, posing a threat to plant growth. Currently, there are over 400 varieties of Bougainvillea with horticultural value internationally. However, research on the differences in salt tolerance among Bougainvillea varieties is still insufficient. Therefore, this study aims to investigate the physiological responses and tolerance differences of various Bougainvillea varieties under different concentrations of salt stress, reveal the effects of salt stress on their growth and physiology, and study the adaptation mechanisms of these varieties related to salt stress. The experimental materials consisted of five varieties of Bougainvillea. Based on the actual salinity concentrations in natural saline-alkali soils, we used a pot-controlled salt method for the experiment, with four treatment concentrations set: 0.0% (w/v) (CK), 0.2% (w/v), 0.4% (w/v), and 0.6% (w/v). After the Bougainvillea plants grew stably, salt stress was applied and the growth, physiology, and salt tolerance of the one-year-old plants were systematically measured and assessed. The key findings were as follows: Salt stress inhibited the growth and biomass of the five varieties of Bougainvillea; the 'Dayezi' variety showed severe salt damage, while the 'Shuihong' variety exhibited minimal response. As the salt concentration and duration of salt stress increase, the trends of the changes in antioxidant enzyme activity and osmotic regulation systems in the leaves of the five Bougainvillea species differ. Membrane permeability and the production of membrane oxidative products showed an upward trend with stress severity. The salt tolerance of the five varieties of Bougainvillea was comprehensively evaluated through principal component analysis. It was found that the 'Shuihong' variety exhibited the highest salt tolerance, followed by the 'Lvyehuanghua', 'Xiaoyezi', 'Tazi', and 'Dayezi' varieties. Therefore, Bougainvillea 'Shuihong', 'Lvyehuanghua', and 'Xiaoyezi' are recommended for extensive cultivation in saline-alkali areas. The investigation focuses primarily on how Bougainvillea varieties respond to salt stress from the perspectives of growth and physiological levels. Future research could explore the molecular mechanisms behind the responses to and tolerance of different Bougainvillea varieties as to salt stress, providing a more comprehensive understanding and basis for practical applications.

Differences in the Suitable Distribution Area between Northern and Southern China Landscape Plants.

Climate change, a global biodiversity threat, largely influences the geographical distribution patterns of species. China is abundant in woody landscape plants. However, studies on the differences in the adaptive changes of plants under climate change between northern and southern China are unavailable. Therefore, herein, the MaxEnt model was used to predict changes in the suitable distribution area (SDA) and dominant environmental variables of 29 tree species under two climate change scenarios, the shared socioeconomic pathways (SSPs) 126 and 585, based on 29 woody plant species and 20 environmental variables in northern and southern China to assess the differences in the adaptive changes of plants between the two under climate change. Temperature factors dominated the SDA distribution of both northern and southern plants. Southern plants are often dominated by one climatic factor, whereas northern plants are influenced by a combination of climatic factors. Northern plants are under greater pressure from SDA change than southern plants, and their SDA shrinkage tendency is significantly higher. However, no significant difference was observed between northern and southern plants in SDA expansion, mean SDA elevation, and latitudinal change in the SDA mass center. Future climate change will drive northern and southern plants to migrate to higher latitudes rather than to higher elevations. Therefore, future climate change has varying effects on plant SDAs within China. The climate change intensity will drive northern landscape plants to experience greater SDA-change-related pressure than southern landscape plants. Therefore, northern landscape plants must be heavily monitored and protected.

Exogenous Salicylic Acid Alleviates NO2 Damage by Maintaining Cell Stability and Physiological Metabolism in Bougainvillea × buttiana 'Miss Manila' Seedlings.

Exogenous substances can alleviate plant damage under adverse conditions. In order to explore whether different concentrations of salicylic acid (SA) can play a role in the resistance of Bougainvillea × buttiana 'Miss Manila' to nitrogen dioxide (NO2) stress and the relevant mechanisms of their effects, different concentrations of SA were applied locally under the control experiment condition of 4.0 µL·L-1 NO2, and the role of SA in alleviating injury was studied. The findings noted a significant increase in metabolic adaptations and antioxidant enzyme activities following 0.25-0.75 mM SA application (p < 0.05), except 1 mM. Superoxide dismutase (SOD) and catalase (CAT) in particular increased by 21.88% and 59.71%, respectively. Such an increase led to effective control of the reduction in photosynthetic pigments and the photosynthetic rate and protection of the structural stability of chloroplasts and other organelles. In addition, the activity of nitrate reductase (NR) increased by 83.85%, and the content of nitrate nitrogen (NO3--N) decreased by 29.23% in nitrogen metabolism. Concurrently, a principal component analysis (PCA) and a membership function analysis further indicated that 0.75 mM SA provided the most notable improvement in NO2 resistance among the different gradients. These findings suggest that 0.25-0.75 mM SA can relieve the stress at 4 µL·L-1 NO2 injury by effectively improving the antioxidant enzyme activity and nitrogen metabolizing enzyme activity, protecting the photosynthetic system and cell structure, but 1 mM SA had the opposite effect. In the future, the specific reasons for inhibition of SA at high concentrations and the comprehensive effects of the application of other exogenous compounds should be further studied.

Combined Transcriptome and Proteome Analysis Provides Insights into Petaloidy in Pomegranate.

Petaloidy leads to a plump floral pattern and increases the landscape value of ornamental pomegranates; however, research on the mechanism of petaloidy in ornamental pomegranates is limited. In this study, we aimed to screen candidate genes related to petaloidy. We performed transcriptomic and proteomic sequencing of the stamens and petals of single-petal and double-petal flowers of ornamental pomegranates. Briefly, 24,567 genes and 5865 proteins were identified, of which 5721 genes were quantified at both transcriptional and translational levels. In the petal and stamen comparison groups, the association between differentially abundant proteins (DAPs) and differentially expressed genes (DEGs) was higher than that between all genes and all proteins, indicating that petaloidy impacts the correlation between genes and proteins. The enrichment results of transcriptome, proteome, and correlation analyses showed that cell wall metabolism, jasmonic acid signal transduction, redox balance, and transmembrane transport affected petaloidy. Nine hormone-related DEGs/DAPs were selected, among which ARF, ILR1, LAX2, and JAR1 may promote petal doubling. Sixteen transcription factor DEGs/DAPs were selected, among which EREBP, LOB, MEF2, MYB, C3H, and trihelix may promote petal doubling. Our results provide transcriptomic and proteomic data on the formation mechanism of petaloidy and a theoretical basis for breeding new ornamental pomegranate varieties.

Study on the Phenotypic Diversity of 33 Ornamental Xanthoceras sorbifolium Cultivars.

Xanthoceras sorbifolium, belonging to the family Sapindaceae, has a beautiful tree shape, elegant leaves, large and many brightly colored flowers, and a long flowering duration. This plant is widely applied in gardens. In this study, 33 cultivars of Xanthoceras sorbifolium were selected from the perspective of ornamental properties, and their phenotypic traits, such as leaves, flowers, and branches, were measured and analyzed, and their phenotypic diversity was comprehensively evaluated using principal component analysis, in order to investigate the phenotypic diversity characteristics of Xanthoceras sorbifolium. The results showed that the genetic diversity index of the qualitative traits varied from 0.14 to 1.50, and that of quantitative traits varied from 1.76 to 2.05. The quantitative traits were more diverse than the qualitative traits. The coefficient of variation of the qualitative traits ranged from 16.90% to 57.96%, and that of quantitative traits ranged from 12.92% to 32.87%. The phenotypic traits of the tested cultivars had relatively rich variation. Furthermore, the level of the phenotypic diversity index of Xanthoceras sorbifolium was not consistent with the level of coefficient of variation, indicating large variation and uneven distribution of variation. Through principal component analysis, 17 quantitative characters were extracted into five principal components, with a cumulative contribution rate of 79.82%, representing the primary information on the quantitative characters of ornamental Xanthoceras sorbifolium cultivars. The F value of the 33 samples ranged from -2.79 to 1.93, and the comprehensive scores of seven cultivars were greater than 1, indicating that these cultivars had rich phenotypic diversity. Therefore, the screening, development, and utilization of fine germplasm resources of Xanthoceras sorbifolium should focus on these cultivars. The 33 cultivars were subsequently clustered into five categories through systematic clustering. The cluster analysis provided references for breeding ornamental Xanthoceras sorbifolium cultivars with different utilization values, such as large white flowers, small red flowers, large red flowers, large orange flowers, and double-petaled flowers.

Complete chloroplast genome and phylogenetic relationship of Nymphaea nouchali (Nymphaeaceae), a rare species of water lily in China.

Nymphaea nouchali is a native species of Chinese water lily with important ornamental, economical, and medicinal purposes. However, due to the serious disturbance by alien biological invasion and human factors, N. nouchali is in an endangered state in China and urgently needs to be protected. Here, we reported the complete chloroplast genome of N. nouchali for the first time, and we found that its plastome is 159 978 bp long, comprising large and small single copies and two inverted repeats (90 001, 19 603, and 50 374 bp, respectively), indicating a typical tetrad structure. In total, 130 genes were identified, including 85 protein-coding genes, 37 transfer RNAs, and 8 ribosomal RNAs. Additionally, 136 simple sequence repeat sites were identified, composed mainly of single nucleotide (46.32%) and trinucleotide (47.05%) sequences. Five highly variable sites (psaI, rps19, ndhF, rps15, and ycf1) with a high Pi value were identified as potential molecular markers. Phylogenetic analysis showed that N. nouchali and N. ampla are closely related, and further validated previous water lily classification results based on morphological characteristics, which divided water lilies into five subgenera: Nymphaea, Brachyceras, Anecphya, Hydrocallis, and Lotos. These results are valuable for the identification and the formulation of protection strategies of N. nouchali, as well as contributing to understanding the evolutionary relationships among Nymphaeaceae species.

Effect of different plant communities on NO2 in an urban road greenbelt in Nanjing, China.

As an important part of urban ecosystems, plants can reduce NO2 concentrations in the air. However, there is little evidence of the effects of different plant communities on NO2 concentrations in street-scale green spaces. We used a multifunctional lifting environmental detector to investigate the impact of environmental factors and small plant communities on NO2 concentrations in street green spaces during the summer and winter in Nanjing, China. The results showed that temperature, atmospheric pressure, and noise were significantly (P < 0.05) correlated with seasonal changes, temperature and humidity significantly (P < 0.01) influenced NO2 concentrations in winter and summer, and the average NO2 concentration in summer was generally higher than in winter. By comparing NO2 concentrations in different plant community structures and their internal spaces, we found that the plant community structure with tree-shrub-grass was more effective in reducing pollution. These findings will help predict the impact of plant communities on NO2 concentrations in urban streets and help city managers and planners effectively reduce NO2 pollution.

Acute NO2 Stress Shortens the Median Survival Period of Bougainvillea glabra 'Elizabeth Angus' by Disrupting Tissue Structure and Photosynthetic Response Centers.

The air pollutant NO2 is one of the major constraints on plant growth, and the ecological value of the ornamental plant Bougainvillea glabra can be weakened by NO2. In this study, an indoor 4 µL·L-1 NO2 simulated fumigation test was conducted with three treatments, CK (normal growth with clean air), T1 (4 µL·L-1 NO2 + 8 h/d), and T2 (4 µL·L-1 NO2 + 24 h/d), which were set up with considerations for time and concentration. The results demonstrated that most of the morphological parameters of B. glabra 'Elizabeth Angus', except for the floral organs, were decreased in the root, stem, leaf, and bract. Continuous fumigation significantly attenuated the growth rate and reduced the water and pigment contents of organs. Excessive NO2 reduced the number and transfer rate of photoelectrons by destroying the photosynthetic reaction center, which in turn weakened photosynthesis, but the plants with intermittent fumigation recovered after fumigation. The Kaplan-Meier (K-M) survival curve displayed median survival periods of 41 and 55.5 h for T1 and T2, respectively, and the morphological structure and most of the indicators of photosynthetic reaction centers changed significantly during stress. Acute injury to B. glabra 'Elizabeth Angus' was caused by 4 µL·L-1 NO2, and B. glabra 'Elizabeth Angus' had limited ability to regulate high concentrations of NO2 acute stress.

Establishing an Efficient Regeneration System for Tissue Culture in Bougainvillea buttiana 'Miss Manila'.

Bougainvillea plants have important ornamental and cultural value, as well as applications, for example, in improving the ecological environment, sterilization and as anti-virals in medicines, etc. Among many varieties, Bougainvillea buttiana 'Miss Manila' is more popular in landscape applications in southern China because of its excellent qualities. However, because of the difficulty of sexual reproduction, low rooting rate of asexual propagation cuttings and low temperature tolerance, its large-scale development is limited. For easy, quick and mass multiplication of such cultivars, tissue culture technique can be put to use. In this study, nodal segments of 'Miss Manila' were used as explants, and a single-factor experiment was carried out with a variety of plant growth regulators and concentrations to establish and optimize a complete tissue culture propagation system. The results showed that the best disinfestation was 75% ethanol treatment for 0.5 min + 0.1% HgCl2 treatment for 9 min, and the survival rate was 86.83%. The best shoot initiation formula was MS + 6-BA (2.5 mg/L) + IBA (0.2 mg/L), and the induction rate was 95.14%. The best formula for shoot proliferation was MS + 6-BA (1.5 mg/L) + NAA (0.1 mg/L), and the proliferation coefficient was 3.13. The best rooting culture formula was 1/2 MS + IBA (2.0 mg/L), and the rooting rate was 81.97%. The survival rate for plantlet refining and transplanting was 93.33%. In this study, a relatively efficient regeneration system for the tissue culture and rapid propagation of Bougainvillea buttiana 'Miss Manila' was established to address the problems of rooting and transplanting of this variety, to benefit research on the industrialized production and regeneration systems of this variety, and to provide a reference for the tissue culture of other varieties of Bougainvillea or other woody plants.

Responses of the growth, photosynthetic characteristics, endogenous hormones and antioxidant activity of Carpinus betulus L. seedlings to different light intensities.

Light is an important ecological factor that affects plant growth, survival and distribution. Carpinus betulus L. is native to central Europe and is used as an ornamental plant with strong adaptability. It is an important tree species for landscaping and timber use. What's more, the antioxidant- and anticancer-related properties of C. betulus leaf extracts are remarkable, that make it a possible raw material for medicine. Light intensity is an important environmental factor affecting the growth and physicochemical changes of C. betulus, but the mechanism of its effect on this species still remains unknown. In this study, the growth, photosynthetic characteristics, endogenous hormones and antioxidant activity responses of C. betulus seedlings to four light intensity gradients (T0: normal light; T1: 75%; T2: 50%; T3: 25% of normal light) were studied after 60 days of shading treatments. The results showed a significant effect of low light intensity on the values of the growth and physiological parameters of C. betulus. The low light intensity caused the inhibition of plant biomass accumulation and the degradation of photosynthetic capacity and stomatal behavior and aggravated the cell membrane lipid peroxidation. However, the plant height growth, leaf area, specific leaf area, photosynthetic pigment content, and contents of GA3 and ABA of C. betulus increased with decreasing light intensity. We found that C. betulus can tolerate mild (T1) and moderate (T2) shading stress by developing photoprotective mechanisms and maintaining relatively high concentrations of organic osmolytes and high antioxidant enzyme activities (superoxide dismutase, peroxidase, catalase and ascorbate peroxidase), but the ability of C. betulus to synthesize osmotic substances and enzymatic antioxidants was impaired under severe shading conditions (T3). Our results suggest that C. betulus can make effective use of low light resources by adjusting its morphology, material distribution, photosynthetic rate and antioxidant enzyme system in suitable low-light environments (50%~75% of normal light); however, light intensity levels that are too low (25% of normal light) will have adverse effects on plant growth. The results of this study provide not only new insights into the response mechanisms of C. betulus to light intensity but also a scientific basis for the cultivation and application of C. betulus in China.

Physiological and biochemical responses of two precious Carpinus species to high-concentration NO2 stress and their natural recovery.

Carpinus betulus and Carpinus putoensis are precious species in the world. Studies on the ecosystem function of the two species are rare. This study investigated the physiological and biochemical responses of C. betulus and C. putoensis to NO2 stress and their natural recovery. C. betulus and C. putoensis seedlings underwent fumigation with 12.0 mg/m3 NO2 for 0, 1, 6, 12, 24, 48, and 72 h, respectively. Then, the plants were allowed to recover at room temperature for 30 d. Physiological and biochemical changes in the leaves were compared between the two species. In terms of peroxidase (POD) activity, the damage response of C. betulus under NO2 stress appeared later than that of C. putoensis. The soluble protein content of C. betulus was noticeably higher than that of C. putoensis, and C. betulus exhibited more stable membrane lipoperoxidation. The tendency of the changes in nitrate reductase of C. betulus was less noticeable than that of C. putoensis. The variation amplitudes of N, K, Mg, Zn and Mn in the leaves of C. putoensis were greater than those of C. betulus. C. putoensis showed more sensitive metabolisms in response to NO2 stress compared with C. betulus. High-concentration NO2 caused damage to C. betulus and C. putoensis was reversible, and both species returned to normal growth via their own metabolism after 30-d recovery. The results of this study may provide useful reference data for quantitative assessment of the ecosystem function of C. betulus and C. putoensis and for their scientific application in urban greening.

Comprehensive Transcriptome Analysis of Rare Carpinus putoensis Plants under NO2 stress.

We evaluated a transcriptome using high-throughput Illumina HiSeq sequencing and related it to the morphology, leaf anatomy, and physiological parameters of Carpinus putoensis putoensis under NO2 stress. The molecular mechanism of the C. putoensis NO2 stress response was evaluated using sequencing data. NO2 stress adversely affected the morphology, leaf anatomy, and total peroxidase (POD) activity. Through RNA-seq analysis, we used NCBI to compare the transcripts with nine databases and obtained their functional annotations. We annotated up to 2255 million clean Illumina paired-end RNA-seq reads, and 250,200 unigene sequences were assembled based on the resulting transcriptome data. More than 89% of the C. putoensis transcripts were functionally annotated. Under NO2 stress, 1119 genes were upregulated and 1240 were downregulated. According to the KEGG pathway and GO analyses, photosynthesis, chloroplasts, plastids, and the stimulus response are related to NO2 stress. Additionally, NO2 stress changed the expression of POD families, and the HPL2, HPL1, and POD genes exhibited high expression. The transcriptome analysis of C. putoensis leaves under NO2 stress supplies a reference for studying the molecular mechanism of C. putoensis resistance to NO2 stress. The given transcriptome data represent a valuable resource for studies on plant genes, which will contribute towards genome annotations during future genome projects.

Taxonomic Identity of Carpinus dayongina Franchet (Betulaceae).

Carpinus polyneura and C. dayongina are recognised as separate species in Flora of China. In this study, the results of an examination of literature, morphological comparison and phenetic clustering of nuclear ITS sequences suggest that C. dayongina is conspecific with C. polyneura. Thus, we propose reducing C. dayongina to a synonym of C. polyneura.