Search

Background Grape hyacinth (Muscari spp.) is one of the most important ornamental bulbous plants. However, its lengthy juvenile period and time-consuming transformation approaches under the available protocols impedes the functional characterisation of its genes in flower tissues. In vitro flower organogenesis has long been used to hasten the breeding cycle of plants but has not been exploited for shortening the period of gene transformation and characterisation in flowers. Results A petal regeneration system was established for stable transformation and function identification of colour gene in grape hyacinth. By culturing on Murashige and Skoog medium (MS) with 0.45 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 8.88 μM 6-benzyladenine (6-BA), during the colour-changing period, the flower bud explants gave rise to regeneration petals in less than 3 months, instead of the 3 years required in field-grown plants. By combining this system with Agrobacterium-mediated transformation, a glucuronidase reporter gene (GUS) was delivered into grape hyacinth petals. Ultimately, 214 transgenic petals were regenerated from 24 resistant explants. PCR and GUS quantitative analyses confirmed that these putative transgenic petals have stably overexpressed GUS genes. Furthermore, an RNAi vector of the anthocyanidin 3-O-glucosyltransferase gene (MaGT) was integrated into grape hyacinth petals using the same strategy. Compared with the non-transgenic controls, reduced expression of the MaGT occurred in all transgenic petals, which caused pigmentation loss by repressing anthocyanin accumulation. Conclusion The Agrobacterium transformation method via petal organogenesis of grape hyacinth took only 3–4 months to implement, and was faster and easier to perform than other gene-overexpressing or -silencing techniques that are currently available.

Understanding how ecological processes mediate community stability as a response to variation in the environment is a central goal in ecology. Four biotic mechanisms – portfolio, selection, insurance, and over-yielding effects – have been suggested to control community stability. While the significance of each of these mechanisms is debated, few studies have elucidated changes in their relative importance along environmental gradients. We used six years of field observations from a natural assembled wetland plant community to explore how stability changed directly or indirectly along an elevation gradient. All four mechanisms mentioned above contributed significantly to the variation in temporal stability (totally 94%), with selection and insurance effects being most important. Generalized multilevel path modelling indicates that the four mechanisms destabilize individual populations and functional groups, even though they have significant stabilizing effects on community productivity. The modelling also suggests that community dominance rather than species diversity largely mediate the indirect effects of elevation on community stability. Increasing elevation indirectly affects stability through changes in community dominance, synchrony, and abundance. Although no significant responses of species diversity were found along elevation gradients, species diversity apparently controlled stability both directly and indirectly through changes in community dominance, synchrony, and abundance. Our study illustrates that understanding the multivariate chains linking the direct, indirect, and interactive effects of elevation gradients on plant communities may help clarifying complex biotic stability mechanisms.

Tripterygiumwilfordii Hook F (TwHF) is one of the most commonly used and effective traditional Chinese herbal medicines against rheumatoid arthritis (RA). Both Tripterygium Glycoside Tablets (TGT) and Tripterygium wilfordii Tablets (TWT) are the representative TwHF-based agents enrolled into the 2019 edition of Medicine Catalog for National Basic Medical Insurance, Injury Insurance, and Maternity Insurance. However, individual differences in TGT/TWT response across patients usually exist in the process of treating RA, implying that the clinical application of the two agents may not be standardized leading to the ineffective treatment and the risk of side effects. Growing evidence show that the bioactive constituents of TwHF may often have toxicity, the package insert of TGT and TWT may not be described in detail, and the therapeutic windows of the two agents are narrow. Thus, it is an urgent task to develop a standardized clinical practice guideline for TGT and TWT in the treatment of RA. In the current study, a group of clinical experts of traditional Chinese medicine and Western medicine in the research field of rheumatism diseases, pharmacists, and methodologists of evidence-based medicine were invited to select the clinical questions, to determine the levels of the evidence and the strength of the recommendations, and to develop the recommendations and good practice points. The guideline is formed based on the combination of clinical research evidence and expert experience (evidence-based, consensus, supplemented by experience). The clinical problems which are supported by clinical evidence may form recommendations, and the clinical problems without clinical evidence may form experts’ suggestions. Both recommendations and experts' suggestions in this guideline summarized the clinical indications, usage, dosage, combined medication, and safety of TGT and TWT against RA systematically and comprehensively, which may offer a professional guidance in the context of the clinical application of the two TwHF-based agents.

The growth and reproduction of plants are highly size-dependent and markedly affected by light intensity. Vallisneria spinulosa S.Z.Yan is a common submersed macrophyte and is widely used in current lake restoration projects. The size of plants to be used to get optimal restoration results is not known, however, and may vary with water clarity. In this study, the effects of different plant sizes (large, medium and small, i.e. approximately 2.40 g, 1.01 g and 0.27 g per plant) on the growth and reproduction of V. spinulosa were tested under three light intensities (high, medium, low, i.e. 25%, 6% and 1% of full sunlight, respectively). Both plant size and light intensity were found to be of great significance for plant growth and reproduction. The relative growth rate (RGR) of biomass was higher for small than for large plants at both high and low light, while the absolute growth rate (AGR) of biomass was higher for large-sized plants at medium and high light intensity. Asexual reproduction of V. spinulosa was positively correlated with plant size and more affected by plant size than by light intensity, while sexual reproduction was most affected by light intensity. RGR and AGR of plant height were highest for medium-sized plants at moderate light intensity. Therefore, the size of submersed macrophytes should be considered when using this species in connection with lake restoration. Verification of our results under field conditions is needed, though, before firm conclusions can be reached about the optimal plant size of V. spinulosa to be used in lake restoration as also other factors like, e.g. periphyton shading and wave actions may play a role.

Background: Hypoxic brain injury is one of the major causes of neurodevelopmental impairment and cardiovascular disability. LncRNA SNHG1 works as a critical factor in hypoxic induced injury, however, the potential mechanism is still not known well.Methods: The expression level of small nucleolar RNA host gene 1 (SNHG1) and miR-140-5p was detected by qRT-PCR. The western blot assay was performed to measure the level of Bcl-XL and apoptosis-related proteins. The target relationship between lncRNA SNHG1 and miR-140-5p, as well as miR-140-5p and Bcl-XL was detected by dual luciferase reporter gene assay. Cell apoptosis was assessed using Annexin V/PI staining by flow cytometry. Cell viability was analyzed by MTT assay.Results: Oxygen glucose deprivation (OGD) treatment inhibited SNHG1 and Bcl-XL expression and enhanced miR-140-5p expression. OGD treatment-induced cell viability inhibition, cell apoptosis promotion were partially abrogated when SH-SY5Y cells were transfected with pcDNA3.1-SNHG1 or miR-140-5p inhibitor. Moreover, luciferase reporter assay revealed that lncRNA SNHG1 bound directly to miR-140-5p, and miR-140-5p directly targeted Bcl-XL. The protective effect of SNHG1 overexpressing on cell apoptosis induced by OGD was attenuated after transfected with miR-140-5p mimic or sh-Bcl-XL in SH-SY5Y cells.Conclusion: LncRNA SNHG1-modulated miR-140-5p inhibition regulates Bcl-XL expression, thereby reducing cell apoptosis and recovering cell viability of SH-SY5Y cells. The results in this study provide novel insight into the mechanism of SNHG1 mediated signaling pathway during hypoxic brain injury.

Interspecific trait variability (e.g., functional diversity) is often correlated with community productivity and thus promotes ecosystem stability. However, we know little about how intraspecific trait variability mediated the effects of environmental gradients on population stability. We applied generalized multilevel path models to investigate multivariate causal hypotheses that environmental effects on population stability of a macrophyte species - Potamogeton maackianus - are mediated by physiological and morphological traits variability. Using a hierarchical nested design, we measured quarterly nine physiological traits, and monthly ten morphological traits from 9720 individuals at 27 plots along water depth gradients in Lake Erhai, China. Results showed that changes in water depth and light availability caused significant shifts in intraspecific trait variability, with cascading direct and indirect effects on temporal stability of P. maackianus. Water depth exerted major control over population stability, both directly and indirectly via shifts in morphological and physiological traits. Light availability affected population stability indirectly by altering morphological traits. Size traits (shoot morphology and root/shoot ratio) and metabolic traits captured the effect of water depth and light availability on population stability with a relative high accuracy. Our study provide a strong support to the hypothesis that intraspecific trait variability mediate the combined effects of variations on water depth and light conditions on the temporal stability of a dominant macrophtye species in Lake Erhai. Maintaining and enhancing trait variability within plant populations may help to buffer negative effects of anthropogenic environmental changes on population stability.

Carbon (C) and nitrogen (N) metabolism are essential for the growth and maintenance of submersed macrophytes. They are affected by nutrient and light availability and seasonal changes. In this study, we investigated the changes of C/N metabolism and growth of three natural submersed macrophyte populations (i.e., each in a different site), Potamogeton maackianus, in response to water depth gradients with 0.5 m interval in a freshwater lake for four seasons. Changes were identified by measuring the biomass of the sampled macrophytes and metabolite contents of soluble carbohydrate (SC), starch, and free amino acid (FAA) in their organs. Results showed that no significant and consistent changes were found in the SC and FAA contents in leaves and stems from 1.0 m to 5.5 m water depth, as compared to previous results of controlled studies, suggesting a relative stable C/N metabolism in response to the depth gradients in studied natural populations of P. maackianus. The metabolite contents with the most variations appeared mainly because of seasonal changes. The metabolite contents of the macrophyte populations were more closely correlated with the chemical parameters than with the light parameters in water columns. No significant correlations were found between the metabolite contents with the light attenuation coefficient ($K$) and Secchi depth (SD) in water columns. The NO3-N and total phosphorus (TP) contents in water columns were positively correlated with the SC and starch contents in leaves, respectively. The NH4-N content in water columns was negatively related to the FAA content in leaves. Metabolite contents had weak correlations with depth gradients and light parameters, probably because of the physiological integration among individuals by developed stolons and the canopy-forming characteristic of this species. Our results highlighted the importance of water quality to the C/N metabolism in natural population of submersed macrophyte. (C) 2017 Elsevier B.V. All rights reserved.

Metals can react with microcystin (MC), which is released from cyanobacterial blooms through various mechanisms; these reactions may mitigate the environmental and health risks of MCs but may also cause harm to aquatic ecosystems and humans. Several studies were conducted, including laboratory tests, ecological simulations, and a field investigation of Poyang Lake. The laboratory studies showed that Fe3+, Cu2+, and Pb2+ stimulated MC photodegradation under high light intensity at the water-sediment interface, which reduced the MC accumulation in the sediment. In the laboratory studies involving the addition of metal ions to lake sediment containing adsorbed MC, MC biodegradation was inhibited by supplementing with high levels of Fe3+, Cu2+, or Pb2+. Fe3+ and Pb2+ promoted MC accumulation in the hydrophyte Eichhornia crassipes at relatively low concentrations, but this effect decreased with increasing high metal concentrations. An ecological survey in Poyang Lake during the dry season demonstrated that high Fe levels can reduce MC accumulation in the sediment, which could be the result of Fe-mediated photodegradation. The results indicate that metals involved in MC transportation and degradation may play an important role in the environmental fate of MC.

Ammonium pulse attributed to runoff of urban surface and agriculture following heavy rain is common in inland aquatic systems and can cause profoundly effects on the growth of macrophytes, especially when combined with low light. In this study, three patterns of NH4-N pulse (differing in magnitude and frequency) were applied to examine their effects on the growth of three submersed macrophytes, namely, Myriophyllum spicatum, Potamogeton maackianus, and Vallisneria natans, in terms of biomass, height, branch/ramet number, root length, leaf number, and total branch length under high and low light. Results showed that NH4-N pulse caused negative effects on the biomass of the submerged macrphytes even on the 13th day after releasing NH4-N pulse. The negative effects on M. spicatum were significantly greater than that on V. natans and P. maackianus. The effects of NH4-N pulse on specific species depended on the ammonium loading patterns. The negative effects of NH4-N pulse on P. maackianus were the strongest at high loading with low frequency, and on V. natans at moderate loading with moderate frequency. For M. spicatum, no significant differences were found among the three NH4-N pulse patterns. Low light availability did not significantly aggregate the negative effects of NH4-N pulse on the growth of the submersed macrophytes. Our study contributes to revealing the roles of NH4-N pulse on the growth of aquatic plants and its species specific effects on the dynamics of submerged macrophytes in lakes.

For submersed macrophytes, carbon (C) and nitrogen (N) metabolism is a central factor affecting growth and survival under low light availability. Three common submersed macrophytes, Vallisneria natans, Potamogeton maackianus, and Potamogeton malaianus, were grown at five water depths (1.0, 2.5, 4.0, 5.5, and 7.0 m) to explore mechanism of low light adaptation in terms of C/N metabolism by examining relative growth rate (RGR) and contents of soluble carbohydrate (SC), starch, total carbon (TC), free amino acid (FAA) and total nitrogen (TN) in leaf, stem and root of the plants. With increasing water depth, P. malaianus, P. maackianus and V. natans initiated dying at 4.0 m, 5.5 m and 7.0 m water depths, respectively. natans showed higher RGR than the other two species. For all the plants, the FAA contents increased with increasing water depth, except for the roots of P. maackianus. The TN contents showed a unimodal curve along water depths with the highest in moderate water depth, except for P. malaianus and-the roots of V. natans. For V. natans and P. maackianus, the C metabolic indices (SC, starch, and TC) showed a unimodal curve along water depths with the lowest in moderate water depth in the leaves and stems, except for TC contents in the leaves of P. maackianus. Compared with P. maackianus and P. malaianus, lower SC and FAA contents and higher starch storage in V. natans contributed to its higher tolerance to deeper depths. The nonlinear changes in metabolite contents along water depths for V. natans and P. maackianus demonstrated complex mechanism for low light adaptation, and thus partly explained the wider ranges of colonizing water depths than P. malaianus. (C) 2015 Elsevier B.V. All rights reserved.

Biochar is a good adsorbent for water pollutants. However, the effects of biochar on aquatic organisms are not well understood. In this study, different amounts of biochar (CK, 0 mg/g; T1, 10 mg/g; T2, 30 mg/g) were added to sediment to study changes in water quality and its impact on three submerged macrophytes (Hydrilla verticillata, Vallisneria natans, and Ceratophyllum demersum) and the sediment microbial community. The results indicated that biochar treatments significantly increased the water pH and conductivity. Compared with the initial values, the total phosphorus (P) contents in the water of the CK, T1, and T2 treatments decreased by 78.5%, 95.0%, and 58.3%, respectively, while the total nitrogen contents increased by 26.26%, −5.81%, and 19.70%, respectively. Compared with those in CK, the relative growth rates of H. verticillata, V. natans, and C. demersum in T1 increased by 28.4%, 163.1%, and 61.3%, respectively, while those in T2 showed no significant difference except that the growth rates of H. verticillata decreased by 17.7%. The P contents of the three submerged macrophytes increased with the increase of biochar addition, except that there was no significant difference between T2 and CK for H. verticillata. Biochar treatments reduced the biomass of total microbial, bacterial, and fungal phospholipid fatty acids in the sediment for H. verticillata and V. natans, and they increased fungal: bacterial ratios in the low-dose biochar treatments for V. natans and C. demersum. This study demonstrates that the addition of biochar to sediment significantly increased the pH and conductivity, and decreased total P contents in the water. Low-dose biochar treatments were more beneficial for water quality improvements and the growth of submerged macrophytes than high-dose biochar.

Anthocyanins are responsible for the different colors of ornamental plants. Grape hyacinth (Muscari armeniacum), a monocot plant with bulbous flowers, is popular for its fascinating blue color. In the present study, we functionally characterized an R2R3-MYB transcription factor gene MaAN2 from M. armeniacum. Our results indicated that MaAN2 participates in controlling anthocyanin biosynthesis. Sequence alignment and phylogenetic analysis suggested that MaAN2 belonged to the R2R3-MYB family AN2 subgroup. The anthocyanin accumulation of grape hyacinth flowers was positively correlated with the expression of MaAN2. And the transcriptional expression of MaAN2 was also consistent with that of M. armeniacum dihydroflavonol 4-reductase (MaDFR) and M. armeniacum anthocyanidin synthase (MaANS) in flowers. A dual luciferase transient expression assay indicated that when MaAN2 was co-inflitrated with Arabidopsis thaliana TRANSPARENT TESTA8 (AtTT8), it strongly activated the promoters of MaDFR and MaANS, but not the promoters of M. armeniacum chalcone synthase (MaCHS), M. armeniacum chalcone isomerase (MaCHI), and M. armeniacum flavanone 3-hydroxylase (MaF3H). Bimolecular fluorescence complementation assay confirmed that MaAN2 interacted with AtTT8 in vivo. The ectopic expression of MaAN2 in Nicotiana tabacum resulted in obvious red coloration of the leaves and much redder flowers. Almost all anthocyanin biosynthetic genes were remarkably upregulated in the leaves and flowers of the transgenic tobacco, and NtAn1a and NtAn1b (two basic helix–loop–helix anthocyanin regulatory genes) were highly expressed in the transformed leaves, compared to the empty vector transformants. Collectively, our results suggest that MaAN2 plays a role in anthocyanin biosynthesis.

Grape hyacinth (Muscari spp.) is a popular ornamental bulbous perennial famous for its blue flowers. To understand the chemical basis of the rich blue colors in this plant, anthocyanin profiles of six blue flowering grape hyacinths as well as one pink and one white cultivar were determined using high-performance liquid chromatography and mass spectrometry. Along with two known compounds, eight putative anthocyanins were identified in the tepals of grape hyacinth for the first time. The accumulation and distribution of anthocyanins in the plant showed significant cultivar and flower development specificity. Violet-blue flowers mainly contained simple delphinidin-type anthocyanins bearing one or two methyl-groups but no acyl groups, whereas white and pink flowers synthesised more complex pelargonidin/cyanidin-derivatives with acyl-moieties but no methyl-groups. The results partially reveal why solid blue, orange or red flowers are rare in this plant in nature. In addition, pelargonidin-type anthocyanins were found for the first time in the genus, bringing more opportunities in terms of breeding of flower color in grape hyacinth.

Summary Through a combination of metabolite analysis with transcriptome sequencing, a new hypothesis was proposed to explain the lack of colour phenotype of the white variant of the blue grape hyacinth., Grape hyacinth (Muscari) is an important ornamental bulbous plant with an extraordinary blue colour. Muscari armeniacum, whose flowers can be naturally white, provides an opportunity to unravel the complex metabolic networks underlying certain biochemical traits, especially colour. A blue flower cDNA library of M. armeniacum and a white flower library of M. armeniacum f. album were used for transcriptome sequencing. A total of 89 926 uni-transcripts were isolated, 143 of which could be identified as putative homologues of colour-related genes in other species. Based on a comprehensive analysis relating colour compounds to gene expression profiles, the mechanism of colour biosynthesis was studied in M. armeniacum. Furthermore, a new hypothesis explaining the lack of colour phenotype of the grape hyacinth flower is proposed. Alteration of the substrate competition between flavonol synthase (FLS) and dihydroflavonol 4-reductase (DFR) may lead to elimination of blue pigmentation while the multishunt from the limited flux in the cyanidin (Cy) synthesis pathway seems to be the most likely reason for the colour change in the white flowers of M. armeniacum. Moreover, mass sequence data obtained by the deep sequencing of M. armeniacum and its white variant provided a platform for future function and molecular biological research on M. armeniacum.

Grape hyacinth (Muscari spp.) is a popular ornamental plant with bulbous flowers noted for their rich blue color. Muscari species have been thought to accumulate delphinidin and cyanidin rather than pelargonidin-type anthocyanins because their dihydroflavonol 4-reductase (DFR) does not efficiently reduce dihydrokaempferol. In our study, we clone a novel DFR gene from blue flowers of Muscari. aucheri. Quantitative real-time PCR (qRT-PCR) and anthocyanin analysis showed that the expression pattern of MaDFR had strong correlations with the accumulation of delphinidin, relatively weak correlations with cyanidin, and no correations with pelargonidin. However, in vitro enzymatic analysis revealed that the MaDFR enzyme can reduce all the three types of dihydroflavonols (dihydrokaempferol, dihydroquercetin, and dihydromyricetin), although it most preferred dihydromyricetin as a substrate to produce leucodelphinidin, the precursor of blue-hued delphinidin. This indicated that there may be other functional genes responsible for the loss of red pelargonidin-based pigments in Muscari. To further verify the substrate-specific selection domains of MaDFR, an assay of amino acid substitutions was conducted. The activity of MaDFR was not affected whenever the N135 or E146 site was mutated. However, when both of them were mutated, the catalytic activity of MaDFR was lost completely. The results suggest that both the N135 and E146 sites are essential for the activity of MaDFR. Additionally, the heterologous expression of MaDFR in tobacco (Nicotiana tabacum) resulted in increasing anthocyanin accumulation, leading to a darker flower color, which suggested that MaDFR was involved in color development in flowers. In summary, MaDFR has a high preference for dihydromyricetin, and it could be a powerful candidate gene for genetic engineering for blue flower colour modification. Our results also make a valuable contribution to understanding the basis of color variation in the genus Muscari.

Purple turnip Brassica rapa ssp. rapa is highly appreciated by consumers but the metabolites and molecular mechanisms underlying the root skin pigmentation remain open to study. Herein, we analyzed the anthocyanin composition in purple turnip (PT) and green turnip (GT) at five developmental stages. A total of 21 anthocyanins were detected and classified into the six major anthocynanin aglycones. Distinctly, PT contains 20 times higher levels of anthocyanins than GT, which explain the difference in the root skin pigmentation. We further sequenced the transcriptomes and analyzed the differentially expressed genes between the two turnips. We found that PT essentially diverts dihydroflavonols to the biosynthesis of anthocyanins over flavonols biosynthesis by strongly down-regulating one flavonol synthase gene, while strikingly up-regulating dihydroflavonol 4-reductase (DFR), anthocyanidin synthase and UDP-glucose: flavonoid-3-O-glucosyltransferase genes as compared to GT. Moreover, a nonsense mutation identified in the coding sequence of the DFR gene may lead to a nonfunctional protein, adding another hurdle to the accumulation of anthocyanin in GT. We also uncovered several key members of MYB, bHLH and WRKY families as the putative main drivers of transcriptional changes between the two turnips. Overall, this study provides new tools for modifying anthocyanin content and improving turnip nutritional quality.

Flower-specific promoters can enable transgenic enhancement of valuable ornamental traits, including flower shape and color. However, the identification of strong, tissue-specific promoters remains a limiting factor. To obtain enhanced flower-specific promoters, we constructed four chimeric promoters (p35S-PCHS-Ω, p35S-LCHS-Ω, pOCS-PCHS-Ω and pOCS-LCHS-Ω) combining the 35S or OCS enhancer fused to a 302 bp CHSA core promoter fragment from petunia (PCHS) or a 307 bp CHS core promoter fragment from lily (LCHS), and also containing an omega element (Ω). Each promoter was fused to the β-glucuronidase (GUS) reporter gene, and we examined the levels and tissue specificity of GUS expression in transgenic Torenia fournieri. p35S-PCHS-Ω and p35S-LCHS-Ω drove strong, constitutive GUS expression in all tissues, especially in colored corollas (p35S-PCHS-Ω) or in colored corollas and roots (p35S-LCHS-Ω). pOCS-PCHS-Ω drove stronger GUS expression in colored corollas than in other tissues but expression was weaker than that of p35S-PCHS-Ω. pOCS-LCHS-Ω drove GUS in colored corollas but also in roots. Among the four chimeric promoters, pOCS-PCHS-Ω exhibited stronger activity only in colored corollas, making it useful for transgenic enhancement of floral traits, such as expressing ‘blue genes’ in lily to produce new lines with blue flowers.

Flavonoid 3′, 5′-hydoxylase (F3′5′H) is a key enzyme for biosynthesis of the blue anthocyanin pigment delphinidin. A number of F3′5′H genes from dicots have been tested for their effects on flower pigmentation; here F3′5′H from a monocot was tested for its effect on delphinidin accumulation in petals. To this end, F3′5′H (PhF3′5′H) from the orchid Phalaenopsis was expressed under the control of the chalcone synthase promoter in petunia flowers. Quantitative RT-PCR showed that PhF3′5′H was expressed mainly in the petal limb; this expression produced an increase in dihydromyricetin and delphinidin and a change in petal color from pink to deeper pink. To increase the accumulation of delphinidin, Hyacinth HyDFR, which encodes dihydroflavonol 4-reductase, and petunia DifF, which encodes a cytochrome b 5 that is required for full activity of F3′5′H were overexpressed. The HyDFR petunia transformants had a deeper color petal limb, increased dihydromyricetin and delphinidin contents and adaxial petals with a number of blue cells. The flowers of the DifF petunia transformants also showed a slight color change. We also tested PhF3′5′H in Lilium oriental ‘Sorbonne’, where transient PhF3′5′H expression by particle bombardment resulted in purple cells in the petals. Production of blue flowers by Phalaenopsis F3′5′H and hyacinth DFR potentially enables manipulation of flower color in ornamental plants, including production of blue flowers.

The inflorescence of the broad-leafed grape hyacinth, Muscari latifolium, shows an interesting, two-tone appearance with the upper flowers being pale blue and the lower ones purple. To elucidate the mechanism of the differential color development, anatomical research was carried out and a cytological study of the colored protoplasts in which the shapes of the cells accumulating anthocyanin were observed by scanning electron microscopy. Next, vacuolar pH was recorded using a pH meter with a micro combination pH electrode, and the sap's metal-ion content was measured by inductively coupled plasma mass spectrometry. The anthocyanin and co-pigment composition was determined by high-performance liquid chromatography (HPLC). Chemical analyses reveal that the difference in metal-ion content of the two parts was not great. The vacuolar pHs of the upper and lower flowers were 5.91 and 5.84, respectively, with the difference being nonsignificant. HPLC results indicate that the dihydroflavonol and flavonol contents are also very similar in the two sorts of flower. However, the upper flowers contained only delphinidin, whereas the lower flowers also contained cyanidin. The total anthocyanin content in the lower flowers was 4.36 mg g(-1), which is approximately seven times higher than in the upper flowers, while the delphinidin content is four times higher. Quantitative real-time PCR analysis established that the two-tone flower was a result of different expressions of the F3'5'H, F3'H and DFR genes, and these lead to different amounts of anthocyanin.

Aimed at identifying the key influencing environmental factors of phytoplankton community, we investigated phytoplankton in June 2010 and February 2011 in Poyang Lake. Species from 58 genuses, 32 families and 7 phyla had been found in the investigations, and the major phyla were Chlorophyta, Bacillariophyta and Cyanophyta. The average abundance of phytoplankton in summer and winter were 35.92×105 cell/L and 26.63×105 cell/L, respectively. Even though the unique hydrodynamic conditions of such a water passing lake (during flood seasons Poyang Lake forming a vast lake, but in dry seasons, lake recedes into a channel) make the water environment unstably, Chlorophyta are able to adapt to the conditions and grow well, therefore, the lake was dominated by Chlorophyta, especially Chlorella and Cosmarium, respectively accounting for 56.38 % and 52.10 % of the total abundance in summer and winter. CCA ordination on 12 sampling sites, 58 genuses of phytoplankton and 10 environmental factors demonstrated that there were great geographic differences of water environmental physicochemical conditions in winter, while in summer, it’s not significant, besides, DO, water temperature and nutrients (nitrogen and phosphate) were the most important environmental factors that regulated the distribution of phytoplankton community.

Ecological processes are generally scale-dependent and there is little consensus about the relative importance of deterministic versus stochastic processes in driving patterns of biological diversity. We investigated how the relationship between functional dispersion and environmental gradients changes with spatial scale in subtropical freshwater lakes. The functional alpha and beta dispersions of all the tested traits were significantly under-dispersed across spatial scales and along environmental gradients. Results showed more functional similarity within communities in leaf dry mass content and flowering duration but less functional turnover among communities in all the tested traits at regional scales (Yunnan-Guizhou plateau and the middle and low reaches of the Yangtze River). The strengths and directions of environmental effects on the functional alpha and beta dispersions depended on the selected traits, diversity metrics and spatial scales. Surprisingly, broad-scale factors - elevation and water transparency - decreased the functional turnover for most traits along the gradients, whereas fine-scale factors - water depth - produced the opposite patterns along the gradient, depending on the trait selected. Our study highlights the dominant role of deterministic assembly processes in structuring the local functional composition and governing the spatial functional turnover of macrophyte communities across multiple spatial scales.

Trait-based approaches have been widely applied to investigate how community dynamics respond to environmental gradients. In this study, we applied a series of maximum entropy (maxent) models incorporating functional traits to unravel the processes governing macrophyte community structure along water depth gradient in a freshwater lake. We sampled 42 plots and 1513 individual plants, and measured 16 functional traits and abundance of 17 macrophyte species. Study results showed that maxent model can be highly robust (99.8%) in predicting the species relative abundance of macrophytes with observed community-weighted mean (CWM) traits as the constraints, while relative low (about 30%) with CWM traits fitted from water depth gradient as the constraints. The measured traits showed notably distinct importance in predicting species abundances, with lowest for perennial growth form and highest for leaf dry mass content. For tuber and leaf nitrogen content, there were significant shifts in their effects on species relative abundance from positive in shallow water to negative in deep water. This result suggests that macrophyte species with tuber organ and greater leaf nitrogen content would become more abundant in shallow water, but would become less abundant in deep water. Our study highlights how functional traits distributed across gradients provide a robust path towards predictive community ecology.

The first enzyme in the flavonoid pathway, chalcone synthase, is encoded by a gene (CHS) whose expression is normally under developmental control. In our previous studies, an 896-bp promoter region of a flower-specific CHS gene was isolated from Lilium orential ‘Sorbonne’, and designated as PLoCHS. Here, the PLoCHS promoter was fused to the β-glucuronidase (GUS) gene to characterize its spatial and temporal expression in Petunia hybrida ‘Dreams Midnight’ using an Agrobacterium-mediated leaf disc transformation method. Our results demonstrated that GUS expression was present in flowers, but reduced or absent in the other tissues (leaf and stem) examined. In petals, GUS activity reached its peak at flower developmental stage 4, and decreased at later stages. Deletion analysis indicated that even a 307-bp fragment of the PLoCHS promoter could still direct flower-specific expression. Further deletion of the region from −261 to −72 bp resulted in weak expression in different organs, including flowers, leaves and stems. This evidence combined with prediction of cis-acting elements in the PLoCHS promoter suggests that the TACPyAT box located in this promoter plays a key role in the regulation of organ-specific expression.