Combined de novo transcriptomic and physiological analyses reveal RyALS3-mediated aluminum tolerance in Rhododendron yunnanense Franch.

Rhododendron (Ericaceae) not only has ornamental value, but also has great medicinal and edible values. Many Rhododendron species are native to acid soils where aluminum (Al) toxicity limits plant productivity and species distribution. However, it remains unknown how Rhododendron adapts to acid soils. Here, we investigated the physiological and molecular mechanisms of Al tolerance in Rhododendron yunnanense Franch. We found that the shoots of R. yunnanense Franch did not accumulate Al after exposure of seedlings to 50 µM Al for 7 days but predominantly accumulated in roots, suggesting that root Al immobilization contributes to its high Al tolerance. Whole-genome de novo transcriptome analysis was carried out for R. yunnanense Franch root apex in response to 6 h of 50 µM Al stress. A total of 443,639 unigenes were identified, among which 1,354 and 3,413 were up- and down-regulated, respectively, by 6 h of 50 µM Al treatment. Both Gene Ontology (GO) enrichment and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that genes involved in "ribosome" and "cytoskeleton" are overrepresented. Additionally, we identified Al-tolerance homologous genes including a tonoplast-localized ABC transporter RyALS3; 1. Overexpression of RyALS3; 1 in tobacco plants confers transgenic plants higher Al tolerance. However, root Al content was not different between wild-type plants and transgenic plants, suggesting that RyALS3; 1 is responsible for Al compartmentalization within vacuoles. Taken together, integrative transcriptome, physiological, and molecular analyses revealed that high Al tolerance in R. yunnanense Franch is associated with ALS3; 1-mediated Al immobilization in roots.

Integrated transcriptome and amino acid profile analyses reveal novel insights into differential accumulation of theanine in green and yellow tea cultivars.

Tea cultivars with yellow- or white-leaf variations have a high economic value due to their high amino acid (especially theanine) concentration. However, the dynamic changes of amino acid components (especially theanine) and related gene expression during new shoot development in these cultivars are still unclear. In this study, 264 tea samples from four representative varieties picked during the harvest period in spring were analyzed for their amino acid profiles. The dynamic change rules of ethylamine and 19 amino acids were summarized in normal green and yellow cultivars during new shoot development. Interestingly, the theanine concentration in the yellow cultivar was significantly higher than that in the green cultivar, and increased gradually as the leaves matured until they reached a maximum in the one bud and three leaves stage. The amino acid concentration in the leaves of the yellow cultivar increased significantly with leaf position, which was generally in contrast to the normal green cultivar. Transcriptome and correlation analyses revealed that CsGS1, CsPDX2, CsGGP5, CsHEMA3 and CsCLH4 might be the key genes potentially responsible for the differential accumulation of theanine in green and yellow tea cultivars. These results provide further information for the utilization and improvement of tea plants.

Hypoglycemic Effect of the N-Butanol Fraction of Torreya grandis Leaves on Type 2 Diabetes Mellitus in Rats through the Amelioration of Oxidative Stress and Enhancement of ß-Cell Function.

Materials and Methods: Sprague-Dawley rats were randomly divided into six groups: control, T2DM, metformin, high-dose BFTL (800 mg/kg), middle-dose BFTL (400 mg/kg), and low-dose BFTL (200 mg/kg). After 4 weeks of BFTL treatment, the correlations of serum indicators with protein expression in tissue were determined, and pathological changes in the liver, kidneys, and pancreas were analyzed. Results: Compared with the results in the T2DM group, serum fasting blood glucose, triglyceride, total cholesterol, malondialdehyde, alanine aminotransferase, and aspartate aminotransferase levels were significantly decreased (p < 0.05), whereas superoxide dismutase and glutathione peroxidase levels were significantly increased (p < 0.05) in the high-, middle-, and low-dose BFTL groups. The treatment also improved oral glucose tolerance. In addition, the pathological changes of the liver, kidney, and pancreas were improved by BFTL treatment. Cytochrome and caspase-3 expression in pancreatic was significantly decreased (p < 0.05) by BFTL treatment, whereas the Bcl-2/Bax ratio was significantly increased (p < 0.05). Discussion and Conclusion. BFTL exerted significant hypoglycemic effect on T2DM model rats, and its mechanism involved the suppression of blood glucose levels and oxidative stress by improving the metabolism of blood lipids and antioxidant capacity, boosting ß-cell function, and inhibiting ß-cell apoptosis.

Purified anthocyanins from Zea mays L. cob ameliorates chronic liver injury in mice via modulating of oxidative stress and apoptosis.

BACKGROUND: Purple corn (Zea mays L.) is one of the main economic crops in China and has been used in the treatment of cystitis, urinary infections and obesity. However, purple corncobs, the by-product remaining after processing and having an intense purple-black color, are normally disposed of as waste or used as animal feed. Therefore, to further expand the medicinal value of purple corncob, its content was analyzed and, after purification, the effect and mechanism of purified purple corncob anthocyanins (PPCCA) on CCl4 -induced chronic liver injury in mice were investigated. RESULTS: It was observed that the total anthocyanin content (TAC) from PPCCA (317.51 ± 9.30 mg cyanidin 3-O-glucoside (C-3-G) g-1 dry weight) was significantly higher than that from the purified purple corn seed anthocyanin (266.73 ± 3.67 mg C-3-G g-1 dry weight), of which C-3-G accounted for 90.6% and 90.4% of the TAC, respectively. Furthermore, compared with the CCl4 group, PPCCA treatment significantly reduced liver index, serum total bilirubin, alanine transaminase, aspartate transaminase and liver malondialdehyde levels, but increased liver superoxide dismutase activity. The pathological changes were also improved, such as more regular arrangement of hepatocytes, less swelling, and fewer vacuoles and apoptotic cells. Additionally, mechanistic studies showed that PPCCA downregulated the expression of Caspase-3, Bax and cytochrome P450 2E1 proteins in the liver and upregulated the expression of Bcl-2. CONCLUSION: These results demonstrated that PPCCA could ameliorate CCl4 -induced chronic liver injury by regulating oxidative stress and hepatocyte apoptosis pathways. © 2021 Society of Chemical Industry.

Complete chloroplast genome of Rhododendron pulchrum, an ornamental medicinal and food tree.

The semi-evergreen azalea, Rhododendron pulchrum, a valuable horticultural and medicinal plant species. Using next-generation sequencing, applying a combination of de novo and reference-guided assembly, we sequenced its complete chloroplast genome. Our study reveals that R. pulchrum have a typical cp genome of 136,249 bp in length, without inverted repeat regions. A total of 73 genes, 42 of which are protein coding genes, 29 tRNA genes, two rRNA genes were identified. The GC content of the whole genome is 35.98%. Phylogenetic analysis indicates that R. pulchrum is closely related to the species of Vaccinium oldhamii and Vaccinium macrocarpon.

Brassinosteroids alleviate high-temperature injury in Ficus concinna seedlings via maintaining higher antioxidant defence and glyoxalase systems.

Although brassinosteroids (BRs) play crucial roles in plant development and stress tolerance, the mechanisms by which they have these effects are poorly understood. Here, we investigated the possible mechanism of exogenously applied BRs on reactive oxygen species (ROS), antioxidant defence and methylglyoxal (MG) detoxification systems in Ficus concinna seedlings grown under high-temperature (HT) stress for 48 h. Our results showed that the activities of ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione S-transferase (GST), glutathione peroxidase (GPX) and glyoxalase II (Gly II) were increased under two levels of HT stress. Compared with control the activities of catalase (CAT) and dehydroascorbate reductase (DHAR) were not changed due to HT stress. The activities of glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and glyoxalase I (Gly I) were increased only at moderate HT stress. Despite these protective mechanisms, HT stress induced oxidative stress in F. concinna seedlings, as indicated by the increased levels of ROS, malondialdehyde (MDA) and MG, and the reductions in chlorophyll levels and relative water content. The contents of reduced glutathione (GSH) and ascorbate (AsA) were not changed under moderate HT stress. Spraying with 24-epibrassinolide (EBR) alone had little influence on the non-enzymatic antioxidants and the activities of antioxidant enzymes. However, EBR pretreatment under HT stress resulted in an increase in GSH and AsA content, maintenance of high redox state of GSH and AsA, and enhanced ROS and MG detoxification by further elevating the activities of SOD, GST, GPX, APX, MDHAR, GR, DHAR, Gly I and Gly II, as evident by lower level of ROS, MDA and MG. It may be concluded that EBR could alleviate the HT-induced oxidative stress by increasing the enzymatic and non-enzymatic antioxidant defence, and glyoxalase systems in F. concinna seedlings.

Nitric oxide functions as a signal in ultraviolet-B-induced baicalin accumulation in Scutellaria baicalensis suspension cultures.

Stress induced by ultraviolet-B (UV-B) irradiation stimulates the accumulation of various secondary metabolites in plants. Nitric oxide (NO) serves as an important secondary messenger in UV-B stress-induced signal transduction pathways. NO can be synthesized in plants by either enzymatic catalysis or an inorganic nitrogen pathway. The effects of UV-B irradiation on the production of baicalin and the associated molecular pathways in plant cells are poorly understood. In this study, nitric oxide synthase (NOS) activity, NO release and the generation of baicalin were investigated in cell suspension cultures of Scutellaria baicalensis exposed to UV-B irradiation. UV-B irradiation significantly increased NOS activity, NO release and baicalin biosynthesis in S. baicalensis cells. Additionally, exogenous NO supplied by the NO donor, sodium nitroprusside (SNP), led to a similar increase in the baicalin content as the UV-B treatment. The NOS inhibitor, Nω-nitro-l-arginine (LNNA), and NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) partially inhibited UV-B-induced NO release and baicalin accumulation. These results suggest that NO is generated by NOS or NOS-like enzymes and plays an important role in baicalin biosynthesis as part of the defense response of S. baicalensis cells to UV-B irradiation.

Effects of potassium supply on limitations of photosynthesis by mesophyll diffusion conductance in Carya cathayensis.

Potassium (K) influences the photosynthesis process in a number of ways; however, the mechanisms underlying the photosynthetic response to differences in K supply are not well understood. Concurrent measurements of gas exchange and chlorophyll fluorescence were made to investigate the effect of K nutrition on photosynthetic efficiency and mesophyll conductance (g(m)) in hickory seedlings (Carya cathayensis Sarg.) in a greenhouse. The results show that leaf K concentrations < 0.7-0.8% appeared to limit the leaf net CO2 assimilation rate (A), and that the relative limitation of photosynthesis due to g(m) and stomatal conductance (g(s)) decreased with increasing supplies of K. However, a sensitivity analysis indicated that A was most sensitive to the maximum carboxylation rate of Rubisco (V(c,max)) and the maximum rate of electron transport (J(max)). These results indicate that the photosynthetic rate is primarily limited by the biochemical processes of photosynthesis (V(c,max) and J(max)), rather than by g(m) and g(s) in K-deficient plants. Additionally, g(m) was closely correlated with g(s) and the leaf dry mass per unit area (M(A)) in hickory seedlings, which indicates that decreased g(m) and g(s) may be a consequence of leaf anatomical adaptation.

cDNA-AFLP analysis of gene expression in hickory (Carya cathayensis) during graft process.

The graft technique is a valid method for propagating plants. A better elucidation of the graft mechanism is helpful in improving the production efficiency and fruit quality in hickory. In this study, cDNA-amplified fragment length polymorphism analysis was used to examine the gene expression in hickory at four time points (at 0, 3, 7 and 14 days) during the graft process. Forty-nine unique genes involved in the graft mechanism were obtained. The expression patterns of these genes were confirmed by real-time reverse transcription-polymerase chain reaction analysis based on 12 selected genes representing different patterns. The 49 genes composed 19 genes of known function, nine genes of unknown function and 21 novel genes. These 19 genes of known functions were involved in the indole-3-acetic acid transport protein, cell cycle, signal transduction, water metabolism, nuclear metabolism, amino acid metabolism, protein metabolism, carbon metabolism and secretion of substances, suggesting that Carya cathayensis Sarg. undergoes a complex metabolism process during the grafting.

Cyclic electron flow around photosystem I is required for adaptation to high temperature in a subtropical forest tree, Ficus concinna.

Dissipation mechanisms of excess photon energy under high-temperature stress were studied in a subtropical forest tree seedling, Ficus concinna. Net CO(2) assimilation rate decreased to 16% of the control after 20 d high-temperature stress, and thus the absorption of photon energy exceeded the energy required for CO(2) assimilation. The efficiency of excitation energy capture by open photosystem II (PSII) reaction centres (F(v)'/F(m)') at moderate irradiance, photochemical quenching (q(P)), and the quantum yield of PSII electron transport (Phi(PSII)) were significantly lower after high-temperature stress. Nevertheless, non-photochemical quenching (q(NP)) and energy-dependent quenching (q(E)) were significantly higher under such conditions. The post-irradiation transient of chlorophyll (Chl) fluorescence significantly increased after the turnoff of the actinic light (AL), and this increase was considerably higher in the 39 degrees C-grown seedlings than in the 30 degrees C-grown ones. The increased post-irradiation fluorescence points to enhanced cyclic electron transport around PSI under high growth temperature conditions, thus helping to dissipate excess photon energy non-radiatively.

Antisense inhibition of Rubisco activase increases Rubisco content and alters the proportion of Rubisco activase in stroma and thylakoids in chloroplasts of rice leaves.

BACKGROUND AND AIMS: Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase (RCA) is a nuclear-encoded chloroplast protein that modifies the conformation of Rubisco, releases inhibitors from active sites, and increases enzymatic activity. It appears to have other functions, e.g. in gibberellin signalling and as a molecular chaperone, which are related to its distribution within the chloroplast. The aim of this research was to resolve uncertainty about the localization of RCA, and to determine whether the distributions of Rubisco and RCA were altered when RCA content was reduced. The monocotyledon, Oryza sativa was used as a model species. METHODS: Gas exchange and Rubisco were measured, and the sub-cellular locations of Rubisco and RCA were determined using immunogold-labelling electron microscopy, in wild-type and antisense rca rice plants. KEY RESULTS: In antisense rca plants, net photosynthetic rate and the initial Rubisco activity decreased much less than RCA content. Immunocytolocalization showed that Rubisco in wild-type and antisense plants was localized in the stroma of chloroplasts. However, the amount of Rubisco in the antisense rca plants was greater than in the wild-type plants. RCA was detected in both the chloroplast stroma and in the thylakoid membranes of wild-type plants. The percentage of RCA labelling in the thylakoid membrane was shown to be substantially decreased, while the fraction in the stroma was increased, by the antisense rca treatment. CONCLUSIONS: From the changes in RCA distribution and alterations in Rubisco activity, RCA in the stroma of the chloroplast probably contributes to the activation of Rubisco, and RCA in thylakoids compensates for the reduction of RCA in the stroma, allowing steady-state photosynthesis to be maintained when RCA is depleted. RCA may also have a second role in protecting membranes against environmental stresses as a chaperone.

Isolation and physiological characteristics of a premature senescence mutant in rice (Oryza sativa L.).

A rice pse(t) (premature senescence, tentatively) mutant line, was isolated from 4,500 independent T-DNA inserted transgenic lines. The symptoms of premature senescence appeared more severely than those of the control plants (Zhonghua 11, japonica) at the last development stage. To characterize the mutant and provide basic information on the candidate genes by mapping to a physical region of 220-kb, experiments were carried out in two phytotrons under controlled temperature of 24 degrees C and 28 degrees C, respectively. The content of chlorophyll, soluble protein and MDA (malondialdehyde), net photosynthesis, the antioxidant enzyme activities of SOD (superoxide dismuase) (EC 1.15.1.1) and POD (peroxidase) (EC 1.11.1.7) and the peptidase activities of leaves were measured from top to bottom according to the leaf positions at the flowering stage. Compared with the control plant, the mutant showed the following characteristics: (1) Higher net photosynthesis rate (P(n)) appeared in the 1st and 2nd leaves, contents of chlorophyll and soluble protein were also higher in the 1st leaf; (2) The activities of SOD, POD and peptidase were higher according to the leaf position from top to bottom; (3) The symptom of premature senescence was accelerated in the mutant at 28 degrees C treatment. The MDA content and the SOD and POD activities between the 24 degrees C and 28 degrees C treatment mutants were not significantly different. Content of chlorophyll and soluble protein of leaves mutant decreased rapidly at 28 degrees C treatment. The results show that pse(t) is sensitive to high temperature. The probable function of PSE(T) is discussed.

Characteristics of photosynthesis in rice plants transformed with an antisense Rubisco activase gene.

Transgenic rice plants with an antisense gene inserted via Agrobacterium tumefaciens were used to explore the impact of the reduction of Rubisco activase (RCA) on Rubisco and photosynthesis. In this study, transformants containing 15% to 35% wild type Rubisco activase were selected, which could survive in ambient CO2 concentration but grew slowly compared with wild type controls. Gas exchange measurements indicated that the rate of photosynthesis decreased significantly, while stomatal conductance and transpiration rate did not change; and that the intercellular CO2 concentration even increased. Rubisco determination showed that these plants had approximately twice as much Rubisco as the wild types, although they showed 70% lower rate of photosynthesis, which was likely an acclimation response to the reduction in Rubsico activase and/or the reduction in carbamylation.

[Construction of expression vector with antisense Rubisco activase gene and its genetic transformation in rice].

In this research, Rubisco activase gene (rca) was amplified using specific primers and inserted into pGEM T-easy vector, and then cut with EcoRI after confirming by sequencing. The fragment was subcloned into pBluescript KS+, digested with the enzyme BamHI and inserted into the binary expression vector pCAMBIA1301, and the resulting construction with antisense rca was named pCAMR02. The pCAMR02 vector was introduced into Agrobacterium tumefaciens strain EHA105 by electroporation and transformed to embryos of rice (Oryza. Sativa L.ssp.japonica) cultivar Zhonghua11 via Agrobacterium tumefaciens system. Plantlets were regenerated in vitro by resistance selection on medium containing various concentrations of hygromycin. Both GUS histochemical assays and PCR amplification demonstrated that antisense rca was integrated into T0 genomes and inherited to T1. The measurement of phenotypes of transgenic rice plants with antisense rca showed that most of them could hardly survive at ambient CO2 concentration, even could not grow. The antisense plants that survived under natural condition were dwarf and grew slower than the wild-type controls, and their contents of RCA and Rubisco changed significantly. These plants generated in this experiment will be used to study the relationship between RCA and Rubisco and their regulation.