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Phenotypic plasticity acts to increase the performance of plants under stress. Leaf morphological plasticity and its causes in different environments are incompletely understood. We measured the leaf morphological parameters of Quercus acutissima Carr. seedlings, including leaf size, leaf shape and venation pattern, assessing the effects of different habitat conditions on leaf morphological plasticity. A field study in forest edge and understory was combined with experiments simulating different light and water conditions. Leaf morphology variations occurred over most of the parameters, and the causes were consistent between the field study and lab experiment. Leaf size decreased with low supply of light and water. Leaf length and width were only affected by leaf area. The leaf petiole did not lengthen under shade stress, suggesting a trade-off relationship between functional tissues and support structures. Leaf shape became narrower in drought and broader in the shade, as reflected in changes in three leaf fractions. Higher vein density played a part in enhancement of mechanical support and water supply. Leaves with more teeth show more active photosynthesis, but are disadvantageous in xeric environments because of higher transpiration. Light was the main factor inducing leaf morphological plasticity. The variations caused by drought were due mainly to the allometry. Our results showed that the leaves of Q. acutissima seedlings respond to different habitats with phenotypic plasticity of morphology, suggesting that this is an important mechanism for seedlings to adapt to broader ecological amplitudes.
Iron deficiency that is induced by high soil pH is a major factor affecting plant growth in calcareous soils and in some areas that have been reclaimed following industrial activities. Since the effects of high soil pH commonly involve Fe deficiency, in this study, we examined whether Fe provided to part of the root system exposed to low pH would alleviate high pH stress in paper birch (Betula papyrifera), trembling aspen (Populus tremuloides) and red-osier dogwood (Cornus stolonifera) seedlings. The plants were grown in a controlled environment growth room in mineral nutrient solution at pH 5 and 9 and provided with either 0 or 40 µM Fe in a split-root system for 8 weeks. At the end of the treatments, plant dry weights, net photosynthesis, transpiration rates, root ferric-chelate reductase activity, and leaf chlorophyll concentrations were measured, and elemental analyses were carried out in young leaves. The results demonstrated high root zone pH affected Fe, P and Zn concentrations in young leaves. In the three considered species, plants with part of their root system exposed to pH 5 had higher dry weights, net photosynthesis, and transpiration rates compared with the plants with the whole root system immersed in pH 9 solution. High root zone pH reduced photosynthesis, transpiration rates, leaf chlorophyll concentrations and the uptake of Fe, P, and Zn in plants. Partial exposure of the root system to low pH and Fe supply reduced leaf chlorosis and partly alleviated the high pH stress in the studied plants by improving Fe uptake, but did not alleviate root growth reductions.
TPSI1/Mt4 family genes are a class of genes induced dramatically by Pi starvation, and are involved in Pi allocation in plants. Its members have been isolated from many plant species, but none has been reported in Brassica napus. Here, we isolated two novel members of TPSI1/Mt4 family from B. napus, which were referred to as BnIPS1;1 and BnIPS1;2. The two genes are 592 and 557 bp long, respectively, and share 91 % sequence identity. They are all intronless, and contain numerous short open reading frames and a conserved 22-nucleotide sequence partially complementary to miR399. Expression analysis revealed that the two genes were induced strongly by Pi starvation and weakly by osmotic stress and salinity. The induction of the two genes is rapid and durative in the absence of Pi and is repressible upon Pi resupply. Furthermore, BnIPS1;1 promoter (-1,459/?42 relative to transcription start site) was isolated and fused to reporter gene GUS. The PBnIPS1;1:GUS construct was introduced into Arabidopsis, and intensive GUS staining was observed in Pi-starved plants. These results further our understanding of TPSI1/Mt4 family genes, and demonstrate that BnIPS1;1 and BnIPS1;2 can be used as tool for investigating Pi-starvation signaling in B. napus and BnIPS1; 1 promoter can be used as inducible promoter for cultivating P-efficient crops.
Background: Calcitonin gene-related peptide (CGRP) can protect against hyperoxia-induced lung injury, making the upregulation of CGRP a potential therapeutic approach for this type of injury. However, the effects of CGRP on the Wnt7b/β-catenin signaling pathway are unclear. In this study, we investigated the roles of CGRP and the Wnt7b/β-catenin signaling pathway in hyperoxia-induced lung injury. Methods: Premature Sprague Dawley (SD) rats were exposed to 21, 40, 60 and 95% oxygen for 3, 7 and 14 days. The animals’ body weights, survival rates and endogenous CGRP levels were measured. Lung samples were harvested for histological analyses and measurements of malondialdehyde (MDA) concentration and total antioxidant capacity (TAOC). We also assessed the MDA concentration and TAOC in the lung tissues after administration of 200 nmol/kg CGRP8–37 (a CGRP antagonist). Finally, alveolar epithelial type II (AEC II) cells were isolated from premature rats, exposed to 21 or 95% oxygen for 3, 7 and 14 days, and treated with 10− 8 mol/l exogenous CGRP. The protein expressions of Wnt7b and β-catenin were assessed using western blotting, and TCF and c-myc mRNA expressions were assessed using qPCR. Results: Rats exposed to 60 and 95% oxygen had significantly lower body weights and survival rates than the 21 and 40% groups, and the decrease was time dependent. Endogenous CGRP was elevated in the lung tissues of premature rats exposed to 95% oxygen. CGRP8–37 induced apparent inflammation in the lung tissue and alveolar structural remodeling. In addition, the expression levels of Wnt7b and β-catenin were markedly increased after exposure for 3 days. They peaked at 7 days, then declined at 14 days. The levels of TCF/c-myc in AEC II cells increased significantly after CGRP treatment when compared with cells that had only undergone hyperoxia. Conclusions: CGRP protected against hyperoxia-induced lung injury in premature rats. This process involves the Wnt7b/β-catenin signaling pathway.
Background: Mishandling of intracellular chloride (Cl−) concentration ([Cl−]i) in cerebrovascular smooth muscle cells is implicated in several pathological processes, including hyperplasia and remodeling. We investigated the effects of ClC-2-mediated Cl− efflux on the proliferation of human brain vascular smooth muscle cells (HBVSMCs) induced by angiotensin II (AngII). Methods: Cell proliferation and motility were determined using the CCK-8, bromodeoxyuridine staining, wound healing and invasion assays. ClC-2, PCNA, Ki67, survivin and cyclin D1 expression, and β-catenin and GSK-3β phosphorylation were examined using western blotting. Histological analyses were performed using hematoxylin and eosin staining and α-SMA staining. Results: Our results showed that AngII-induced HBVSMC proliferation was accompanied by a decrease in [Cl− ]i and an increase in ClC-2 expression. Inhibition of ClC-2 by siRNA prevented AngII from inducing the efflux of Cl− . AngII-induced HBVSMC proliferation, migration and invasion were significantly attenuated by ClC-2 downregulation. The inhibitory effects of ClC-2 knockout on HBVSMC proliferation and motility were associated with inactivation of the Wnt/β-catenin signaling pathway, as evidenced by inhibition of β-catenin phosphorylation and nuclear translocation, and decrease of GSK-3β phosphorylation and survivin and cyclin D1 expression. Recombinant Wnt3a treatment markedly reversed the effect of ClC-2 knockdown on HBVSMC viability. An in vivo study revealed that knockdown of ClC-2 with shRNA adenovirus ameliorated basilar artery remodeling by inhibiting Wnt/β-catenin signaling in AngII-treated mice. Conclusion: This study demonstrates that blocking ClC-2-mediated Cl− efflux inhibits AngII-induced cerebrovascular smooth muscle cell proliferation and migration by inhibiting the Wnt/β-catenin pathway. Our data indicate that downregulation of ClC-2 may be a viable strategy in the prevention of hyperplasia and remodeling of cerebrovascular smooth muscle cells.
Topping, the critical cultural practice, is a kind of damage occurred tobacco shoot, which can obviously promote roots growth. Up to now, the mechanism regulating roots growth after topping is still unclear. In our previous works, miR171d was identified as a major topping responsive miRNA in tobacco roots, and its target gene was predicted to belong to the GRAS gene family. In the present study, NtGRAS-R1, a novel GRAS transcription regulator, was firstly cloned from tobacco roots. NtGRAS-R1 contained an open reading frame of 1527 bp encoding a 508-amino acids protein, and its molecular mass and isoelectric point was 56.199 KD and 5.24, respectively. GRAS proteins in tobacco can be grouped into nine subfamilies, including HAM, SCR, LAS, DLT, DELLA, SCL, LISCL, SHR and PAT1. NtGRAS-R1 is highly homologous with NtGRAS44 and belongs to HAM subfamily. The expression of NtGRAS-R1 can be detected in tobacco roots, leaves and stems, and its expression level was highest in tobacco roots. The analysis of transgenic tobacco showed that NtGRAS-R1 was involved in several biology processes, such as rooting, germination and root development, which will be helpful to explore the roles of NtGRAS-R1 in response to tobacco topping.
In this study, we investigated responses of the mid-successional species Acer truncatum Bunge and the late-successional species Quercus variabilis Blume to three solar illumination conditions: (1) constant low light (CL), (2) constant high light (CH) and (3) low light first and high light afterwards (LH). The last treatment was to simulate a canopy opening. Both species exhibited increases in biomass, totally and in part, and decreases in leaf water content, specific leaf area and chlorophyll concentrations in LH treatment compared to CL treatment. For A. truncatum, exposure to high light condition (LH) increased crown area, and decreased root to shoot ratio, stem mass ratio and leaf perimeter. However, for Q. variabilis, LH treatment increased stem diameter at ground height, effective quantum yield, photochemical quenching and decreased maximum photosystem II quantum yield. The biomass allocation pattern did not change in Q. variabilis among three light conditions. With respect to newly developed leaves, no significant differences were found in leaf size of Q. variabilis between LH treatment and CH treatment while that of A. truncatum decreased in LH treatment. All chlorophyll fluorescence parameters in newly developed oak leaves in LH treatment increased compared to those of CH treatment while no difference was found for A. truncatum between LH and CH treatment. A. truncatum displayed a greater overall plasticity than Q. variabilis although the oak seedlings have a greater plasticity with respect to chlorophyll concentration and chlorophyll fluorescence parameters. A. truncatum should be a better candidate for vegetation recovery, especially in places with heterogeneous light conditions.
Leaf morphological, physiological and biochemical characteristics of Robinia pseudoacacia L. seedlings were studied under different stress conditions. The plants were subjected to drought and shade stress for one month. Leaf inclination, chlorophyll fluorescence and chlorophyll content were measured at the first day (shortterm stress) and at the end of the stress period (long-term stress) and in the recovery period. Leaf inclination was affected mainly by light; a low level of irradiance caused leaves to be arranged horizontally. Diurnal rhythmicity was lost after the long-term stress, but resumed, in part, in the recovery period. Drought stress caused leaves to tilt more obviously and decreased damage to the photosystem. Sun avoiding movement in a single leaf and sun tracking movement in the whole plant coexisted. Significant physiological changes occurred under different conditions of light. Increased energy dissipation and light capture were the main responses to high and low level of irradiance, respectively, and these were reflected by changes of chlorophyll fluorescence and chlorophyll content. Phenotypic plasticity in the leaflet enhanced the protective response to stress. These adaptive mechanisms may explain better survival of R. pseudoacacia seedlings in the understory, especially during the drought periods, and made it to be the preponderant reforestation species in Shandong Province of China.
The study pays attention to disturbances in early successional communities of wetland vegetation. We conducted artificial disturbances in a community of Suaeda salsa and Phragmites australis in the Yellow River Delta (China). Eight types of disturbances combining mowing treatments with species treatments were applied. Removal of the standing litters of P. australis or not was defined as mowing treatments, and removal of two species solo or both was defined as species treatments. We sampled 80 quadrats from the treatments plots at different intervals after the disturbance to investigate plant height, abundance, aboveground biomass, the distance between plants to reflect the effect of disturbance on composition, structure, productivity, and function of the plant communities. The strategies of seedling emergence and height growth differed as the canopy changed. Biomass contribution of different species, combined with disturbance intensity, was the main factors that affected the productivity. Homogeneity of disturbance was better for maintaining the functions of plant community in compared with the competitiveness (C), stress-tolerance (S) and ruderality (C-S-R) signatures with the control. Facilitations were reflected by the stagger arrangements in relative growth rates of the two species and in plant-plant interactions calculated by a modified function of competition. Adapting to symmetric disturbance and developing facilitative interactions are important requirements for early succession terrestrial vegetation to establish and stabilize in the seriously saline environments of wetlands.
Leaf traits scaling relationships were compared in different months (May, June, July and August) in a temperate shrub species, Vitex negundo Linn. var. heterophylla (Franch.) Rehd. Leaf traits variation and the impact of environment were also studied. Our results showed that specific leaf area (SLA), leaf nitrogen content (Nmass), leaf phosphorus content (Pmass) and leaf chlorophyll content (Chlmass) were positively correlated, and all of the pairwise relationships showed a common standardized major axis (SMA) slope in different months. The variation of SLA, chlorophyll a content (Chlmass a), chlorophyll b content (Chlmass b), Chlmass a/b and maximum quantum yield of PSII (Fv/Fm) were mainly habitat-dependent, while the variation of Pmass and Nmass/Pmass were mainly month-dependent. Redundancy analysis (RDA) was used to further explore the relationships between leaf traits and environmental factors. We found that plants under shade (the relative photosynthetic photon flux density was about 10%) developed light-capturing behavior at leaf level (higher SLA) and cellular level (higher Chlmass/Nmass and lower Chlmass a/b). The increased Nmass/Pmass with month was mainly resulted from the large absorption of soil nitrogen and the decrease of soil pH. In a word, our study indicates that species may not have fundamentally different carbon capture strategies in different months. Variation of different leaf traits are related to different environmental factors in the field.
Plant cell walls primarily comprise lignin, which performs functions of mechanical support, water transport, and stress responses. Lignin biosynthesis pathway proceeds through metabolic grid featuring complexity and diversity in enzymatic reaction. Cinnamate-4-hydroxylase (C4H, EC 1.14.13.11) is the gene encoding enzyme that catalyzes the second step of phenylpropanoid pathway responsible for biosynthesis of lignin. A full-length cDNA of C4H (designated as GbC4H), which spanned 1816-bp with a 1518-bp open reading frame encoding a 505-amino-acid protein, was cloned from Ginkgo biloba. A GbC4H genomic DNA fragment, spanning 3249-bp, was cloned and found to contain two exons and one intron. GbC4H protein showed high similarities with other plant C4Hs to include conserved domains of cytochrome P450 family. GT-1, W-box, and Myb/Myc recognition sites involved in stress response were detected in a 1265-bp upstream promoter region of GbC4H. Phylogenetic analysis suggested the common evolutionary ancestor shared by plant C4Hs including the gymnosperm enzyme. pET-28a-GbC4H plasmid was constructed and expressed in Escherichia coli strain BL21. Enzymatic assay revealed that recombinant GbC4H protein catalyzes conversion of trans-cinnamic acid to p-coumaric acid. Expression analyses in different organs showed high expression of GbC4H in stems and roots, whereas low expressions was found in fruits, carpopodium, and petioles. Further analysis indicated linear correlation of lignin contents with transcript levels of GbC4H among different tissues. GbC4H transcription was increased by treatments with UV-B, cold, salicylic acid, and abscisic acid, indicating the possible role of GbC4H in response to stresses and hormonal signal. Understanding of GbC4H function could benefit molecular breeding and reinforcement of defense mechanisms in Ginkgo.
Plant physiology and biochemistry are both affected by salinity, which is an important abiotic stressor. In this study, we identified transcript-derived fragments (TDFs) in response to salt stress in black locust (Robinia pseudoacacia L.) using cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis. Seventy-four TDFs were identified in the leaves of two-year-old plants after NaCl treatment (500 mM for 0, 5, 10 and 15 days). Based on the gene ontology (GO) terminology, 30 TDFs shared high homology with known genes and were classified into 6 groups: metabolism-related factors, defense-related proteins, transcription factors, stress and signal transductionrelated factors and energy-related factors. Eight TDFs were selected for further study, and their expression patterns in the leaves were verified by real-time polymerase chain reaction (RT-PCR) at different stages of salt stress. Our data provide a theoretical basis for research on the mechanisms of salt tolerance in woody plants.
Drought, salinity and cold are the major environmental factors impacting on survival and productivity of Tibetan hulless barley in Tibetan Plateau of China. Tibetan hulless barley cultivar, Tibetan Heiqingke No. 1, has developed a strong tolerance and adaptation to stresses in relation to the wild barley. The differences of dehydrin gene transcription and translation between Tibetan Heiqingke No. 1 and the wild barley under drought, salinity and low temperature stresses were investigated in the present study to figure out the putative mechanism of stress tolerance of Tibetan Heiqingke No. 1. The leaf relative water contents (RWCs) decreased more slowly in Tibetan hulless barley than the wild barley under osmotic and low temperature conditions. Electrolyte leakage, malondialdehyde and H₂O₂ contents increased faster in wild barley than those of Tibetan hulless barley, which indicated that cells of wild barley received more damages than Tibetan hulless barley. Furthermore, the expression of several dehydrin genes, belonging to four different classifications respectively, was also investigated. Polyclonal antibodies against dehydrins were obtained from rabbit after prokaryotic expression and purification of TDHN4, a dehydrin protein from Tibetan hulless barley. With these antibodies and dehydrin gene fragments, western blotting analysis and RT-PCR showed that Tibetan Heiqingke No. 1 accumulated higher abundance of dehydrins than stresssensitive wild barley under all stress conditions.
Tobacco-specific nitrosamines (TSNAs) are carcinogenic chemicals found in tobacco plants. The increasing health consciousness of individuals had led to an increased interest in research on reducing TSNAs content. The aim of this study was to use a pot experiment in which exogenous substances were applied to burley tobacco to dissect the mechanism of TSNAs production. The results indicated that spraying the exogenous substances IAA, NAA, SA and combination thereof on burley tobacco after topping decreased TSNAs content by 2.69–29.4 % in upper leaves and 0.23–39.3 % in middle leaves without affecting total sugar, total nitrogen, potassium and chlorine contents. The application of exogenous substances could down-regulate expression of the NR gene and the activity of the NR enzyme, resulting in less accumulation of the TSNAs precursor nitrite. The exogenous substances significantly reduced nicotine accumulation, which was consistent with low enzyme activities and the down-regulated expressions of genes involved in nicotine biosynthesis, especially significant in the case of quinolinate phosphoribosyltransferase. These results suggested that the application of exogenous substances on burley tobacco after topping could reduce TSNAs content which may be attributed to the regulation of exogenous substances on nitrite and nicotine. This also implies one potential improvement to agronomic practices aimed at controlling the accumulation of TSNAs in burley tobacco.
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