Coupled hydraulics and carbon economy underlie age-related growth decline and revitalisation of sand-fixing shrubs after crown removal.

Crown removal revitalises sand-fixing shrubs that show declining vigour with age in drought-prone environments; however, the underlying mechanisms are poorly understood. Here, we addressed this knowledge gap by comparing the growth performance, xylem hydraulics and plant carbon economy across different plant ages (10, 21 and 33 years) and treatments (control and crown removal) using a representative sand-fixing shrub (Caragana microphylla Lam.) in northern China. We found that growth decline with plant age was accompanied by simultaneous decreases in soil moisture, plant hydraulic efficiency and photosynthetic capacity, suggesting that these interconnected changes in plant water relations and carbon economy were responsible for this decline. Following crown removal, quick resprouting, involving remobilisation of root nonstructural carbohydrate reserves, contributed to the reconstruction of an efficient hydraulic system and improved plant carbon status, but this became less effective in older shrubs. These age-dependent effects of carbon economy and hydraulics on plant growth vigour provide a mechanistic explanation for the age-related decline and revitalisation of sand-fixing shrubs. This understanding is crucial for the development of suitable management strategies for shrub plantations constructed with species having the resprouting ability and contributes to the sustainability of ecological restoration projects in water-limited sandy lands.

Hypoglycemic oligostilbenes from the stems of Caragana sinica.

Six new oligostilbenes, carastilphenols A-E (1-5) and (-)-hopeachinol B (6), with three reported oligostilbenes were obtained from the stems of Caragana sinica. The structures of compounds 1-6 were determined by comprehensive spectroscopy analysis, and their absolute configurations were determined by electronic circular dichroism calculations. Thus, natural tetrastilbenes were determined as absolute configuration for the first time. Also, we did several pharmacological essays. In the antiviral tests, compounds 2, 4 and 6 showed moderate anti-coxsackie virus B3 type (CVB3) effect on Vero cells activities in vitro with IC50 values of 19.2 âˆ¼ 69.3 µM; and compounds 3 and 4 showed different levels of anti-respiratory syncytial virus (RSV) effect on Hep2 cells activities in vitro with IC50 values of 23.1 and 33.3 µM, respectively. As for hypoglycemic activity, compounds 6-9 (10 µM) showed the inhibition of α-glucosidase in vitro with IC50 values of 0.1 âˆ¼ 0.4 µM; and compound 7 showed significant inhibition (88.8%, 10 µM) of protein tyrosine phosphatase 1B (PTP1B) with IC50 value of 1.1 µM in vitro.

Lignocellulose conversion of ensiled Caragana korshinskii Kom. facilitated by Pediococcus acidilactici and cellulases.

To explore the biofuel production potential of Caragana korshinskii Kom., Pediococcus acidilactici and an exogenous fibrolytic enzyme were employed to investigate the fermentation profile, structural carbohydrates degradation, enzymatic saccharification and the dynamics of bacterial community of C. korshinskii silage. After 60 d of ensiling, all additives increased the fermentation quality. The highest lactic and acetic acids and lowest non-protein nitrogen (NPN) and ammonia nitrogen (NH3 -N) were observed in P. acidilactici and Acremonium cellulase (PA + AC) treated silage. Additionally, all additives significantly increased the ferulic acid content and fibre degradability with the highest values obtained from PA + AC silage. The bacterial community in all silages was dominated by P. acidilactici throughout the entire fermentation process. The bacterial community was also modified by the silage additives exhibiting a relatively simple network of bacterial interaction characterized by a lower bacterial diversity in P. acidilactici (PA) treated silage. The highest 6-phospho-beta-glucosidase abundance was observed in PA-treated silage at the mid-later stage of ensiling. PA treatment exhibited lower structural carbohydrates degradation but performed better in lignocellulose conversion during enzymatic saccharification. These results indicated that pretreating C. korshinskii improved its silage quality and potential use as a lignocellulosic feedstock for the production of bio-product and biofuel.

Transcriptome-based identification and expression profiling of AP2/ERF members in Caragana intermedia and functional analysis of CiDREB3.

BACKGROUND: The AP2/ERF transcription factor family plays important roles in regulation of plant growth and development as well as the response of plants to stress. However, there are currently few studies focusing on the function of the AP2/ERF-type transcription factors in Caragana intermedia Kuang et H. C. Fu. Here, the expression pattern of AP2/ERF transcription factors family in different tissues and under four stress treatments were evaluated, and the function of CiDREB3 was examined. METHODS AND RESULTS: In this study, the genes encoding the AP2/ERF family of transcription factors were screened from the C. intermedia drought transcriptome database and subjected to bioinformatic analysis using the online tool and software. The expression pattern of the members of AP2/ERF transcription factors in C. intermedia were detected via quantitative real-time PCR (qRT-PCR). The function of CiDREB3 on growth, development and drought tolerance was evaluated by transgenic Arabidopsis. As a result, 22 sequences with complete ORFs were obtained and all sequences were divided into 13 sub-groups. Most of the AP2/ERF transcription factors exhibited tissue-specific expression and were induced by cold, heat, NaCl and mannitol treatments. Furthermore, heterologous expression of CiDREB3 altered the morphology of the transgenic Arabidopsis thaliana L. Heynh and improved its drought tolerance during seedlings development. CONCLUSIONS: Taken together, the results of the present study helped to better understand the function of the AP2/ERF family transcription factors in response to multiple abiotic stresses and uncovered the role of CiDREB3 in affecting the morphology and abiotic stress tolerance of Arabidopsis.

Functional characterization of an unobtrusive protein, CkMT4, in re-establishing desiccation tolerance in germinating seeds.

Desiccation tolerance (DT) is gradually lost during seed germination, while it can be re-established by pre-treatment with polyethylene glycol (PEG) and/or abscisic acid (ABA). Increasing knowledge is available on several stress-related proteins in DT re-establishment in herb seeds, but limited information exists on novel proteins in wood seeds. This study aimed to investigate the role of metallothionein CkMT4, a protein species with the highest fold increase in abundance in Caragana korshinskii seeds on PEG treatment. The fluctuation in mRNA levels of CkMT4 during seed development was consistent with the changes in DT, and the expression of CkMT4 could be up-regulated by ABA. Besides metal-binding capacity, CkMT4 might supply Cu2+/Zn2+ to superoxide dismutase (SOD) under high redox potential provided by PEG treatment for excess reactive oxygen species (ROS) scavenging. The overexpression of CkMT4 in yeast results in enhanced oxidation resistance. Experimentally, this study demonstrated the overexpression of CkMT4 in Arabidopsis seeds benefited the re-establishment of DT and enhanced the activity of SOD. On the whole, these findings suggested that CkMT4 facilitated the re-establishment of DT in C. korshinskii seeds mainly through diminishing excess ROS, which put the mechanism underlying the re-establishment of DT in xerophytic wood seeds into a new perspective.

Nutritive value and ruminal degradation of seven Chinese herbs as forage for Tan sheep.

Tan sheep is an indigenous ovine breed of China known for its high meat quality and pleasing taste. Seven herbs of traditional Chinese medicine, namely, Ephedra sinica, Glycyrrhiza uralensis, Caragana korshinskii, Allium mongolicum, Thymus vulgaris, Astragalus membranaceus, and Lespedeza bicolor are commonly grazed by Tan sheep. It has been widely believed that these herbs are of high nutritive value, which may significantly contribute to the high meat quality and distinct flavor of Tan sheep. However, the nutritive values of these herbs have not been evaluated to date. In this study, samples of the seven herbs were collected from the steppe of Yanchi County of Ningxia Autonomous Region of China. The dry matter (DM), crude protein (CP), ether extract (EE), ash (Ash), calcium (Ca), phosphorus (P), neutral detergent fiber (NDF), and acid detergent fiber (ADF) of these herbs were measured using locally cultivated alfalfa as the standard forage. Digestion of the dry matter, neutral detergent fiber, acid detergent fiber, and crude protein in the rumen of Tan sheep was examined using the nylon bag method, in order to evaluate their feeding nutritional value. Our results show that all the seven herbs meet the nutritional needs of ruminants based on the standard forage alfalfa. However, Ephedra, Glycyrrhiza, Caragana, Allium, Astragalus, and Lespedeza have higher nutritive value than Thymus (P < 0.05). According to the ruminal degradation rates of dry matter, neutral detergent fiber, acid detergent fiber, and crude protein, the nutritive value of Caragana, Allium, and Lespedeza is higher than that of Ephedra, Glycyrrhiza, Astragalus, and Thymus (P < 0.05). The overall nutritive value of Allium is the highest among the seven herbs and therefore Allium is recommended to better meet the nutritional needs of Tan sheep.

Neuraminidase Inhibitors from the Roots of Caragana sinica.

Two new oligostilbenes, caragasinins D and E, along with four known compounds, kobophenol A, α-viniferin, wistin, and 5-hydroxy-2-[(4-hydroxyphenyl)acetyl]-3-methoxybenzoic acid, were isolated from the roots of Caragana sinica. These compounds were spectroscopically analyzed for their structures and configurations and compared with existing data. The configurations of caragasinins D and E were elucidated by 1 H-NMR spectroscopy, CD spectroscopy, and time-dependent density-functional theory simulated ECD spectral data. All six compounds were evaluated for their inhibitory activity against neuraminidase (NA) from Clostridium perfringens. Among the tested compounds, 5-hydroxy-2-[(4-hydroxyphenyl)acetyl]-3-methoxybenzoic acid demonstrated statistically significant NA inhibitory activity, which was comparable to the positive control, mangiferin.

Alteration in isotopic composition of gross rainfall as it is being partitioned into throughfall and stemflow by xerophytic shrub canopies within water-limited arid desert ecosystems.

Isotopic composition of gross rainfall has been extensively used as a conservative tracer to track water movement and other hydrological processes in vegetated ecosystems. Recent studies from forest ecosystems, however, demonstrated that vegetation canopies can alter the isotopic composition of rainwater during rainfall partitioning into throughfall and stemflow, likely leading to errors and biases in aforementioned studies. No known studies, to date, had investigated this topic in shrub-dominated arid and semi-arid ecosystems where water is typically the driving factor in ecological, hydrological and biogeochemical processes. In this study, event-based gross rainfall, the throughfall and stemflow induced by shrubs of Caragana korshinskii were measured and samples were collected within a water-limited arid desert ecosystem of northern China, and their water stable isotopes (18O and 2H) were also analyzed in the laboratory. We mainly aimed to investigate whether there is an isotopic enrichment or depletion in stemflow and throughfall in comparison to gross rainfall, and to evaluate the possible underlying mechanisms. Our results indicated an enrichment of both isotopes in stemflow, while a general more depletion in throughfall than in gross rainfall, which is presumably affected by a combinative effects of canopy evaporation, isotopic exchange, and selective canopy storage. Deuterium excess of stemflow were found to be significantly higher (P < 0.05) than that of gross rainfall and throughfall. Moreover, we detected the pronounced "amount effect", with a significant (P < 0.05) negative relationship between isotopic composition and the amount of gross rainfall, throughfall, and stemflow, respectively. Our study is expected to contribute to an improved understanding of physical processes and water routing in shrub canopies within vast arid desert ecosystems.

Analysis of Metabolite Accumulation Related to Pod Color Variation of Caragana intermedia.

Caragana intermedia, a leguminous shrub widely distributed in cold and arid regions, is rich in secondary metabolites and natural active substances, with high nutritional and medical values. It is interesting that the pods of C. intermedia often show different colors among individual plants. In this study, 10-, 20- and 30-day-old red and green pods of C. intermedia were used to identify and characterize important metabolites associated with pod color. A total 557 metabolites, which could be classified into 21 groups, were detected in the pod extracts using liquid chromatography coupled with ESI-triple quadrupole-linear ion trap mass spectrometer (LC-ESI-MS/MS). Metabolomics analysis revealed significant differences in 15 groups of metabolites between red and green pods, including amino acids, nucleotide derivatives, flavonoids, and phytohormones. Metabolic pathway analysis showed that the shikimic acid and the phytohormone metabolic pathways were extraordinarily active in red pods, and the difference between red and green pods was obvious. Moreover, red pods showed remarkable flavonoids, cytokinins, and auxin accumulation, and the content of total flavonoids and proanthocyanidins in 30-day-old red pods was significantly higher than that in green pods. This metabolic profile contributes to valuable insights into the metabolic regulation mechanism in different color pods.

iTRAQ-based quantitative proteomic analysis reveals pathways associated with re-establishing desiccation tolerance in germinating seeds of Caragana korshinskii Kom.

Mature seeds of Caragana korshinskii Kom. lose desiccation tolerance (DT) during germination. However, DT can be re-induced by polyethylene glycol (PEG) or abscisic acid (ABA), and this provides a novel system to analyse the mechanism of DT. Although the global gene expression during the re-establishment of DT treated by PEG or ABA has been characterized, the changes in protein species abundance are still unknown. Here, we used an iTRAQ-based quantitative proteomics approach to perform a comparative analysis of protein abundances in germinated seeds of PEG-treated, ABA-treated and untreated radicles in response to the re-establishment of DT. A total of 3009 proteins were quantified; compared with the untreated samples, 274 and 261 differentially abundant protein species were found in PEG- and ABA-treated samples, respectively. The observed protein abundance patterns showed a clear overlap in seeds with ABA-and PEG-induced re-establishment of DT. During the re-establishment of DT, the expression of auxin-repressed protein and metallothionien-like protein was induced, and the content of proline increased significantly; however, proteins related to cell proliferation were found to be expressed at lower levels, and the glycolytic pathway was partially inhibited. These results provide promising insights into the molecular mechanisms underlying the re-establishment of DT in germinated seeds. BIOLOGICAL SIGNIFICANCE: The major challenge for seed storage is the complexity of the tolerance to desiccation. Re-establishment of desiccation tolerance (DT) in germinating seed as a model was used for DT researching. Although several researches have been focusing on the transcriptome of DT re-establishment, little proteomic information remained to be available. In the present study, the iTRAQ-based quantitative proteomic analysis was employed to identify the differentially accumulated protein species from radicles of germinating C. korshinskii seedling after ABA-, PEG- treatment or not. The data obtained not only provides basic information about proteome, but also characterize the quantitative changes that occur in the abundance of proteins species during DT re-establishment. These results are helpful understanding the mechanism of DT re-establishment induced by PEG or ABA.

Identification of the WRKY gene family and functional analysis of two genes in Caragana intermedia.

BACKGROUND: WRKY transcription factors, one of the largest families of transcriptional regulators in plants, play important roles in plant development and various stress responses. The WRKYs of Caragana intermedia are still not well characterized, although many WRKYs have been identified in various plant species. RESULTS: We identified 53 CiWRKY genes from C. intermedia transcriptome data, 28 of which exhibited complete open reading frames (ORFs). These CiWRKYs were divided into three groups via phylogenetic analysis according to their WRKY domains and zinc finger motifs. Conserved domain analysis showed that the CiWRKY proteins contain a highly conserved WRKYGQK motif and two variant motifs (WRKYGKK and WKKYEEK). The subcellular localization of CiWRKY26 and CiWRKY28-1 indicated that these two proteins localized exclusively to nuclei, supporting their role as transcription factors. The expression patterns of the 28 CiWRKYs with complete ORFs were examined through quantitative real-time PCR (qRT-PCR) in various tissues and under different abiotic stresses (drought, cold, salt, high-pH and abscisic acid (ABA)). The results showed that each CiWRKY responded to at least one stress treatment. Furthermore, overexpression of CiWRKY75-1 and CiWRKY40-4 in Arabidopsis thaliana suppressed the drought stress tolerance of the plants and delayed leaf senescence, respectively. CONCLUSIONS: Fifty-three CiWRKY genes from the C. intermedia transcriptome were identified and divided into three groups via phylogenetic analysis. The expression patterns of the 28 CiWRKYs under different abiotic stresses suggested that each CiWRKY responded to at least one stress treatment. Overexpression of CiWRKY75-1 and CiWRKY40-4 suppressed the drought stress tolerance of Arabidopsis and delayed leaf senescence, respectively. These results provide a basis for the molecular mechanism through which CiWRKYs mediate stress tolerance.

Natural neuro-inflammatory inhibitors from Caragana turfanensis.

Because of the critical role of over-activated microglia in the progress of neurodegenerative diseases, it has been selected as a potential therapeutic target for drug discovery. In order to find natural neuroinflammatory inhibitors, we carried out a bioactivity-oriented phytochemical research of Caragana turfanensis Kom. (Krassn.), which is a folk medicine widely distributed in Xinjiang. As a result, a new coumarin lactone caraganolide A (1) and 35 known components were characterized from the effective extract of C. turfanensis. Furthermore, their anti-neuroinflammatory effects were evaluated in LPS-induced BV2 microglial cells using Griess assay to determine the release of nitric oxide (NO). Compounds 1, 2, 4-6, 9, 13-15, 20, 29 and 30 exhibited significant inhibitory activities and no obvious cytotoxicities were observed at their effective concentrations. It is noteworthy, the new compound caraganolide A (1) (IC50 1.01±1.57µM) and 3',7,8-trihydroxy-4'-methoxyisoflavone (5) (IC506.87±2.23µM) exhibited more excellent action than that of positive control minocycline (IC50 9.07±0.86µM).

Modulating role of ROS in re-establishing desiccation tolerance in germinating seeds of Caragana korshinskii Kom.

In close agreement with visible germination, orthodox seeds lose desiccation tolerance (DT). This trait can be regained under osmotic stress, but the mechanisms are poorly understood. In this study, germinating seeds of Caragana korshinskii Kom. were investigated, focusing on the potential modulating roles of reactive oxygen species (ROS) in the re-establishment of DT. Germinating seeds with 2 mm long radicles can be rendered tolerant to desiccation by incubation in a polyethylene glycol (PEG) solution (-1.7 MPa). Upon PEG incubation, ROS accumulation was detected in the radicles tip by nitroblue tetrazolium chloride staining and further confirmed by confocal microscopy. The PEG-induced re-establishment of DT was repressed when ROS scavengers were added to the PEG solution. Moreover, ROS act downstream of abscisic acid (ABA) to modulate PEG-mediated re-establishment of DT and serve as a new inducer to re-establish DT. Transcriptomic analysis revealed that re-establishment of DT by ROS involves the up-regulation of key genes in the phenylpropanoid-flavonoid pathway, and total flavonoid content and key enzyme activity increased after ROS treatment. Furthermore, DT was repressed by an inhibitor of phenylalanine ammonia lyase. Our data suggest that ROS play a key role in the re-establishment of DT by regulating stress-related genes and the phenylpropanoid-flavonoid pathway.

Soil, Leaf and Root Ecological Stoichiometry of Caragana korshinskii on the Loess Plateau of China in Relation to Plantation Age.

Caragana korshinskii, a leguminous shrub, a common specie, is widely planted to prevent soil erosion on the Loess Plateau. The objective of this study was to determine how the plantation ages affected soil, leaf and root nutrients and ecological stoichiometry. The chronosequence ages of C. korshinskii plantations selected for this study were 10, 20 and 30 years. Soil organic carbon (SOC) and soil total nitrogen (STN) of C. korshinskii plantations significantly increased with increase in the chronosequence age. However, soil total phosphorous (STP) was not affected by the chronosequence age. The soil C: N ratio decreased and the soil C: P and N: P ratios increased with increasing plantation age. The leaf and root concentrations of C, N, and P increased and the ratios C: N, C: P, and N: P decreased with age increase. Leaf N: P ratios were >20, indicating that P was the main factor limiting the growth of C. korshinskii. This study also demonstrated that the regeneration of natural grassland (NG) effectively preserved and enhanced soil nutrient contents. Compared with NG, shrub lands (C. korshinskii) had much lower soil nutrient concentrations, especially for long (>20 years) chronosequence age. Thus, the regeneration of natural grassland is an ecologically beneficial practice for the recovery of degraded soils in this area.

Carbon Storage Patterns of Caragana korshinskii in Areas of Reduced Environmental Moisture on the Loess Plateau, China.

Precipitation patterns are influenced by climate change and profoundly alter the carbon sequestration potential of ecosystems. Carbon uptake by shrubbery alone accounts for approximately one-third of the total carbon sink; however, whether such uptake is altered by reduced precipitation is unclear. In this study, five experimental sites characterised by gradual reductions in precipitation from south to north across the Loess Plateau were used to evaluate the Caragana korshinskii's functional and physiological features, particularly its carbon fixation capacity, as well as the relationships among these features. We found the improved net CO2 assimilation rates and inhibited transpiration at the north leaf were caused by lower canopy stomatal conductance, which enhanced the instantaneous water use efficiency and promoted plant biomass as well as carbon accumulation. Regional-scale precipitation reductions over a certain range triggered a distinct increase in the shrub's organic carbon storage with an inevitable decrease in the soil's organic carbon storage. Our results confirm C. korshinskii is the optimal dominant species for the reconstruction of fragile dryland ecosystems. The patterns of organic carbon storage associated with this shrub occurred mostly in the soil at wetter sites, and in the branches and leaves at drier sites across the arid and semi-arid region.

Impacts of leguminous shrub encroachment on neighboring grasses include transfer of fixed nitrogen.

Shrub encroachment induced by global change and human disturbance strongly affects ecosystem structure and function. In this study, we explore the degree to which invading leguminous shrubs affected neighboring grasses, including via the transfer of fixed nitrogen (N). We measured N concentrations and natural abundance (15)N of shoot tissues from three dominant grasses from different plant functional groups across seven distances along a local transect (up to 500 cm) to the leguminous shrub, Caragana microphylla. C. microphylla did transfer fixed N to neighboring grasses, but the amount and distance of N transferred were strongly species-specific. Shoot N concentrations decreased significantly with distance from C. microphylla, for a rhizomatous grass, Leymus chinensis, and a bunchgrass, Achnatherum sibiricum. However, N concentrations of another bunchgrass, Stipa grandis, were higher only directly underneath the shrub canopy. Shoot δ(15)N values of L. chinensis were enriched up to 500 cm from the shrub, but for S. grandis were enriched only below the shrub canopy. In contrast, δ(15)N of A. sibiricum did not change along the 500-cm transect. Our results indicated the rhizomatous grass transferred fixed N over long distances while bunchgrasses did not. The presence of C. microphylla increased the shoot biomass of L. chinensis but decreased that of S. grandis and A. sibiricum. These findings highlight the potential role of nutrient-acquisition strategies of neighboring grasses in moderating the interspecific variation of fixed N transfer from the leguminous shrub. Overall, leguminous shrubs have either positive or negative effects on the neighboring grasses and dramatically affect plant community composition and structure.

Two NAC transcription factors from Caragana intermedia altered salt tolerance of the transgenic Arabidopsis.

BACKGROUND: Plants are continuously challenged by different environment stresses, and they vary widely in their adjustability. NAC (NAM, ATAF and CUC) transcription factors are known to be crucial in plants tolerance response to abiotic stresses, such as drought and salinity. ANAC019, ANAC055, and ANAC072, belong to the stress-NAC TFs, confer the Arabidopsis abiotic stress tolerance. RESULTS: Here we isolated two stress-responsive NACs, CiNAC3 and CiNAC4, from Caragana intermedia, which were induced by ABA and various abiotic stresses. Localization assays revealed that CiNAC3 and CiNAC4 localized in the nuclei, consistent with their roles as transcription factors. Histochemistry assay using Pro(CiNAC4)::GUS transgenic Arabidopsis showed that the expression of the GUS reporter was observed in many tissues of the transgenic plants, especially in the root vascular system. Overexpression of CiNAC3 and CiNAC4 reduced ABA sensitivity during seed germination, and enhanced salt tolerance of the transgenic Arabidopsis. CONCLUSIONS: We characterised CiNAC3 and CiNAC4 and found that they were induced by numerous abiotic stresses and ABA. GUS histochemical assay of CiNAC4 promoter suggested that root, flower and local damaged tissues were the strongest stained tissues. Overexpression assay revealed that CiNAC4 play essential roles not only in promoting lateral roots formation, but also in responding to salinity and ABA treatment of Arabidopsis.

Changes in spatial patterns of Caragana stenophylla along a climatic drought gradient on the Inner Mongolian Plateau.

Few studies have investigated the influence of water availability on plant population spatial patterns. We studied changes in the spatial patterns of Caragana stenophylla along a climatic drought gradient within the Inner Mongolian Plateau, China. We examined spatial patterns, seed density, "nurse effects" of shrubs on seedlings, transpiration rates and water use efficiency (WUE) of C. stenophylla across semi-arid, arid, and intensively arid zones. Our results showed that patches of C. stenophylla populations shifted from a random to a clumped spatial pattern towards drier environments. Seed density and seedling survival rate of C. stenophylla decreased from the semi-arid zone to the intensively arid zone. Across the three zones, there were more C. stenophylla seeds and seedlings underneath shrub canopies than outside shrub canopies; and in the intensively arid zone, there were almost no seeds or seedlings outside shrub canopies. Transpiration rates of outer-canopy leaves and WUE of both outer-canopy and inner-canopy leaves increased from the semi-arid zone to the intensively arid zone. In the intensively arid zone, transpiration rates and WUE of inner-canopy leaves were significantly lower and higher, respectively, than those of outer-canopy leaves. We conclude that, as drought stress increased, seed density decreased, seed proportions inside shrubs increased, and "nurse effects" of shrubs on seedlings became more important. These factors, combined with water-saving characteristics associated with clumped spatial patterns, are likely driving the changes in C. stenophylla spatial patterns.

Kobophenol A inhibits sodium nitroprusside-induced cardiac H9c2 cell death through suppressing activation of JNK and preserving mitochondrial anti-apoptotic Bcl-2 and Mcl-1.

Sodium nitroprusside (SNP) releases nitric oxide (NO), a powerful vasodilator, and thus widely used in intensive care unit for treating hypertension emergency. However, cardiac toxicity after SNP administration is a clinical problem. For finding a natural compound that suppressing SNP-induced cardiac toxicity, we tested the protective potential of kobophenol A (Kob A), purified from the root of Caragana sinica, against the toxic effects of SNP. The severe cardiac H9c2 cell death was induced by SNP (2 mM) treatment. Kob A ameliorated SNP-induced cardiac H9c2 cell death, and this protective effect of Kob A may be related to the inhibition of c-Jun NH2-terminal kinase (JNK) and p38 mitogen-activated protein (MAP) kinase activation following SNP administration. In addition, the downregulation of cellular Bcl-2 and Mcl-1 levels by SNP exposure was strongly abrogated in the presence of Kob A. These biological properties of Kob A might provide insights into developing new cardioprotectant against SNP-induced cardiac cell death.

Reference gene selection for qRT-PCR in Caragana korshinskii Kom. under different stress conditions.

Caragana korshinskii Kom., which is widely distributed in the northwest China and Mongolia, is an important forage bush belonging to the legume family with high economic and ecological value. Strong tolerance ability to various stresses makes C. korshinskii Kom. a valuable species for plant stress research. In this study, suitable reference genes for quantitative real-time reverse transcription PCR (qRT-PCR) were screened from 11 candidate reference genes, including ACT, GAPDH, EF1α, UBQ, TUA, CAP, TUB, TUB3, SKIP1, SKIP5-1 and SKIP5-2. A total of 129 samples under drought, heat, cold, salt, ABA and high pH treatment were profiled, and software such as geNORM, NormFinder and BestKeeper were used for reference gene evaluation and selection. Different suitable reference genes were selected under different stresses. Across all 129 samples, GAPDH, EF1α and SKIP5-1 were found to be the most stable reference genes, and EF1α+SKIP5-1 is the most stable reference gene combination. Conversely, TUA, TUB and SKIP1 were not suitable for using as reference genes owing to their great expression variation under some stress conditions. The relative expression levels of CkWRKY1 were detected using the stable and unstable reference genes and their applicability was confirmed. These results provide some stable reference genes and reference gene combinations for qRT-PCR under different stresses in C. korshinskii Kom. for future research work, and indicate that CkWRKY1 plays essential roles in response to stresses in C. korshinskii.