叶施NO对NaCl胁迫下红砂幼苗叶片和根系中氮代谢酶及营养物质的影响

以当年生红砂(Reaumuria soongorica)幼苗为材料,采用盆栽实验,考察叶面喷施不同浓度(0、0.01、0.10、0.25、0.50、1.00 mmol·L^-1)NO供体硝普钠 (SNP) 对NaCl(300 mmol·L^-1)胁迫下红砂根、叶中可溶性蛋白、游离氨基酸和硝态氮含量,以及谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、硝酸还原酶(NR)活性的影响,并采用主成分分析和隶属函数法筛选NO对NaCl胁迫缓解效应的氮代谢指标和最佳NO浓度,以探讨外源NO对NaCl 胁迫下红砂缓解效应的氮代谢响应机制。结果表明：(1)在300 mmol·L^-1 NaCl胁迫处理下,红砂幼苗根、叶中可溶性蛋白、硝态氮含量以及GS、GOGAT、NR活性均比对照显著下降。(2)外源NO能显著提高盐胁迫下红砂叶、根中GS、GOGAT、NR活性和硝态氮含量,增加根中可溶性蛋白和游离氨基酸含量。(3)NR和GOGAT活性可用于评价NO对NaCl胁迫下红砂幼苗的缓解作用,外源NO(SNP)对红砂幼苗在NaCl胁迫下的缓解效果强弱表现为0.25 mmol·L^-1> 0.50 mmol·L^-1> 0.10 mmol·L^-1> 1.00 mmol·L^-1> 0.01 mmol·L^-1。研究发现,300 mmol·L^-1 NaCl胁迫显著抑制了红砂幼苗氮代谢,外源NO(SNP)有助于提高盐胁迫下红砂NR活性,加快硝态氮转化为铵态氮,促进红砂叶片和根中GS/GOGAT对转化物的同化,从而增强红砂幼苗的耐盐性,并以0.25 mmol·L^-1SNP处理时缓解作用最佳;NR和GOGAT活性可作为NO缓解盐胁迫的评价指标。     

The chromosome number, karyotype and genome size of the desert plant diploid Reaumuria soongorica (Pall.) Maxim

Extreme drought and salt resistant plant Reaumuria soongorica is of great potential for revealing genetic bases unique to naturally stress-tolerant plants. A preliminary genome survey, including chromosome number, karyotype, chromosomal localization of 45S rDNA loci and genome size was conducted with R. soongorica collected from Lanzhou, China. Chromosome counting showed that R. soongorica is diploid with chromosome number of 22. Karyotypical analysis illustrated that the chromosomes size ranges from 3.38 to 5.51 μm, and the chromosomal formula is 2n = 2x = 22 = 4 m + 14sm + 4st. Fluorescence in situ hybridization revealed that four pairs of 45SrDNA signals were detected at the end of R. soongorica chromosomes. The flow cytometry analysis indicated that the mean C value of R. soongorica is 0.806 pg with predicted genome size of about 778 Mb. The results indicate that the extreme drought and salt resistance of R. soongorica was not attributed to a big and complicate genome and also offer some clues in resolving the problems of taxonomy and evolution in Tamaricaceae.     

干旱胁迫下红砂幼苗非结构性碳水化合物动态变化特征

该试验以荒漠区主要建群种红砂幼苗为研究对象,设置适宜水分(CK)、轻度干旱(MD)、中度干旱(SD)和重度干旱(VSD)4个胁迫处理(即田间持水量的80%、60%、40%和20%),采用盆栽控水试验,分别测定干旱胁迫15、30、45和60 d时红砂幼苗的叶、茎、粗根和细根中非结构碳水化合物(NSC)及其组分的含量,分析不同胁迫强度下不同干旱持续时间红砂幼苗NSC的动态变化及各组分差异,以揭示红砂NSC对干旱胁迫的响应机制。结果表明:(1)干旱胁迫强度和胁迫持续时间对红砂幼苗不同器官NSC及其组分均有显著影响,其中胁迫持续时间对NSC动态变化的影响尤为显著。(2)干旱胁迫初期,红砂叶中的NSC含量呈下降趋势,而茎中的NSC含量呈上升趋势,粗根和细根中NSC含量在各胁迫处理下基本保持稳定。(3)干旱胁迫后期,红砂叶和茎中的可溶性糖、淀粉和NSC含量逐渐增加,而粗根和细根中的淀粉和NSC含量呈下降趋势(中度干旱除外),且这一时期重度干旱处理下各器官可溶性糖和NSC的含量明显高于CK。研究发现,重度干旱胁迫能显著诱导提高红砂幼苗不同器官中的NSC含量,并通过分解根中淀粉和增加叶片中可溶性糖含量的方式来调节细胞渗透势平衡,以维持细胞活力,进而保持红砂在干旱胁迫后期的存活。     

Reference Gene Selection for Quantitative Real-Time PCR Normalization in Reaumuria soongorica

Despite its superiority for evaluating gene expression, real-time quantitative polymerase chain reaction (qPCR) results can be significantly biased by the use of inappropriate reference genes under different experimental conditions. Reaumuria soongorica is a dominant species of desert ecosystems in arid central Asia. Given the increasing interest in ecological engineering and potential genetic resources for arid agronomy, it is important to analyze gene function. However, systematic evaluation of stable reference genes should be performed prior to such analyses. In this study, the stabilities of 10 candidate reference genes were analyzed under 4 kinds of abiotic stresses (drought, salt, dark, and heat) within 4 accessions (HG010, HG020, XGG030, and XGG040) from 2 different habitats using 3 algorithms (geNorm, NormFinder, and BestKeeper). After validation of the ribulose-1,5-bisphosphate carboxylase/oxygenase large unite (rbcL) expression pattern, our data suggested that histone H2A (H2A) and eukaryotic initiation factor 4A-2 (EIF4A2) were the most stable reference genes, cyclophilin (CYCL) was moderate, and elongation factor 1α (EF1α) was the worst choice. This first systematic analysis for stably expressed genes will facilitate future functional analyses and deep mining of genetic resources in R. soongorica and other species of the Reaumuria genus.     

盐胁迫下荒漠共生植物红砂与珍珠的根茎叶中离子吸收与分配特征

西北荒漠地区C3小灌木红砂(Reaumuria soongorica)和C4半灌木珍珠猪毛菜(Salsola passerina)在特定环境下混生在一起,分布面积广阔。以采自腾格里沙漠边缘荒漠地带的天然野生珍珠猪毛菜和红砂群落的幼苗为材料,经0、100、200、300、400mmol/L NaCl盐溶液共同胁迫10 d,检测它们的含水量、主要矿质离子在根茎叶的含量与分布,揭示二者耐盐的共生协同的离子平衡适应机制。试验结果发现,珍珠猪毛菜叶片具有"吸钾排钠的"的耐盐特征,红砂叶片具备"吸钠排钾"的特征,吸收利用无机矿质离子具备互补效应。二者耐盐Cl、Ca和Si离子吸收与累积能力存在很大差异:随着盐胁迫程度加剧,红砂的根茎叶中Cl离子含量持续增加,并且为珍珠猪毛菜的2—5倍;珍珠猪毛菜根中Ca离子含量为红砂的2—3倍,但含量变化不显著;红砂根中Si离子含量迅速降低后稳定,并且是珍珠猪毛菜根的3—5倍,其他器官变化差异较小。因此,红砂与珍珠猪毛菜的共培养盐胁迫下根中吸收的离子侧重不同,红砂以Na、Cl、Si为主,珍珠猪毛菜以K、Ca为主。随着盐胁迫的程度加强,离子选择吸收系数S k,Na的变化趋势降低,表明二者叶部对Na的选择性减小,K的选择性吸收积累增大,增强了它们的抗盐性,最终使叶片所受盐害减小。总之红砂与珍珠猪毛菜共生的耐盐离子稳态机制显著不同,离子吸收与分布具有互补互利的效应。     

Environmental stresses induce health-promoting phytochemicals in lettuce

Plants typically respond to environmental stresses by inducing antioxidants as a defense mechanism. As a number of these are also phytochemicals with health-promoting qualities in the human diet, we have used mild environmental stresses to enhance the phytochemical content of lettuce, a common leafy vegetable. Five-week-old lettuce (Lactuca sativa L.) plants grown in growth chambers were exposed to mild stresses such as heat shock (40 °C for 10 min), chilling (4 °C for 1 d) or high light intensity (800 μmol m^?2 s^?1 for 1 d). In response to these stresses, there was a two to threefold increase in the total phenolic content and a significant increase in the antioxidant capacity. The concentrations of two major phenolic compounds in lettuce, chicoric acid and chlorogenic acid, increased significantly in response to all the stresses. Quercetin-3-O-glucoside and luteolin-7-O-glucoside were not detected in the control plants, but showed marked accumulations following the stress treatments. The results suggest that certain phenolic compounds can be induced in lettuce by environmental stresses. Of all the stress treatments, high light produced the greatest accumulation of phenolic compounds, especially following the stress treatments during the recovery. In addition, key genes such as phenylalanine ammonia-lyase (PAL), l-galactose dehydrogenase (l-GalDH), and γ-tocopherol methyltransferase (γ-TMT) involved in the biosynthesis of phenolic compounds, ascorbic acid, and α-tocopherol, respectively, were rapidly activated by chilling stress while heat shock and high light did not appear to have an effect on the expression of PAL and γ-TMT. However, l-GalDH was consistently activated in response to all the stresses. The results also show that these mild environmental stresses had no adverse effects on the overall growth of lettuce, suggesting that it is possible to use mild environmental stresses to successfully improve the phytochemical content and hence the health-promoting quality of lettuce with little or no adverse effect on its growth or yield.     

Antioxidant responses of wheat plants under stress

Currently, food security depends on the increased production of cereals such as wheat (Triticum aestivum L.), which is an important source of calories and protein for humans. However, cells of the crop have suffered from the accumulation of reactive oxygen species (ROS), which can cause severe oxidative damage to the plants, due to environmental stresses. ROS are toxic molecules found in various subcellular compartments. The equilibrium between the production and detoxification of ROS is sustained by enzymatic and nonenzymatic antioxidants. In the present review, we offer a brief summary of antioxidant defense and hydrogen peroxide (H₂O₂) signaling in wheat plants. Wheat plants increase antioxidant defense mechanisms under abiotic stresses, such as drought, cold, heat, salinity and UV-B radiation, to alleviate oxidative damage. Moreover, H₂O₂ signaling is an important factor contributing to stress tolerance in cereals.     

Lipotoxicity in renal proximal tubular cells: Relationship between endoplasmic reticulum stress and oxidative stress pathways

Hyperlipidemia in the general population has been linked to the development of chronic kidney disease with both oxidative and endoplasmic reticulum stress implicated. Physiological levels (50-300 µmol/L) of saturated fatty acids such as palmitic acid (PA) cause cytotoxicity in vitro. We investigated cell type- and stimulus-specific signaling pathways induced by PA in renal proximal tubular cells and whether oxidative stress leads to ER stress or vice versa and which pathways predominate in signaling for PA-induced apoptosis and necrosis. NRK-52E cells were incubated with PA or hydrogen peroxide (H₂O₂) combined with SP600125 which blocks c-Jun N-terminal kinase (JNK) activation; salubrinal, which maintains eukaryotic initiation factor 2α in its phosphorylated state and the antioxidant EUK-134 - a superoxide dismutase mimetic with catalase activity. We found that (i) PA causes both oxidative and ER stress leading to apoptosis which is mediated by phosphorylated JNK; (ii) oxidant-induced apoptosis generated by H₂O₂ involves ER stress signaling and CHOP expression; (iii) the ER stress mediated by PA is largely independent of oxidative stress; (iv) in contrast, the apoptosis produced by PA is mediated partly via oxidative stress. PA-mediated cell signaling in renal NRK-52E cells therefore differs from that identified in neuronal, hepatic and pancreatic beta cells.     

Enhancement of PPAR-β activity by repetitive low-grade H2O2 stress protects human umbilical vein endothelial cells from subsequent oxidative stress-induced apoptosis

Repetitive stress has been shown to up-regulate antioxidant defense and increase survival after subsequent oxidative injury. The up-regulation of antioxidant defense has been identified as an underlying cause of the apoptosis-inhibitory effects exerted by repetitive stress. However, it remains unclear what the important signaling mechanisms are by which cells preexposed to low-grade stress deal with apoptosis-inducing stress. In this study, we repetitively stressed human umbilical vein endothelial cells (HUVECs) through multiple exposures to a low dose (30 μM) of H₂O₂ in culture for 4 weeks. We then examined the effects of repetitive stress on PPAR-β expression and activity as well as the role of PPAR-β in the protective potency of repetitive stress. Our results show that repetitive stress enhances PPAR-β expression and activity, thereby inhibiting oxidative stress-induced apoptosis. Further, PPAR-β-directed antisense oligonucleotides reduced the PPAR-β protein content, enhanced the H₂O₂-mediated apoptosis, and ablated the protective effect of repetitive low-grade H₂O₂ stress. The specific PPAR-β agonist L-165041 significantly potentiated the apoptosis induced by H₂O₂ (p < 0.05) and increased the protective effect of repetitive stress. These findings indicate that repetitive low-grade H₂O₂ stress protects HUVECs from subsequent oxidative stress-induced apoptosis by enhancing PPAR-β expression and activity.     

Lipoic acid prevents liver metabolic changes induced by administration of a fructose-rich diet

Background

To evaluate whether co-administration of R/S-α-lipoic acid can prevent the development of oxidative stress and metabolic changes induced by a fructose-rich diet (F).

Methods

We assessed glycemia in the fasting state and during an oral glucose tolerance test, triglyceridemia and insulinemia in rats fed with standard diet (control) and fructose without or with R/S-α-lipoic acid. Insulin resistance and hepatic insulin sensitivity were also calculated. In liver, we measured reduced glutathione, protein carbonyl groups, antioxidant capacity by ABTS assay, antioxidant enzymes (catalase and superoxide dismutase 1 and 2), uncoupling protein 2, PPARδ and PPARγ protein expressions, SREBP-1c, fatty acid synthase and glycerol-3-phosphate acyltransferase-1 gene expression, and glucokinase activity.

Results

R/S-α-lipoic acid co-administration to F-fed rats a) prevented hyperinsulinemia, hypertriglyceridemia and insulin resistance, b) improved hepatic insulin sensitivity and glucose tolerance, c) decreased liver oxidative stress and increased antioxidant capacity and antioxidant enzymes expression, d) decreased uncoupling protein 2 and PPARδ protein expression and increased PPARγ levels, e) restored the basal gene expression of PPARδ, SREBP-1c and the lipogenic genes fatty acid synthase and glycerol-3-phosphate acyltransferase, and f) decreased the fructose-mediated enhancement of glucokinase activity.

Conclusions

Our results suggest that fructose-induced oxidative stress is an early phenomenon associated with compensatory hepatic metabolic mechanisms, and that treatment with an antioxidant prevented the development of such changes.

General significance

This knowledge would help to better understand the mechanisms involved in liver adaptation to fructose-induced oxidative stress and to develop effective strategies to prevent and treat, at early stages, obesity and type 2 diabetes mellitus.