Metabolism of reactive nitrogen species in pea plants under abiotic stress conditions

Nitric oxide (*NO) is a key signaling molecule in different physiological processes of animals and plants. However, little is known about the metabolism of endogenous *NO and other reactive nitrogen species (RNS) in plants under abiotic stress conditions. Using pea plants exposed to six different abiotic stress conditions (high light intensity, low and high temperature, continuous light, continuous dark and mechanical wounding), several key components of the metabolism of RNS including the content of *NO, S-nitrosothiols (RSNOs) and nitrite plus nitrate, the enzyme activities of l-arginine-dependent nitric oxide synthase (NOS) and S-nitrosogluthathione reductase (GSNOR), and the profile of protein tyrosine nitration (NO(2)-Tyr) were analyzed in leaves. Low temperature was the stress that produced the highest increase of NOS and GSNOR activities, and this was accompanied by an increase in the content of total *NO and S-nitrosothiols, and an intensification of the immunoreactivity with an antibody against NO(2)-Tyr. Mechanical wounding, high temperature and light also had a clear activating effect on the different indicators of RNS metabolism in pea plants. However, the total content of nitrite and nitrate in leaves was not affected by any of these stresses. Considering that protein tyrosine nitration is a potential marker of nitrosative stress, the results obtained suggest that low and high temperature, continuous light and high light intensity are abiotic stress conditions that can induce nitrosative stress in pea plants.     

Progress and challenges for abiotic stress proteomics of crop plants

Plants are continually challenged to recognize and respond to adverse changes in their environment to avoid detrimental effects on growth and development. Understanding the mechanisms that crop plants employ to resist and tolerate abiotic stress is of considerable interest for designing agriculture breeding strategies to ensure sustainable productivity. The application of proteomics technologies to advance our knowledge in crop plant abiotic stress tolerance has increased dramatically in the past few years as evidenced by the large amount of publications in this area. This is attributed to advances in various technology platforms associated with MS‐based techniques as well as the accessibility of proteomics units to a wider plant research community. This review summarizes the work which has been reported for major crop plants and evaluates the findings in context of the approaches that are widely employed with the aim to encourage broadening the strategies used to increase coverage of the proteome     

Physiological aspects of raffinose family oligosaccharides in plants: protection against abiotic stress

Abiotic stresses resulting from water deficit, high salinity or periods of drought adversely affect plant growth and development and represent major selective forces during plant evolution. The raffinose family oligosaccharides (RFOs) are synthesised from sucrose by the subsequent addition of activated galactinol moieties donated by galactinol. RFOs are characterised as compatible solutes involved in stress tolerance defence mechanisms, although evidence also suggests that they act as antioxidants, are part of carbon partitioning strategies and may serve as signals in response to stress. The key enzyme and regulatory point in RFO biosynthesis is galactinol synthase (GolS), and an increase of GolS in expression and activity is often associated with abiotic stress. It has also been shown that different GolS isoforms are expressed in response to different types of abiotic stress, suggesting that the timing and accumulation of RFOs are controlled for each abiotic stress. However, the accumulation of RFOs in response to stress is not universal and other functional roles have been suggested for RFOs, such as being part of a carbon storage mechanism. Transgenic Arabidopsis plants with increased galactinol and raffinose concentrations had better ROS scavenging capacity, while many sugars have been shown in vitro to have antioxidant activity, suggesting that RFOs may also act as antioxidants. The RFO pathway also interacts with other carbohydrate pathways, such as that of O‐methyl inositol (OMI), which shows that the functional relevance of RFOs must not be seen in isolation to overall carbon re‐allocation during stress responses.     

Glutathione reductase from pea leaves: response to abiotic stress and characterization of the peroxisomal isozyme

The glutathione reductase (GR; EC 1.6.4.2) isozyme present in peroxisomes has been purified for the first time, and its unequivocal localization in these organelles, by immunogold electron microscopy, is reported. The enzyme was purified c. 21-fold with a specific activity of 9523 units mg(-1) protein, and a yield of 44 microg protein kg(-1) leaves was obtained. The subunit size of the peroxisomal GR was 56 kDa and the isoelectric point was 5.4. The enzyme was recognized by a polyclonal antibody raised against total GR from pea (Pisum sativum) leaves. The localization of GR in peroxisomes adds to chloroplasts and mitochondria where GR isozymes are also present, and suggests a multiple targeting of this enzyme to distinct cell compartments depending on the metabolism of each organelle under the plant growth conditions. The expression level of GR in several organs of pea plants and under different stress conditions was investigated. The possible role of peroxisomal GR under abiotic stress conditions, such as cadmium toxicity, high light, darkness, high temperature, wounding and low temperature, is discussed.     

Influence of abiotic stress signals on secondary metabolites in plants

Plant secondary metabolites are unique sources for pharmaceuticals, food additives, flavors, and industrially important biochemicals. Accumulation of such metabolites often occurs in plants subjected to stresses including various elicitors or signal molecules. Secondary metabolites play a major role in the adaptation of plants to the environment and in overcoming stress conditions. Environmental factors viz. temperature, humidity, light intensity, the supply of water, minerals, and CO₂ influence the growth of a plant and secondary metabolite production. Drought, high salinity, and freezing temperatures are environmental conditions that cause adverse effects on the growth of plants and the productivity of crops. Plant cell culture technologies have been effective tools for both studying and producing plant secondary metabolites under in vitro conditions and for plant improvement. This brief review summarizes the influence of different abiotic factors include salt, drought, light, heavy metals, frost etc. on secondary metabolites in plants. The focus of the present review is the influence of abiotic factors on secondary metabolite production and some of important plant pharmaceuticals. Also, we describe the results of in vitro cultures and production of some important secondary metabolites obtained in our laboratory.     

Molecular cloning,characterization and expression analysis of BcHHP3 under abiotic stress in Pak-choi (Brassica rapa ssp. Chinensis)

In this study, BcHHP3 was isolated from Pak-choi; it has an open-reading frame (ORF) of 1044 base pairs, encoding 347 amino acids, a molecular weight of 39.35?kDa, isoelectric point (pI) of 9.08, an instability index of 48.35, and grand average of hydropathicity of 0.382. Multi-alignment and phylogenetic analysis showed that BcHHP3 bears a high similarity to AtHHP2. As predicted by SOMPA and SWISS-MODEL databases, the structure of the BcHHP3 protein is relatively stable and highly conservative. Real-time quantitative polymerase chain reaction (qRT-PCR) analysis showed that BcHHP3 was induced to co-express under cold and abscisic acid (ABA) stresses. The BcHHP3-GFP fusion protein was localized on the cell membrane and nuclear membrane. This work might be useful for future analysis of other HHP-like genes in Pak-choi.     

辣椒OSCA基因家族的全基因组鉴定及不同胁迫条件下表达分析

钙通透性阳离子通道蛋白(OSCA)在渗透胁迫感应中发挥重要的作用.本研究利用辣椒(Capsicum an-nuum L.)全基因组信息,鉴定出14个OSCA基因(CaOSCA),染色体定位及同源性分析表明,它们分别位于1、2、4、6、7、8、9、12号染色体上,亚细胞定位预测表明其编码产物均位于质膜上.系统进化树分析这...     

Isolation and properties of lactate dehydrogenase from germinating pea plants

Lactate dehydrogenase (LDH) was isolated from pea seedlings by means of protamine sulphate and (NH₄)₂SO₄ fractionation and chromatography on DEAE-cellulose and Sephadex G-150. The enzyme had a MW of ca 145 500. The kinetic properties studied were the lactate oxidation pH optimum (9·1) and the pyruvate reduction pH optimum (7·1). K_m values were determined for four natural substrates (Lactate, pyruvate, NAD⁺ and NADH) and for other acids (glycollate, α-ketoglutarate and glyoxylate). The K_i value was determined for p-chloromercuribenzoate (PCMB) which is a noncompetitive inhibitor of LDH from pea plants, and the course of irreversible inhibition of the enzyme by iodoacetamide (IA) and n-ethylmaleimide (NEMI) was studied. Preincubation of LDH with the coenzyme protects against PCMB inhibition, indicating the important role of the sulfhydryl group in the active site.     

Silicon: a duo synergy for regulating crop growth and hormonal signaling under abiotic stress conditions

Abiotic stresses, such as salinity, heavy metals and drought, are some of the most devastating factors hindering sustainable crop production today. Plants use their own defensive strategies to cope with the adverse effects of these stresses, via the regulation of the expression of essential phytohormones, such as gibberellins (GA), salicylic acid (SA), jasmonates (JA), abscisic acid (ABA) and ethylene (ET). However, the efficacy of the endogenous defensive arsenals of plants often falls short if the stress persists over an extended period. Various strategies are developed to improve stress tolerance in plants. For example, silicon (Si) is widely considered to possess significant potential as a substance which ameliorate the negative effects of abiotic stresses, and improves plant growth and biomass accumulation. This review aims to explain how Si application influences the signaling of the endogenous hormones GA, SA, ABA, JA and ET during salinity, wounding, drought and metal stresses in crop plants. Phytohormonal cross talk plays an important role in the regulation of induced defences against stress. However, detailed molecular and proteomic research into these interactions is needed in order to identify the underlying mechanisms of stress tolerance that is imparted by Si application and uptake.     

Comparative functional analysis of three abiotic stress-inducible promoters in transgenic rice

To identify minimal effective promoters for driving abiotic stress-inducible transgene expression in rice, we selected promoter elements of three stress-responsive genes, viz. rab16A coding for dehydrin, OsABA2 coding for zeaxanthin epoxidase, and a gene coding for a hypothetical protein (HP1) based on the presence of ABA-, salt- and drought-responsive cis-acting elements. These were translationally fused to the gusA reporter gene and introduced into rice to study their effect on heterologous gene expression. The OsABA2 promoter was found to be the most effective and desirable promoter among the three in terms of driving a low constitutive transgene expression under normal conditions and high induction in response to ABA, salt and drought stress, the highest being a 12-fold induction in response to ABA. The rab16A and HP1 promoters resulted in high levels of constitutive expression. While induction of GUS activity was generally two- to threefold for all the treatments in roots for both the promoters, induction in leaves was generally insignificant, the exceptions being rab16A in response to continuous salt stress and HP1 in response to water deficit. It was also observed that the three promoters, in general, resulted in lower constitutive expression, but higher induction in roots as compared to leaves. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. R. Wu: Deceased. This paper is humbly dedicated to the memory of Professor Ray Wu.     

非生物胁迫下耐辐射异常球菌drB0118基因功能分析

【目的】鉴定和确定被预测为编码干燥相关蛋白的耐辐射异常球菌(Deinococcus radiodurans) drB0118基因功能,探讨该基因对盐、渗透和氧化胁迫抗性的作用。【方法】构建drB0118基因缺失突变株(ΔB0118),通过氯化钠、D-山梨糖醇和过氧化氢等胁迫冲击实验及氧化胁迫条件下qRT-PCR分析,研究drB0118突变对非生物胁迫反应及氧化胁迫相关基因表达的影响。【结果】drB0118突变导致菌株对NaCl和D-sorbitol胁迫的抗性降低;对氧化胁迫(H2O2)敏感;qRT-PCR分析显示,drB0118突变引起氧化胁迫抗性基因pod和oxyR分别下调4倍和10倍。【结论】D. radiodurans中drB0118参与了盐、渗透和氧化等多种非生物胁迫反应。