Molybdenum application enhances antioxidant enzyme activity and COR15a protein expression under cold stress in wheat

Under cold stress, reactive oxygen species (ROS) are considered the main source of damage to plant cells. Mechanisms of ROS scavenging in wheat are very important during stress and the antioxidant enzymes superoxide dismutase, Catalase and Glutathione peroxidase are key to facilitating ROS scavenging. Molybdenum (Mo) is involved in many plant physiological and biochemical processes including antioxidant enzymes. This study reports research to investigate the effect of Mo application in enhancing antioxidant enzymes in two wheat cultivars. The results confirmed that antioxidant defense is important in wheat that is exposed to abiotic stress and that changes in activities of antioxidant enzymes occurred during exposure of plants to low non-freezing temperatures and by adding Mo. Mo application had a positive effect on gene expression of both Cbf14 and COR15a protein expression, indicating upregulation of the stress response regulon. In addition, Mo enhanced antioxidant enzymes activity and improved frost tolerance.     

Effect of drought on chlorophyll content and antioxidant enzyme activities in leaves of three wheat cultivars varying in productivity

Seedlings of three wheat varieties (Triticum aestivum L.)—highly productive cv. Ballada, moderately productive cv. Belchanka, and low productive cv. Beltskaya—were exposed to progressive soil drought (cessation of watering for 3, 5, and 7 days) and then analyzed for chlorophyll content and activities of ferredoxin-NADP⁺ oxidoreductase (FNR) and antioxidant enzymes, namely, glutathione reductase (GR) and ascorbate peroxidase (AscP). In addition, the proline content, and the extent of lipid peroxidation were examined. In the first period of water limitation, the water loss from leaves was slight for all wheat cultivars, which is characteristic of drought-resistant varieties. After 7-day drought the leaf water content decreased by 5.2–6.8%. The total chlorophyll content expressed per unit dry weight increased insignificantly during the first two periods of drought but decreased by 13–15% later on. This decrease was not accompanied by changes in chlorophyll a/b ratio. The plant dehydration did not induce significant changes in FNR activity. Activities of GR and AscP in leaves of wheat cultivars Ballada and Belchanka increased on the 3rd and 5th days of drought. Owing to the coordinated increase in GR and AscP activities, the lipid peroxidation rate remained at nearly the control level observed in water-sufficient plants. When the dehydration period was prolonged to 7 days, activities of GR and AscP in wheat cultivars reduced in parallel with the increase in malonic dialdehyde (MDA) content, indicating that the antioxidant enzyme defense system was weakened and lipid peroxidation enhanced. Unlike Ballada and Belchanka, the wheat cv. Beltskaya did not exhibit the increase in GR and AscP activities during progressive soil drought. The increase in MDA content by 16% in this cultivar was only observed after a 7-day drought period. The proline content in leaves of all wheat cultivars increased substantially during drought treatment. Thus, in wheat cultivars examined, different responses of the defense systems were mobilized to implement plant protection against water stress. The activities of antioxidant enzyme defense system depended on wheat cultivar, duration of drought, and the stage of leaf development.     

β-Aminobutyric acid increases abscisic acid accumulation and desiccation tolerance and decreases water use but fails to improve grain yield in two spring wheat cultivars under soil drying

A pot experiment was conducted to investigate the effect of the non-protein amino acid, β-aminobutyric acid (BABA), on the homeostasis between reactive oxygen species (ROS) and antioxidant defence during progressive soil drying, and its relationship with the accumulation of abscisic acid (ABA), water use, grain yield, and desiccation tolerance in two spring wheat (Triticum aestivum L.) cultivars released in different decades and with different yields under drought. Drenching the soil with 100 μM BABA increased drought-induced ABA production, leading to a decrease in the lethal leaf water potential (Ψ) used to measure desiccation tolerance, decreased water use, and increased water use efficiency for grain (WUE(G)) under moderate water stress. In addition, at severe water stress levels, drenching the soil with BABA reduced ROS production, increased antioxidant enzyme activity, and reduced the oxidative damage to lipid membranes. The data suggest that the addition of BABA triggers ABA accumulation that acts as a non-hydraulic root signal, thereby closing stomata, and reducing water use at moderate stress levels, and also reduces the production of ROS and increases the antioxidant defence enzymes at severe stress levels, thus increasing the desiccation tolerance. However, BABA treatment had no effect on grain yield of wheat when water availability was limited. The results suggest that there are ways of effectively priming the pre-existing defence pathways, in addition to genetic means, to improve the desiccation tolerance and WUE(G) of wheat.     

尿素施用量对小麦根际土壤微生物数量及土壤酶活性的影响

在大田高产条件下,研究了不同尿素施用量下两种不同穗型小麦品种“兰考矮早8”和“豫麦49—198”根际微生物数量和土壤酶活性的变化。结果表明：微生物数量随小麦生育时期的进行呈规律性变化,其中微生物总量在拔节期和抽穗期时数量较多。尿素施用量对小麦根际微生物数量和酶活性均产生一定的影响,并且处理间差异达到显著水平。两个小麦品种根际微生物总量、细菌、放线菌、真菌数量随着尿素施用量的增加呈先上升后下降的趋势,但两个品种根际微生物数量最高时的尿素量略有差异。同一生育时期,随着尿素施用量的提高,土壤蛋白酶、过氧化氢酶呈先增加后降低的变化趋势,以他（391kg／hm^2）或T3（586kg／hm^2）处理的酶活性较高,T4（782kg／hm^2）处理的酶活性略有降低;脲酶活性则呈上升趋势,以T4（782kg／hm^2）处理的脲酶活性最大。表明适宜尿素施用量有益于小麦根际微生物数量和酶活性的提高,过高则微生物数量和酶活性下降。     

Changes in activities of antioxidant enzymes and their gene expression during leaf development of sweetpotato

To understand the functions of antioxidant enzymes during leaf development in sweetpotato, we investigated the activities of several antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX), ascorbate peroxidase (APX) and catalase (CAT). Significant increases were observed in the activities of SOD, POX and APX during the late stage of leaf development, whereas CAT activity increased during the early developmental stage. By RT-PCR analysis, various POX and APX genes showed differential expression patterns during leaf development. Four POX genes swpa3, swpa4, swpa6, swpb4 and one APX gene swAPX1 exhibited high levels of gene expression during the senescence stage of leaf development, but two POX genes, swpa1 and swpa7 were preferentially expressed at both the mature green and the late senescence stages of leaf development. These results indicate that hydrogen peroxide (H₂O₂)-related antioxidant enzymes are differentially regulated in the process of leaf development of sweetpotato.     

小麦光温敏雄性不育系BS366铜锌超氧化物歧化酶基因的克隆及表达分析

超氧化物歧化酶(SOD,superoxide dismutase)是植物中一种主要的抗氧化酶,在植物应对逆境胁迫及抗衰老中起重要作用。本研究从基因芯片数据中筛选获得小麦Cu/Zn-SOD基因的EST序列,通过序列比对后拼接得到小麦Cu/Zn-SOD的候选基因,利用PCR技术在小麦光温敏雄性不育材料BS366中克隆并获得该基因。通过对Cu/Zn-SOD基因序列进行生物信息学分析,结果表明,该基因拥有连续且完整的开放阅读框,长495bp,编码164个氨基酸。氨基酸序列分析发现,该蛋白具有保守的Cu/Zn-SOD功能结构域与典型的Cu/Zn-SOD三维结构,且定位于细胞质中。通过同源进化分析表明,该蛋白与二穗短柄草(Brachypodium distachyon(L.)Beauv.)和大麦(Hordeum vulgare L.)的Cu/Zn-SOD蛋白亲缘关系较近,相似度分别为89%和94%。利用实时荧光定量PCR技术对其在小麦不同组织的表达特异性及不同逆境胁迫下的表达模式进行分析,结果表明,该基因在根、茎、叶、雌蕊、雄蕊、颖壳中均有表达,属于组成型表达,且在小麦的地上部含叶绿体的组织中含量较高;同时受多种胁迫诱导,可能参与了多种胁迫诱导调控途径。通过对该基因在不同育性环境中BS366育性转换期花药中的表达模式分析,发现可育环境下,在小孢子母细胞时期和减数分裂期的表达量分别约为对照的8倍与16倍;而不育环境下,该基因表达水平无明显变化。因而推测,小麦Cu/Zn-SOD基因可能参与了光温敏雄性不育系BS366的育性调控。本研究为深入研究Cu/Zn-SOD基因在小麦中的作用机理奠定了重要基础。     

Photosynthesis, antioxidant activities and transcriptional responses in two sugarcane (Saccharum officinarum L.) cultivars under salt stress

This study analyzes changes in gene expression and the biochemical and physiological properties of the antioxidant system in the leaves of two sugarcane cultivars under salt stress. In both salt-stressed cultivars, no alteration in the foliar nitrogen content was observed; however, there was a reduction in the phosphorus and potassium levels and an increase in the sodium and chloride concentrations. There was also a reduction in gas exchange on the third day under salt stress. Although the content of soluble sugars remained stable in both species, there was a decrease in free amino acids. However, only the RB872552 cultivar displayed a lower leaf protein content compared to the control. The salt stress resulted in higher superoxide dismutase and l-ascorbate peroxidase activities, but only for the RB92579 cultivar. On the other hand, both cultivars were able to maintain lower malondialdehyde contents than the control plants. The gene expression analysis revealed down-regulated expression levels, including the levels of those enzymes linked to higher activities under salt stress. Our results showed that gene induction and leaf antioxidative cycle enzyme activity do not occur at the same time. The variations in gene expression and physiological responses are also discussed.     

The response of Mo-hydroxylases and abscisic acid to salinity in wheat genotypes with differing salt tolerances

The differential responses of the wheat cultivars Shi4185 and Yumai47 to salinity were studied. The higher sensitivity of Yumai47 to salinity was linked to a greater growth reduction under salt stress, compared to more salt-tolerant Shi4185. Salinity increased the Na⁺, proline and superoxide anion radical (O₂ ^?) contents in both cultivars. Leaf Na⁺ content increased less in the more salt-tolerant cultivar Shi4185 than salt-sensitive Yumai47. The proline content increased more significantly in Shi4185 than Yumai47; on the contrary, superoxide anion radical content increased less in Shi4185 than Yumai47. This data indicated that wheat salinity tolerance can be increased by controlling Na⁺ transport from the root to shoot, associated with higher osmotic adjustment capability and antioxidant activity. Although salinity increased aldehyde oxidase (AO) activity and abscisic acid (ABA) content in the leaves and roots of both cultivars following the addition of NaCl to the growth medium, AO and ABA increased more in the salt-sensitive cultivar Yumai47 than the more salt-tolerant cultivar Shi4185. Xanthine dehydrogenase (XDH) activity in the leaves of both cultivars increased with increasing concentrations of NaCl; however, leaf XDH activity increased more significantly in Yumai47 than Shi4185. Root XDH activity in Shi4185 decreased with increasing NaCl concentrations, whereas salinity induced an increased root XDH activity in Yumai47. The involvement of AO and XDH enzymatic activities and altered ABA content in the response mechanisms of wheat to salinity are discussed herein.     

Influence of silicon on antioxidant mechanisms and lipid peroxidation in chickpea (Cicer arietinum L.) cultivars under drought stress

Abstract

The effects of exogenous silicon (Si) on leaf relative water content (RWC), and the growth, Si concentrations, lipid peroxidation (MDA), lipoxygenase (LOX) activity, proline and H₂O₂ accumulation, non-enzymatic antioxidant activity (AA) and the activity of some antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX) in shoots of ten chickpea cultivars grown under drought were investigated. Drought stress decreased the growth of all the cultivars while applied Si improved the growth at least five of the 10 chickpea cultivars. Silicon applied to the soil at 100 mg kg^?1 significantly increased Si concentrations of the cultivars and counteracted the deleterious effects of drought in 5 of the ten chickpea cultivars by increasing their RWC. In most cultivars tested H₂O₂, proline and MDA content and LOX activity were increased by drought whereas application of Si decreased their levels. APX activity was increased by drought but it was depressed by Si. In general, SOD and CAT activities of the cultivars were decreased by drought. Depending on cultivars, the CAT activity was decreased, and increased or unchanged in response to applied Si, while the SOD activity of the cultivars increased or unchanged by Si. The non-enzymatic antioxidant activity of the cultivars was also increased by Si. These observations implied an essential role for Si in minimizing drought stress-induced limitation of the growth and oxidative membrane damage in chickpea plants.     

Responses of growth and antioxidant systems in <Emphasis Type="Italic">Carthamus</Emphasis><Emphasis Type="Italic">tinctorius</Emphasis> L. under water deficit stress

An investigation was carried out to find out the extent of changes occurred in two safflower (Carthamus tinctorius L.) cultivars in response to water deficit stress. Two safflower cultivars namely IL.111 and Isfahan were used for the study. Thirty days after sowing, plants were grown under soil moisture corresponding to 100, 85, 70 and 55% field capacity for next 30 days. Water deficit treatments significantly decreased the shoot length, shoot dry matter, root dry matter, relative growth rate, leaf relative water content (LRWC) and leaf water potential (Ψ_W), whereas root length, root-to-shoot ratio, lipid peroxidation and antioxidant compounds such as ascorbic acid (AA), α-tocopherol (α-Toc) and reduced glutathione (GSH) and superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), and peroxidase (POX, EC 1.11.1.7) activities were increased. Water deficit stressed plants maintained higher levels of compounds and scavenging enzymes. Significant differences were observed between cultivars and irrigation levels treatments. The cv. IL.111 could be considered more tolerant to water stress than cv. Isfahan, registering greater biomass, LRWC and leaf water potential (Ψ_W), associated with high antioxidant activity.     

Insect-resistant Bt-maize response to the short-term non-target mite-pest infestation and soil drought

Transgenic lepidopteran insect-resistant maize expressing the cry1Ab gene (Bt) and its non-transgenic counterpart at the 12-leaf-stage (V₁₂) were infested by the two-spotted spider mite or dehydrated by cessation of soil watering to check Bt-maize capacity to respond to other stresses than those assured by the presence of Cry protein. Since the antioxidant enzymes are key components of plant defence against biotic and abiotic stresses, the engagement of leaf superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) in response to 6-day mite feeding and soil drought has been investigated. The reduction of leaf hydration and soluble protein content in the fully expanded 8th leaf was independent of genotype and more pronounced in response to water cessation than mite infestation. Similarly, the changes in enzyme activities depended more on the kind of stress than the presence of the transgene. Water shortage in the soil enhanced the activity of all enzymes, whereas mite feeding decreased the activity of SOD and CAT, and markedly increased POX in the 8th leaf of both cultivars. In mite-infested leaves of the non-transgenic plant, the CAT activity remained unaffected, whereas decreased in leaves of Bt maize due to the hampered activity of CAT-2. In comparison to the control, all enzyme activity in the 10th non-infested leaf of mite-infested non-transgenic maize decreased, whereas it changed in the 10th leaf of Bt maize in the same way as in the 8th mite-infested leaf. The results suggest that SOD, CAT and POX can strongly confer short-term drought-stress response in both maize cultivars, whereas POX is the only responsive enzyme in mite-infested Bt maize.     

Silicon increases boron tolerance and reduces oxidative damage of wheat grown in soil with excess boron

The effect of silicon on the growth, boron concentrations, malondialdehyde (MDA) content, lipoxygenase (LOX) activity, proline (PRO) and H₂O₂ accumulation, and the activities of major antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)] and non-enzymatic antioxidants (AA) of wheat grown in soil originally with toxic B concentrations were investigated. Applied of 5.0 and 10.0 mM Si to the B toxic soil significantly increased Si concentration of the wheat and counteracted the deleterious effects of B on shoot growth. The contents of PRO, H₂O₂, MDA, and LOX activity of wheat grown in B toxic soil were significantly reduced by Si treatments. Compared with control plants, the activities of SOD, CAT, APX and content of AA were decreased by applied Si. Based on the present work, it can be concluded that Si alleviates B toxicity of wheat by preventing oxidative membrane damage and also translocation of B from root to shoot and/or soil to plant.     

大气-土壤-小麦生态系统中铅的分布和迁移规律研究

研究铅在国道附近大气－土壤－小麦生态系统中的分布及其迁移规律。结果表明,大气中铅浓度与汽车流量成正比,而与风速、温度等相关性不明显;距公路越远,土壤及小麦中铅的含量越高,含是与距离呈一定的匀相关性,５ｍ及６０ｍ为其转折点;铅在土壤中由上上及在小麦体中由根向茎、穗的迁移较小;小麦各器官中铅含量大小依次为根〉叶〉穗〉茎〉籽。叶片、穗尚从大气中直接部分铅。在小麦不同生长阶段中,各器官积累铅量不同,其积累     

盐胁迫对冬小麦幼苗干物质分配和生理生态特性的影响

以冬小麦(Triticum aestivum L.)品种周麦18(Zhoumai18)和豫麦49(Yumai49)为材料,采用盆栽培养,研究100、250、350 mmol/L和450 mmol/L NaCl 浓度胁迫下,小麦幼苗干物质分配、根系特征、叶绿素含量、游离脯氨酸含量和根系活力变化规律.结果表明,随着盐分浓度的增加,两个品种小麦的叶面积、地上干重以及根的长度显著减小;根系干重、根直径、根表面积、根体积、根系活力以及叶绿素含量呈先上升后下降趋势,在250 mmol/L NaCl处理下达最大.叶绿素a/b随NaCl浓度升高而上升.随盐分浓度变化周麦18叶片游离脯氨酸含量高而变化幅度大,450mmol/L处理组的含量高于对照组1.5倍.供试品种冬小麦耐盐阈值为250～350 mmol/L.     

Comparative changes in the antioxidant system in the flag leaf of early and normally senescing near-isogenic lines of wheat (Triticum aestivum L.)

Key message

The antioxidant system was significantly inhibited in the early aging line than the near-isogenic normal aging line during senescence.

Abstract

The antioxidant system plays pivotal roles in removal of reactive oxygen species (ROS) produced during leaf senescence. To explore its roles in leaf senescence of wheat (Triticum aestivum L.), the concentrations of antioxidants, activities, and gene expression of antioxidant enzymes were evaluated in flag leaves of the early aging line (EAL) and the near-isogenic normal aging line (NL) during senescence. The results showed that the total chlorophyll and soluble protein in the EAL declined earlier and faster, while more malondialdehyde and ROS accumulated compared with the NL. The activities of superoxide dismutase, catalase, ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase were lower in the EAL than in the NL across multiple measuring dates. Additionally, the EAL had less amounts of reduced ascorbate and glutathione as well as lower reduction state with the progression of senescence. Concomitantly, the gene expression of antioxidant enzymes in the EAL was also significantly repressed relative to those in the NL during natural senescence. Taken together, the earlier onset and faster rate of senescence in the EAL could be a result of an imbalance of ROS production and ROS-scavenging antioxidant system, which provided valuable hints toward understanding leaf senescence of wheat.     

Higher salinity tolerance cultivars of winter wheat relieved senescence at reproductive stage

In order to probe into the potential of relieving senescence in the new cultivars, we have compared gas exchanges and related physiological parameters of two cultivars of winter wheat (DK961, salt-tolerant; JN17, salt-sensitive) under a series of NaCl concentrations. Through out the whole reproductive period, it was noted that net photosynthetic rate, stomatal conductance, pigment contents, ions contents, leaf area index, leaf area duration, leaf relative water content and dry matter accumulation of spikes decreased in both cultivars with saline concentrations increasing. However, the salt-tolerant cultivars showed none significant reductions in those parameters compared with control under 0.3% and 0.5% salt concentrations, with only considerable decrease happening when soil salt concentration exceeded to 0.7%. Sharply contrast to salt-tolerant cultivars, the salt-sensitive cultivars appeared linear reductions in physiological parameters under a series of salt concentrations (0.3%, 0.5% and 0.7%), with the photosynthetic duration being evidently shorter. Significantly positive correlations among K⁺/Na⁺, reproductive growth period and total growth period were noted in salt-sensitive cultivars, however, none significant relations appeared among those parameters in salt-tolerant cultivars, indicating a strong tolerant behavior happened. Our result suggested that higher salinity tolerance cultivars of winter wheat could relieve senescence at the reproductive stage.     

根域限制下水氮供应对膜下滴灌棉花根系及叶片衰老特性的影响

在新疆的气候生态条件下, 选用北疆2个棉花(Gossypium hirsutum)主栽品种‘新陆早13号’和‘新陆早33号’为供试材料, 设置限根(RR)与对照(CK)处理, 每个处理设置4个水氮水平: 水氮亏缺(W₀N₀)、水分亏缺(W₀N₁)、氮素亏缺(W₁N₀)与水氮适量(W₁N₁), 组成再裂区试验方案。采用管栽方法, 通过人工改变根系垂直生长深度和水氮供应, 在棉花产量形成期测定根系及叶片抗氧化保护酶系活性、生物量累积及分配等, 探讨根域限制及水氮供应对棉花根系生长及叶片衰老的影响机理。结果表明: 根域限制条件下, 棉花根系生物量、根系与叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)与过氧化氢酶(CAT)活性、棉株总生物量、根冠比均低于对照, 而地上部生物量与籽棉产量显著高于对照。水氮供应能有效地调节根系及叶片的生长, 不同水氮处理间棉花根系与叶片抗氧化保护酶系活性、叶绿素含量、地上部生物量及籽棉产量均表现为W₁N₁ > W₀N₁ > W₁N₀ > W₀N₀, 根冠比与根系生物量的表现与之相反。根域限制与水氮供应表现出互作优势, 根域限制下适量水氮供应处理的地上部生物量与籽棉产量均明显高于其他处理, 根冠比较低。因此, 在棉花根系生长受限的条件下, 优化生育期间水氮供应, 可以增强根系及叶片的抗氧化保护酶系活性、增加光合产物向地上部的分配比例、增加产量, 是进一步挖掘膜下滴灌棉花增产潜力的有效途径。     

Zinc efficiency is correlated with enhanced expression and activity of zinc-requiring enzymes in wheat

Zinc (Zn) is an essential micronutrient for plants. The ability of plants to maintain significant yields under low Zn is termed Zn efficiency (ZE) and its genetic and mechanistic basis is still not well understood. Previously, we showed that root Zn uptake did not play a role in ZE. In the current study, Zn-efficient and -inefficient wheat (Triticum aestivum) genotypes were grown for 13 d in chelate buffer nutrient solutions at low (0.1 pM), sufficient (150 pM), and high (1 microM) Zn(2+) activities and analyzed for root-to-shoot translocation of Zn, subcellular leaf Zn distribution, and activity and expression of the Zn-requiring enzymes in leaves. No correlation between ZE and Zn translocation to the shoot was found. Furthermore, total and water-soluble concentrations of leaf Zn were not associated with ZE, and no differences in subcellular Zn compartmentation were found between Zn-efficient and -inefficient genotypes. However, the expression and activity of the Zn-requiring enzymes copper (Cu)/Zn superoxide dismutase (SOD) and carbonic anhydrase did correlate with differences in ZE. Northern analysis suggested that Cu/ZnSOD gene expression was up-regulated in the Zn-efficient genotype, Kirgiz, but not in inefficient BDME. Under Zn deficiency stress, the very Zn-efficient genotype Kirgiz and moderately Zn-efficient Dagdas exhibited an increased activity of Cu/ZnSOD and carbonic anhydrase when compared with Zn-inefficient BDME. These results suggest that Zn-efficient genotypes may be able to maintain the functioning of Zn-requiring enzymes under low Zn conditions; thus, biochemical Zn utilization may be an important component of ZE in wheat.