干旱高温胁迫下转基因水稻的生理变化

用PEG-6000、38℃及PEG-6000+38℃胁迫处理携带谷胱甘肽转移酶(GST)和过氧化氢酶(CAT1)的转基因水稻和非转基因水稻(Oryza sativa L.cv中花11), 并比较分析了二者在胁迫下的生理指标和抗氧化酶活性变化.结果显示,在单一PEG及PEG和高温复合胁迫下,植株生长、光合参数和相对含水量的降低幅度及H2O2和MDA(malon dialdehyde)的积累量在非转基因水稻与转基因水稻之间都有显著差异.在这2种胁迫下,转基因水稻的可溶性糖含量及CAT和POD (Peroxidase) 活性与非转基因水稻也有显著差异.这些结果表明,谷胱甘肽转移酶(GST)和过氧化氢酶(CAT1)的表达减轻了转基因水稻在单一干旱及干旱+高温复合胁迫下的伤害.     

Improved Performance of Transgenic Glycinebetaine-Accumulating Rice Plants under Drought Stress

Plasmid DNA (pChlCOD), containing the selectable hygromycin phosphotransferase hpt gene for hygromycin B resistance and the Arthrobacter globiformis codA gene for choline oxidase which catalyzes the direct conversion of choline to glycinebetaine, was delivered into rice plants using Agrobacterium-mediated gene transfer via scutellum-derived calli. Southern, Northern and Western blot analyses demonstrated that the foreign gene had been transferred, integrated into rice chromosomal DNA and expressed. Drought test indicated that glycinebetaine acts as an osmoprotectant and its production in transgenic rice plant helped the cells to maintain osmotic potential and increased root growth, and thus enhanced the ability of the plants to tolerate water deficit This revised version was published online in July 2006 with corrections to the Cover Date.     

不同抗性苹果砧木叶片抗坏血酸代谢对干旱胁迫的响应

以抗旱性强的苹果砧木新疆野苹果[Malus sieversii (Ledeb.) Roem.]和抗旱性弱的平邑甜茶[M.hupehensis (Panlp.) Reld.]叶片为试材,在20%PEG-6000模拟干旱胁迫条件下,研究了抗坏血酸(AsA)含量、氧化还原状态、合成和代谢相关酶活性对干旱胁迫的响应。结果显示,在正常水分条件下,新疆野苹果和平邑甜茶叶片间AsA含量及代谢相关酶活性差异不显著。干旱胁迫过程中,二者之间叶片AsA、谷胱甘肽(GSH)、总谷胱甘肽和总抗坏血酸含量变化趋势基本一致,但新疆野苹果叶片中的含量增加幅度明显大于平邑甜茶;且与平邑甜茶相比,新疆野苹果叶片具有较高的L-半乳糖酸-1,4-内酯脱氢酶(GalLDH)、脱氢抗坏血酸还原酶(DHAR)和谷胱甘肽还原酶(GR)活性及AsA/DHA(脱氢抗坏酸)和GSH/GSSG(氧化型谷胱甘肽)比率。研究表明,干旱胁迫下,新疆野苹果能够维持较高的AsA合成和再生能力,并具有维持高AsA水平特性,这可能与其具有较强的抗旱性相关。     

Dissecting Maize Productivity: Ideotypes Associated with Grain Yield under Drought Stress and Well-watered Conditions

To increase maize (Zea mays L.) yields in drought‐prone environments and offset predicted maize yield losses under future climates, the development of improved breeding pipelines using a multi‐disciplinary approach is essential. Elucidating key growth processes will provide opportunities to improve drought breeding progress through the identification of key phenotypic traits, ideotypes, and donors. In this study, we tested a large set of tropical and subtropical maize inbreds and single cross hybrids under reproductive stage drought stress and well‐watered conditions. Patterns of biomass production, senescence, and plant water status were measured throughout the crop cycle. Under drought stress, early biomass production prior to anthesis was important for inbred yield, while delayed senescence was important for hybrid yield. Under well‐watered conditions, the ability to maintain a high biomass throughout the growing cycle was crucial for inbred yield, while a stay‐green pattern was important for hybrid yield. While new quantitative phenotyping tools such as spectral reflectance (Normalized Difference Vegetation Index, NDVI) allowed for the characterization of growth and senescence patterns as well as yield, qualitative measurements of canopy senescence were also found to be associated with grain yield.     

Melatonin Promotes Rice Seed Germination under Drought Stress by Regulating Antioxidant Capacity

Drought stress is a serious threat to the germination of plant seeds and the growth of seedlings. Melatonin has been proven to play an important role in alleviating plant stress. However, its effect on seed germination under drought conditions is still poorly understood. Therefore, we studied the effects of melatonin on rice seed germination and physiological characteristics under drought stress. Rice seeds were treated with different concentrations of melatonin (i.e., 0, 20, 100, and 500 μM) and drought stress was simulated with 5% polyethylene glycol 6000 (PEG6000). The results showed that 100 μM melatonin can effectively improve the germination potential, rate and index; the vigor index of rice seeds; and the length of the shoot and root. In addition, that treatment also increased the activity of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), and reduced the content of malondialdehyde (MDA). The grey relational grade between the shoot MDA content and the melatonin seed-soaking treatment was the highest, which could be useful for evaluating the effect of melatonin on drought tolerance. Two-way analysis of variance showed that the effect of single melatonin treatment on rice seeds was more significant than that of single drought stress and interaction treatment of drought and melatonin (p < 0.05). The subordinate function results showed that 100 μM melatonin significantly improved the germination and physiological indexes of rice seeds and effectively alleviated the adverse effects of drought stress on rice seedlings. The results helped to improve the understanding of the morphological and physiological involvement of melatonin in promoting seed germination and seedling development under drought stress.     

Elucidating Rice Cell Metabolism under Flooding and Drought Stresses Using Flux-Based Modeling and Analysis

Rice (Oryza sativa) is one of the major food crops in world agriculture, especially in Asia. However, the possibility of subsequent occurrence of flood and drought is a major constraint to its production. Thus, the unique behavior of rice toward flooding and drought stresses has required special attention to understand its metabolic adaptations. However, despite several decades of research investigations, the cellular metabolism of rice remains largely unclear. In this study, in order to elucidate the physiological characteristics in response to such abiotic stresses, we reconstructed what is to our knowledge the first metabolic/regulatory network model of rice, representing two tissue types: germinating seeds and photorespiring leaves. The phenotypic behavior and metabolic states simulated by the model are highly consistent with our suspension culture experiments as well as previous reports. The in silico simulation results of seed-derived rice cells indicated (1) the characteristic metabolic utilization of glycolysis and ethanolic fermentation based on oxygen availability and (2) the efficient sucrose breakdown through sucrose synthase instead of invertase. Similarly, flux analysis on photorespiring leaf cells elucidated the crucial role of plastid-cytosol and mitochondrion-cytosol malate transporters in recycling the ammonia liberated during photorespiration and in exporting the excess redox cofactors, respectively. The model simulations also unraveled the essential role of mitochondrial respiration during drought stress. In the future, the combination of experimental and in silico analyses can serve as a promising approach to understand the complex metabolism of rice and potentially help in identifying engineering targets for improving its productivity as well as enabling stress tolerance.Rice (Oryza sativa) is one of the prominent food crops in the world and is considered a major staple food in many Asian countries: each person consumes more than 100 kg of rice per year, on average (Nelson, 2011). Since the Green Revolution in 1960, there has been a progressive increase in the yield of rice. However, the growing population and adverse climatic changes pose huge challenges to sustaining the growing demand for rice. Moreover, of several abiotic stresses, flooding stress tremendously limits rice productivity (Xu et al., 2006), particularly in the rain-fed lowlands of Southeast Asia. In general, when plants are water-logged by flooding, they experience a lower oxygen availability (hypoxia) or a total absence of oxygen (anoxia), thus severely impairing energy generation through reduced/eliminated mitochondrial respiration. Nevertheless, rice is unique in its ability to survive up to 2 weeks in complete submergence conditions by prolonged flood (Jackson and Ram, 2003; Bailey-Serres and Voesenek, 2008). Rice seeds can germinate and grow up to the coleoptile even in anoxia through its distinctive metabolic adaptations. Under such conditions, predominant amounts of energy required for survival are produced by fermentative pathways, especially ethanolic fermentation (Atwell et al., 1982; Alpi and Beevers, 1983; Perata and Alpi, 1993; Guglielminetti et al., 1995; Gibbs et al., 2000; Magneschi and Perata, 2009). Besides flooding stress, the photosynthetic carbon-fixing capacity of rice and other C₃ plants is limited by an energetically wasteful process, photorespiration. It is initiated by the partial replacement of oxygen instead of CO₂ into the bifunctional enzyme Rubisco, which is the first reaction in the CO₂-fixing pathway. This oxygenase side reaction of Rubisco produces one molecule of 3-phosphoglycerate (3-PGA) and one molecule of 2-phosphoglycolate (2-PG) for each molecule of oxygen fixed. Of these, 2-PG is a wasteful by-product, requiring sufficient amounts of energy for its salvation into 3-PGA through a series of enzymatic steps known as “the photorespiratory cycle” (Jordan and Ogren, 1984). In addition, the amount of CO₂ available for photosynthesis is drastically reduced under drought conditions owing to the partial/full closure of leaf stomata, resulting in increased photorespiration (de Carvalho, 2008). Thus, photorespiration has always been considered as a major target for crop improvement in C₃ plants such as rice to increase its productivity.Over the last few decades, several efforts have been made to understand the cellular metabolism of rice by conventional experimental approaches. However, its metabolic adaptations under different stress conditions remain poorly characterized. Thus, with recent advancements in the modern genomic era, a systematic approach is required to improve our understanding of the metabolic changes of rice. In the postgenomic era, it can be achieved by utilizing the abundant high-throughput “omics” data sets in combination with in silico metabolic modeling. To this end, the development of a predictive in silico model based on the available biochemical, genomic, and regulatory information, and its subsequent examination by the well-established computational framework of constraints-based flux analysis, also known as flux balance analysis (FBA), is highly desired.Interestingly, flux-based metabolic modeling of plant systems has recently gained more attention following its tremendous success in elucidating the metabolic capabilities of myriad microbial and mammalian species and rationally engineering them to achieve desirable phenotypes (Edwards et al., 2002; Lee et al., 2005; Lewis et al., 2012). In this regard, metabolic network models for several plants, such as Arabidopsis (Arabidopsis thaliana; Poolman et al., 2009; de Oliveira Dal’Molin et al., 2010a; Saha et al., 2011; Chung et al., 2012; Mintz-Oron et al., 2012), barley (Hordeum vulgare; Grafahrend-Belau et al., 2009), rapeseed (Brassica napus; Hay and Schwender, 2011; Pilalis et al., 2011), maize (Zea mays; Saha et al., 2011), and a general C₄ plant model (de Oliveira Dal’Molin et al., 2010b) have already been developed, and some of them are even in genome scale. Nevertheless, to the best of our knowledge, the metabolic model of rice is not available to date. Therefore, in this study, we reconstructed a combined metabolic/regulatory network model representing the central metabolism of rice cells and subsequently utilized it for characterizing the cellular behavior in two different tissue types: germinating seeds and photorespiring leaves. In addition, we also simulated their metabolic states during abiotic stresses and investigated how they acclimatize to such conditions.     

Rhizospheric Bicarbonate Improves Glucose Metabolism and Stress Tolerance of Broussonetia papyrifera L. Seedlings under Simulated Drought Stress

Russian Journal of Plant Physiology - Heavy concentrations of bicarbonates present throughout karst ecosystems necessitate identification of the role of $${text{HCO}}_{3}^{ - }$$ in plant growth,...     

Polyamine Metabolism and Osmotic Stress : I. Relation to Protoplast Viability

Cereal leaves subjected to the osmotica routinely used for protoplast isolation show a rapid increase in arginine decarboxylase activity, a massive accumulation of putrescine, and slow conversion of putrescine to the higher polyamines, spermidine, and spermine (HE Flores, AW Galston 1984 Plant Physiol 75: 102). Mesophyll protoplasts from these leaves, which have a high putrescine:polyamine ratio, do not undergo sustained division. By contrast, in Nicotiana, Capsicum, Datura, Trigonella, andVigna, dicot genera that readily regenerate plants from mesophyll protoplasts, the response of leaves to osmotic stress is opposite to that in cereals. Putrescine titer as well as arginine and ornithine decarboxylase activities decline in these osmotically stressed dicot leaves, while spermidine and spermine titers increase. Thus, the putrescine:polyamine ratio in Vigna protoplasts, which divide readily, is 4-fold lower than in oat protoplasts, which divide poorly. We suggest that this differing response of polyamine metabolism to osmotic stress may account in part for the failure of cereal mesophyll protoplasts to develop readily in vitro.     

外源一氧化碳对干旱胁迫下水稻幼苗抗氧化系统的影响

以水稻(Oryza sativa L.) 品种‘D奇宝优1号'幼苗为材料,采用20%聚乙二醇(PEG-6000)模拟水分胁迫,研究外源一氧化碳(CO)对干旱胁迫下水稻幼苗抗氧化能力的影响,以探索CO提高水稻幼苗抗旱性的机制.结果显示,CO供体高铁血红素(Hematin,H)显著降低干旱胁迫下水稻幼苗叶片质膜相对透性和丙二醛(MDA)含量,提高脯氨酸和可溶性蛋白的含量,不同程度地促进叶片中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)的活性,而CO清除剂血红蛋白(Hemoglobin,Hb)则逆转CO供体对干旱胁迫下水稻幼苗氧化损伤的缓解效应.由此表明,外源CO能通过调整保护酶活性和渗透调节物质含量来提高水稻幼苗的抗氧化能力,有效增强其抗旱性.     

Xanthine Dehydrogenase Involves in the Response of Photosystem and Reactive Oxygen Metabolism to Drought Stress in Rice

Xanthine dehydrogenase (XDH) is a crucial enzyme involved in purine metabolism. Although the essential role of XDH is well studied in leguminous plants and Arabidopsis, the importance of this enzyme remains uncertain in rice. To evaluate how biochemistry indicators respond to XDH down-regulation and up-regulation in rice seedings during drought stress, RNA interference (RNAi) and CDS over-expression were used to generate transgenic lines of Nipponbare (Oryza sativa L.) in which OsXDH, the gene for XDH were silenced and over-expression. When the XDH-suppressed line was subjected to drought stress, the chlorophyll content and chlorophyll fluorescence parameters were markedly reduced in conjunction with significantly O ₂ ^? enhanced production rate and MDA accumulation. This drought-hypersensitive biochemistry indicators was reversed in XDH-intensified line. Meanwhile, the XDH activity and its downstream metabolites were induced by drought stress. These observations support the notion that xanthine dehydrogenase was involved in regulating photosystem and reactive oxygen metabolism in drought stress in rice seedling.     

Dissecting Long-Term Glucose Metabolism Identifies New Susceptibility Period for Metabolic Dysfunction in Aged Mice

Metabolic disorders, like diabetes and obesity, are pathogenic outcomes of imbalance in glucose metabolism. Nutrient excess and mitochondrial imbalance are implicated in dysfunctional glucose metabolism with age. We used conplastic mouse strains with defined mitochondrial DNA (mtDNA) mutations on a common nuclear genomic background, and administered a high-fat diet up to 18 months of age. The conplastic mouse strain B6-mt^FVB, with a mutation in the mt-Atp8 gene, conferred β-cell dysfunction and impaired glucose tolerance after high-fat diet. To our surprise, despite of this functional deficit, blood glucose levels adapted to perturbations with age. Blood glucose levels were particularly sensitive to perturbations at the early age of 3 to 6 months. Overall the dynamics consisted of a peak between 3–6 months followed by adaptation by 12 months of age. With the help of mathematical modeling we delineate how body weight, insulin and leptin regulate this non-linear blood glucose dynamics. The model predicted a second rise in glucose between 15 and 21 months, which could be experimentally confirmed as a secondary peak. We therefore hypothesize that these two peaks correspond to two sensitive periods of life, where perturbations to the basal metabolism can mark the system for vulnerability to pathologies at later age. Further mathematical modeling may perspectively allow the design of targeted periods for therapeutic interventions and could predict effects on weight loss and insulin levels under conditions of pre-diabetic obesity.     

干旱胁迫下转基因甘薯块根膨大期水分利用效率和生理代谢特征

以转Cu/Zn-SOD和APX基因及其非转基因甘薯进行盆栽试验,在甘薯块根膨大期进行正常供水(田间最大持水量的80%)、中度缺水(田间最大持水量的60%)和重度缺水(田间最大持水量的40%)3种水分处理,分别测定转基因植株和对照植株在薯块膨大期的第20天和第70天的抗氧化酶系统、可溶性糖含量、光合系统之间的差异,以及在不同水分胁迫处理下产量和水分利用效率之间的差异。以此研究外源基因的超表达是否可以提高甘薯的产量及水分利用效率。结果显示:(1)转基因甘薯(TS)的SOD、APX活性以及可溶性糖含量均高于非转基因对照株(NT),但POD活性低于NT;TS和NT植株的APX活性、可溶性糖含量、净光合速率以及蒸腾速率均随干旱胁迫加重呈递减趋势。(2)干旱胁迫70d时,TS和NT植株光合参数均较胁迫20d时降低,且TS和NT间的净光合速率没有明显差异。(3)TS和NT两株系的块根产量在中度胁迫下最高而在重度胁迫下最低,而TS具有较高的块根产量且在重度胁迫下产量降低幅度较小。(4)TS的气孔导度和蒸腾速率显著低于NT,且TS的水分利用效率较NT更高。研究表明,Cu/Zn-SOD和APX基因可以显著增加干旱胁迫下甘薯块根膨大期的SOD、APX活性和可溶性糖含量,提高其水分利用效率,从而减轻干旱胁迫对产量的影响。     

干旱胁迫下腐植酸对燕麦叶片非结构性碳水化合物代谢的影响

以燕麦品种‘燕科2号’为试验材料,采用盆栽方式,分别在正常供水(75%田间持水量)、中度干旱胁迫(60%田间持水量)和重度干旱胁迫(45%田间持水量)3个水分条件下喷施腐植酸(HA)和等量清水(CK),对燕麦叶片中非结构性碳水化合物(NSC)含量及相关酶活性和籽粒产量进行测定,以明确腐植酸在干旱胁迫下对燕麦叶片非结构性碳水化合物代谢变化的影响,探讨HA对燕麦耐旱性的影响及其作用机制。结果表明：(1)随着土壤水分含量的减少,燕麦叶片中蔗糖和淀粉含量逐渐显著降低,蔗糖合成酶(SS)、蔗糖磷酸合成酶(SPS)活性显著降低,而酸性转化酶(S AI)和淀粉水解酶(α GC)活性显著提高。(2)燕麦叶片可溶性总糖和还原糖含量随着土壤水分含量的减少表现出先升高后降低的变化趋势,导致籽粒产量显著下降,且干旱胁迫程度越重变化幅度越大。(3)叶面喷施HA能不同程度提升中度和重度干旱胁迫下燕麦叶片中上述非结构性碳水化合物含量,并调节相关酶活性,显著提高籽粒产量,并在重度胁迫下的效果更佳。研究发现,腐植酸可以通过调控燕麦叶片NSC的代谢来响应干旱胁迫,降低叶片细胞渗透势,有效缓解干旱胁迫造成的损伤,增强植株耐旱性。     

Physiological Mechanism of Drought-Resistant Rice Coping With Drought Stress

Journal of Plant Growth Regulation - Drought stress is one of the major threats to rice production. The weakening of leaf photosynthesis due to drought is the main reason for the reduction of grain...     

干旱胁迫对宁夏枸杞叶片蔗糖代谢及光合特性的影响

以宁夏枸杞品种‘宁杞1号’为材料,采用防雨棚内盆栽控水法研究了轻度干旱(LD)、中度干旱(MD)、重度干旱(SD)和正常灌水(CK)4种土壤水分条件下枸杞叶片糖分含量、蔗糖代谢酶活性和光合参数的响应规律,为宁夏枸杞在干旱地区高产栽培提供参考依据。结果显示:(1)干旱胁迫降低了枸杞青果期叶片蔗糖和淀粉含量,轻度干旱胁迫可提高枸杞色变期叶片果糖和蔗糖含量,而成熟期叶片淀粉含量则随着干旱加重而升高。(2)干旱胁迫降低了青果期枸杞叶片中性转化酶(NI)、蔗糖磷酸合成酶(SPS)和蔗糖合成酶(SS)的活性,降低了色变期和成熟期叶片酸性转化酶(AI)和SS的活性,但提高了色变期和成熟期叶片NI的活性。(3)随着干旱胁迫的加剧,叶片净光合速率(Pn)、蒸腾速率(Tr)和气孔导度(Gs)呈降低趋势,瞬时水分利用率(WUE)呈现:LD>CK>SD>MD的变化趋势。研究认为,轻度干旱胁迫能促进宁夏枸杞成熟期叶片蔗糖积累和水分利用率提高,有利于更多光合同化物输送到果实中。