Sensing and signalling in response to oxygen deprivation in plants and other organisms

AIMS AND SCOPE: All aerobic organisms require molecular di-oxygen (O2) for efficient production of ATP though oxidative phosphorylation. Cellular depletion of oxygen results in rapid molecular and physiological acclimation. The purpose of this review is to consider the processes of low oxygen sensing and response in diverse organisms, with special consideration of plant cells. CONCLUSIONS: The sensing of oxygen deprivation in bacteria, fungi, metazoa and plants involves multiple sensors and signal transduction pathways. Cellular responses result in a reprogramming of gene expression and metabolic processes that enhance transient survival and can enable long-term tolerance to sub-optimal oxygen levels. The mechanism of sensing can involve molecules that directly bind or react with oxygen (direct sensing), or recognition of altered cellular homeostasis (indirect sensing). The growing knowledge of the activation of genes in response to oxygen deprivation has provided additional information on the response and acclimation processes. Conservation of calcium fluxes and reactive oxygen species as second messengers in signal transduction pathways in metazoa and plants may reflect the elemental importance of rapid sensing of cellular restriction in oxygen by aerobic organisms.     

Molecular and biochemical characterisation of Mycobacterium smegmatis alcohol dehydrogenase C

The gene encoding of an alcohol dehydrogenase C (ADHC) from Mycobacterium smegmatis was cloned and sequenced. The protein encoded by this gene has 78% identity with Mycobacterium tuberculosis and Mycobacterium bovis BCG ADHC. The M. smegmatis ADHC was purified from M. smegmatis and the kinetic parameters of this enzyme showed that using NADPH as electron donor it has a strong preference for aliphatic and aromatic aldehyde substrates. Like the M. bovis BCG ADHC, this enzyme is more likely to act as an aldehyde reductase than as an alcohol dehydrogenase. The discovery of such an ADHC in a fast-growing, and easily engineered mycobacterial species opens the way to the utilisation of this M. smegmatis enzyme as a convenient model for the study of the physiological role of this alcohol dehydrogenase in mycobacteria.     

Enzymatic responses of Drosophila melanogaster to long- and short-term exposures to ethanol

The effects of environmental ethanol on larva-to-pupa survival and on the activities of four enzymes were investigated in three Drosophila melanogaster strains. The strains had different allelic combinations at the Odh and Aldox loci on their third chromosomes, but they all carried the Adh ^S -Gpdh ^F allelic combination on the second chromosome. Replicates of each of the strains were exposed to three different ethanol treatments: (i) no ethanol in the medium (control); (ii) 5% ethanol for a single generation (short-term exposure); (iii) 5% ethanol for 20 generations (long-term exposure). In all experiments, the activities of four enzymes (ADH, ODH, GPDH and AOX) were measured in larvae, pupae and adults. The results showed that (i) the larval and adult metabolic responses to environmental ethanol were different; (ii) enzyme activity changes under short-term exposure differed from those measured under long-term exposure; (iii) the activities of the allozymes common to all strains (ADH-S and GPDH-F), differed depending on the genetic background. Changes in larva-to-pupa survival were seen when the larvae of control and exposed lines of the three strains were confronted with various concentrations of ethanol. In all three strains, the exposed lines had significantly higher initial survival rate and ethanol tolerance than the control lines. Strain-specific differences were observed in the ethanol tolerance of both types of line. Received: 26 November 1996 / Accepted: 14 February 1997     

CHO cell responses to low oxygen: regulation of oxygen consumption and sensitization to oxidative stress

We have investigated the regulation of oxygen consumption and modulation of glutathione levels in CHO-K1 cells under oxygen-limiting conditions. We report here suppression of oxygen consumption and alteration of the supply-dependent relationship as a consequence of prolonged hypoxic or anoxic exposure. The suppression is characterized by an increase in the value of P(o(2)/50) (the oxygen tension at which oxygen consumption is half maximal). Under prolonged anoxia there is also a decrease in the cells' potential to use oxygen. Elevated glucose consumption under low oxygen conditions may contribute to the suppression in respiration. The glutathione concentration remains constant throughout hypoxic exposure but may decrease by as much as 40% under anoxia. The glutathione level in hypoxic and anoxic cells increases by two- and four-fold, respectively, over that of the control cells when exposed to a cytotoxic level of oxygen (93%). This suggests that anoxic and hypoxic exposure sensitizes CHO cells to oxidative stress. (c) 1992 John Wiley & Sons, Inc.     

Effects of extended and repeated exposures to low temperature on mortality of the peach-potato aphid Myzus persicae

Abstract.

• 1 The survival of adult and first-instar Myzus persicae reared at 20°C and 10°C was investigated after brief (1 min) exposure in the absence of plant material to temperatures between −5°C and −25°C, and extended exposures on plants of 1–10 days at a constant 5°C, 3°C and −5°C and a 24 h cycling regime between 5°C (18 h) and −5°C (6 h).
• 2 Life stage, rearing temperature, period of exposure and temperature regime all had a significant effect on the ability of aphids to survive cold. The effects of life stage and rearing temperature were most noticeable following exposure to cycling temperatures and extended exposures at −5°C, and least apparent after 1 min exposures at lower sub-zero temperatures.
• 3 Mortality following exposure to temperatures cycling between −5°C and 5°C was greater than that at 3°C (the mean of the cycling temperatures) and less than at a constant −5°C, suggesting that when temperatures fluctuate by a few degrees around 0°C the minimum temperature may affect survival to a greater extent than the mean.
• 4 These results suggest that an overwintering population of acclimated M.persicae would persist without significant mortality after a period of 7–10 days with −5°C frosts each night.

     

低温胁迫下6种木兰科植物的生理响应及抗寒相关基因差异表达

分析了6种木兰科植物对低温胁迫的生理响应及耐寒相关调控基因HSP90和WRKY33的差异表达,为木兰科植物抗寒机理的研究和抗寒品种的选育提供理论基础。结果表明,6种木兰科植物的低温LT50在-10.64—-22.06℃,从高到低依次为红花深山含笑、峨眉含笑、杂交含笑、阔瓣含笑、六瓣含笑和乐东拟单性木兰;低温过程中,6种木兰科植物叶片可溶性蛋白(SP)、游离脯氨酸(Pro)含量、超氧化物歧化酶(SOD)和过氧化物歧化酶(POD)活性呈先升高后降低的趋势,可溶性糖(SS)和丙二醛含量(MDA)则不断积累;筛选出REC、MDA、SP、SS和Pro作为6种木兰科植物抗寒性评价的关键指标;聚类分析将6种木兰科植物在抗寒性能上分为强、中、弱三类,分别为乐东拟单性木兰和六瓣含笑,阔瓣含笑、杂交含笑和峨眉含笑,以及红花深山含笑。对HSP90、WRKY33基因的差异表达分析表明,2个基因在6种木兰科植物中的相对表达量呈先升高后降低的趋势,在临近各树种LT 50时,2个基因的表达被强烈抑制且后期表达量不可逆。0℃时,2个基因的表达量差异不显著;-5℃时,2个基因开始被激活,表达量增加;-10℃时,HSP90、WRKY33基因在红花深山含笑叶片中的表达量较-5℃时分别下调了0.76倍和0.68倍,而在其他5个树种中的表达被进一步激活;-15℃时,HSP90和WRKY33基因在抗寒性中等的阔瓣含笑、杂交含笑、峨眉含笑中亦被强烈抑制,较-10℃时分别下调了0.38倍、0.33倍、0.32倍和0.71倍、0.72倍、0.74倍,在抗寒性强的乐东拟单性木兰和六瓣含笑中的表达被进一步激活;-20℃以后,2个基因在6个树种中的表达均被强烈抑制,但在抗寒性最强的乐东拟单性木兰中的表达量仍高于其他5个树种。抗寒基因的激活与表达是影响植物抗寒性的重要因素,抗寒性不同的树种对低温的应答机制明显不同。抗寒性越强的树种越能快速启动低温应答机制,激活抗寒相关基因的表达,进而调整生理生化活动以抵御和适应冷应力。不抗寒树种中抗寒基因的表达则受到抑制,降低了其对低温逆境的耐受能力。     

Molecular cloning and differential expression of an aldehyde dehydrogenase gene in rice leaves in response to infection by blast fungus

Aldehyde dehydrogenase (ALDH) superfamily is a group of enzymes metabolizing endogenous and exogenous aldehydes. Using differential display RT-PCR and cDNA library screening, a full-length aldehyde dehydrogenase cDNA (ALDH7B7) was isolated from rice leaves infected by incompatible race of blast fungus Magnaporthe grisea. The deduced amino acid sequence consists of 509 amino acid residues and shares 74∼81% identity with those of ALDH7Bs from other plants. ALDH7B7 expression was induced by blast fungus infection, ultraviolet, mechanical wound in rice leaves and was not detected in untreated rice organs. This gene has also been found to be inducible after exogenous phytohormones application, such as salicylic acid, methyl ester of jasmonic acid and abscisic acid. The function of ALDH7B7 in the interaction process between blast fungus and rice is discussed.     

The metabolic and biochemical responses of tropical whitespotted bamboo shark Chiloscyllium plagiosum to alterations in environmental temperature

The capacity of tropical whitespotted bamboo sharks Chiloscyllium plagiosum to metabolically compensate, at both the whole‐animal and biochemical levels, to prolonged exposure to temperatures higher (30° C) and lower (20 and 15° C) than their native temperature (24·5° C) was examined. As expected, whitespotted bamboo shark oxygen consumption increased upon exposure to 30° C and decreased at 20 and 15° C. Initial changes in oxygen consumption were maintained even after months at the experimental temperature, indicating that whitespotted bamboo sharks did not compensate metabolically to the experimental temperatures. Maximal activities and thermal sensitivity of citrate synthase and lactate dehydrogenase from whitespotted bamboo shark white locomotor muscle were similar between control animals maintained at 24·5° C and those maintained at 15° C, indicating that cold‐exposed animals did not compensate at the biochemical level. Similarly, lactate dehydrogenase activity did not change following prolonged exposure to 30° C. White muscle from whitespotted bamboo sharks maintained at 30° C had significantly lower citrate synthase activity than did control animals. This result was surprising given the lack of metabolic compensation at the whole‐animal level. Overall, whole‐animal oxygen consumption measurements supported the hypothesis that animals from thermally stable environments lacked the capacity to metabolically compensate to altered temperatures. Enzymatic results, however, suggested that the metabolic potential of muscle could change following temperature acclimation even in the absence of metabolic compensation at the whole‐animal level.     

三种植物线粒体基因低温差异表达比较分析

线粒体作为植物细胞的能量代谢中心,在植物响应逆境胁迫中有重要的作用。该研究基于雪莲(Saussurea involucrata)、番茄(Lycopersicon esculentum)和拟南芥(Arabidopsis thaliana)三种不同低温耐受性植物的低温转录组。通过blast比对和数据库检索筛选相关物种的线粒体基因,使用PlantCARE在线网站分析启动子,使用mega7软件对系统发育树构建分析。结果表明:通过差异表达基因筛选,分别在雪莲、拟南芥、番茄中筛选出2、24、15个显著差异表达基因,主要包括线粒体核糖体亚基和电子传递链各复合体亚基,其中部分基因的低温差异表达情况如NAD1和NAD5,可能与植物的低温适应性有关;通过表达模式的聚类分析,雪莲与拟南芥在基因的表达模式上相对于番茄更为相近,且不同类别的基因表达模式在不同物种间差异较大;雪莲与其他菊科植物的呼吸链相关基因的蛋白序列具有很大差异,与万年藓(Climacium dendroides)、牛舌藓(Anomodon minor)等高山植物的进化距离较近。在整体上拟南芥、番茄和雪莲三种植物线粒体基因在低温响应上具有很大差...     

Phenological responses to temperature of an annual and a perennial Lesquerella species

BACKGROUND AND AIMS: The annual Lesquerella fendleri, native to the south-western desert of United States and Mexico, and the perennial L. mendocina, native to Argentina, may have potential as new crops for cold-arid environments. The introduction of a new crop requires an understanding of environmental influences on growth and development, particularly temperature, which has been recognized as the main factor affecting the rate of development in crops. The objective of this study was to examine differences in the phenology of L. fendleri and L. mendocina and in the response to temperature in both vegetative and reproductive phases. METHODS: Plants of each species were grown at a range of constant temperatures under controlled conditions and developmental responses were analysed and quantified. KEY RESULTS: The rate of development of L. fendleri increased linearly with temperature in the phase from emergence (EM) to floral bud appearance (FBA) over the range 9-20 degrees C, and for the phase from FBA to first flower open (FL) over the range 9-24 degrees C. In contrast, the rate of development of L. mendocina was insensitive to temperature during the phase EM to FBA. In the phase FBA to FL, L. mendocina had a lower sensitivity to temperature than L. fendleri. In addition, L. fendleri exhibited a quantitative response to supra-optimal temperatures (reducing rate of development with further increases in temperature) whereas L. mendocina showed a qualitative response, with development ceasing to progress at temperatures above the optimum. CONCLUSIONS: This differential behaviour at high temperatures could explain the biennial habit found for L. mendocina sown during late spring under field conditions, whereas it behaves as an annual when sown in autumn-winter. The possibility is discussed of using this information for establishing the coincidence of critical stages with environmental conditions that can limit potential and actual yield through agronomic practices.     

Adaptation to low temperature and regulation of gene expression in antarctic psychrotrophic bacteria

Exposure to extremes of temperatures cause stresses which are sometimes lethal to living cells. Microorganisms in nature, however, are extremely diverse and some of them can live happily in the freezing cold of Antarctica. Among the cold adapted psychrotrophs and psychrophiles, the psychrotrophic bacteria are the predominant forms in the continental Antarctica. In spite of living in permanently cold area, the antarctic bacteria exhibit, similar to mesophiles, ‘cold-shock’ response albeit at a much lower temperatures, e.g., at 0–5°C. However, because of permanently cold condition and the long isolation of the continent, the microorganisms have acquired new adaptive features in the membranes, enzymes and macromolecular synthesis. Only recently these adaptive modifications are coming into light due to the efforts of various laboratories around the world. However, a lot more is known about adaptive response to low temperature in mesophilic bacteria than in antarctic bacteria. Combined knowledge from the two systems is providing useful clues to the understanding of basic biology of low temperature growing organisms. This article will provide an overview of this area of research with a special reference to sensing of temperature and regulation of gene expression at lower temperature.     

达氏鳇不同发育期胚胎对低温的耐受研究

研究了达氏鳇12个发育期胚胎经过不同低温（2 ℃、3 ℃、5 ℃、7 ℃和8 ℃）处理12 h、24 h、2 d、3 d、6 d、10 d、15 d、20 d和30 d后的孵化率和仔鱼成活率.结果表明,卵黄栓期、隙状胚孔期、神经管闭合期胚胎在2～8 ℃水温下,处理24 h后孵化率为0;卵裂期、囊胚早期、原肠中期胚胎在2～8 ℃水温下,处理3 d后孵化率低于30%;囊胚晚期、原肠早期、眼基期、尾芽期、心跳期和尾达头部期胚胎在5～8 ℃水温下,处理3 d后孵化率、仔鱼成活率超过70%;随低温处理时间延长,胚胎和仔鱼的死亡率增加,处理时间与孵化率、仔鱼成活率呈负相关;囊胚晚期、原肠早期、眼基期胚胎在5 ℃水温下耐受力较强,处理10 d后孵化率、仔鱼成活率超过70%.本研究表明,达氏鳇胚胎发育过程中囊胚晚期、原肠早期和眼基期胚胎可以在某一低温下进行短期保存,其孵化率、仔鱼成活率与常温（16～17 ℃）下没有显著差异.这对于达氏鳇胚胎（受精卵）的长途运输有重要意义.     

Genetics of gene expression responses to temperature stress in a sea urchin gene network

Stress responses play an important role in shaping species distributions and robustness to climate change. We investigated how stress responses alter the contribution of additive genetic variation to gene expression during development of the purple sea urchin, Strongylocentrotus purpuratus, under increased temperatures that model realistic climate change scenarios. We first measured gene expression responses in the embryos by RNA‐seq to characterize molecular signatures of mild, chronic temperature stress in an unbiased manner. We found that an increase from 12 to 18 °C caused widespread alterations in gene expression including in genes involved in protein folding, RNA processing and development. To understand the quantitative genetic architecture of this response, we then focused on a well‐characterized gene network involved in endomesoderm and ectoderm specification. Using a breeding design with wild‐caught individuals, we measured genetic and gene–environment interaction effects on 72 genes within this network. We found genetic or maternal effects in 33 of these genes and that the genetic effects were correlated in the network. Fourteen network genes also responded to higher temperatures, but we found no significant genotype–environment interactions in any of the genes. This absence may be owing to an effective buffering of the temperature perturbations within the network. In support of this hypothesis, perturbations to regulatory genes did not affect the expression of the genes that they regulate. Together, these results provide novel insights into the relationship between environmental change and developmental evolution and suggest that climate change may not expose large amounts of cryptic genetic variation to selection in this species.     

冬小麦拔节期不同茎蘖对低温胁迫的反应及抗冻性评价

以小麦济南17和山农8355为材料,在低温胁迫条件下,测定了不同茎蘖功能叶和叶鞘超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性及丙二醛(MDA)和可溶性蛋白含量,并利用主成分分析、聚类分析对其抗冻性进行综合评价.结果表明： 低温胁迫条件下,小麦拔节期不同茎蘖功能叶和叶鞘中SOD、POD和CAT活性均不同程度地上升,MDA和可溶性蛋白含量则不同程度地上升或下降.利用主成分分析和聚类分析,将济南17不同茎蘖分为3类：主茎和一级分蘖Ⅰ、Ⅱ属强抗冻蘖组,一级分蘖Ⅲ、Ⅳ和二级分蘖Ⅰp属中度抗冻蘖组,二级分蘖Ⅱp属弱抗冻蘖组;将山农8355不同茎蘖分为3类：主茎和一级分蘖Ⅰ、Ⅱ、Ⅲ属强抗冻蘖组,一级分蘖Ⅳ和二级分蘖Ⅰp属中度抗冻蘖组,二级分蘖Ⅱp属弱抗冻蘖组.表明冬小麦拔节期不同茎蘖存在抗冻性差异,且低位蘖较高位蘖抗冻.
     

盐穗木甜菜碱醛脱氢酶基因（BADH）的克隆及其在盐胁迫下的表达分析

根据我们实验室已发表的植物甜菜碱醛脱氢酶基因（BADH）的同源保守区设计引物,通过RT—PCR扩增获得了由1503个核苷酸组成的盐穗木BADH基因开放阅读框,推测该基因编码500个氨基酸,分子量约为54．49kDa的多肽。推测的盐穗木BADH氨基酸序列中包含一段甜菜碱醛脱氢酶中高度保守的十肽序列（VTLELGGKSP）以及与酶功能有关的半胱氨酸残基（Cys）。序列比对结果显示,盐穗木BADH与盐地碱蓬、中亚滨藜、盐爪爪以及菠菜等的核苷酸序列同源性在81％以上,与水稻的同源性也达到68％。半定量RT—PCR分析结果表明,盐穗木BADH基因的表达受盐胁迫诱导,推测BADH可能与盐穗木具有较强的耐盐能力有关。