Signal transduction and biotechnology in response to environmental stresses

Providing sufficient food to burgeoning population from the steadily shrinking arable land seems to be very difficult in near future and is one of the foremost challenges for plant scientists. In addition, there are several biotic and abiotic stresses which frequently encounter crop plants during various stages of life cycle, resulting in considerable yield losses. Environmental stresses, including drought, flooding, salinity, temperature (both low and high), high radiation, and xenobiotics induce toxicity, membrane damage, excessive reactive oxygen species (ROS) production, reduced photosynthesis, and altered nutrient acquisition. Several indigenous defence mechanisms (physiological and molecular) are triggered in plants on exposure to environmental cues. Enhancement of resistance of crop plants to environmental stresses has been the topic of prime interest for agriculturalists and plant scientists since long. Development of water and salinity stress-tolerant crops through genetic engineering provides an avenue towards the reclamation of farmlands that have been lost due to salinity and lack of irrigation water/rainfall. Understanding the complexity of stress tolerance mechanisms in orthodox or model plants at the genetic and molecular levels improves feasibility of enhancing tolerance of sensitive crop plants.     

Isolation and functional analysis of cotton universal stress protein promoter in response to phytohormones and abiotic stresses

The 949 bp promoter fragment upstream from the translation initiation site of the GUSP gene encoding a universal stress protein was isolated from the genomic DNA of Gossypium arboreum. Some putative cis-acting elements involved in stress responses including E-box, ABRE, DPBF-box, and MYB-core elements were found in the promoter region. In an Agrobacterium-mediated transient expression assay, strong activation of the GUSP full promoter region occurred in tobacco leaves following dehydration, abscisic acid, salt, heavy metal, gibberellic acid and dark treatments. Deletion analysis of the promoter revealed that the dehydration, abscisic acid and salt responses were affected by the deletion between −208 and −949 bp and showed 2–4-fold induction. However, in response to dark, gibberellic acid and heavy metals the induction was only 2-fold. These findings further our understanding of the regulation of GUSP expression. This is an important study as no report of this universal stress protein promoter is available in literature.     

Different protein kinase families are activated by osmotic stresses in Arabidopsis thaliana cell suspensions. Involvement of the MAP kinases AtMPK3 and AtMPK6

Five Ca(2+)-independent protein kinases were rapidly activated by hypoosmotic stress, moderate or high hyperosmolarity induced by several osmolytes, sucrose, mannitol or NaCl. Three of these kinases, transiently activated by hypoosmolarity, recognised by anti-phosphorylated mitogen-activated protein (MAP) kinase antibodies, sensitive to a MAP kinase inhibitor and inactivated by the action of a tyrosine phosphatase, corresponded to MAP kinases. Using specific antibodies, two of the MAP kinases were identified as AtMPK6 and AtMPK3. The two other protein kinases, durably activated by high hyperosmolarity, did not belong to the MAP kinase family. Activation of AtMPK6 and AtMPK3 by hypoosmolarity depended on upstream protein kinases sensitive to staurosporine and on calcium influx. In contrast, these two transduction steps were not involved in the activation of the two protein kinases activated by high hyperosmolarity.     

Protein response of insect cells to bioreactor environmental stresses

Protein expression of Spodoptera frugiperda (Sf9) insect cells was characterized upon exposure to environmental stresses typically present in bioreactors including heat shock, oxygen deprivation, shear stress, change of pH, and salinity or ethanol shock. This study fills the void in knowledge as to how bioreactor hydrodynamics, anoxia, small changes in pH as well as salinity alterations due to pH control or exposure to ethanol used in asepsis treatments affect protein expression in Sf9 cells. Heat shock at 43 degrees C induced proteins at 83 kDa, 68-78 kDa and six small heat shock proteins (hsps) at 23-15.5 kDa. Anaerobic conditions in CO2 atmosphere reduced significantly the normal protein synthesis and induced a small subset of heat shock proteins at 70 kDa. Oxygen deprivation in nitrogen atmosphere transiently induces the 70 kDa proteins and had minor effects on the normal protein synthesis. Exposure to increased salinity or ethanol concentration failed to trigger the stress response, but may extensively inhibit the induction of normal proteins even though there was a negligible change in cell viability. Shear stress that had a major reducing effect on cell viability did not change the protein synthesis profile of Sf9 cells. Both long and short term exposures to small pH changes had negligible effects on protein synthesis.     

Autophagy in response to environmental stresses: New monitoring perspectives

Autophagy, an evolutionarily conserved process of cellular homeostasis in all eukaryotes, has been heavily implicated in many aspects of human health and diseases. However, its pivotal roles, particularly in stress and adaptive responses in other species in the environment, have perhaps not received the attention they deserve. Autophagy processes may underlie important ecological phenomena such as coral bleaching, as well as various forms of responses and adaptations to environmental forcing and deterioration. Investigating and assessing autophagy responses in the contexts of environmental stresses and ecological changes would therefore be important. Such investigations in indicator organisms could provide valuable parameters for ecosystems health assessment. Understanding autophagy responses in ecologically important species could also be useful in efforts of species and biodiversity conservation.     

Sweet corn response to combined nitrogen and salinity environmental stresses

To define the nature of the combined response curve of sweet corn (Zea mays L.) plants to nitrogen and salinity, a lysimeter study was designed to follow water and solute budgets with combinations of the two variables over wide ranges of 0.5–7.5 dS m^–1 and 0–150% of local N-fertilization recommendations. Patterns of water-use efficiency, N content, N uptake, and shoot dry-matter yield indicated the predominance of environmental interactions over Cl-nitrate physiological antagonism. At low salinities, the leaf N content, N uptake, and yield increased with increased N fertilization up to 45% of local N-fertilization recommendations, nitrogen was efficiently stripped from the percolating water and practically no nitrate was leached. At higher N fertilization the amount of leached N increased linearly with increased N input, and N uptake and yield were independent of N rates, levelling off at increased values for decreased salinities. The Liebig–Sprengel and Mitscherlich–Baule models were evaluated against measured data; both achieved similar values for the system's inherent N, the salinity level corresponding with zero-yield, and the predicted yields, which were highly correlated with the experimental data (R ² > 0.9). It is suggested that both models can be used successfully in mechanistic-based plant–soil solution models to predict yield, water and nutrient needs, and the resulted N leaching.     

水稻OsERF96应答病原菌的表达及启动子的功能分析

前人研究发现水稻Os ERF96(ethylene responsive factor 96)可应答白叶枯病病原菌的侵染,但其功能及表达调控机制仍不清楚,本研究进一步分析了该基因在水稻应答稻瘟病病原菌侵染及外源激素(SA和Me JA)处理下的转录情况,并分析了其启动子的诱导表达活性。结果表明:相对于对照组,Os ERF96在接种稻瘟病后1~4 d表达量显著上调,以第1天最高,此后逐渐下降,外源SA处理后Os ERF96表达量持续上调;利用Os ERF96启动子驱动GUS的转基因株系分析了Os ERF96启动子的诱导活性,结果表明GUS在根、茎和叶均有不同程度组成型表达,稻瘟病菌接种后4~7 d GUS活性持续升高。GUS活性定量表明,稻瘟病菌和SA处理条件下均出现了升高。综上所述,Os ERF96可应答白叶枯病或稻瘟病病原菌的浸染,其启动子是一个对病原菌侵染产生应答的诱导性启动子。     

细菌sRNA对环境胁迫的响应机制

细菌小RNA (Small RNAs,sRNAs)是一类长度大约在40?400个核酸之间,不编码蛋白质的RNA,在细菌适应环境方面起重要的调节作用。当环境中温度、营养、外膜蛋白、pH、铁等条件改变时,sRNA常常通过连接双组分信号转导系统和调节蛋白,来传递压力信号并调节应激响应,其作用方式一般是通过碱基互补配对的方式与靶mRNA结合,从而调控靶mRNA的翻译和稳定性;或直接与靶标蛋白质结合,调节靶标蛋白质的生物活性。本文总结了细菌在多种环境压力下,sRNA的调控响应机制。     

Cloning and expression pattern of heat shock protein genes from the endoparasitoid wasp, Pteromalus puparum in response to environmental stresses

Six heat shock protein (HSP) genes from five HSP families in the parasitoid, Pteromalus puparum, were evaluated for their response to temperature (-15 ~ 3°C , and 30 ~ 42°C for 1 h), heavy metals (0.5 ~ 5 mM Cd(2+) and Cu(2+) for 24 h and 60 h), and starvation (24 h). Compared with other insect HSPs, all conserved motifs are found in P. puparum HSPs, and they are very similar to those of the recently sequenced ectoparasitoid Nasonia vitripennis. The temporal gene expression patterns indicated that these six HSP genes were all heat-inducible, of which hsp40 was the most inducible. The temperatures for maximal HSP induction at high and low temperature zone were 36 or 39°C and -3°C, respectively. In the hot zone, all HSP genes have the same initial temperature (33°C) for up-regulation. Low concentrations of Cd(2+) for a short-term promoted the expression of all HSP genes, but not high concentrations or long-term treatments. Cu(2+) stress for 24 h increased expression of nearly all HSP. Four HSP genes changed after starvation. We infer that all six HSP genes are sensitive to heat. This may help understand the absence of P. puparum during the summer and winter. The expression profiles of six HSP genes in P. puparum under heavy metal stress indicates that HSP is a short-term response to cellular distress or injury induced by Cd(2+) and Cu(2+).     

原绿球藻对环境胁迫的生理和分子响应机制

原绿球藻(Prochlorococcus)作为海洋丰度最高的浮游植物,对海洋生态系统的物质循环和能量流动起着重要的驱动作用。原绿球藻生长和光合作用活性容易受到环境胁迫的影响,进而影响整个海洋生态系统的稳定性。因此,研究原绿球藻应对环境胁迫的响应机制具有重要的生态意义。原绿球藻主要通过分化出不同的生态型来适应不同光照和营养盐的海洋环境,但仍然会很难快速适应各种突如其来的海洋环境变化。本文从原绿球藻应对环境胁迫的角度,探讨了其生理和分子响应机制的最新研究进展,包括光系统I循环电子传递在光照变化时发挥的重要作用,通过RNA快速响应而调控基因表达应对环境胁迫,以及在辅助异养细菌的保护下应对活性氧的胁迫等。本文也展望了原绿球藻对环境胁迫响应的生理和分子机制的未来研究方向,旨在为原绿球藻抗逆机制的深入研究提供参考。     

几种白念珠菌高铁还原酶基因的功能研究

【目的】高铁还原酶在白念珠菌铁离子获得过程中发挥着重要作用。通过研究高铁还原酶基因的功能和表达调控,揭示其不同的环境应答策略。【方法】通过Northern杂交的方法分析不同低铁环境对高铁还原酶基因表达水平的影响。构建高铁还原酶基因缺失菌株,探究基因缺失后对细胞表面高铁还原酶活力和生长状况的影响。利用激光共聚焦显微镜观察Frp1蛋白的细胞学定位。【结果】酸性条件显著上调FRE10基因的表达水平,而碱性条件能显著提高FRE2基因的表达量。在酸性条件下,FRE10基因的缺失会显著地下调细胞表面高铁还原酶活力。在碱性条件下,fre2Δ/Δ缺失菌株表现出严重缺陷的生长能力和显著降低的表面高铁还原酶活力。细胞学定位实验发现Frp1蛋白位于液泡中。【结论】FRE2和FRE10基因的表达模式主要是酸碱依赖性的。Fre2是碱性条件下高铁还原酶活力的主要贡献者。Frp1蛋白位于液泡中,在液泡内储存铁的活化和转运过程中可能发挥重要作用。     

Expression analysis of five maize MAP kinase genes in response to various abiotic stresses and signal molecules

Mitogen-activated protein kinase (MAPK) cascades are universal signal transduction modules in eukaryotes. Plant MAPK cascades are complicated networks and play vital roles in signal transduction induced by biotic and abiotic stresses. In this paper, expression patterns of MAPKs in maize roots treated with low-temperature, osmotic stresses, wounding, plant hormones and UV-C irradiation were investigated. Semi-quantitative RT-PCR reveals that the expression of MAPKs in maize roots which treated with low-temperature in light or in low light are inducible. The expression patterns of MAPKs in maize roots with treatments of CaCl₂, SA, GA and wounding are approximately the same. A detailed time course experiment shows that the expression patterns of ZmSIMK are different with treatments of PEG and NaCl, respectively. These results suggest that the expression patterns of MAPKs are complicated and the signal pathways are interlaced into a network in maize roots.