Comparative Physiological and Proteomic Analysis Reveals the Leaf Response to Cadmium-Induced Stress in Poplar (Populus yunnanensis)

Excess amounts of heavy metals are important environmental pollutants with significant ecological and nutritional effects. Cdmium (Cd) is of particular concern because of its widespread occurrence and high toxicity. We conducted physiological and proteomic analyses to improve our understanding of the responses of Populus yunnanensis to Cd stress. The plantlets experienced two apparent stages in their response to Cd stress. During the first stage, transiently induced defense-response molecules, photosynthesis- and energy-associated proteins, antioxidant enzymes and heat shock proteins (HSPs) accumulated to enhance protein stability and establish a new cellular homeostasis. This activity explains why plant photosynthetic capability during this period barely changed. During the second stage, a decline of ribulose-1, 5-bisphosphate carboxylase (RuBisCO) and HSP levels led to imbalance of the plant photosynthetic system. Additionally, the expression of Mitogen-activated protein kinase 3 (MPK3), Mitogen-activated protein kinase 6 (MPK6) and a homeobox-leucine zipper protein was higher in the second stage. Higher expression of caffeoyl-CoA O-methyltransferase (CCoAOMT) may regulate plant cell wall synthesis for greater Cd storage. These genes may be candidates for further research and use in genetic manipulation of poplar tolerance to Cd stress.     

Identification of proteins regulated by ABA in response to combined drought and heat stress in maize roots

We adopted a proteomics approach to identify and analyze the differential expression of maize root proteins associated with abscisic acid (ABA) regulation under combined drought and heat stress. Using mass spectrometry, we identified 22 major proteins that were significantly up-regulated under combined drought and heat stress. These 22 proteins were classified into 6 functional categories: disease/defense (8), metabolism (3), cell growth/division (3), signal transduction (2), transporters (2) and unclassified (4). Our previous reports showed that ABA regulates the expression of several small heat-shock proteins (sHSPs) in maize leaves subjected to the combination of drought and heat stress; however, no sHSPs were identified among the root proteins up-regulated in this study. RT-PCR and western blot analyses were used to identify six known sHSPs. The maize roots were pretreated with 100 μM of ABA, and subsequently, the expression of the 22 up-regulated proteins and 6 sHSPs was examined. 11 proteins were up-regulated in an ABA-dependent manner, 13 proteins were up-regulated in an ABA-independent manner, and 4 proteins were up-regulated but inhibited by ABA. The up-regulated proteins are interesting candidates for further physiological and molecular investigations of combination stress tolerance in maize.     

Heat shock protein 70 regulates the abscisic acid-induced antioxidant response of maize to combined drought and heat stress

We investigated the interaction between heat shock protein 70 (HSP70) and abscisic acid (ABA)-induced antioxidant response of maize to the combination of drought and heat stress. First, the increased activities of enzymes, including superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR) and catalase (CAT), induced by drought were less than those by heat or combined drought and heat stress, except some individual cases (e.g. CAT in leaves, GR in roots). Second, both HSP70 synthesis and H₂O₂ production increased prominently under drought, heat or their combination stress; the increase in leaves induced by drought and heat combination was the highest, followed by heat and by drought, while the increase in roots had not visible difference. Third, either in leaves or roots, pretreatment with ABA inhibitor, HSP70 inhibitor and H₂O₂ scavenger, significantly arrested the stress-induced increase of antioxidant enzyme activities, and ABA inhibitor and H₂O₂ scavenger obviously suppressed HSP70 synthesis, while HSP70 inhibitor slightly heightened H₂O₂ accumulation. Finally, 100 μM ABA significantly enhanced the activities of antioxidant enzymes, HSP70 expression and H₂O₂ production under stresses in comparison with ABA-deficient mutant vp5 maize plants without pretreatment. Thus, ABA-induced H₂O₂ production enhances the HSP70 synthesis and up-regulates the activities of antioxidant enzymes, resulting in the suppression of cellular reactive oxygen species (ROS) levels. Our results suggest that HSP70 may play a crucial role in ABA-induced antioxidant defense of maize to drought and heat combination.     

Photosynthesis and antioxidative defense mechanisms in deciphering drought stress tolerance of crop plants

Crop plants are regularly exposed to an array of abiotic and biotic stresses, among them drought stress is a major environmental factor that shows adverse effects on plant growth and productivity. Because of this these factors are considered as hazardous for crop production. Drought stress elicits a plethora of responses in plants resulting in strict amendments in physiological, biochemical, and molecular processes. Photosynthesis is the most fundamental physiological process affected by drought due to a reduction in the CO₂ assimilation rate and disruption of primary photosynthetic reactions and pigments. Drought also expedites the generation of reactive oxygen species (ROS), triggering a cascade of antioxidative defense mechanisms, and affects many other metabolic processes as well as affecting gene expression. Details of the drought stress-induced changes, particularly in crop plants, are discussed in this review, with the major points: 1) leaf water potentials and water use efficiency in plants under drought stress; 2) increased production of ROS under drought leading to oxidative stress in plants and the role of ROS as signaling molecules; 3) molecular responses that lead to the enhanced expression of stress-inducible genes; 4) the decrease in photosynthesis leading to the decreased amount of assimilates, growth, and yield; 5) the antioxidant defense mechanisms comprising of enzymatic and non-enzymatic antioxidants and the other protective mechanisms; 6) progress made in identifying the drought stress tolerance mechanisms; 7) the production of transgenic crop plants with enhanced tolerance to drought stress.     

钼在桃树干旱胁迫响应中的作用解析

以毛桃(Amygdalus persica)实生苗为试材, 研究干旱胁迫下, 钼酸铵处理对钼辅因子硫化酶编码基因(LOS5/ABA3)表达量、脱落酸(ABA)含量及抗旱相关生理指标的影响。结果表明, 干旱胁迫下, 喷施不同浓度钼酸铵处理毛桃实生苗叶片, 其含水量及叶绿素和脯氨酸含量显著高于对照, 且以0.04%钼酸铵处理效果最好; 电解质渗漏率显著低于对照。干旱胁迫下, 与对照相比, 喷施0.04%钼酸铵的毛桃实生苗叶片中LOS5/ABA3表达量显著提高; ABA含量、水分利用效率和净光合速率均高于对照, 蒸腾速率低于对照, 且差异显著; 叶片抗氧化酶活性显著升高, MDA含量显著降低; 离体处理的叶片质量损失减缓, 且差异显著。研究表明毛桃实生苗在干旱胁迫下喷施钼酸铵可通过上调钼辅因子硫化酶编码基因的表达水平, 提高叶片中ABA和脯氨酸含量及抗氧化酶活性, 从而缓解干旱胁迫下的细胞膜氧化伤害, 降低叶片失水速率, 减轻干旱胁迫对毛桃实生苗的伤害。     

Comparative physiological and proteomic responses to drought stress in two poplar species originating from different altitudes

Cuttings of Populus kangdingensis C. Wang et Tung and Populus cathayana Rehder were examined during a single growing season in a greenhouse for comparative analysis of their physiological and proteomic responses to drought stress. The said species originate from high and low altitudes, respectively, of the eastern Himalaya. Results revealed that the adaptive responses to drought stress vary between the two poplar species. As a consequence of drought stress, the stem height increment and leaf number increment are more significantly inhibited in P. cathayana compared with P. kangdingensis. On the other hand, in response to drought stress, more significant cellular damages such as reduction in leaf relative water content and CO₂ assimilation rate, increments in the contents of malondialdehyde and hydrogen peroxide and downregulation or degradation of proteins related to photosynthesis occur in P. cathayana compared with P. kangdingensis. On the other hand, P. kangdingensis can cope better with the negative impact on the entire regulatory network. This includes more efficient increases in content of solute sugar, soluble protein and free proline and activities of antioxidant enzymes, as well as specific expressions of certain proteins related to protein processing, redox homeostasis and sugar metabolism. Morphological consequences as well as physiological and proteomic responses to drought stress between species revealed that P. kangdingensis originating from a high altitude manifest stronger drought adaptation than did P. cathayana originating from a low altitude. Functions of various proteins identified by proteomic experiment are related with physiological phenomena. Physiological and proteomic responses to drought stress in poplar may work cooperatively to establish a new cellular homeostasis, allowing poplar to develop a certain level of drought tolerance.     

Antioxidant responses of wheat plants under stress

Currently, food security depends on the increased production of cereals such as wheat (Triticum aestivum L.), which is an important source of calories and protein for humans. However, cells of the crop have suffered from the accumulation of reactive oxygen species (ROS), which can cause severe oxidative damage to the plants, due to environmental stresses. ROS are toxic molecules found in various subcellular compartments. The equilibrium between the production and detoxification of ROS is sustained by enzymatic and nonenzymatic antioxidants. In the present review, we offer a brief summary of antioxidant defense and hydrogen peroxide (H₂O₂) signaling in wheat plants. Wheat plants increase antioxidant defense mechanisms under abiotic stresses, such as drought, cold, heat, salinity and UV-B radiation, to alleviate oxidative damage. Moreover, H₂O₂ signaling is an important factor contributing to stress tolerance in cereals.     

Abscisic acid cross-talking with hydrogen peroxide and osmolyte compounds may regulate the leaf rolling mechanism under drought

Leaf rolling observed in some crops such as maize, rice, wheat and sorghum is an indicator of decreased water status. Moderate leaf rolling not tightly or early increases the photosynthesis and grain yield of crop cultivars under environmental stresses. Moreover, the effects of exogenous abscisic acid (ABA) on stomatal conductance, water status and synthesis of osmotic compounds are a well-known issue in plants subjected to water deficit. However, it is not clear how the cross-talk of ABA with H₂O₂ and osmolyte compounds affects the leaf rolling mechanism. Regulation mechanism of leaf rolling by ABA has been first studied in maize seedlings under drought stress induced by polyethylene glycol 6000 (PEG 6000) in this study. ABA treatment under drought stress reduced hydrogen peroxide (H₂O₂) content and the degree of leaf rolling (%) while the treatment-induced ABA synthesis, osmolyte levels (proline, polyamine and total soluble sugars) and some antioxidant enzyme activities in comparison to the plants that were not treated with ABA. Furthermore, exogenous ABA up-regulated the expression levels of arginine decarboxylase (ADC) and pyrroline-5-carboxylate synthase (P5CS) genes and down-regulated polyamine oxidase (PAO), diamine oxidase (DAO) and proline dehydrogenase (ProDH) gene expressions. When endogenous ABA content was decreased by the treatment of fluoridone (FLU) that is an ABA inhibitor, leaf rolling degree (%), H₂O₂ content and antioxidant enzyme activities increased, but osmolyte levels, ADC and P5CS gene expressions decreased. Finally, the treatment of ABA to maize seedlings exposed to drought stress resulted in the stimulation of the antioxidant system, osmotic adjustment and reduction of leaf rolling. We concluded that ABA can be a signal compound cross-talking H₂O₂, proline and polyamines and thus involved in the leaf rolling mechanism by providing osmotic adjustment. The results of this study can be used to provide data for the molecular breeding of maize hybrids with high grain yield by means of moderately rolled leaves.     

Arbuscular mycorrhizal fungi modulates dynamics tolerance expression to mitigate drought stress in Ephedra foliata Boiss

Arbuscular mycorrhizal fungi (AMF) are one of the most important drivers of soil ecosystem dynamics. AMF have the potential to improve plant growth and development by modulating key hormonal pathways, which result in decreasing the adverse impact of abiotic stress, such as drought. Pot experiments were conducted in this study to investigate the ability of AMF to ameliorate the adverse impact of drought in Ephedra foliate. Non-inoculated AMF E. foliate (Ef) plants, exhibited reduced growth in response to drought stress with a concomitant lowering of chlorophyll pigments, relative to non-stressed and AMF inoculated plant. AMF inoculated E. foliate showed improved nitrogen metabolism by positively regulating nitrate and nitrite reductase activity which results in greater ammonium availability for the synthesis of amino acids. Inoculation with AMF also increased antioxidant enzyme activity, ascorbic acid contents, and reduction in glutathione level. This resulted in significant amelioration of oxidative damage to plant membranes by restricting the excess generation of reactive oxygen species (ROS), such as hydrogen peroxide. Greater content of proline, glucose, and total soluble protein in AMF-inoculated plants provided further benefit to E. foliate plants and their ability to withstand drought stress, and also evident by a greater level of sucrose phosphate synthase activity. AMF significantly enhanced the uptake of essential nutrients like K, Mg, and Ca. Importantly, higher concentrations of plant hormones, including indole acetic acid (IAA), indole butyric acid (IBA), gibberellic acid (GA), and abscisic acid (ABA), were maintained in AMF-inoculated Ef plants. AMF inoculation also boosted phosphorous metabolism by increasing alkaline and acid phosphatase enzyme activity. In summary, AMF-inoculation of Ef plants significantly reduced the deleterious effect of drought stress by up-regulating the antioxidant defense system, synthesis of osmolytes, and maintaining phytohormone levels.     

Influence of Exogenous Glycine Betaine and Abscisic Acid on Papaya in Responses to Water-deficit Stress

The effects of exogenous foliar glycine betaine (GB) and abscisic acid (ABA) on papaya responses to water stress were investigated under distinct water regimes. Papaya seedlings (Carica papaya L. cultivar “BH-65”) were pretreated with GB or ABA and subsequently subjected to consecutive periods of drought, rehydration, and a second period of drought conditions. Results indicated that water stress induced ABA, jasmonic acid (JA), and proline accumulation but did not modify malondialdehyde (MDA) concentration. In addition, water deprivation reduced photosynthetic rate, stomatal conductance, relative water content (RWC), leaf fresh weight, and increased leaf abscission. GB applied prior to drought imposition decreased the impact of water stress on ABA, JA, proline accumulation, leaf water status, growth, and photosynthetic performance. However, ABA-pretreated plants did not show alteration of most of these parameters under water stress conditions when compared with non-pretreated plants except a clear induction of JA accumulation. Taken together, the data suggest that GB may modulate ABA, JA, and proline accumulation through the control of stomatal movement and the high availability of compatible solutes, leading to improvement of leaf water status, growth, and photosynthetic machinery function. In contrast, exogenous ABA did not stimulate papaya physiological responses under drought, but interestingly ABA in combination with drought could induce progressive JA synthesis, unlike drought alone, which induces a transitory JA increase and may trigger endogenous ABA accumulation. The data also suggest that irrespective of the pretreatments, papaya did not suffer oxidative damage.     

Proline accumulation is inhibitory to Arabidopsis seedlings during heat stress

The effect of proline (Pro) accumulation on heat sensitivity was investigated using transgenic Arabidopsis (Arabidopsis thaliana) plants ectopically expressing the Δ(1)-pyrroline-5-carboxylate synthetase 1 gene (AtP5CS1) under the control of a heat shock protein 17.6II gene promoter. During heat stress, the heat-inducible expression of the AtP5CS1 transgene was capable of enhancing Pro biosynthesis. Twelve-day-old seedlings were first treated with heat at 37 °C for 24 h to induce Pro and then were stressed at 50 °C for 4 h. After recovery at 22 °C for 96 h, the growth of Pro-overproducing plants was significantly more inhibited than that of control plants that do not accumulate Pro, manifested by lower survival rate, higher ion leakage, higher reactive oxygen species (ROS) and malondialdehyde levels, and increased activity of the Pro/P5C cycle. The activities of antioxidant enzymes superoxide dismutase, guaiacol peroxidase, and catalase, but not those of glutathione reductase and ascorbate peroxidase, increased in all lines after heat treatment, but the increase was more significant in Pro-overproducing seedlings. Staining with MitoSox-Red, reported for being able to specifically detect superoxide formed in mitochondria, showed that Pro accumulation during heat stress resulted in elevated levels of ROS in mitochondria. Interestingly, exogenous abscisic acid (ABA) and ethylene were found to partially rescue the heat-sensitive phenotype of Pro-overproducing seedlings. Measurement of ethylene and ABA levels further confirmed that these two hormones are negatively affected in Pro-overproducing seedlings during heat stress. Our results indicated that Pro accumulation under heat stress decreases the thermotolerance, probably by increased ROS production via the Pro/P5C cycle and inhibition of ABA and ethylene biosynthesis.