6-苄（基）腺嘌呤和抗坏血酸对渗透胁迫时杨树光合作用光抑制的影响

研究了6-BA和A5A对渗透胁迫时杨树幼苗叶片光合作用光抑制和活性氧代谢的影响．结果表明,渗透胁迫时杨树叶片净光合速率(Pn)和表观量子效率(AQY)降低,光合作用光抑制加剧,超氧化物歧化酶(SOD)活性升高,抗坏血酸过氧化物酶(APX)活性降低,O2产生加快,H2O2和膜脂过氧化产物丙二醛(MDA)含量升高．6-BA和A5A预处理使胁迫时叶片SOD和APx活性升高。O2生成减少。H2O22和MDA含量降低,同时缓解了光合作用的光抑制．相关分析表明,杨树叶片活性氧水平和MDA含量与Pn和AQY呈负相关．胁迫时杨树叶片活性氧的积累与光合作用光抑制有一定关系,6-BA和A5A对光抑制的缓解作用与其对活性氧清除系统的促进作用有关。     

Identification of cell-wall stress as a hexose-dependent and osmosensitive regulator of plant responses

Development, abiotic and biotic stress each affect the physical architecture and chemical composition of the plant cell wall, making maintenance of cell-wall integrity an important component of many plant processes. Cellulose biosynthesis inhibition (CBI) was employed to impair the functional integrity of the cell wall, and the plant's response to this specific stress was characterized in an Arabidopsis seedling model system. CBI caused changes in the expression of genes involved in mechanoperception, the response to microbial challenge, and lignin and cell-wall polysaccharide biosynthesis. Following CBI, activation of a UDP- d -xylose 4-epimerase gene correlated with increases in arabinose and uronic acid content in seedling cell walls. Activation of pathogen response genes, lignin deposition and lesion formation were dependent on externally supplied sugars and were suppressed by osmotic support. Lignin deposition in the root elongation zone caused by CBI was reduced in atrbohd (NADPH oxidase) mutant seedlings but increased in jasmonic acid resistant1 ( jar1-1 ) mutant seedlings. Phytohormone measurements showed that CBI-induced increases in jasmonic (JA) and salicylic acids were dependent on sugar availability and prevented by osmotic support. We show that CBI activates responses commonly attributed to both abiotic and microbial challenges. Glucose/sucrose and turgor pressure are critical components in maintenance of cell-wall integrity and the regulation of induced responses, including JA biosynthesis. Lignin deposition induced by CBI is regulated by JAR1-1 and NADPH oxidase-dependent signalling processes. Our results identify components of the mechanism that mediates the response to impairment of cell-wall integrity in Arabidopsis thaliana .     

Acetobixan,an Inhibitor of Cellulose Synthesis Identified by Microbial Bioprospecting

In plants, cellulose biosynthesis is an essential process for anisotropic growth and therefore is an ideal target for inhibition. Based on the documented utility of small-molecule inhibitors to dissect complex cellular processes we identified a cellulose biosynthesis inhibitor (CBI), named acetobixan, by bio-prospecting among compounds secreted by endophytic microorganisms. Acetobixan was identified using a drug-gene interaction screen to sift through hundreds of endophytic microbial secretions for one that caused synergistic reduction in root expansion of the leaky AtcesA6^prc1-1 mutant. We then mined this microbial secretion for compounds that were differentially abundant compared with Bacilli that failed to mimic CBI action to isolate a lead pharmacophore. Analogs of this lead compound were screened for CBI activity, and the most potent analog was named acetobixan. In living Arabidopsis cells visualized by confocal microscopy, acetobixan treatment caused CESA particles localized at the plasma membrane (PM) to rapidly re-localize to cytoplasmic vesicles. Acetobixan inhibited ¹⁴C-Glc uptake into crystalline cellulose. Moreover, cortical microtubule dynamics were not disrupted by acetobixan, suggesting specific activity towards cellulose synthesis. Previous CBI resistant mutants such as ixr1-2, ixr2-1 or aegeus were not cross resistant to acetobixan indicating that acetobixan targets a different aspect of cellulose biosynthesis.     

Cytosolic invertases contribute to cellulose biosynthesis and influence carbon partitioning in seedlings of Arabidopsis thaliana

In plants, UDP‐glucose is the direct precursor for cellulose biosynthesis, and can be converted into other NDP‐sugars required for the biosynthesis of wall matrix polysaccharides. UDP‐glucose is generated from sucrose by two distinct metabolic pathways. The first pathway is the direct conversion of sucrose to UDP‐glucose and fructose by sucrose synthase. The second pathway involves sucrose hydrolysis by cytosolic invertase (CINV), conversion of glucose to glucose‐6‐phosphate and glucose‐1‐phosphate, and UDP‐glucose generation by UDP‐glucose pyrophosphorylase (UGP). Previously, Barratt et al. (Proc. Natl Acad. Sci. USA, 106, 2009 and 13124) have found that an Arabidopsis double mutant lacking CINV1 and CINV2 displayed drastically reduced growth. Whether this reduced growth is due to deficient cell wall production caused by limited UDP‐glucose supply, pleiotropic effects, or both, remained unresolved. Here, we present results indicating that the CINV/UGP pathway contributes to anisotropic growth and cellulose biosynthesis in Arabidopsis. Biochemical and imaging data demonstrate that cinv1 cinv2 seedlings are deficient in UDP‐glucose production, exhibit abnormal cellulose biosynthesis and microtubule properties, and have altered cellulose organization without substantial changes to matrix polysaccharide composition, suggesting that the CINV/UGP pathway is a key metabolic route to UDP‐glucose synthesis in Arabidopsis. Furthermore, differential responses of cinv1 cinv2 seedlings to exogenous sugar supplementation support a function of CINVs in influencing carbon partitioning in Arabidopsis. From these data and those of previous studies, we conclude that CINVs serve central roles in cellulose biosynthesis and carbon allocation in Arabidopsis.     

Regulation of respiration in plants: a role for alternative metabolic pathways

Respiratory metabolism includes the reactions of glycolysis, the tricarboxylic acid cycle and the mitochondrial electron transport chain, but is also directly linked with many other metabolic pathways such as protein and lipid biosynthesis and photosynthesis via photorespiration. Furthermore, any change in respiratory activity can impact the redox status of the cell and the production of reactive oxygen species. In this review, it is discussed how respiration is regulated and what alternative pathways are known that increase the metabolic flexibility of this vital metabolic process. By looking at the adaptive responses of respiration to hypoxia or changes in the oxygen availability of a cell, the integration of regulatory responses of various pathways is illustrated.     

根系渗透胁迫时杨树光合作用光抑制与活性氧的关系

为更多地了解自然条件下活体叶片的光抑制,研究了渗透胁迫时杨树无性系幼苗叶片的光抑制与活性氧代谢的关系．结果表明,随胁迫时间的延长和胁迫强度的增大,杨树叶片O2^-生成加快,H2O2和丙二醛(MDA)含量增多,超氧物歧化酶(SOD)活性升高,过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性降低,活性氧代谢失衡,光合作用的光抑制加剧．用二乙基二硫代氨基甲酸铜盐抑制SOD活性,或用甲基紫精加速O2^-的生成,亦可使杨树叶片发生光抑制．渗透胁迫时杨树无性系幼苗清除H2O2能力降低,限制了叶片通过Mehler反应耗散过剩光能,防御光破坏作用的发挥;光抑制的发生与活性氧的积累有关．     

Evidence for a role of salicylic acid in the oxidative damage generated by NaCl and osmotic stress in Arabidopsis seedlings.

Previous studies have shown that salicylic acid (SA) is an essential component of the plant resistance to pathogens. We now show that SA plays a role in the plant response to adverse environmental conditions, such as salt and osmotic stresses. We have studied the responses of wild-type Arabidopsis and an SA-deficient transgenic line expressing a salicylate hydroxylase (NahG) gene to different abiotic stress conditions. Wild-type plants germinated under moderate light conditions in media supplemented with 100 mM NaCl or 270 mM mannitol showed extensive necrosis in the shoot. In contrast, NahG plants germinated under the same conditions remained green and developed true leaves. The lack of necrosis observed in NahG seedlings under the same conditions suggests that SA potentiates the generation of reactive oxygen species in photosynthetic tissues during salt and osmotic stresses. This hypothesis is supported by the following observations. First, the herbicide methyl viologen, a generator of superoxide radical during photosynthesis, produced a necrotic phenotype only in wild-type plants. Second, the presence of reactive oxygen-scavenging compounds in the germination media reversed the wild-type necrotic phenotype seen under salt and osmotic stress. Third, a greater increase in the oxidized state of the glutathione pool under NaCl stress was observed in wild-type seedlings compared with NahG seedlings. Fourth, greater oxidative damage occurred in wild-type seedlings compared with NahG seedlings under NaCl stress as measured by lipid peroxidation. Our data support a model for SA potentiating the stress response of the germinating Arabidopsis seedling.     

植物盐胁迫应答蛋白质组学分析

土壤盐渍化是限制植物生长和分布的关键因素之一,揭示植物盐胁迫应答的分子机理是借助分子生物学手段提高植物耐盐性的基础.近年来,人们利用高通量蛋白质组学技术分析了拟南芥、水稻等19种植物的盐胁迫应答蛋白质表达图谱.从植物类群(盐生植物和甜土植物)、组织器官(根、地上部分/茎、胚根和胚轴、叶片、花序和配子体)、细胞(悬浮培养细胞、愈伤组织细胞和单细胞生物)和亚细胞结构(叶绿体、质膜和质外体)几方面整合分析了植物盐胁迫应答蛋白质组表达模式特征,主要特征包括:(1)盐生植物通过全面调节细胞骨架重塑、离子转运和区隔化、渗透平衡、活性氧(ROS)清除、信号转导、光合作用和能量代谢等信号与代谢网络体系,获得相对较高的抗/耐盐能力;(2)植物地上部分(叶片、茎、配子体)或光合组织细胞(悬浮培养细胞、愈伤组织细胞和单细胞盐藻)通过调节参与光合作用、碳和能量代谢、ROS清除过程蛋白质的表达模式应对盐胁迫环境;(3)植物地下部分(根、胚根)通过调控信号转导和离子转运相关蛋白质感知/传递盐胁迫信号并维持离子平衡;(4)花序中参与渗透调节、转录调控、蛋白质加工和ROS清除的蛋白质在盐胁迫条件下变化显著;(5)叶绿体通过调控参与光合作用、蛋白质加工和周转,以及氧化还原系统平衡等过程应对盐胁迫;(6)质外体中参与细胞壁代谢、胁迫防御和信号转导过程的蛋白质受盐胁迫影响明显;(7)细胞膜中参与维持膜结构稳定、物质/离子运输和信号转导过程的蛋白质对植物盐胁迫应答具有重要作用.这些分析为深入研究植物耐盐的分子机制提供了重要信息.     

NaCl胁迫抑制玉米幼苗光合作用的可能机理

用NaCl 100mmol/L处理玉米幼苗,216 h内不同时间分别测叶片光合速率及其它生理指标,结果表明随处理时间的延长,光合速率下降,气孔导度减小,细胞间隙CO2浓度先降低后升高(图1);叶片MDA(图5a)、Cl-、Pi(图4)及可溶性糖(图2b)含量增加;质膜透性增大(图2a);Chla/Chl b(图3b)及Fv/Fm(图5b)减小;MDH、PEPC酶活性降低(图6).细胞间隙CO2浓度的变化,说明光合速率下降的原因短时间内以渗透胁迫产生的气孔限制因素为主,长时间时以非气孔因素为主.Cl-多对细胞产生离子毒害,Pi的增多可竞争性地抑制RuBP羧化酶的活性.MDA的增加表明活性氧增多,活性氧通过使酶失活和膜伤害抑制细胞生长,使细胞内糖利用减少,可溶性糖含量增加,进而反馈性地抑制光合作用.活性氧还可以漂白叶绿素、增强光抑制及使与光合作用有关的酶失活,而抑制光合作用.因此,NaCl胁迫下光合作用降低的原因是多因素共同作用的结果,短时间内以气孔限制因素为主,长时间时以非气孔因素为主,在非气孔因素中活性氧的增加是主导因素.     

Comparative proteomic analysis reveals the positive effect of exogenous spermidine on photosynthesis and salinity tolerance in cucumber seedlings

Key message

Our results based on proteomics data and physiological alterations proposed the putative mechanism of exogenous Spd enhanced salinity tolerance in cucumber seedlings.

Abstract

Current studies showed that exogenous spermidine (Spd) could alleviate harmful effects of salinity. It is important to increase our understanding of the beneficial physiological responses of exogenous Spd treatment, and to determine the molecular responses underlying these responses. Here, we combined a physiological analysis with iTRAQ-based comparative proteomics of cucumber (Cucumis sativus L.) leaves, treated with 0.1 mM exogenous Spd, 75 mM NaCl and/or exogenous Spd. A total of 221 differentially expressed proteins were found and involved in 30 metabolic pathways, such as photosynthesis, carbohydrate metabolism, amino acid metabolism, stress response, signal transduction and antioxidant. Based on functional classification of the differentially expressed proteins and the physiological responses, we found cucumber seedlings treated with Spd under salt stress had higher photosynthesis efficiency, upregulated tetrapyrrole synthesis, stronger ROS scavenging ability and more protein biosynthesis activity than NaCl treatment, suggesting that these pathways may promote salt tolerance under high salinity. This study provided insights into how exogenous Spd protects photosynthesis and enhances salt tolerance in cucumber seedlings.

     

The molecular basis of shoot responses of maize seedlings to Trichoderma harzianum T22 inoculation of the root: a proteomic approach

Trichoderma spp. are effective biocontrol agents for several soil-borne plant pathogens, and some are also known for their abilities to enhance systemic resistance to plant diseases and overall plant growth. Root colonization with Trichoderma harzianum Rifai strain 22 (T22) induces large changes in the proteome of shoots of maize (Zea mays) seedlings, even though T22 is present only on roots. We chose a proteomic approach to analyze those changes and identify pathways and genes that are involved in these processes. We used two-dimensional gel electrophoresis to identify proteins that are differentially expressed in response to colonization of maize plants with T22. Up- or down-regulated spots were subjected to tryptic digestion followed by identification using matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry and nanospray ion-trap tandem mass spectrometry. We identified 91 out of 114 up-regulated and 30 out of 50 down-regulated proteins in the shoots. Classification of these revealed that a large portion of the up-regulated proteins are involved in carbohydrate metabolism and some were photosynthesis or stress related. Increased photosynthesis should have resulted in increased starch accumulation in seedlings and did indeed occur. In addition, numerous proteins induced in response to Trichoderma were those involved in stress and defense responses. Other processes that were up-regulated were amino acid metabolism, cell wall metabolism, and genetic information processing. Conversely, while the proteins involved in the pathways noted above were generally up-regulated, proteins involved in other processes such as secondary metabolism and protein biosynthesis were generally not affected. Up-regulation of carbohydrate metabolism and resistance responses may correspond to the enhanced growth response and induced resistance, respectively, conferred by the Trichoderma inoculation.     

Variations in the natural abundance of oxygen-18 and deuterium in plant carbohydrates

Variations in the natural abundance of ¹⁸O and ²H in plant cellulose are influenced by the isotopic composition of the water directly involved in metabolism—the metabolic water fraction. The isotopic distinction between the metabolic source water and total tissue water must reflect the formation of isotopic gradients within the tissue that are influenced by the rate of water turnover, by properties of the water conducting system and by environmental conditions. It seems that the ¹⁸O abundance in the metabolic water is conserved in cellulose with a relatively constant isotope effect. The relationship of the ²H abundance between metabolic water and cellulose is more complex. Hydrogen incorporated into photosynthetic products during primary reduction steps is highly depleted in ²H. However, a large proportion of these hydrogens are subsequently replaced by exchange with water, leading to ²H enrichment during heterotrophic metabolism. Deciphering the oxygen isotope ratio of cellulose could help in providing insights into the carbon and oxygen fluxes exchanged between plants and the atmosphere. This is because the ¹⁸O abundance in cellulose records the ¹⁸O abundance in the metabolic water, which in turn, controls the oxygen isotopic signatures of the CO₂ and O₂ released by plants into the atmosphere. The hydrogen isotope effects associated with carbohydrate metabolism provide insights into the autotrophic state of a plant tissue. This is because the hydrogen isotope ratio of carbohydrates must reflect the net effects of the two opposing isotope effects associated with photosynthesis and heterotrophic metabolism.     

拟南芥RPP1A参与幼苗生长的蛋白质组学分析

核糖体蛋白不仅参与蛋白质合成,而且参与植物生长发育的调控.利用拟南芥核糖体磷酸蛋白P1(ribosomal phosphoprotein P1,RPP1)家族基因RPP1A缺失突变体rpp1a研究RPP1A缺失对幼苗蛋白质表达水平的影响,揭示其参与调控幼苗生长的作用机制.表型分析发现,与野生型WT相比,RPP1A缺失导...