Ospdr9, which encodes a PDR-type ABC transporter, is induced by heavy metals, hypoxic stress and redox perturbations in rice roots

Little is known about the role of pleiotropic drug resistance (PDR)-type ATP-binding (ABC) proteins in plant responses to environmental stresses. We characterised ospdr9, which encodes a rice ABC protein with a reverse (ABC-TMS(6))(2) configuration. Polyethylene glycol and the heavy metals Cd (20 microM) and Zn (30 microM) rapidly and markedly induced ospdr9 in roots of rice seedlings. Hypoxic stress also induced ospdr9 in rice roots, salt stress induced ospdr9 at low levels but cold and heat shock had no effect. The plant growth regulator jasmonic acid, the auxin alpha-naphthalene acetic acid and the cytokinin 6-benzylaminopurine triggered ospdr9 expression. The antioxidants dithiothreitol and ascorbic acid rapidly and markedly induced ospdr9 in rice roots; the strong oxidant hydrogen peroxide also induced ospdr9 but at three times lower levels. The results suggested that redox changes may be involved in the abiotic stress response regulation of ospdr9 in rice roots.     

Upland rice and lowland rice exhibited different PIP expression under water deficit and ABA treatment

Aquaporins play a significant role in plant water relations. To further understand the aquaporin function in plants under water stress, the expression of a subgroup of aquaporins, plasma membrane intrinsic proteins （PIPs）, was studied at both the protein and mRNA level in upland rice （Oryza sativa L. cv. Zhonghan 3） and lowland rice （Oryza sativa L. cv. Xiushui 63） when they were water stressed by treatment with 20% polyethylene glycol （PEG）. Plants responded differently to 20% PEG treatment. Leaf water content of upland rice leaves was reduced rapidly. PIP protein level increased markedly in roots of both types, but only in leaves of upland rice after 10 h of PEG treatment. At the mRNA level, OsPIP1,2, OsPIP1,3, OsPIP2;1 and OsPIP2;5 in roots as well as OsPIP1,2 and OsPIP1;3 in leaves were significantly up-regulated in upland rice, whereas the corresponding genes remained unchanged or down-regulated in lowland rice. Meanwhile, we observed a significant increase in the endogenous abscisic acid （ABA） level in upland rice but not in lowland rice under water deficit. Treatment with 60 μM ABA enhanced the expression of OsPIP1;2, OsPIP2;5 and OsPIP2;6 in roots and OsPIP1;2, OsPIP2;4 and OsPIP2;6 in leaves of upland rice. The responsiveness of PIP genes to water stress and ABA were different, implying that the regulation of PIP genes involves both ABA-dependent and ABA-independent signaling oathways during water deficit.     

Ammonium Ion, Ethylene, and Abscisic Acid in Polyethylene Glycol-treated Rice Leaves

Polyethylene glycol (PEG)-treatment decreased chlorophyll and protein contents and increased NH₄ ⁺ content due to decreased glutamine synthetase activity in detached rice leaves. PEG-treatment also increased abscisic acid (ABA) content and decreased ethylene production. Addition of fluridone, an inhibitor of ABA biosynthesis, reduced ABA content in rice leaves but did not prevent chlorophyll and protein loss in rice leaves induced by PEG. Silver thiosulfate, an inhibitor of ethylene action, was effective in preventing PEG-promoted chlorophyll and protein loss, but had no effect on PEG-induced NH₄ ⁺ accumulation. The current results suggest that NH₄ ⁺ accumulation in rice leaves induced by PEG increases leaf sensitivity to ethylene, which in turn results in an enhancement of chlorophyll and protein loss. This revised version was published online in July 2006 with corrections to the Cover Date.     

Activation of abscisic acid biosynthesis in the leaves of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> in response to water deficit

It is well known that endogenous abscisic acid (ABA) levels increase rapidly in response to drought stress and that this induces stomatal closure. In Arabidopsis thaliana, ABA levels increased rapidly in the leaves and roots when intact wild-type whole plants were exposed to drought stress. However, if the leaves and roots were separated and exposed to drought independently, the ABA level increased only in the leaves. These results suggest that, under our experimental conditions, ABA is synthesized mainly in the leaves in response to drought stress and that some of the ABA accumulated in the leaves is transported to the roots. Tracer experiments using isotope-labeled ABA indicate that the movement of ABA from leaves to roots is activated by water deficit in the roots. We also demonstrate that the endogenous ABA level in the leaves increased only when the leaves themselves were exposed to drought stress, suggesting that leaves play a major role in the production of ABA in response to acute water shortage.     

Gene expression changes in response to drought stress in <Emphasis Type="Italic">Citrullus colocynthis</Emphasis>

Citrullus colocynthis (L.) Schrad, closely related to watermelon, is a member of the Cucurbitaceae family. This plant is a drought-tolerant species with a deep root system, widely distributed in the Sahara-Arabian deserts in Africa and the Mediterranean region. cDNA amplified fragment length polymorphism (cDNA-AFLP) was used to study differential gene expression in roots of seedlings in response to a 20% polyethylene glycol-(PEG8000) induced drought stress treatment. Eighteen genes which show similarity to known function genes were confirmed by quantitative relative (RQ) real-time RT-PCR to be differentially regulated. These genes are involved in various abiotic and biotic stress and developmental responses. Dynamic changes with tissue-specific pattern were detected between 0 and 48 h of PEG treatment. In general, the highest induction levels in roots occurred earlier than in shoots, because the highest expression was detected in roots following 4 and 12 h, in shoots following 12 and 48 h of drought. These drought-responsive genes were also affected by the plant hormones abscisic acid (ABA), salicylic acid (SA), or jasmonic acid (JA), indicating an extensive cross-talk between drought and plant hormones. Collectively, these results will be useful to explore the functions of these multiple signal-inducible genes for unveiling the relationship and crosstalk between different signaling pathways.     

O-acetylserine (thiol) lyase activity in Phragmites and Typha plants under cadmium and NaCl stress conditions and the involvement of ABA in the stress response

The roles of O-acetylserine (thiol) lyase (OASTL, EC 4.2.99.8) and abscisic (ABA) acid in stress responses to NaCl and cadmium treatments were investigated in Typha latifolia L. and Phragmites australis (Cav.) Trin. ex Steudel plants. OASTL activity increased under stress (25-300 microM Cd, 100mM NaCl, 1 microM ABA) in both Typha and Phragmites mainly in roots, contributing substantially to satisfy the higher demand of cysteine for adaptation and protection. The earliest significant responses in intact roots were recorded after 12-24 h of Cd treatments, but different levels of stimulation were also observed after 3 and 7 days of exposure. The OASTL activity responses of Phragmites to salinity (100mM NaCl) were higher than those of Typha. Cysteine synthesis in Typha is much higher than in Phragmites, which supports the efficiency of the thiol-metabolism-based protection shown in Typha. Exogenous ABA increased OASTL activity in both species. Cd treatments led to increased ABA levels in roots. Phragmites showed higher ABA levels compared to Typha. The increase of ABA content indicates the involvement of this phytohormone in early stress responses, while the stimulation of OASTL following the ABA application suggests that ABA has a role in an OASTL activation pathway.     

Abscisic acid root and leaf concentration in relation to biomass partitioning in salinized tomato plants

Salinization is one of the most important causes of crop productivity reduction in many areas of the world. Mechanisms that control leaf growth and shoot development under the osmotic phase of salinity are still obscure, and opinions differ regarding the Abscisic acid (ABA) role in regulation of biomass allocation under salt stress. ABA concentration in roots and leaves was analyzed in a genotype of processing tomato under two increasing levels of salinity stress for five weeks: 100 mM NaCl (S10) and 150 mM NaCl (S15), to study the effect of ABA changes on leaf gas exchange and dry matter partitioning of this crop under salinity conditions. In S15, salinization decreased dry matter by 78% and induced significant increases of Na⁺ and Cl⁻ in both leaves and roots. Dry matter allocated in different parts of plant was significantly different in salt-stressed treatments, as salinization increased root/shoot ratio 2-fold in S15 and 3-fold in S15 compared to the control. Total leaf water potential (Ψ_w) decreased from an average value of approximately −1.0 MPa, measured on control plants and S10, to −1.17 MPa in S15. In S15, photosynthesis was reduced by 23% and stomatal conductance decreased by 61%. Moreover, salinity induced ABA accumulation both in tomato leaves and roots of the more stressed treatment (S15), where ABA level was higher in roots than in leaves (550 and 312 ng g⁻¹ fresh weight, respectively). Our results suggest that the dynamics of ABA and ion accumulation in tomato leaves significantly affected both growth and gas exchange-related parameters in tomato. In particular, ABA appeared to be involved in the tomato salinity response and could play an important role in dry matter partitioning between roots and shoots of tomato plants subjected to salt stress.     

水分胁迫下山黧豆中ABA及ODAP的积累研究

用PEG、PEG+ABA、ABA分别处理15d龄的山黧豆幼苗,取其叶片为实验材料,测定内源ABA、ODAP、MDA和H₂O₂含量以及几种抗氧化酶活性,结果表明,与对照相比处理材料叶片中ABA和ODAP含量显著增加;外源ABA的加入降低了PEG胁迫引起的MDA和H₂O₂含量的增加,延缓了PEG胁迫引起的CAT活性的衰减,提高了GR活性.用外源ABA长时间处理山黧豆,发现叶片中ABA含量显著增加,随后出现ODAP的积累;ABA处理初期(0～3d)对叶片中活性氧代谢影响不大,随着ABA处理时间的延长(7～15d),可引起叶片中SOD、POD、CAT、GR活性的降低,MDA、H₂O₂含量的增加,表明ABA确实可促进ODAP的积累.     

The effect of drought on levels of abscisic acid,cytokinins, gibberellins and ethylene in aeroponically-grown sunflower plants

Abscisic acid (ABA), cytokinins and gibberellin-like substances (GAs) were extracted from the roots and shoots of 17-day-old sunflower seedlings which had been droughted or were unstressed. Plants were grown in an aeroponic chamber which allowed for good control over degree of water stress and easy access to roots. Following methanolic extraction of lyophilized material, cytokinins were separated from the acidic growth-regulators on a cellulose PO₄ cationic exchange column. The cytokinins were analysed by paper chromatography and HPLC and the soybean hypocotyl section assay. Semipurified acidic regulators were chromatographed on SiO₂ columns and HPLC and aliquots assayed with the dwarf rice cv. Tan-ginbozu bioassay for GAs. Fractions known to contain ABA were purified by sequential reverse-phase HPLC of the acid and then of the methyl ester forms followed by quantitation as Me-ABA on GLC-EC. ABA losses were measured by using an internal standard [³H]-ABA). Ethylene production was also monitored in stressed and unstressed seedlings.The effect of drought on GAs and ethylene was minimal. The ABA levels were markedly higher in droughted plants. Stressed roots had 32 times more ABA than controls. The levels of cytokinins in the shoots of droughted plants were about half those in unstressed shoots, and qualitative differences occurred in the roots. Under stress a large peak of activity was present similar to zeatin glucoside which was not present in the unstressed condition. The results are discussed in relation to drought-effects on metabolism.     

PEG and ABA trigger methyl jasmonate accumulation to induce the MEP pathway and increase tanshinone production in Salvia miltiorrhiza hairy roots

Tanshinones, a group of active ingredients in Salvia miltiorrhiza, are derived from at least two biosynthetic pathways, which are the mevalonate (MVA) pathway in the cytosol and the 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway in the plastids. Abscisic acid (ABA) and methyl jasmonate (MJ) are two well-known plant hormones induced by water stress. In this study, effects of polyethylene glycol (PEG), ABA and MJ on tanshinone production in S. miltiorrhiza hairy roots were investigated, and the role of MJ in PEG- and ABA-induced tanshinone production was further elucidated. The results showed that tanshinone production was significantly enhanced by treatments with PEG, ABA and MJ. The mRNA levels of 3-hydroxy-3-methylglutaryl co-enzyme A reductase (HMGR), 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) and 1-deoxy-d-xylulose 5-phosphate synthase (DXS), as well as the enzyme activities of HMGR and DXS were stimulated by all three treatments. PEG and ABA triggered MJ accumulation. Effects of PEG and ABA on tanshinone production were completely abolished by the ABA biosynthesis inhibitor [tungstate (TUN)] and the MJ biosynthesis inhibitor [ibuprofen (IBU)], while effects of MJ were almost unaffected by TUN. In addition, MJ-induced tanshinone production was completely abolished by the MEP pathway inhibitor [fosmidomycin (FOS)], but was just partially arrested by the MVA pathway inhibitor [mevinolin (MEV)]. In conclusion, a signal transduction model was proposed that exogenous applications of PEG and ABA triggered endogenous MJ accumulation by activating ABA signaling pathway to stimulate tanshinone production, while exogenous MJ could directly induce tanshinone production mainly via the MEP pathway in S. miltiorrhiza hairy roots.     

水稻H3.2型组蛋白基因RH3.2A的克隆与盐胁迫下的表达分析

组蛋白H3与其他类型的组蛋白分子H2A,H2B,H4共同构成了真核生物核小体的八聚体核心。研究发现组蛋白H3的多种翻译修饰,如甲基化、乙酰化、磷酸化等在调控基因转录过程种发挥了重要的作用。本研究从盐胁迫处理的水稻幼苗组织中分离了一个新的水稻组蛋白H3基因RH3．2A,编码具有136个氨基酸残基的多肽,与多种植物的组蛋白H3蛋白具有高度的氨基酸一致性。多序列比较发现,除了基因结构差异之外,还有3个位置的氨基酸残基（32、88、91）在H3．1与H3．2型组蛋白H3中存在差异。研究了RH3．2A基因在高盐和ABA胁迫下的表达,结果发现在水稻根部RH3．2A基因受高盐的强烈诱导,而在叶片RH3．2A基因的表达则不受高盐诱导,此外RH3．2A基因也受外源ABA的诱导,结合启动子分析的结果,我们认为RH3．2A基因可能参与了依赖于ABA的高盐胁迫应答反应。文章讨论了植物组蛋白H3基因在高盐胁迫应答反应中可能的作用。     

Inducible Expression of Translation Elongation Factor 1A Gene in Rice Seedlings in Response to Environmental Stresses

Differences of gene expression between salinity-stressed and control rice ( Oryza sativa L. ssp. indica) cultivar "Zhaiyeqing 8" were compared using differential display PCR (DD-PCR) technique. Sequence analysis of one salt-inducible cDNA clone revealed that this clone represented a new member of rice translation elongation factor lA (eEF1A) gene family and was tentatively named REF1A. Northern blot hybridization using REF1A fragment as a probe was performed to investigate the expression of rice translation elongation factor lA gene in response to various environmental factors. It was observed that expression of the eEF1A gene in rice shoots was dramatically induced by salinity stress or exogenous application of abscisic acid (ABA). The induction of this gene by ABA stress occurred more quickly than that by salinity stress. In addition, expression of rice translation elongation factor lA gene was also induced by drought (15% PEG6000), cold (4 ℃ ) or heat-shock (37 ℃ ) stresses. The results suggested that the induction of translation elongation factor lA gene expression by environmental stresses might reflect the general adaptive response of rice plants to the adverse circumstances.     

5个毛果杨PtrZFP基因的鉴定和表达分析

ZFP转录因子是植物中的一类具有指环结构域的转录因子。从毛果杨中鉴定出5条ZFP基因（命名为PtrZFP1-5）,对其特性和表达模式进行了分析,以期初步了解这些基因是否能对胁迫做出应答。对PtrZFP1-5基因进行生物学分析,进一步利用qRT-PCR技术分析NaCl、PEG₆₀₀₀和ABA胁迫处理后毛果杨根、茎和叶中5条基因的表达情况。PtrZFP1-5基因编码蛋白氨基酸残基数为258~338 aa,编码蛋白的分子量为27.7~37.3 kDa,理论等电点为4.87~8.61,5个基因不均等的分布在毛果杨基因组的3条染色体上。qRT-PCR结果显示,0.2 mol·L^-1 NaCl、15%（w/v）PEG6000和100 μmol·L^-1 ABA胁迫处理后,5个PtrZFP基因在毛果杨根、茎和叶中的表达模式明显不同。PtrZFP1基因在3种胁迫后毛果杨中均被明显的上调表达;PtrZFP2基因在盐、渗透和ABA胁迫处理后,叶中的表达都明显被抑制;PtrZFP3基因受到干旱胁迫时在根中的响应最为明显;而叶和茎中,表达量在大部分胁迫的大部分时间点无明显改变。PtrZFP4基因也能在根和茎中对干旱胁迫做出明显应答。PtrZFP5基因在经受盐和ABA胁迫后,在叶中的表达受到明显抑制。PtrZFP1-5这5个基因至少能在一种器官中对一种胁迫处理做出应答,但参与的胁迫应答类型和机制可能不同。     

Expression of ASCORBATE PEROXIDASE 8 in roots of rice (Oryza sativa L.) seedlings in response to NaCl

Reactive oxygen species are thought to play an important role in NaCl stress. Therefore, the expression patterns of the gene family encoding the H(2)O(2)-scavenging enzyme ascorbate peroxidase (APx; EC1.11.1.11) were analysed in roots of etiolated rice (Oryza sativa L.) seedlings in response to NaCl stress. Applying semi-quantitative RT-PCR, the mRNA levels were quantified for two cytosolic (OsAPx1 and OsAPx2), two peroxisomal (OsAPx3 and OsAPx4), and four chloroplastic (OsAPx5, OsAPx6, OsAPx7, and OsAPx8) isoforms identified in the rice genome. NaCl at 150 mM and 200 mM increased the expression of OsAPx8 and the activities of APx, but had no effect on the expression of OsAPx1, OsAPx2, OsAPx3, OsAPx4, OsAPx5, OsAPx6, and OsAPx7 in rice roots. However, NaCl at 300 mM up-regulated OsAPx8 expression, increased APx activity, and down-regulated OsAPx7 expression, but had no effect on the expression of OsAPx1, OsAPx2, OsAPx3, OsAPx4, OsAPx5, and OsAPx6. The accumulation of abscisic acid (ABA) in response to NaCl was observed in rice roots. Exogenously applied ABA also specifically enhanced the expression of OsAPx8 in rice roots. The accumulation of ABA in rice roots in response to NaCl was inhibited by fluridone (Flu), an inhibitor of carotenoid biosynthesis. Flu treatment also suppressed NaCl-enhanced OsAPx8 expression and APx activity. The effect of Flu on the expression of OsAPx8 and increase in APx activity was reversed by the application of ABA. It appears that NaCl-enhanced expression of OsAPx8 in rice roots is mediated through an accumulation of ABA. Evidence is provided to show that Na(+) but not Cl(-) is required for enhancing OsAPx8 expression, APx activity, and ABA accumulation in rice roots treated with NaCl. H(2)O(2) treatment resulted in an enhancement of OsAPx8 induction but no accumulation of ABA. Diphenylene iodonium treatment, which is known to inhibit NaCl-induced accumulation of H(2)O(2) in rice roots, did not suppress OsAPx8 induction and ABA accumulation by NaCl. It appears that H(2)O(2) is not involved in the regulation of NaCl-induced OsAPx8 expression in rice roots.