Distinct signalling pathways for induction of MAP kinase activities by cadmium and copper in rice roots

Plant growth is severely affected by toxic concentrations of heavy metals. On characterizing the heavy metal-induced signalling pathways, the effects of cadmium (CdCl2) and copper (CuCl2) on MBP (myelin basic protein) kinase activities in Oryza sativa L. cv. TNG67 were analysed and it was found that Cd2+-induced 42 kDa MBP kinase has the characteristics of a mitogen-activated protein (MAP) kinase. This study confirmed that the 42 kDa kinase-active band contains, at least, the activities of OsMPK3 and OsMPK6. Then, the heavy metal signal transduction pathways leading to MAP kinase activation in rice roots were examined. Pretreatment with sodium benzoate, a hydroxyl radical scavenger, attenuated Cd2+- or Cu2+-induced MAP kinase activation. The Cd2+-, but not Cu2+-, induced MAP kinase activities were suppressed by diphenylene iodonium (DPI), an NADPH oxidase inhibitor, and Cd2+ induced NADPH oxidase-like activities, suggesting that NADPH oxidases may be involved in Cd2+-induced MAP kinase activation. Using a Ca2+ indicator, it was demonstrated that Cd2+ and Cu2+ induce Ca2+ accumulation in rice roots. The Cd2+- and Cu2+-induced MAP kinase activation required the involvement of Ca2+-dependent protein kinase (CDPK) and phosphatidylinositol 3-kinase (PI3 kinase) as shown by the inhibitory effect of a CDPK antagonist, W7, and a PI3 kinase inhibitor, wortmannin, respectively. Furthermore, bongkrekic acid (BK), a mitochondrial permeability transition pore opening blocker, suppressed Cd2+-, but not Cu2+-, induced MAP kinase activation, indicating that Cd2+-induced MAP kinase activities are dependent on the functional state of mitochondria. Collectively, these findings imply that Cd2+ and Cu2+ may induce MAP kinase activation through distinct signalling pathways. Moreover, it was found that the 42 kDa MAP kinase activities are higher in Cd-tolerant cultivars than in Cd-sensitive cultivars. Therefore, the Cd-induced 42 kDa MAP kinase activation may confer Cd tolerance in rice plants.     

缺铁水稻根转录本微点阵分析

水稻不仅是非常重要的粮食作物 ,也是用于研究的模式植物之一 .由于水稻基因组测序的完成 ,用功能基因组学的现代方法来研究缺铁相关基因的表达调控是最高效的方法之一。在前期工作的基础上 ,精心设计了缺铁和EDTA鳌合二价铁诱导5天的水稻根实验 ,并进行了转录水平的微点阵 (microarray)分析。但只获得了第 5天的结果。在 10 5 31个水稻cDNA芯片图谱中 ,缺铁和加铁比较发现了 4 5 1个差异点。对缺铁诱导的 4 5 1个差异cDNA逐一地进行NCBI (美国国家生物技术信息中心 )的BLAST(局部定位排列搜索工具 )数据库检索、分析和归类。发现其中缺铁与加铁 ( -Fe/Fe -EDTAratio)之间的相对表达水平(REL)在 2 - 9.175之间的缺铁诱导上调基因为 2 0 3个 ,缺铁诱导的下调基因为 2 4 8个。对每一类上调基因都逐一地进行了NCBI-PubMed的文献检索。利用国际网络数据库进行了功能鉴定。     

水稻根系响应镉胁迫的蛋白质差异表达

为探讨水稻根系对镉胁迫的分子生理响应,以抗镉水稻PI312777和镉敏感水稻IR24为材料,设置Cd~(2+)浓度为0、50和100μmol/L的水培试验,处理7 d后分析了水稻根系的蛋白质差异表达。结果表明,在镉胁迫下水稻PI312777和IR24根系有18个蛋白质发生了差异表达,其中的12个得到MALDI-TOF/MS鉴定。这些鉴定的蛋白功能可分四类:(1)与活性氧(ROS)胁迫相关的过氧化物酶(POD)、蛋氨酸腺苷转移酶(MAT)、类萌发素蛋白前体;(2)与谷胱甘肽(GSH)合成相关的S-腺苷甲硫氨酸合成酶(SAMS)、谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH);(3)与逆境胁迫相关的ABA胁迫诱导蛋白含HVA22域蛋白、ABA-胁迫-成熟诱导蛋白5(ASR5);(4)与细胞分裂调控相关的GTP结合核蛋白Ran-2。镉胁迫下SAMS和GTP结合核蛋白Ran-2在两种水稻根系均发生上调表达;MAT、POD、类萌发素蛋白前体和GS发生下调表达;依赖NADP-GDH、GDH和磷酸甘油酸变位酶在IR24根部均发生下调表达,在PI312777根部仅在100μmol/L Cd~(2+)处理发生下调表达;含HVA22域蛋白在PI312777根部上调表达,在IR24根部发生下调表达;ASR5在PI312777根部上调表达,在IR24根部的表达无显著差异;100μmol/L Cd~(2+)胁迫下60S酸性核糖体蛋白P0在水稻PI312777根部表达下调,在IR24根部表达上调。可见,镉胁迫使水稻根部ROS增加,形成氧化胁迫反应,造成毒害作用,而水稻根通过调节SAMS和GS提高GSH合成降低镉毒害。ASR5和HVA22蛋白等逆境胁迫蛋白的表达差异则是水稻品种间抗性差异的重要原因之一。     

Association mapping of cadmium, copper and hydrogen peroxide tolerance of roots and translocation capacities of cadmium and copper in Arabidopsis thaliana

Association mapping analysis of Cd, Cu and H ₂O ₂ tolerance, judged by relative root length (RRL: % of root length in stress condition relative to that in control condition), and Cd and Cu translocation ratios (amount of metal in the shoot to the total) were performed using 90 accessions of Arabidopsis thaliana . Using 140 SNPs that were distributed across the genome, association mapping analysis was performed with a haploid setting by the Q + K method, which minimizes detection of false associations by combining the Q-matrix of the structured association (Q) with kinship (K) to control for the population structure. Six, five and five significant (−log ₁₀ P -value is 1.3 ≥) linkages were detected between the SNPs and Cd, Cu and H₂O₂ resistant RRLs, respectively. In addition, six significant linkages were identified with translocation capacities of Cd and Cu. Among those detected loci, two each of Cu and Cd tolerance RRLs were collocated with those of H₂O₂ tolerance RRL, while one locus each was detected by Cu and Cd tolerance RRLs that collocated with their translocation ratios. These results suggested that these factors might partly explain the phenotypic variation of tolerance RRLs to Cd and Cu of Arabidopsis thaliana . Finally, using a different approach to analyze interactions between individual phenotypes, namely clustering analysis, we found an expected segregation of resistant SNPs (single-nucleotide polymorphisms) of the multiple RRLs in the typical accession groups carrying multiple traits. Almost none of the loci detected by association mapping analysis were linked to the loci of previously identified critical genes regulating the traits, suggesting that this could be useful to identify complex architecture of genetic factors determining variation among multiple accessions.     

Cardiac and ventilatory responses ofCrangon crangon to cadmium,copper and zinc

The acute (30 min) responses of heart and scaphognathite activities ofCrangon crangon on exposure to concentrations of 1–20 mg Cd, Cu or Zn 1⁻¹ are increased beat frequencies. The relative magnitude of response(Δ f) is linearly related to immediate pretreatment frequency(f) and standardised responses(Δ fs) are given forf values of 70 and 100 beats min⁻¹ for hearts and scaphognathites, respectively.Δ fs values for each organ are also linearly related to test concentration for each metal. Qualitative changes to organ activities described include an increased incidence of scaphognathite reversals in concentrations of 5.0 mg Cu 1⁻¹ and in 20.0 mg Zn 1⁻¹. Chronic (13 days) exposure to incipient lethal levels of the test metals produced increases in scaphognathite rates of the Cd-treated animals and in heart and scaphognathite rates of the coppertreated animals. The general applicability of these methods to studies of pollution stress in decapods is discussed.     

Comparison of antioxidant responses to cadmium and lead in Bruguiera gymnorrhiza seedlings

Seedlings of mangrove plant Bruguiera gymnorrhiza cultured in sand with Hoagland’s nutrient solution were treated with 1 to 30 mM Cd(NO₃)₂ or Pb(NO₃)₂ for 2 months. In all Cd/Pb treatments, the malondialdehyde content increased while the chlorophyll content declined. Peroxidase (POD) and superoxide dismutase (SOD) activities in roots increased at moderate Cd/Pb concentrations (1–10 mM), whereas decreased at higher concentrations (20–30 mM). Catalase (CAT) activity in roots was inhibited by 1–10 mM Cd but enhanced by 1–10 mM Pb. The activities of POD, SOD and CAT in leaves were less affected by Cd and Pb than in roots. A new SOD and three CAT isoenzymes were induced by Pb. In contrast, no additional SOD and CAT isoenzymes were induced by Cd.     

Short term signaling responses in roots of young soybean seedlings exposed to cadmium stress

In the present study, the expression of fourteen genes involved in various signal transduction pathways was examined in young soybean (Glycine max) seedlings exposed to cadmium at two concentrations (10 mg L⁻¹ and 25 mg L⁻¹) for short time periods (3, 6 and 24 h). The results show that cadmium causes induction of genes encoding proteins involved in ethylene and polyamines metabolism, nitric oxide generation, MAPK cascades and regulation of other genes’ expression. The bioinformatic analysis of promoter sequences of Cd-inducible genes revealed that their promoters possess several regulative motifs associated with the plant response to stress factors and abscisic acid and ethylene signaling. The involvement of ethylene in the response of soybean seedlings to cadmium stress was further confirmed by the real-time analysis of ethylene production during 24 h of CdCl₂ treatment. The role of the described signaling elements in transduction of the cadmium signal in young soybean seedlings is discussed.     

不同耐性水稻幼苗根系对镉胁迫的形态及生理响应

采用水培试验,以两个耐镉性不同的水稻品种为材料,研究了不同浓度镉胁迫对水稻幼苗根系形态、根系活力、游离脯氨酸含量及抗氧化酶活性的影响。结果表明:低于5 μmol/L Cd胁迫对2个水稻品种总根长、根表面积、根体积、根干重、根系活力无明显影响,在1 μmol/L Cd时,甚至起促进作用。随Cd浓度增加表现出一定的抑制效应,秀水63在10 μmol/L Cd胁迫下根系形态、根系活力明显受到抑制,而秀水09在25 μmol/L Cd胁迫下明显受抑。随Cd胁迫浓度的增加,游离脯氨酸含量、丙二醛(MDA)含量、超氧化物歧化酶(SOD)、过氧化物酶(POD)活性均呈上升趋势,两品种相比,秀水09的游离脯氨酸含量、SOD和POD增幅大于秀水63,而MDA含量增幅小于秀水63;过氧化氢酶(CAT)活性变化表现为先上升后下降,10-100 μmol/L Cd胁迫下秀水63根系中CAT活性明显低于秀水09。总之,水稻对Cd毒害响应存在明显的品种差异,且Cd胁迫下根系生理响应的差异是品种间耐性差异的重要原因之一。     

热带地区不同水稻品种对土壤镉胁迫的生理生化响应

以热带地区主栽品种博优225和矮糯为材料,采用盆栽试验研究了不同水稻品种对土壤外源重金属Cd胁迫的耐性机理。结果表明:在各浓度镉胁迫下,2个品种水稻都是根累积的镉含量要高于茎叶和稻壳、糙米,即根>茎叶>稻壳>糙米;随着镉胁迫浓度的增加,细胞膜受损的伤害程度越大,矮糯质膜透性略高于博优225;随着Cd浓度的升高,水稻叶片丙二醛(MDA)、脯氨酸含量逐渐增加,且博优225的增加幅度明显大于矮糯,超氧化物歧化酶(SOD)和过氧化物酶(POD)活性均呈上升趋势,且博优225明显高于矮糯;过氧化氢酶(CAT)活性呈下降趋势;表明同样条件下博优225受Cd毒害更严重。