Germin-Like Polypeptides Increase in Barley Roots during Salt Stress

The 26 kilodalton, isoelectric point 6.3 and 6.5 (Gs1 and Gs2) polypeptides that increase in barley (Hordeum vulgare L.) roots during salt stress were isolated and identified. Both Gs1 and Gs2 had high sequence similarity to germin, a protein that increases significantly in germinating wheat seeds. Like germin, Gs1 and Gs2 were resistant to proteases and were glycosylated. Immunoblots were probed with antibodies to Gs1 and Gs2 to determine the distribution of these polypeptides among organs and cell-free fractions. Gs1 and Gs2 were present in roots and coleoptiles, but absent from leaves. In roots, Gs1 and Gs2 were present in the mature region, but not the tip. Gs1 and Gs2 increased in roots, but decreased in coleoptiles in response to salt stress. Gs1 and Gs2 were distributed among the soluble, microsomal, and cell wall fractions of roots, but the majority of Gs1 and Gs2 was present in the soluble fraction. Although Gs1 and Gs2 were heat stable, their synthesis was not affected by abscisic acid treatment. Gs2 accumulated during abscisic acid treatment, whereas Gs1 did not. However, a 25.5 kilodalton, isoelectric point 6.1 polypeptide that was immunologically related to Gs1 did accumulate with abscisic acid treatment.     

Apoptosis-Like Cell Death in Barley Roots under Salt Stress

Salt stress-induced cell death was investigated in barley roots.Cleavage of nuclear DNA was observed 1 h after salt stress.Oligonucleosomal fragments of DNA were detected electrophoretically8 h after salt stress. These phenomena indicate that apoptosis-likecell death can occur under salt stress. (Received February 12, 1997; Accepted July 3, 1997)     

盐胁迫下大麦根系木质部压力的自调节现象

用植物木质部压力探针测定的结果表明,水培大麦幼苗根的木质部压力在环境条件恒定不变时始终保持波动,并且在受到轻度的盐胁迫和当盐胁迫解除时表现出高度的自调节现象.这种波动和自调节现象将对植物水势的测定和根的径向反射系数的测定产生很大的影响,并可能与植物的抗盐性有关.小麦根在同样条件下未表现出上述现象.     

多胺浸种改善盐胁迫大麦根系液泡膜功能的机理

研究了0.1 mmol/L 腐胺 (Put) 和0.5 mmol/L 亚精胺 (Spd) 浸种对200 mmol/L NaCl胁迫下大麦(Hordeum vulgare L.)幼苗生长速率、干物质积累、离子分布、液泡膜蛋白结合多胺含量以及液泡膜膜脂组分与功能的影响.结果表明,Put和Spd浸种均可缓解盐胁迫对大麦幼苗的盐害,促进生长和干物质积累,降低大麦幼苗体内[Na+]/[K+].与盐处理的对照植株相比,Put和Spd浸种均可提高大麦幼苗根系液泡膜磷脂含量,降低糖脂结合半乳糖含量,而膜上非共价结合多胺含量Spd+PAx (一种未知多胺) 与 Put+Dap (二氨基丙烷)之比((Spd+PAx)/(Put+Dap))、共价和非共价结合多胺总量均上升.统计分析结果表明,液泡膜非共价结合多胺(Spd+PAx)/(Put+Dap)与H+-ATPase和H+-PPase活性呈显著正相关关系.     

多胺浸种改善盐胁迫大麦根系液泡膜功能的机理

研究了 0 .1mmol/L腐胺 (Put)和 0 .5mmol/L亚精胺 (Spd)浸种对 2 0 0mmol/LNaCl胁迫下大麦 (HordeumvulgareL .)幼苗生长速率、干物质积累、离子分布、液泡膜蛋白结合多胺含量以及液泡膜膜脂组分与功能的影响。结果表明 ,Put和Spd浸种均可缓解盐胁迫对大麦幼苗的盐害 ,促进生长和干物质积累 ,降低大麦幼苗体内 [Na ]/[K ]。与盐处理的对照植株相比 ,Put和Spd浸种均可提高大麦幼苗根系液泡膜磷脂含量 ,降低糖脂结合半乳糖含量 ,而膜上非共价结合多胺含量Spd PAx (一种未知多胺 )与Put Dap (二氨基丙烷 )之比 ( (Spd PAx) / (Put Dap) )、共价和非共价结合多胺总量均上升。统计分析结果表明 ,液泡膜非共价结合多胺 (Spd PAx) / (Put Dap)与H _ATPase和H _PPase活性呈显著正相关关系。     

Salt Stress Induced Nuclear and DNA Degradation in Meristematic Cells of Barley Roots

Root growth of barley (Hordeum vulgare L., cv. Akashinriki)was inhibited by 200 raM NaCl, when 1 mM CaCl₂ was present inthe hydroponic culture solution. Increasing the CaCl₂ up to10 mM partially prevented this inhibition. However, inhibitionalso occurred with 100 mM NaCl in the presence of 0.1 mM CaCl₂.The nuclei of meristematic cells in roots in which growth hadbeen inhibited by salt stress were studied after staining withDAPI (4',6-diamino-2-phenylindol). Nuclear deformation of thecells occurred with 12 h of salt stress with 500 mM NaCl, andwas followed by degradation. The nuclear degradation was alsoobserved when the roots were exposed to more than 300 mM NaClfor 24 h. Biochemical analysis revealed that nuclear degradationwas accompanied by apoptosis-like DNA fragmentation. The intracellularmechanisms of nuclear degradation in cells after salt stressare discussed. ¹Emertius professor, Okayama University.     

Effect of Aluminium on Oxidative Stress Related Enzymes Activities in Barley Roots

The impact of aluminium stress on activities of enzymes of the oxidative metabolism: superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD), NADH peroxidase (NADH-POD) and oxalate oxidase (OXO) was studied in barley (Hordeum vulgare L. cv. Alfor) root tips. SOD appeared to be involved in detoxification mechanisms at highly toxic Al doses and after long Al exposure. POD and APX, H₂O₂ consuming enzymes, were activated following similar patterns of expression and exhibiting significant correlation between their elevated activities and root growth inhibition. The signalling role of NADH-POD in oxidative stress seems to be more probable than that of OXO, which might be involved in Al toxicity mechanism. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genome-Wide Identification of the F-box Gene Family and Expression Analysis under Drought and Salt Stress in Barley

The F-box protein-encoding gene family plays an essential role in plant stress resistance. In present study, 126 non-redundant F-box genes were identified in barley (Hordeum vulgare L., Hv). The corresponding proteins contained 165– 887 amino acid residues and all were amphiphilic, except 5 proteins. Phylogenetic analysis of F-box protein sequences in barley and stress-related F-box protein sequences in wheat and Arabidopsis thaliana (At) was used to classify barley F-box genes are divided into 9 subfamilies (A–I). A structure-based sequence alignment demonstrated that F-box proteins were highly conserved with a total of 10 conserved motifs. In total, 124 F-box genes were unevenly distributed on 7 chromosomes; another 2 genes have not been anchored yet. The gene structure analysis revealed high variability in the number of exons and introns in F-box genes. Comprehensive analysis of expression profiles and phylogenetic tree analysis, a total of 12 F-box genes that may be related to stress tolerance in barley were screened. Of the 12 detected F-box genes, 8 and 10 were upregulated after drought and salt stress treatments, respectively, using quantitative real-time polymerase chain reaction (qRT-PCR). This study is the first systematic analysis conducted on the F-box gene family in barley, which is of great importance for clarifying this family’s bioinformatic characteristics and elucidating its function in barley stress resistance. These results will serve as a theoretical reference for subsequent research on molecular regulation mechanisms, genetic breeding, and improvement.     

多胺对盐胁迫下黄瓜SOS2基因家族表达的影响

该研究基于黄瓜基因组数据库,利用生物信息学和荧光定量PCR等方法,对黄瓜SOS2基因家族进行全基因组鉴定,并对其表达特性进行系统分析。结果表明：(1)在黄瓜基因组中,共鉴定到 15个SOS2基因(CsSOS2 1~CsSOS2 15),且不均匀分布在5条染色体上;亚细胞定位显示SOS2蛋白主要位于细胞质膜。(2)系统进化分析显示,CsSOS2 2和CsSOS2 6与AtSOS2亲缘关系更近。(3)顺式作用元件预测显示,SOS2基因启动子序列主要含有干旱诱导和防御应激响应元件。(4)结构分析显示,SOS2蛋白所含保守基序的排列顺序完全一致,且主要含有STKc和NAF保守结构域,这些结构可能在基因响应盐胁迫过程中起调控作用。(5)荧光定量试验表明,SOS2基因家族主要在黄瓜的根和叶片响应盐胁迫时上调表达,其中,盐胁迫处理1 d时有5个基因上调表达,并随处理时间延长盐胁迫下的基因表达有所下调;增施多胺显著上调了CsSOS2 1~CsSOS2 5、CsSOS2 8、CsSOS2 9、CsSOS2 12和CsSOS2 15在不同组织中的表达,说明盐胁迫下多胺能诱导黄瓜SOS2基因家族的表达,进而参与植物耐盐分子网络的调控。     

Effect of Salt Stress on Ion Channel Selective Permeability of Plasmalemma in Sorghum Roots

100 mmol/L, 200 mmol/L and 300 mmol/L NaC1 was used in proper order to treat three-day old seedlings of Sorghum vulgare Pets. The plasma membrane of roots was isolated and purified by aqueous biphasic partition device. The plasmalemma was incorporated into planar bilayer lipid membrane and ion channels were measured by electrical methods. Ion selective permeabilities (PK: PNa) were assayed in asymmetrical solutions containing 100 mmol/L NaC1 in Cis chamber and 100 mmol/L KC1 in Tran with chamber and were calculated from Goldman-Hodgkin-Kaltz equation. PK: PNa was 3.5 in plasmalemma of control roots and 1.5 in plasmalemma of salt stressed roots. It showed that the changes of ion selective permeability was very significant under salt stress. The importance of the change of ion selective permeability is discussed.     

Germin Gene Expression Is Induced in Wheat Leaves by Powdery Mildew Infection

Germin gene expression is induced in wheat (Triticum aestivum L.) leaves by powdery mildew (Erysiphe graminis f. sp. tritici) infection. Germin is a protein marker for early cereal development and is an oxalate oxidase, an enzyme that catalyzes the conversion of oxalate to CO2 and H2O2. The induction of germin gene expression by powdery mildew infection is consistent with the importance of H2O2 to plant defense and identifies a mechanism for the elevation of H2O2 levels in wheat leaves. Germin mRNA levels increased 2 d after inoculation of seedlings with powdery mildew and continued to increase throughout an 8-d time course. The increase in accumulation of germin mRNA was accompanied by an increase in the germin oligomer, which reached maximal levels by d 6. An increase in oxalate oxidase activity paralleled germin oligomer accumulation. Germin gene expression was induced in a relatively resistant cultivar (Bobwhite) as well as in a susceptible cultivar (Cheyenne), suggesting that the induction of germin gene expression is an indicator of powdery mildew infection rather than cultivar resistance.     

Accumulation of Microsomal Polypeptides in Barley Roots during Aluminium Stress

Accumulation of two peripheral membrane polypeptides (20 and 28 kDa) in roots of Al-sensitive (cv. Alfor) and Al-resistant (cv. Bavaria) barley cultivars were analysed during Al stress. Both cultivars were subjected to Al concentration ranging from 0 to 150 µM for 24, 48, 72 and 96 h. Accumulation of both polypeptides was determined 24 h after exposure of plants to Al and content of both polypeptides showed only small depedence upon Al concentration and duration of Al treatment. Although, based on root growth test, Bavaria showed significantly greater resistance to Al than Alfor, analysis of 20 and 28 kDa polypeptide pattern has not revealed significant difference between the two cultivars. However, accumulation of 20 and 28 kDa polypeptides in Alfor was selectively induced by Al treatment because different pH of the root media (pH 3.5 to 6.5) or application of other metals (Cu, Co, or Cd) failed to induce these two bands. On the other hand, accumulation of these polypeptides in Bavaria was induced not only by Al, but also by Cd and in a lesser extent by Co treatment.     

高钙逆境对拟南芥根部基因表达的影响

土壤钙含量过高会对植物正常生长发育形成严重的威胁.这是中国西南喀斯特区域农业生产和环境保护的重要限制因素.了解植物对高钙胁迫的响应机制,可以促进喀斯特地区可持续农业发展和生态环境保护.为揭示植物对高钙胁迫相应的分子机理,本研究充分利用拟南芥(Arabidpsis thaliana)丰富的生物信息资源,系统比较和分析了对照和高钙胁迫处理情况下拟南芥根部的基因表达情况.40 mmol/L CaCl2处理明显抑制了拟南芥根部生长.转录组分析表明,高钙处理下,拟南芥根部乙烯、脱落酸和茉莉酸途径表达上调,其中,茉莉酸合成、响应和代谢途径相关基因表达均上调,说明这3个激素相关途径,对植物应对高钙胁迫具有重要的作用,尤其是茉莉酸相关途径.     

盐胁迫下大麦根系多胺代谢与其耐盐性的关系

研究了0～300mmol/LNaCl对大麦(Hordeum-vulgare-L.)幼苗生长速率、根系游离和结合态多胺含量以及多胺生物合成关键酶活性的影响。结果表明,在0～200mmol/L NaCl处理下精氨酸脱羧酶（ADC）、多胺氧化酶(PAO)以及转谷酰胺酶(Tgase)活性明显提高,而在300 mmol/L NaCl处理下活性下降。与之对应,游离腐胺（Put）含量随处理盐浓度的提高一直呈上升趋势,亚精胺(Spd)和在根系内检测到的未知多胺(Pax)在低浓度盐处理时含量上升,随盐浓度的提高含量下降。盐处理前后精胺(Spm)含量变化不明显。低浓度盐处理时游离态（Spd+Pax）/Put上升,随盐浓度的提高比值明显下降。结合态Put、Spd和Pax含量以及结合态多胺总量均在低浓度盐处理时上升,随盐浓度的提高含量明显下降。统计分析显示,大麦相对生长速率与游离态（Spd+Pax）/Put和结合态多胺含量间均呈极显著正相关关系,与游离态多胺和结合态多胺的比值间均呈显著负相关关系,上述结果说明盐胁迫下大麦体内游离态Spd、Pax与Put以及结合态形式之间的平衡与大麦耐盐性关系密切,游离态Put向Spd 、Pax以及结合态形式转化均有利于大麦耐盐性的提高.