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.     

药物诱导的玉米根尖细胞凋亡

同时应用ＤＮＡ Ｌａｄｄｅｒｉｎｇ、ＤＮＡ Ｇｅｌ Ｂｌｏｔ以及基于染色体涂片 原位末端标记技术,从染色体、细胞核和ＤＮＡ不同水平对细胞毒素类药和的放线菌Ｄ、放线菌酮和秋水仙碱诱导的玉米（Ｚｅａ ｍａｙｓ Ｌ．）根尖分生组织细胞死亡作了检测。结果表明：同动物中一样,这些药物诱导的玉米根尖分生组织细胞死亡也具有ＤＮＡ Ｌａｄｄｅｒ、染色质和细胞核浓缩等典型的调亡特征,说明这些细胞毒素类药物能够诱导植     

Effect of Salt Stress on Germin Gene Expression in Barley Roots

Germin gene expression in barley (Hordeum vulgare L.) seedlings responds to developmental and environmental cues. During seed germination, germin mRNA levels were maximal 2 d after the start of imbibition in control seedlings and declined to low levels by 6 d. When seeds were sown in the presence of 200 mM NaCl, germin mRNA levels were also maximal after 2 d, but NaCl treatment, which slowed seedling growth, prolonged germin gene expression for an additional 1 d. In 4-d-old seedlings, germin mRNA levels were highest in roots and higher in the vascular transition region than in shoots. In roots of 6-d-old seedlings, germin gene expression was regulated by salt shock and plant growth regulators. Induced germin mRNA levels were maximal 8 h after treatment with NaCl, salicylate, methyl salicylate, or methyl jasmonate and 4 h after treatment with abscisic acid and indoleacetic acid. Like germin mRNA, dehydrin mRNA levels were maximal 8 h after NaCl treatment. In contrast, peroxidase mRNA levels declined to less than control levels within 30 min of treatment. Germin gene expression is regulated developmentally by salt stress and by treatments with plant hormones. Since germin is an oxalate oxidase, these result imply that oxalate has important roles in plant development and homeostasis.     

Active H Efflux from Cells of Low-salt Barley Roots during Salt Accumulation

Measurements were made of net H⁺ loss from low-salt barley roots accumulating salt. Comparison of rates of loss from roots in different concentrations of KCl showed that H⁺ loss increased in the same way as the Mechanism II component of salt uptake. This H⁺ loss appeared to be coupled to salt uptake and was not due to increased respiration or metabolic breakdown of sugars. In view of the large negative potential of the cells (−60 millivolts), it is suggested that the H⁺ loss is due to an outward proton transport process.     

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/L NaCl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300mol/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。     

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.     

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。     

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.     

低温胁迫诱导玉米根尖细胞凋亡的形态和生化证据

近来体外培养细胞体系研究的结果表明 ,一定条件的低温胁迫可导致植物细胞凋亡。本文利用DNA梯状图谱 (DNAladdering)以及染色体涂片的TUNEL(terminaldeoxynucleotidyltransferase mediateddUTPnickend labeling)原位检测 ,从原位以及生化水平对低温胁迫下的玉米根尖细胞死亡作了研究 ,形态和生化方面的证据表明 ,同体外一样 ,低温胁迫也能诱导体内植物细胞凋亡。同时对低温胁迫下植物细胞凋亡的形态及生化变化特征和变化次序做了初步观察。实验结果对植物抗低温的生理机制提供了新的研究思路     

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.     

Intracellular pH and Proton-Transport in Barley Root Cells under Salt Stress: in Vivo 31P-NMR Study

Salt stress-induced changes of intracellular pH and in levelsof phosphorous compounds were monitored in intact root tipsof barley seedlings (Hordeum vulgare cv. Akashin-riki) by invivo ³¹P-nuclear magnetic resonance (NMR) spectroscopy. Vacuolaralkalization was observed after treatment with both 300 and500 mM NaCl. Much of the observed apparent alkalization of thecytoplasm was eliminated when the effect of Na⁺ ions on thetitration curve was considered. Within 1 h after the initiationof salt stress, levels of glucose-6-phosphate and UDP-glucosedecreased markedly, and such decreases might lead directly orindirectly to cell death. Simultaneous measurements of the externaland intracellular pH revealed the promotion of external acidificationand internal alkalization during salt stress. Possible mechanismsof Na⁺/H⁺ antiport at the tonoplast and the role of proton-pumpin the plasma membrane are discussed. ³Present address: Shijonawate Gakuen Women's Junior College,Daito, Osaka, 574 Japan.     

SO2对蚕豆根尖和叶尖细胞遗传损伤作用的研究

采用微核实验技术,研究大气污染物SO₂对蚕豆根尖和叶尖细胞的遗传损伤效应。结果表明2.80和28 mg·m^-3的SO₂熏气可以诱发蚕豆根尖和叶尖细胞损伤,导致根尖细胞有丝分裂指数下降,根尖和叶尖间期细胞微核率增高,并具有时间效应和浓度效应。经水恢复培养后,根尖分裂细胞数增多,微核率降低,说明恢复培养能够缓解高浓度SO₂对根尖细胞的遗传损伤。用石蜡层隔断SO₂在根部水中的溶解后,根尖细胞微核率低于叶尖细胞微核率,而在非隔断组中则相反,说明SO₂在水中的溶解是产生毒性效应的重要原因。高浓度SO₂熏气对蚕豆根尖和叶尖细胞具有遗传学毒性,由于根尖分生区具有较高的分裂指数和微核率,对环境SO₂毒性的反应更灵敏,蚕豆根尖微核实验更适于对环境SO₂的监测。     

F-actin is a Nuclear and Chromosomal Component of the Meristematic Cells of Allium sativum

It is known that actin functionates in the form of F-actin. However, the presence of Factin in eukaryotic nuclei and chromosomes has not been well established. The authors labeled meristematic cells of Allium sativum L. with rabbit anti-chicken actin antibody and FITC-conjugated goat anti-rabbit IgG antibody and observed with fluorescence microscopy. Both the nuclei and chromosomes showed prominent yellow-green fluorescence, indicating the presence of actin in them. Fluorescence examination with TR1TC-conjugated phalloidin demonstrated prominent red fluorescence in the intact interphase cells, cytoplasm-free interphase nuclei, prophase and metaphase chromosomes as well as the daughter nuclei at telophase indicating the presence of F-actin; but the fluorescence was absent or very weak in the cells exposed to cytochalasin D before fixation. When double labeling of the anti-actin antibody and phalloidin was applied, the same nuclei and chromosomes were found to emanate yellow-green fluorescence representing actin at the excitation wavelength of F1TC, and red fluorescence representing F-actin at the excitation wavelength of TRITC, respectively. The FITC fluorescence and TRITC fluorescence shared the same distribution among the nuclei and chromosomes. These results indicate that F-actin is a component of the nuclei and chromosomes of the meristematic cells of A. sativum. It also suggests that F-actin may be the major existing form of actin in them.     

SO2对蚕豆根尖和叶尖细胞遗传损伤作用的研究

采用微核实验技术,研究大气污染物SO2对蚕豆根尖和叶尖细胞的遗传损伤效应。结果表明2.80和28 mg.m-3的SO2熏气可以诱发蚕豆根尖和叶尖细胞损伤,导致根尖细胞有丝分裂指数下降,根尖和叶尖间期细胞微核率增高,并具有时间效应和浓度效应。经水恢复培养后,根尖分裂细胞数增多,微核率降低,说明恢复培养能够缓解高浓度SO2对根尖细胞的遗传损伤。用石蜡层隔断SO2在根部水中的溶解后,根尖细胞微核率低于叶尖细胞微核率,而在非隔断组中则相反,说明SO2在水中的溶解是产生毒性效应的重要原因。高浓度SO2熏气对蚕豆根尖和叶尖细胞具有遗传学毒性,由于根尖分生区具有较高的分裂指数和微核率,对环境SO2毒性的反应更灵敏,蚕豆根尖微核实验更适于对环境SO2的监测。