Effect of Oxygen on the Rate of Photorespiration in Detached Tobacco Leaves

The rate of carbon dioxide exchange in both light and darkness by detached tobacco leaves placed at various oxygen concentrations was measured by an Infra-Red CO₂ Analyzer and a Clark oxygen electrode. It was observed that during illumination oxygen had two different effects. One was to stimulate carbon dioxide evolution and the other to inhibit carbon dioxide absorption. Concentration of carbon dioxide at compensation point was found to be a linear function of oxygen concentration and this has been explained as due mainly to an increased evolution of carbon dioxide. Such an evolution during illumination has been called photorespiration. Increased concentrations of oxygen also had a stimulating effect on the magnitude of the initial post-illumination burst of carbon dioxide in darkness, but no effect on the subsequent steady rates. These data have been explained as due to the suspension of regular respiration in darkness and its replacement by a different process, tentatively called photorespiration. A second effect of oxygen was to reduce the efficiency (called “carboxylation efficiency”) with which a leaf was able to remove carbon dioxide from the atmosphere.     

Effects of Inorganic Nitrogen Sources and Physical Factors on the Formation of Ubiquinone by Tobacco Plant Cells in Suspension Culture

In order to obtain basic information on ubiquinone (UQ) formation by BY-2 cells in suspension culture, effects of inorganic phosphate and nitrogen sources and such physical factors as initial pH, light irradiation, shaking condition and temperature were investigated. The concentration of phosphate and nitrogen sources had no marked effect, but the ratio of ammonium nitrogen to nitrate nitrogen was significantly effective on UQ formation. The UQ content in BY-2 cells tends to increase at higher ratios of ammonium nitrogen. The increase in the UQ content was recognized at higher concentrations of 2, 4-d, especially with lower concentrations of sucrose. Physical factors had no marked effect on UQ formation except temperature. The UQ content was considerably raised at higher temperatures.     

Inhibition of Tobacco Mosaic Virus Reproduction in Isolated Tobacco Protoplasts by Means of Pancreatic Ribonuclease

Treatment of protoplasts with RNase solutions (1 — 100 μg/ml) sharply reduced their ability to support TMV reproduction. Treatment of protoplasts with RNase both prior to and after inoculation with the virus, and also treatments of varying duration, had approximately the same effect on inhibiting TMV reproduction; however, the effect became sharply intensified with higher RNase concentrations. The authors assume that the RNase inhibitive effect in protoplasts is ensured by the specific structure of this relatively small positively-charged macromolecule, not by its enzymic properties. The RNase molecules apparently have a damaging effect on the plasmalemma and possibly on other cell membranes with which the initial stages of TMV reproduction are associated. The development of the RNase damaging effect can be prevented by bivalent metal ions in the incubation medium, especially by calcium and manganese. The protective effect of cations is presumably caused not only by the charge, but by their steric configuration since K⁺ and Mg⁺⁺ are ineffective in protecting the protoplasts.     

藏东南地区植烟土壤环境研究

研究表明,土壤温度对藏东南地区烤烟产、质量具有关键影响.烟苗假植、适时垄栽、全程覆膜及非灌溉条件下中耕调控,增温保墒效果显著,烟株生长发育正常,烤烟土壤温度及水分生态适应性基本适宜.植烟土壤土质轻粗、土体疏松,有利于烟根烟碱合成和吸钾量的提高.pH6.2～7.3范围内,烤烟产、质量不受pH单一因子的影响.植烟土壤有效氮、磷(P2O57.1～12.2mg/kg)、钾严重失衡(14.6∶1∶13.6),充分发挥磷肥促进早发、加强生长、提前成熟的作用,对高海拔的藏东南地区烤烟优质适产具有特殊意义.     

Response of Peanut,Corn, Tobacco,and Soybean to Criconemella ornate

The relative susceptibility of four field crops to Criconemella ornata differed greatly in microplot tests. As few as 178 freshly-introduced C. ornata/500 cm³ of soil stunted peanut. In contrast, this nematode had no effect on the growth of corn or soybean. Large populations remaining after culture of peanut or corn enhanced the growth of tobacco. A problem of comparing the effects of a freshly introduced population of this nematode with large residual populations was encountered. Freshly extracted, greenhouse-grown inoculum caused the typical "yellows disease" on peanut, whereas much greater residual population densities following a poor host (tobacco) had little effect on the growth of peanut. It is suggested that many of the nematodes in the field following a poor host are dead. Peanut supported greater reproduction (up to 970-fold) than did other crops tested. Corn was intermediate, with a population increase as great as 264-fold; soybean and tobacco failed to maintain initial population densities.     

Carbohydrate Accumulation Affects the Redox State of Ascorbate in Detached Tobacco Leaves

A decreased utilization of NADPH for CO₂ fixation as a result of carbohydrate accumulation in chlorotic leaves is generally believed to be associated with an increase in oxidative stress. Molecular oxygen may serve as an alternative electron acceptor of photosynthesis under these conditions. In order to test this hypothesis mature leaves of tobacco plants (Nicotiana tabacum) were detached, fed with glucose (50 mM) via the petiole, and used to study the effect of carbohydrate accumulation on the pigment content and the components of the superoxide dismutase-ascorbate-glutathione cycle. During a period of five days the concentration of total soluble carbohydrates increased substantially in leaves supplied with glucose in comparison with control leaves supplied only with water. This increase was accompanied by a twofold decrease in the chlorophyll content. In detached water-fed leaves the levels of most of the antioxidative components increased, whereas glucose feeding had no or only little additional effect on the activities of the protective enzymes, but caused a 1.6- and 4-fold increase in the contents of glutathione and ascorbate, respectively. In relation to the total foliar ascorbate pool, the amount of reduced ascorbate increased from about 30 % to 60 % upon feeding with glucose. These results do not support the idea that the accumulation carbohydrates per se cause an increased rate of superoxide production which necessitates increased activities of antioxidative enzymes. It rather appears that glucose-fed leaves have an increased reductive capacity that can be released via glutathione into the ascorbate pool, thereby, regulating the redox state of ascorbate.     

Effect of Salicylic Acid and Phenylserine on the Hypersensitive Reaction of Tobacco to Tobacco Mosaic Virus

Injection of leaves of tobacco (Nicotiana tahacum cv. ‘Xanthi’ nc) with salicylic acid (SA) or phenylsene (PS) had an effect on the local lesion development caused by tobacco mosaic virus (TMV), depending upon the concentration used and the time interval between injection and challenge inoculation. Maximum reduction in lesion size was obtained with 0.75 mM SA or with 8 mM PS. Concentrations higher than 1 mM SA or 25 mM PS damaged the leaf tissue, PS being far less toxic than SA. The leaves responded rapidly to injection with SA or PS. A time interval of only 1 h between injection and TMV inoculation reduced the lesion size significantly. Isolated tobacco cell walls incubated with SA yielded carbohydrate fractions capable of reducing lesion size significantly after injection. Cell walls incubated without SA or with PS did not yield active carbohydrate fractions.     

自然条件下滤减UV-B辐射对烤烟光合色素含量的影响

在自然环境中,以烟草栽培品种K326为材料,通过覆盖不同透明薄膜滤减UV—B辐射,研究100％（CK）、75％（T1）、50％（T2）、35％（T3）UV—B辐射透过率处理下,不同强度UV—B辐射对烟草光合色素含量的影响。结果表明：烤烟三类光合色素对UV—B辐射有不同响应。类胡萝卜素对UV—B辐射响应较敏感。成熟初期,类胡萝卜素含量与UV—B辐射强度变化具有较好的正相关性,而chl a和chl b含量基本与UV—B辐射强度呈反向变化关系。成熟后期,由于UV—B辐射累积效应,光合色素含量变化没有明显规律。现蕾期至成熟采烤烟初期,chl a：chl b与UV—B辐射的反向变化关系较明显,后期则无明显规律,其含量的下降与UV—B辐射的累积效应有关。     

Preconditioning of Tobacco and Bean Leaves to Heat Shock by High Temperature or NaCl

The present paper examines whether leaves subjected to heat or salt pretreatment are better adapted to heat shock than untreated leaves. Such adaptation could result from either acquired thermo-stability and/or better recovery. The rates of metabolic processes such as ¹⁴CO₂ fixation, ¹⁴C-leucine incorporation into the TCA insoluble fraction, ³²P incorporation into phospholipids and ³⁶Cl efflux were used as indicators of adaptability. It was found that pretreatments (heat or salt) protect leaves only slightly against the short-term effects of a heat treatment, but have a pronounced effect on recovery. Cytokinin level was reduced by heat shock, and as in the case of metabolic response, pretreatment had no effect on this reduction.     

The Effects of Aspirin and Polyacrylic Acid on the Multiplication and Spread of TMV in Different Cultivars of Tobacco with and without the N-gene

Aspirin treatment of leaves of Nicotiana tabacum cv. Samsun at 20°C induced PR-proteins and reduced the amount of tobacco mosaic virus (TMV) accumulated 7 days after inoculation. However, at 32°C both the amount of PR-proteins induced and the reduction of TMV accumulated were less. Polyacrylic acid did not induce PR-proteins, and caused little or no reduction in the amount of TMV accumulated at 20°C. In cv. Samsun NN at 32°C. aspirin induced the PR-proteins and reduced the spread of TMV to surrounding tissue as treasured by the size of lesions produced on subsequent transfer to 20°C. Polyacrylic acid did not induce PR-proteins in Samsun NN and had no effect on the spread of TMV. In cv. Xanthi-ne, at 32°C aspirin and polyacrylic acid induced PR-proteins and reduced the spread of TMV. At 35°C, polyacrylic acid induced little or no PR-proteins and did not affect the spread of TMV.     

Effect of Glyoxylate on the Sensitivity of Net Photosynthesis to Oxygen (the Warburg Effect) in Tobacco

The addition of glyoxylate to tobacco (Nicotiana tabacum) leaf discs inhibited glycolate synthesis and photorespiration and increased net photosynthetic ¹⁴CO₂ fixation. This inhibition of photorespiration was investigated further by studying the effect of glyoxylate on the stimulation of photosynthesis that occurs when the atmospheric O₂ level was decreased from 21 to 3% (the Warburg effect). The Warburg effect is usually ascribed to the increased glycolate synthesis and metabolism that occurs at higher O₂ concentrations. Photosynthesis in control discs increased from 59.1 to 94.7 micromoles of CO₂ per gram fresh weight per hour (a 60% increase) when the O₂ level was lowered from 21 to 3%, while the rate for discs floated on 15 millimolar glyoxylate increased only from 82.0 to 99.7 micromoles of CO₂ per gram fresh weight per hour (a 22% increase). The decrease in the O₂ sensitivity of photosynthesis in the presence of glyoxylate was explained by changes in the rate of glycolate synthesis under the same conditions.

The rate of metabolism of the added glyoxylate by tobacco leaf discs was about 1.35 micromoles per gram fresh weight per hour and was not dependent on the O₂ concentration in the atmosphere. This rate of metabolism is about 10% the amount of stimulation in the rate of CO₂ fixation caused by the glyoxylate treatment on a molar carbon basis. Glyoxylate (10 millimolar) had no effect on the carboxylase/oxygenase activity of isolated ribulose diphosphate carboxylase. Although the biochemical mechanism by which glyoxylate inhibits glycolate synthesis and photorespiration and thereby decreases the Warburg effect is still uncertain, these results show that cellular metabolites can regulate the extent of the Warburg effect.

     

Ethylene Stimulation During the Hypersensitive Reaction of Soybean to Tobacco Necrosis Virus under Different Photoperiods

Abstract The production of stress ethylene was increased in soybean leaves hypersensitively responding to tobacco necrosis virus, independently of photoperiod. However, only little increase occurred under continuous darkness, whereas most occurred under continuous darkness, whereas most occurred under continuous light. Ethylene stimulation paralleled accumulation of 1-aminocyclopropane-1-carboxylic (ACC) and its conversion to ethylene. Continuous darkness substantially inhibited viral antigen accumulation but not lesion area in comparison to continuous light. Ethylene release, viral lesion area and antigen accumulation were substantially increased when darkened leaf tissues were fed with glucose, this suggesting that dark inhibition was due to energy and/or, metabolic depletion. Co²⁺ and aminoethoxyvinylglycine, which completely inhibited stress ethylene, and ACC, which conspicuously increased it, had no effect on both viral lesion and antigen accumulation.
These results indicate that stress ethylene developing during a HR to virus does not affect the localizing mechanism operating during it.     

新型植物免疫诱抗剂ZNC对烟草的促生抗病效果

免疫诱抗剂智能聪(ZNC)是一种宛氏拟青霉乙醇提取物,具有促进植物生长提高植物抗逆的能力。本文在烟草不同生长期喷施ZNC,分析其诱导烟草抗病效果及对烟草生长发育的影响。结果表明,3个浓度的ZNC均对病毒病表现出良好防效,且减轻了根茎类病害和后期叶斑类病害的发生程度,其中500 ng/mL ZNC抗病毒病、赤星病等病害的效果最好,抗病毒效果可达57.07%,表明该诱抗剂对烟草主要病害都有较好的诱抗效果。同时,50 ng/mL和500 ng/mL处理分别较CK增产6.00%和5.41%,且500 ng/mL还增加了总糖、还原糖、总氮、K2O等指标的含量,表现出了一定的增产提质效果。试验还表明,尽管1000 ng/mL ZNC表现出抗病效果,但对烟草生长表现出显著的抑制作用,说明该诱抗剂在一定的剂量范围内对烟草有较好促生抗病效果,仍需要进一步试验来确定。     

黄瓜花叶病毒衣壳蛋白存在于被其侵染的烟草叶绿体中

用原生质体法制备出高纯度的完整叶绿体经SDS-PAGE电泳,银染后,发现黄瓜花叶病毒(CMV)侵染的烟草病叶叶绿体蛋白质图谱和健叶叶绿体相比,多出一条染色较弱的迁移率与CMV衣壳蛋白质相同的带,经Western转移,用CMV游离衣壳蛋白亚基抗血清进行斑点酶联(Immunoblot)检测,证明这条带就是CMV衣壳蛋白质。健康叶绿体中加入去掉叶绿体的病叶汁液而制备出的叶绿体中无CMV衣壳蛋白质,说明这不是在叶绿体提纯过程中得到的假象,即衣壳蛋白质存在于被CMV侵染的完整叶片叶绿体中。这个结果否认了以往报道的CMV衣壳蛋白质不存在于烟草叶绿体中的结论。另外还发现,叶绿体中的衣壳蛋白质浓度与叶片症状严重程度呈正相关。     

Studies on the Residual Respiration not Inhibited by KCN Plus Hydroxamic Acid in Tobacco Callus Cultures

A residual respiration not inhibited by KCN plus hydroxamic acid had been observed in many plant organs and tissues. The relative O2 uptake of it was 20–30% of total respiration in tobacco callus cultures. However, there is no report concerning the nature of the residual respiration and its localization in cell. The object of this study is to elucidate the characteris- tics of this residual respiration and its localization in cell. The experimental results are as follows: 1. The additions of glycolate and glyoxylate cause a marked rise in residual respiration not inhibited by KCN (or NaN3) plus m-CLAM. 2. The O2 uptake induced by glycolate and the residual respiration is inhibited by the addition of α-hydroxy ethanesulfonate. 3. The mitochondrial respiration is completely inhibited by KCN plus m-CLAM, but no effect by adding of glycolate. 4. Oxidation reactions of glycolate and glyoxylate in supernatant are observed after mitochondria are removed. Based on the above results, it is suggested that the residual respiration not inhibited by KCN plus m-CLAM in tobacco callus cultures is primarily catalyzed by glycolic acid oxidase localized within microbodies.     

The Effects of 2-5A on Protein Synthesis in Wheat Germ Extracts and Tobacco Protoplasts

Abstract

Nonphosphorylated 2-5A inhibited translation and caused RNA degradation in wheat germ extract, whereas 3-5A had no effect. Protein synthesis inhibition by 2-5A was observed in tobacco protoplasts. 70 kD 2-5A-binding protein was found in potato leaf extracts by chemical crosslinking.     

Interaction between a Nanovirus-like Component and the Tobacco Curly Shoot Virus／Satellite Complex

The biological role of DNA1, a nanovirus-like component shown to be associated with the begomovirus/satellite complex, has not yet been identified. Here, we demonstrated that DNA1 of Tobacco curly shoot virus isolate Y35 (TbCSV-Y35) attenuated leaf-curling symptoms induced by TbCSV-Y35 or TbCSV-Y35 plus Y35 DNAβ in the early stage of symptom development and induced leaf cluster at a later stage of symptom development in Nicotiana benthamiana plants. The leaf disc assay demonstrated that TbCSV-Y35 DNA1 replicated autonomously. Southern blot analysis revealed that TbCSV-Y35 DNA1 reduced viral DNA accumulation. Viral DNA accumulation was not reduced when plants were co-inoculated with TbCSV-Y35 DNAβ, but the TbCSV-Y35 DNAβ level was dramatically reduced in the presence of TbCSV-Y35 DNA1. To determine whether the interaction between TbCSV/satellite complex and DNA1 had isolate specificity, DNA1 of TbCSV isolate Y132 was cloned and sequenced. It was found to have 75% nucleotide sequence identity with TbCSV-Y35 DNA1. Infectivity tests showed that TbCSV-Y132 DNA1 had no effect on the symptoms induced by TbCSV-Y35 or TbCSV-Y35 and Y35 DNAβ in N. benthamiana plants, although Y 132 DNA1 could replicate in these plants.     

磷脂酶D对ParA1诱导的烟草的细胞死亡和过氧化氢及莨菪亭积累的影响

利用药理学方法,研究了烟草寄生疫霉(Phytophthora parasitica)分泌的蛋白激发子ParA1诱导烟草悬浮细胞后,磷脂酶D对ParA1诱导的过敏细胞死亡和其它防卫反应的影响.用100nmol/LParA1处理烟草悬浮细胞后能够诱导细胞死亡、过氧化氢和莨菪亭的积累.磷脂酶D抑制剂正丁醇能够抑制ParA1诱导的这些防卫反应,仲丁醇所起的抑制作用比正丁醇小,正丁醇和仲丁醇产生的抑制效果具有浓度依赖效应.而叔丁醇不能抑制ParA1诱导的这些反应.结果表明,磷脂酶D参与了ParA1诱导烟草悬浮细胞的信号传导过程.     

Improved Performance of Transgenic Fructan-Accumulating Tobacco under Drought Stress

Fructans are polyfructose molecules produced by approximately 15% of the flowering plant species. It is possible that, in addition to being a storage carbohydrate, fructans have other physiological roles. Owing to their solubility they may help plants survive periods of osmotic stress induced by drought or cold. To investigate the possible functional significance of fructans, use was made of transgenic tobacco (Nicotiana tabacum) plants that accumulate bacterial fructans and hence possess an extra sink for carbohydrate. Biomass production was analyzed during drought stress with the use of lines differing only in the presence of fructans. Fructan-producing tobacco plants performed significantly better under polyethylene-glycol-mediated drought stress than wild-type tobacco. The growth rate of the transgenic plants was significantly higher (+55%), as were fresh weight (+33%) and dry weight (+59%) yields. The difference in weight was observed in all organs and was particularly pronounced in roots. Under unstressed control conditions the presence of fructans had no significant effect on growth rate and yield. Under all conditions the total nonstructural carbohydrate content was higher in the transgenic plants. We conclude that the introduction of fructans in this non-fructan-producing species mediates enhanced resistance to drought stress.     

Effects of Singlet Oxygen Quenchers and pH on the Bacterially Induced Hypersensitive Reaction in Tobacco Suspension Cell Cultures

Suspension cell cultures of Nicotiana tabacum L. inoculated with the incompatible pathogen Pseudomonas syringae pv pisi undergo a hypersensitive reaction. Addition of the singlet oxygen quencher bixin to cell suspensions had no effect on hypersensitive cell death. Addition of the singlet oxygen quencher 1,4-diazabicyclo octane (DABCO) increased the medium pH and delayed the onset of cell death. This delay was eliminated when cell suspensions were buffered, and could also be induced by increasing medium pH with KOH. Bixin and DABCO also did not suppress the hypersensitive reaction in tobacco leaves. These data do not support a role for singlet oxygen in the hypersensitive reaction. Medium pH, however, appears to be a critical factor in cell suspension cultures.