重铬酸钾对蚕豆根尖细胞致畸效应的研究

以蚕豆根尖为材料,研究重铬酸钾对蚕豆根尖细胞的致畸效应。采用蚕豆根尖细胞的微核试验和染色体畸变试验方法,以不同浓度的重铬酸钾为诱变剂,测定蚕豆根尖细胞的微核率和染色体畸变率。结果表明:重铬酸钾能诱发较高频率的微核率,即在一定浓度范围内,其微核率随重铬酸钾处理浓度的升高而增加,但高于一定浓度后反而呈下降趋势;不同浓度的重铬酸钾均使蚕豆根尖细胞有丝分裂指数增大;重铬酸钾还能诱导蚕豆根尖细胞产生较高频率的染色体畸变,且产生多种类型的染色体畸变。结论是重铬酸钾对蚕豆根尖细胞具有明显的致畸效应。     

The Development of Primary Root Nodules on Vicia faba L. Grown at Two Temperatures

The development of primary root nodules on the field bean (Viciafaba L.) grown at 10 and 18 °C was examined. The sequenceof anatomical changes observed was the same in both temperatureregimes. Nodules developed much more slowly at 10 °C andthe nodules were much larger when corresponding anatomical changesoccurred. Primary root nodules eventually ceased growth in bothtemperature regimes but ultimate nodule volume was nearly twiceas great as 10 °C as at 18 °C. The larger size did notcompensate for the lower specific nitrogenase activity of cold-grownnodules: nitrogen fixation (acetylene reduction) rates on awhole plant basis were much lower at 10 °C than at 18 °C.There was no difference in the total number of primary rootnodules in the two temperature regimes but their distributionwas biased towards the upper part of the root and the epicotylat 18 °C. Vicia faba L., field bean, nodulation, nitrogen fixation, temperature     

Isotopic Studies of Nitrogen Fixation in Non-Legume Root Nodules

Isotopic studies are presented on six of the eight recognizednon-legume nodule-forming genera of Angiosperms. Evidence hasbeen obtained of the occurrence of fixation of atmospheric nitrogenin the root nodules of Casuarina, Ceanothus, and Shepherdia,now examined isotopically for the first time. In further studiesof the nitrogen-fixing nodules of Hippopha and Alnus it is shownthat the fixation continues for a longer period after detachmentfrom the plant than is the case with legume nodules, and thatthe enrichment in ¹³N which they (and also detached Casuarinanodules) finally achieve after exposure to excess of the freeisotope considerably exceeds that shown by detached legume nodules.Fixation in detached Myrica nodules was not clearly affectedby reduction in oxygen supplied until the proportion of oxygenwas less than 5 per cent.     

Verapamil-Sensitive Calcium Transporter in the Peribacteroid Membrane of Symbiosomes from Vicia faba Root Nodules

Based on electron microscopic studies and visualization of calcium with the Ca indicator pyroantimonate, it was established that a prolonged incubation of the bean (Vicia faba L.) root nodules and isolated symbiosomes in EGTA-containing buffer depletes calcium in these nitrogen-fixing units. Other experiments demonstrated that the induction of calcium deficit in symbiosomes both in vivo and in vitro substantially decreases their nitrogenase activity. The addition of verapamil and ruthenium red, well-known inhibitors of Ca²⁺ channels, to the suspension of root nodules largely prevented both the EGTA-induced calcium efflux from the symbiosomes and the decrease in their nitrogenase activity. Similar effects of verapamil were also observed on isolated symbiosomes. The treatment of isolated symbiosomes with valinomycin in the presence of K⁺ induced a rapid efflux of Ca²⁺ from symbiosomes; this efflux was strongly inhibited by verapamil. The results present evidence for the existence in the peribacteroid membrane of a Ca²⁺-transporting system that exports Ca²⁺ from the symbiosomes.     

Nitrogen Metabolism of Vicia faba

Vicia faba plants were grown for four and six weeks without externally supplied nitrogen. Some nitrogen was transported to the plant axis from the cotyledons throughout this period, but the amount available was insufficient to support maximum shoot growth. During this period the protein content of the shoot declined whilst the free amino acids, especially aspartic acid, glutamic acid, histamine and the combined pool for threonine, serine, asparagine and glutamine and ammonia, increased in amount. In contrast to the shoot the protein content of the root increased as did their free amino acid content, but the increase in the latter was less than in the shoot and only the combined value for threonine, serine, asparagines and glutamine increased significantly. During tbe last two weeks growth, some soluble non-amino acid compound appeared to donate nitrogen to the pool of free amino acids in the root and shoot.     

Oxygen Diffusion Pathways and Nitrogen Fixation in Legume Root Nodules

The theoretical analysis presented in this paper suggests thatthe nature of the diffusion pathway from the surface of a noduleto the infected zone depends on the morphology of the nodule;in particular the cross-sectional area of the intercellularspaces in the inner cortex. If the diffusion barrier containscontinuous pathways there will be no pressure difference betweenthe atmosphere and the infected zone. The conditions under whichthe intercellular spaces of the inner cortex could be air-filled,water-filled, or a combination of both are explored. An experimentto resolve this issue is suggested. Information obtained usingcryo-scanning electron microscopy and oxygen electrode datahave been used to illustrate various points. Expressions arederived for the diffusion resistance of the nodule and its componentparts. To provoke further discussion a simple mechanism forthe control of diffusion is described in general terms. Oxygen, diffusion resistance, nodule, nitrogen fixation, soybean, Glycine max. (L.) Merr. cv. Fiskeby V, Trifolium repens L. cv. Blanca, lucerne, Medicago saliva L. cv. Europe     

Inhibition of Nitrogen Fixation in Non-Legume Root Nodules by Hydrogen and Carbon Monoxide

The effect of the presence of hydrogen and of carbon monoxideon the fixation of nitrogen in detached root nodules of non-legumeshas been studied, fixation being measured by the use of ¹⁵N.Parallel tests on legumes (pea and soya bean) have been included.Fixation in the nodules of Casuarina, Alnus, and Myrica is inhibitedin the presence of substantial proportions of hydrogen, to adegree resembling that shown in legumes. Fixation in Alnus andMyrica is arrested in the presence of small proportions of carbonmonoxide, and here again the sensitiveness is of the same orderas in legumes.     

硫酸铜对蚕豆根尖细胞有丝分裂的影响

以蚕豆根尖为材料,研究硫酸铜对蚕豆根尖细胞的遗传毒性效应。采用蚕豆根尖细胞的微核试验方法和染色体畸变试验方法,以不同浓度的硫酸铜为诱变剂,测定蚕豆根尖细胞的有丝分裂指数、微核率和染色体畸变率。结果表明:不同浓度的硫酸铜均能使蚕豆根尖细胞有丝分裂指数明显增加,即5个实验组的分裂指数均明显高于对照组(P<0.01或P<0.001);不同浓度的硫酸铜对蚕豆根尖细胞有丝分裂各期百分数的影响有异;能诱发较高频率的微核率,即在一定浓度范围内,其微核率随硫酸铜处理浓度的升高而增加,但随着硫酸铜浓度的进一步升高而呈下降趋势;硫酸铜还能诱导染色体产生多种类型的畸变,染色体畸变率随硫酸铜处理浓度的升高而增加,随着硫酸铜浓度的进一步升高而呈下降趋势,但均明显高于对照组(P<0.001)。结论是硫酸铜对蚕豆根尖细胞具有明显的遗传毒性效应。     

乙酸铜对蚕豆根尖细胞致畸效应

采用蚕豆根尖细胞的微核试验和染色体畸变试验方法,以不同浓度的乙酸铜为诱变剂,选择不同的处理时间,测定蚕豆根尖细胞的有丝分裂指数、微核率和染色体畸变率。结果表明：乙酸铜能诱发较高频率的微核率,处理6h、12h时微核率均随着乙酸铜浓度的升高而增加,具有明显的剂量效应;处理24h时在实验浓度范围内,其微核率随乙酸铜浓度的升高而增加,但高于一定浓度后反而呈下降趋势。不同浓度的乙酸铜在不同处理时间均使蚕豆根尖细胞有丝分裂指数增大。乙酸铜还能诱导蚕豆根尖细胞产生较高频率的染色体畸变,且产生多种类型的染色体畸变。因此,乙酸铜对蚕豆根尖细胞具有明显的致畸效应。     

铬渣堆场渗滤液对蚕豆根尖细胞微核的影响

利用蚕豆根尖微核技术监测化工铬渣堆场渗出液的遗传毒性,结果表明,化工铬渣渗出液含有较高浓度的Cr6+,能明显诱导蚕豆根尖细胞微核的形成,具有较强的遗传毒性。当铬渣渗出液中Cr6+浓度为186.6460 mg/L时,微核率达到最大。铬渣渗出液所诱导的微核率与阴性对照组相比有极显著差异。     

Effect of Fusicoccin on Dark CO(2) Fixation by Vicia faba Guard Cell Protoplasts

When Vicia faba guard cell protoplasts were treated with fusicoccin, dark ¹⁴CO₂ fixation rates increased by as much as 8-fold. Rate increase was saturated with less than 1 micromolar fusicoccin. Even after 6 minutes of dark ¹⁴CO₂ fixation, more than 95% of the incorporated radioactivity was in stable products derived from carboxylation of phosphoenolpyruvate (about 50% and 30% in malate and aspartate, respectively). The relative distribution of ¹⁴C among products and in the C-4 position of malate (initially more than 90% of [¹⁴C]malate) was independent of fusicoccin concentration. After incubation in the dark, malate content was higher in protoplasts treated with fusicoccin. A positive correlation was observed between the amounts of ¹⁴CO₂ fixed and malate content.

It was concluded that (a) fusicoccin causes an increase in the rate of dark ¹⁴CO₂ fixation without alteration of the relative fluxes through pathways by which it is metabolized, (b) fusicoccin causes an increase in malate synthesis, and (c) dark ¹⁴CO₂ fixation and malate synthesis are mediated by phosphoenolpyruvate carboxylase.

     

Variability among Rhizobium Strains Originating from Nodules of Vicia faba

Rhizobium strains from nodules of Vicia faba were diverse in plasmid content and serology. Results of multilocus gel electrophoresis and restriction fragment length polymorphism indicated several deep chromosomal lineages among the strains. Linkage disequilibrium among the chromosomal types was detected and may have reflected variation of Rhizobium strains in the different geographical locations from which the strains originated. An investigation of pea strains with antibodies prepared against fava bean strains and restriction fragment length polymorphism analyses, targeting DNA regions coding for rRNA and nodulation, indicated that Rhizobium strains from V. faba nodules were distinguishable from those from Pisum sativum, V. villosa, and Trifolium spp.     

Nitrogen Fixation (C(2)H(2) Reduction) by Broad Bean (Vicia faba L.) Nodules and Bacteroids under Water-Restricted Conditions

Water potentials of leaves and nodules of broad bean (Vicia faba L.) cultivated on a sandy mixture were linearly and highly (r² = 0.99) correlated throughout a water deprivation of plants. A decrease of 0.2 megapascal of the nodule water potential (Ψ_nod) induced an immediate 25% inhibition of the highest level of acetylene reduction of broad bean nodules attached to roots. This activity continued to be depressed when water stress increased, but the effect was less pronounced. Partial recovery of optimal C₂H₂ reduction capacity of mildly water stressed nodules (Ψ_nod = −1.2 megapascals) was possible by increasing the external O₂ partial pressure up to 60 kilopascals. The dense packing of the cortical cells of nodules may be responsible for the limitation of O₂ diffusion to the central tissue. Bacteroids isolated from broad bean nodules exhibited higher N₂ fixation activity with glucose than with succinate as an energy-yielding substrate. Bacteroids from stressed nodules appeared more sensitive to O₂, and their optimal activity declined with increasing nodule water deprivation. This effect could be partly due to decreased bacteroid respiration capacity with water stress. Water stress was also responsible for a decrease of the cytosolic protein content of the nodule and more specifically of leghemoglobin. The alteration of the bacteroid environment appears to contribute to the decline in N₂ fixation under water restricted conditions.     

甲硝唑诱导蚕豆根尖细胞微核的研究

本实验研究了甲硝唑诱导蚕豆根尖细胞微核的效应。实验表明：(1)浓度为0.1、6、12、40和500 mg/L甲硝唑均能使蚕豆根尖细胞微核率显著增加,且蚕豆根尖细胞微核率和甲硝唑浓度之间存在明显的剂量-效应关系;(2)甲硝唑能引起DNA损伤,具有分子诱变剂的性能。     

羊奶果不同发育阶段根瘤的细胞结构及固氮、吸氢活性

比较羊奶果根瘤三个不同发育阶段的显微,亚显微结构和固氮,吸氢活性的差异。探讨了根瘤结构与功能的关系。结果表明：早期侵染方式为皮层细胞间隙侵染,此期的内生菌是一种分枝,具隔膜的菌丝体,早期侵染细胞有脂体存在。成熟根瘤含菌细胞明显多于幼瘤和衰老瘤。成熟根瘤具有大量泡囊,成熟泡囊具分隔,双层壁结构。衰老瘤泡囊分隔消失,不呈双层壁结构。成熟根瘤的固氮,吸氢活性明显高于幼瘤和衰老瘤。     

Micronuclei and Radiosensitiviy in the Root Meristem of Vicia faba

Percentages of cells with micronuclei in four regions of theroot meristems of Vicia faba are used as measures of sensitivityto acute X-irradiation. There are two peaks in these percentages,occurring at about four and eight days after 360 rads and twoand six days after 180 rads. Two peaks exist, probably becausethe radiation delays cells that were in G₁ much more than cellsthat were in G₂ in reaching the first post-irradiation mitosisand consequently in displaying micronuclei in the followinginterphase. The relative heights of the two peaks thereforereflect the relative numbers of cells in G₁ and G₂ as well asthe relative sensitivity of the two phases to chromosomal damage. The cells of the quiescent centre are injured least by the radiationas they are mostly held at Gt. The meristem thus obeys the lawof Bergonié and Tribondeau, but differs from that ofZea in that the meristematic cells of the cap initials and steleimmediately adjacent to the quiescent centre resemble the quiescentcentre much more closely than the stele 250 µ away inthe numbers of micronuclei produced. This is consistent withthe differences already known between the two species concerningrates of division in the different regions of the meristem andthe behaviour of the meristem after severe radiation injury.     

Changes in the Light Scattering by Symbiosomes from Vicia faba Root Nodules as Indicator of Ion and Metabolite Transport across the Peribacteroid Membrane

Passive transport of ions and metabolites across the peribacteroid membrane (PBM) was investigated on symbiosome preparations isolated from the broad bean (Vicia faba L.) root nodules and suspended in a potassium-free medium. Optical density of the symbiosome suspension at 546 nm was monitored as an indicator of light-scattering changes. Depolarization of the PBM with tetraphenylphosphonium cation (TPP⁺) caused an increase in light scattering of symbiosome suspension. This effect was enhanced after adding a K⁺ ionophore valinomycin to the incubation medium. A similar effect was observed after supplementing the symbiosome suspension with nigericin, a K⁺/H⁺ antiporter. Similar experiments on bacteroid suspensions prepared from isolated symbiosomes did not reveal any appreciable changes in light scattering in the presence of the same membrane-active substances. The light scattering by symbiosome suspensions decreased after adding malate or succinate, while the subsequent addition of centimolar concentrations of K⁺ substantially accelerated this process. Light scattering by the symbiosome suspension was insensitive to the addition of glutamate, a substance normally impermeant through the PBM of legume root nodules. These results suggest that the changes in light scattering by symbiosomes reflect the osmotically induced changes of symbiosome volume. These volume changes were assigned to alteration of the peribacteroid space (PBS). The incubation of symbiosomes in a potassium-free medium acidified their the PBS; this acidification was accelerated by valinomycin, carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone (FCCP), and nigericin, and it was abolished in the presence of comparatively high concentrations of K⁺ in the incubation medium. The results indicate a relatively high permeability of the PBM to K⁺ ions.     

镉离子对蚕豆根尖细胞分裂的影响

蚕豆根尖经孙同浓度（0.1-10.0ppm）的CdCl2溶液处理24-48小时后,根的生长受到不同程度的抑制。对根尖细胞分裂的毒害则表现为低毒C-有丝分裂和高毒的染色体断裂、粘连和液化。此外,对Cd++的毒害机理进行了简略讨论。     

EDTA对蚕豆根尖细胞微核的诱变作用

为探讨EDTA对高等植物细胞的诱变作用 ,以及验证EDTA对金属离子的螯合作用 ,本试验用不同浓度的EDTA处理蚕豆根尖细胞 ,进行微核试验 ,并以Pb(NO3 ) 2 作为阳性对照 ,以无离子水作为阴性对照 ,发现EDTA能使蚕豆根尖细胞微核率有明显增加。由此证明EDTA对高等植物细胞有一定的诱变作用     

Nitrogen Fixation and ATPase of Root Nodules from Sesbania cannabina with Chemical and Physical Treatments

The nitrogenase activity of root nodules from Sesbania cannabina plants treated with Na2S2O4, DTT and trypsin was increased by 108%–114%, 106%–117% and 103%–119%, respectively. EM observation showed that the density of ATP-hydrolase as the marker of lead phosphate particles which were distributed on the peribacteroid membrane was much more significant than that of the control, but the bacteroids in peribacteroid membrane did not have ATP-hydrolase particles present. Dark treatment of the same age plants accelerated the nodule senescence and the ATP-hydrolase particles most densified on the peribacteroid membrane of the nodules, meanwhile, dense ATP-hydrolase particles also appeared in a number of degenerative bacteroids. This again confirms the conformation change of ATP-ase in bacteroids from ATP synthetase to ATP-hydrolase and its relation to nitrogen fixation with the senescence of nodules. The comparison of ATP-hydrolase particle density on the peribacteroid membrane of the nodule cells with different treatmemts are carried out and the role of the ATP-hydrolase on the peribacteroid membrane in substance transportation are discussed.