An investigation of herbicide interaction with the H+-ATPase activity of plant plasma membranes

This study has investigated the activity of several herbicide classes at the plant plasma membrane. Two-phase partitioning was used to prepare highly purified plasma membrane vesicles from the monocotyledon weed black-grass (Alopecurus myosuroides Huds.) and the dicotyledon crop sugar beet (Beta vulgaris L. cv. Celt). The purity of the plasma membrane H⁺-ATPase activity was characterised with respect to inhibitors, pH and substrate specificity. In both species, contamination of the plasma membrane by tonoplast fragments was largely eliminated and chlorophyll was absent. In addition, the plasma membrane H⁺-ATPase from black-grass and sugar beet exhibited high vanadate sensitivity and a sharp pH profile around 6·5. Subsequently, H⁺-ATPase activity was assayed in the presence (100 μM ) and absence of four graminicide classes and auxin-type herbicides. Graminicides, including the aryloxyphenoxypropionate diclofop-methyl and the thiocarbamate triallate, inhibited H⁺-ATPase activity by 50–80% in both species. However, other graminicides, including the cyclohexanediones and the chloroacetamide alachlor, had no affect. Similarly, auxin-type herbicides such as 2,4-D and MCPA did not inhibit H⁺-ATPase activity. Results are discussed in relation to the proposed mode of action of these herbicides. © 1998 SCI     

Effect of SS-toxin,a metabolite of <Emphasis Type="Italic">Stemphylium solani</Emphasis>, on H<Superscript>+</Superscript>-ATPase activity and standard redox system in plasma membranes from seedlings leaves of garlic (<Emphasis Type="Italic">Allium sativum</Emphasis>)

Leaf blight of garlic (Allium sativum) is a severe disease in garlic-growing regions. SS-toxin is a newly described non-proteinaceous toxin produced by the phytopathogenic fungus, Stemphylium solani, the cause of garlic leaf blight. In this study, the effects of SS-toxin on the H⁺-ATPase activity and standard redox activity in the plasma membranes isolated by aqueous polymer two-phase partitioning from garlic seedling leaves were studied in vitro. The H⁺-ATPase activity, NADH oxidation rate and Fe(CN)₆³⁻ reduction rate of the plasma membranes isolated from susceptible and resistant cultivars were all inhibited in a dose-dependent manner. Our results suggest that, under in vitro conditions, the plasma membrane H⁺-ATPase and standard redox system can be both the cellular targets of SS-toxin.     

Properties and functions of proton pumps in higher plants

The properties of the major H⁺-pumps in higher plant cells are described: these are the H⁺-ATPase of the plasma membrane and the H⁺-ATPase and the H⁺-pyrophosphatase of the tonoplast. They are distinguished by their structure, mechanism of action, kinetic properties, and location within the cell. The uses and value of isolated and purified membrane vesicles in transport studies are discussed. A particular example is given by the use of plasma membrane vesicles, purified by phase partition, to study proton/ sucrose co-transport in Ricinus cotyledons. Current evidence for the interference with these pumps by growth regulators, toxins and xenobiotics is outlined.     

黄腐酸对雾培马铃薯幼苗抗旱性的影响

试验以马铃薯(Solanum tuberosum)幼苗为研究对象,采用雾培装置进行不同黄腐酸用量和干旱胁迫处理(分别为CK,不加黄腐酸和PEG; 10％ PEG;0.01％ FA+ 10％ PEG;0.03％ FA+ 10％ PEG;0.05％ FA+ 10％ PEG和0.10％ FA+ 10％ PEG),分析其对马铃薯生长发育及抗性生理的影响。结果表明,在干旱胁迫下,雾培马铃薯幼苗的成活率、叶绿素含量、膜稳定指数、根系活力、匍匐茎数、结薯率和产量呈下降趋势,而丙二醛(MDA)、过氧化氢(H₂O₂)、脯氨酸、可溶性糖含量和过氧化氢酶(CAT)活性及超氧阴离子(O^-₂)产生速率呈上升趋势,过氧化物歧化酶(SOD)和过氧化物酶(POD)活性先增加后下降,说明雾培马铃薯幼苗在干旱胁迫下生长发育受到显著抑制,正在遭受逆境胁迫。经黄腐酸处理后,幼苗的成活率、匍匐茎数、结薯率和产量则得到了提高,而叶绿素含量、膜稳定指数和根系活力仍呈下降趋势,但较单一干旱胁迫的下降趋势缓慢,而MDA含量、SOD和POD活性及产生速率呈先升高后下降的趋势,脯氨酸、可溶性糖、H₂O₂含量及CAT活性呈上升趋势,其中H₂O₂含量上升趋势较单一干旱胁迫缓慢,说明黄腐酸处理后使干旱胁迫下雾培马铃薯幼苗的生长发育得到了促进,减轻了干旱胁迫对幼苗造成的伤害,提高了植株的整体抗旱性,而这种抗性与黄腐酸浓度呈明显相关性,0.05％ FA对植株的的保护效应最显著,而0.10％ FA则协同PEG加剧了对幼苗的伤害作用。     

Ion flux kinetics in blue light‐grown field dodder (Cuscuta campestris) seedlings

Previous studies have revealed that blue light stimulates coiling and prehaustoria development in young de‐etiolated dodder seedlings prior to host attachment. In this study, the Ca²⁺ and H⁺ flux kinetics that are associated with blue light‐mediated coiling and prehaustoria development in de‐etiolated dodder seedlings were measured uninvasively using the ion‐selective, vibrating microelectrode ion flux system. The findings showed that blue light induces rapid transient changes in the net Ca²⁺ and H⁺ fluxes. A clearly pronounced lag of about several minutes was observed between the blue light‐induced changes in the Ca²⁺ and H⁺ fluxes, with H⁺ being a leading ion. The addition of 1 mol L^?1 vanadate, a known blocker of the plasma‐membrane H⁺‐ATPase, to the bathing solution completely prevented both the H⁺ and the Ca²⁺ flux responses and inhibited coiling and prehaustoria development, suggesting a causal relationship between the blue light‐induced changes in the net Ca²⁺ and H⁺ fluxes and seedling movements. It is concluded that the plasma‐membrane H⁺‐ATPase plays a key role in a dodder's parasitic mode of growth and its adaptive response to the environment.     

工业废水对黄瓜幼苗生长及叶片抗氧化系统的影响

采用以甘肃省境内某地区化肥厂、造纸厂的工业废水以及此两厂的混合废水作溶剂的培养液(分别记作废液1、废液2和废液3)培养黄瓜幼苗,研究了其对黄瓜幼苗的生长及其叶组织中活性氧清除系统的影响。结果发现,(1)培养5 d后,生长在废液1中的幼苗,叶片组织中的几种抗氧化酶除APX的活性显著增加外(P<0.01),CAT、SOD和GR活性均无明显变化(P>0.05)。生长在废液2中的CAT和GR的活性基本未变,APX和SOD的活性分别在P<0.01和P<0.05的水平上增加。在废液3中,CAT和SOD及APX的活性分别在P<0.05和P<0.01的水平上增加,而GR活性降低(P<0.05)。(2)培养13 d后,无论在哪种废水中,黄瓜幼苗的伸长生长和干物质积累及叶片组织中APX、SOD、CAT和GR活性、GSH和ASA含量均明显降低(P<0.05或P<0.01),H2O2、O2、MDA含量和电解质泄漏率明显增加(P<0.05或P<0.01)。结果表明,化肥厂、造纸厂的工业废水以及此两厂的混合废水对黄瓜幼苗叶组织中的抗氧化系统有明显的破环作用,最终影响幼苗的生长。     

植物根围促生细菌菌剂增强玉米干旱耐受性研究

选用植物根围促生细菌(PGPR)菌剂BBS处理玉米幼苗,研究干旱胁迫下玉米幼苗叶片相关生理特性及基因表达差异。结果表明,干旱胁迫15 d后,BBS处理组玉米幼苗的萎蔫程度显著低于对照组;处理组植株叶片MDA含量仅为对照组的74.35%,叶片脯氨酸和可溶性糖含量分别是对照组的3.39倍和1.07倍;处理组植株叶片ZmP5CS1的表达水平及SOD活性均显著高于对照组;BBS处理组植株叶片在干旱胁迫过程中一直保持较低的H_2O_2含量。研究得出,BBS处理能够有效减缓干旱胁迫对玉米幼苗造成的伤害,进而加速正常浇水后的恢复。处理组植株叶片NCED1和ZmDREB2.7的高水平表达也表明干旱胁迫下玉米幼苗与BBS的互作过程可能有依赖ABA信号途径及不依赖ABA信号途径的共同参与。     

盐胁迫下宁夏枸杞根系Na~+、K~+平衡及抑制剂对其影响的研究

以宁夏枸杞为试验材料,利用非损伤微测技术及原子吸收分光光度法,研究NaCl胁迫下枸杞根系中Na~+、K~+吸收转运及质膜Na~+/H~+逆向转运蛋白抑制剂阿米洛利和质膜H~+-ATPase抑制剂矾酸钠对宁夏枸杞根系离子平衡的影响。结果表明:随着NaCl胁迫浓度的增加,枸杞根系中Na~+含量总体升高,与对照差异不显著;K~+含量呈先升后降趋势;Na~+/K~+呈先降后升趋势,且低于对照;Na~+外排先增加后减少,K~+外排降低,且显著高于对照。随着胁迫时间的延长,枸杞根系中Na~+、K~+含量增加,Na~+外排逐渐减慢,K~+外排增加;阿米洛利与矾酸钠均显著降低了盐诱导下的Na~+和H~+内流。NaCl胁迫下宁夏枸杞根系借助于其质膜上的Na~+/H~+逆向运输系统,将胞内过量的Na~+排出胞外,同时抑制K~+外排,保持细胞内较低的Na~+/K~+,维持胞内Na~+、K~+平衡,从而表现出较强的耐盐性。     

磁化水对不同土壤水分下黄瓜幼苗生长、光合和养分吸收的影响

采用二因素二水平完全试验设计,研究了砂培条件下磁化处理（分别为0.3T磁场强度处理的磁化蒸馏水和蒸馏水处理）对不同土壤水分条件下（正常水分处理：田间持水量的80%~85%;干旱处理：田间持水量的40%~45%,于黄瓜4~5叶期维持7 d）黄瓜幼苗生长、水分关系、光合作用和养分吸收的影响。结果表明,磁化水浇灌显著降低黄瓜幼苗的生长和水分利用,其中茎粗、地上部生物量、根生物量、总生物量、根体积、根表面积、整株耗水量和整株水分利用效率分别降低6.7%、8.9%、19.1%、9.9%、22.1%、18.5%、6.2%和10.9%;磁化水浇灌导致黄瓜幼苗叶SPAD值降低了3.7%,叶气孔导度和蒸腾速率则分别增加了21.8%和17.5%,叶瞬时水分利用效率降低了17.7%,但对净光合速率影响不大;磁化水浇灌使PSII最大光化学效率和电子传递速率分别降低了5.2%和18.6%,同时增加叶片中K含量。除叶片K含量外,干旱条件下磁化水对黄瓜生长和生理代谢影响不大,表明磁化水的作用依赖于土壤水分条件。     

黄瓜抗细菌性角斑病诱抗药剂的筛选及抗性机制研究

为筛选诱导黄瓜抗细菌性角斑病(angular leaf spot, ALS)的最佳诱抗药剂,选取11种商品诱抗剂,研究其室内诱导黄瓜抗细菌性角斑病的作用效果。结果显示11种诱抗剂的预处理均能有效提高黄瓜植株的长势,提高黄瓜对细菌性角斑病的抗性,表现为叶片病斑面积显著下降。其中药剂信号施康乐(SK)能有效减小叶片病斑面积,防效最好,而药剂顶花增瓜嫩直长(DH)处理的植株叶片病斑面积最大。为了进一步明确诱抗剂的作用机制,对诱抗剂SK和DH处理后的抗性相关基因SOD、SSU以及PR1-1a的相对表达量进行了初步分析。结果表明,SK和DH处理后3个基因的相对表达量均显著提高,SK处理的相对表达量显著高于DH处理。综上所述,合理施用信号施康乐可以诱导黄瓜幼苗提高对细菌性角斑病的抗性,对于黄瓜细菌性角斑病的早期预防和减少农药使用具有重要意义。     

Effect of chitosan on growth and plasma membrane properties of Rhizopus stolonifer (Ehrenb.:Fr.) Vuill

The effect of chitosan (0.5, 1.0 and 2.0 mg ml⁻¹) on mycelium growth, potassium efflux, pH of the incubation medium, and activity of plasma membrane H⁺-ATPase of Rhizopus stolonifer was evaluated. The mycelium growth of R. stolonifer was reduced on media amended with chitosan at 1.0 and 2.0 mg ml⁻¹. The highest antifungal index (65%) was obtained on media containing chitosan at 2.0 mg ml⁻¹. Potassium efflux induced by the addition of the three chitosan concentrations was demonstrated. The highest potassium effluxes were observed with addition of chitosan at 2.0 mg ml⁻¹. R. stolonifer grown 72 h on Potato Dextrose Broth without chitosan generated a decrease in the medium’s pH from 5.4 to 4.8. However, the addition of chitosan at all concentrations caused increases on pH of medium cultures. The highest pH increases were observed on media amended chitosan at 2.0 mg ml⁻¹ resulting in the raising of pH level to 6.9. Additionally, there was an important inhibitory effect of chitosan (1.0 and 2.0 mg ml⁻¹) on H⁺-ATPase activity in the plasma membrane of R. stolonifer (32.70%).     

木霉分生孢子和厚垣孢子对黄瓜幼苗生理特性及枯萎病防效的影响

采用前期筛选出的对黄瓜枯萎病菌有较好拮抗作用的3株木霉菌,即哈茨木霉Trichoderma harzianum 809、拟康氏木霉Trichoderma pseudokoningii 886和棘孢木霉Trichoderma asperellum 525,利用盆栽试验,测定了木霉菌孢子不同类型施用对黄瓜幼苗生长、生理特性...     

Effects of dp-B on ATPase activity of insect plasma membrane

Inhibition of ATPase from aphid plasma membrane, sarcoplasmic reticulum (SR), and brain synaptosomal plasma membrane of locust, by 1,5-diphenyl-2-penten-1-one (dp-B), was studied. Results demonstrated that dp-B exhibited the similar effects on ATPase found in the three membranes amongst which the plasma membrane Ca²⁺–Mg²⁺-ATPase is the primary target. The effects of dp-B on Ca²⁺–Mg²⁺-ATPase activity were bi-directional: the enzyme enhanced hydrolyzation when dp-B or Ca²⁺ concentrations were low but inhibited significantly when at high concentrations. The inhibition of Ca²⁺–Mg²⁺-ATPase by dp-B from brain synaptosomal plasma membrane of insect is higher than that from other organizational plasma membranes of the insect. Dp-B inhibiting Ca²⁺- and CaM-requiring ATPase activity may be an important mechanism by which the insect gets poisoned (for example aphid).     

木霉菌对黄瓜幼苗生长和膜脂过氧化指标的影响及对枯萎病的防治效果

本文开展了3种生防木霉菌,包括棘孢木霉Trichoderma asperellum 525、哈茨木霉T.harzianum 610和拟康氏木霉T.pseudokoningii 886防治黄瓜枯萎病的盆栽试验,研究这3种木霉菌对黄瓜幼苗生长、膜脂过氧化指标的影响及对黄瓜枯萎病的防治效果。结果表明,木霉菌对黄瓜枯萎病的田间防治效果均达到78%以上,且以棘孢木霉525的田间防治效果最高,达到81.53%。与只接种枯萎病病原菌的对照相比,3株木霉菌单独接种或与黄瓜枯萎病病原菌同时接种均可以显著提高黄瓜幼苗株高、茎粗、叶面积、根体积、地上部鲜重、地下部鲜重,显著提高黄瓜叶片超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）、多酚氧化酶（PPO）活性,显著降低质膜透性和丙二醛（MDA）含量,其中以拟康氏木霉886单独接种促进效果最强。研究表明,3种木霉菌通过促进黄瓜幼苗生长,增强植物抗氧化酶活性,降低质膜透性和丙二醛含量,从而提高对黄瓜枯萎病的抗性。     

Osmoregulatory function in ducks following ingestion of the organophosphorus insecticide fenthion

Salt gland function and osmoregulation in aquatic birds drinking hyperosmotic water has been suggested to be impaired by organophosphorus insecticides. To test this hypothesis, adult black ducks (Anas rubripes) were provided various regimens of fresh or salt (1.5% NaCl) water before, during, and after ingestion of mash containing 21 ppm fenthion. Ducks were bled by jugular venipuncture after 1, 7, and 12 days of treatment, and were then killed. Brain and salt gland acetylcholinesterase activities were substantially inhibited (44–61% and 14–36%) by fenthion. However, salt gland weight and Na⁺-K⁺-ATPase activity, and plasma Na⁺, Cl^?, and osmolality, were uniformly elevated in all groups receiving salt water including those ingesting fenthion. In a second study, salt gland Na⁺-K⁺-ATPase activity in mallards (A. platyrhynchos) was not affected after in vitro incubation with either fenthion oxon at concentrations ranging from 0.04 to 400 μM, but was reduced in the presence of 40 and 400 μM DDE (positive control). These findings suggest than environmentally realistic concentrations of organophosphorus insecticides do not markedly affect osmoregulatory function in adult black ducks.     

木霉不同施用方式对黄瓜膜脂过氧化作用、保护性酶活性及枯萎病防效的影响

采用3株对黄瓜枯萎病菌有较好拮抗作用的木霉,棘孢木霉Trichoderma asperellum 525、哈茨木霉T.harzianum 610和拟康氏木霉T.pseudokoningii 886,研究木霉不同施用方式对黄瓜幼苗叶片膜脂过氧化作用、保护性酶活性及防治枯萎病效果的影响。结果显示,所有木霉处理均能够提高黄瓜幼苗的株高和茎粗,增加幼苗地上部和地下部鲜重,对黄瓜枯萎病的防效均达到64%以上。播种后8~14 d,各处理的黄瓜幼苗叶片质膜透性和丙二醛(MDA)含量不同程度下降,保护性酶活性不同程度的提高,其中以先接种拟康氏木霉886,2 d后接种病原菌的T886-F处理的提高程度最显著。播种后14 d,该处理的叶片质膜透性和MDA含量比只接种病原菌的F处理分别下降了37.02%和14.80%,同时,叶片的SOD、POD、CAT、PPO活性分别比F增加了46.82%、34.93%、18.75%和11.63%。以上结果说明,本研究的3株木霉均能通过改善黄瓜幼苗叶片膜脂过氧化能力,增加保护酶活性,促进幼苗生长,防治黄瓜枯萎病。在实际应用中,提前施入木霉菌剂,有利于促进黄瓜的生长、提高幼苗抗病性、提高病害防治效果。本研究结果为木霉菌剂的高效使用、保障黄瓜安全生产提供指导。     

Plant growth enhancement and disease control byTrichoderma harzianum in vegetable seedlings grown under commercial conditions

Trichoderma harzianum was applied to cucumber and pepper seedlings as a peat-bran preparation incorporated into the propagative mixture in a commercial production nursery. On marketing day (after 18 and 30 days for cucumber and pepper, respectively), significant increases of 23.8% and 17.2% in seedling height, 96.1% and 50% in leaf area, and 24.7% and 28.6% in plant dry weight were observed in cucumber and pepper seedlings, respectively, as compared to their non-treated counterparts.Trichoderma-treated seedlings were much more developed and vigorous and had higher chlorophyll contents. No significant differences were found in N, P or K content between treatments. Cucumber seedlings were then transplanted to a commercial greenhouse and analyzed over two successive growth cycles following soil fumigation with methyl bromide (500 kg/ha). Results revealed theTrichoderma-treated plants to be more resistant to damping-off disease. During the first cycle, immediately after soil fumigation, no damping-off was observed with either treatment, except in border beds where 4% of the non-treated plants died, as compared to no damping-off in theTrichoderma-treated plants. During the second growing cycle however, significant reductions in damping-off of 67% and 52% were obtained in middle and border beds, respectively, as compared to the non-treated controls.     

Lack of prevention of chlorsulfuron-induced inhibition by amino acids

The amino acids valine and isoleucine, which had been indicated by Ray (1984) as antidotes against growth inhibition of pea plants induced by chlorsulfuron, were ineffective in preventing the damage induced by chlorsulfuron on the plasma membrane of maize roots as measured by K⁺ uptake. In addition, in seedlings of two corn genotypes and three pea genotypes grown in culture solution containing chlorsulfuron, growth inhibition was not alleviated by valine and isoleucine. However, in agreement with Ray's results, acetolactate synthase, the key enzyme of the biosynthesis of valine and isoleucine, was inhibited in vitro by chlorsulfuron in both maize and pea. It is suggested that the chlorsulfuron mechanism of action involves several sites.     

Inhibitory effects of BHC isomers on Na+-K+-ATPase,yeast growth,and nerve conduction

The biological effects of benzene hexachloride (BHC, 1,2,3,4,5,6-hexachlorocyclohexane) isomers, such as toxicity against yeasts, inhibition of beef brain Na⁺-K⁺-ATPase, and blocking action on conduction in cockroach nerve, were determined and compared with those shown by homologous alcohols and other molecules. Each type of biological activity correlated well with the physicochemical properties of the test compounds such as hydrophobicity (as defined by their partition coefficients in 1-octanol/H₂O). The insecticidal action of γ-BHC showed little correspondence to Na⁺-K⁺-ATPase inhibition or to nerve blocking in insects.     

土壤干旱对冬小麦幼苗生长和叶片生理特性的影响

采用盆栽试验对不同土壤干旱下冬小麦幼苗生长和生理特性进行了初步研究。结果表明,幼苗株高、叶面积、叶干重、茎干重、根干重、总生物量、叶水势、叶片相对含水率均随土壤水分的减少而呈降低趋势;根冠比、叶片水分饱和亏、可溶性糖含量、脯氨酸含量、质膜透性、MDA含量以及叶片SOD、POD活性均呈增加趋势。这说明,在干旱胁迫下,冬小麦幼苗一方面通过改变生物量分配策略,较多地提高根比重,以利于根系吸水增加抗旱性;另一方面,通过改变叶片的生理特性以提高抗旱性。