Purification,regulation and cloning of a glutathione transferase (GST) from maize resembling the auxin-inducible type-III GSTs

The glutathione transferases (GSTs) from maize (Zea mays L.) with activities toward the chloroacetanilide herbicide metolachlor and the diphenyl ether herbicide fluorodifen were fractionated into two pools based on binding to affinity columns. Pool 1 GSTs were retained on Orange A agarose and were identified as isoenzymes Zea mays (Zm) GST I-I, Zm GST I-II and Zm GST I-III, which have been described previously. Pool 2 GSTs selectively bound to S-hexyl-glutathione-Sepharose and were distinct from the pool 1 GSTs, being composed of a homodimer of 28.5 kDa subunits, termed Zm GST V-V, and a heterodimer of the 28.5 kDa polypeptide and a 27.5 kDa subunit, termed Zm GST V-VI. Using an antibody raised to Zm GST V-VI, a cDNA expression library was screened and a Zm GST V clone identified showing sequence similarity to the type-III auxin-inducible GSTs previously identified in tobacco and other dicotyledenous species. Recombinant Zm GST V-V showed high GST activity towards the diphenyl ether herbicide fluorodifen, detoxified toxic alkenal derivatives and reduced organic hydroperoxides. Antibodies raised to Zm GST I-II and Zm GST V-VI were used to monitor the expression of GST subunits in maize seedlings. Over a 24 h period the Zm GST I subunit was unresponsive to chemical treatment, while expression of Zm GST II was enhanced by auxins, herbicides, the herbicide safener dichlormid and glutathione. The Zm GST V subunit was more selective in its induction, only accumulating significantly in response to dichlormid treatment. During development Zm GST I and Zm GST V were expressed more in roots than in shoots, with Zm GST II expression limited to the roots.     

Herbicide-quinone competition in the acceptor complex of photosynthetic reaction centers from Rhodopseudomonas sphaeroides: a bacterial model for PS-II-herbicide activity in plants

A select group of herbicides that inhibit photosystem II also act at the acceptor side of the reaction center (RC) from the photosynthetic bacterium Rhodopseudomonas sphaeroides, with much the same relative specificity as in plants. These include the triazines and some phenolic compounds. The proposal that herbicides inhibit the electron transfer from the primary quinone (QA) to the secondary quinone (QB) by competing for the secondary quinone binding site--the B-site--[5], is tested here with terbutryn, the most potent of the triazines. Competition between terbutryn and ubiquinone (Q-10) was observed using the kinetics of the back-reaction as a measure of inhibition. The model includes binding equilibria before and after flash activation. The binding constants for the preflash (dark) equilibria, for reaction centers in 0.14% lauryl dimethylamine-N-oxide (LDAO), were KDi = 0.8 microM terbutryn, KDq = 2 microM Q-10; both are detergent-concentration dependent. After flash activation, binding equilibrium is not fully restored on the time scale of the back-reaction because terbutryn unbinds slowly. This gives rise to biphasic decay kinetics from which koff for terbutryn was estimated to be 3 sec-1. Titrations of the rate of the slow back reaction indicated that the post-flash equilibrium is less sensitive to inhibitor, in a manner that is independent of the much stronger binding of the semiquinone, Q-B, and indicative of a direct effect of the redox state of QA on the affinity of the B-site for ligands. However, the effects on KLi and KDq could not be separated: either KLi greater than KDi or KLq less than KDq. Some triazine-resistant mutants have been isolated and are described. All appear to be herbicide binding site mutants. Whole cells and photosynthetic membrane vesicles (chromatophores) exhibit a 10-50-fold increase in resistance to triazines due, in large part, to an increase in the rate of unbinding (koff). The modifications of the binding site appear to diminish the affinity of the B-site for ubiquinone as well as terbutryn. It is concluded that bacterial RCs are a useful model for the study of herbicide activity and specificity.     

二氯喹啉酸对稗草生长发育及生长素含量的影响

通过外施二氯喹啉酸、生长素处理,研究二氯喹啉酸对稗草Echinochloa crusgalli生长特性、蛋白浓度、过氧化物酶及超氧化物歧化酶活性、生长素含量的影响。结果表明,800 g·hm-2二氯喹啉酸处理24 h未对S型稗草的生长产生明显抑制,而二氯喹啉酸单独处理及外施IAA的二氯喹啉酸处理均能显著增加R型稗草鲜重。在不施药、二氯喹啉酸单独处理、二氯喹啉酸与IAA共同处理的条件下,R型稗草比S型稗草的蛋白浓度分别提高23.5%、29.8%和64.0%,R型稗草表现出较高的抗逆境耐受能力。经二氯喹啉酸处理后,两种生物型稗草SOD活性上升,POD活性下降。施外源IAA能提高S型稗草POD活性,而对R型稗草SOD、POD活性影响不显著。提高生长素含量可显著促进R型稗草的生长。     

Hydrilla tuber formation in response to single and sequential bensulfuron methyl exposures at different times

Hydrilla (Hydrilla verticillata (L.f.) Royle) grown in outdoor tanks was exposed to bensulfuron methyl concentrations of 25, 50, or 100 ppb on June 16, August 20 or October 15; 50 ppb June 16 and August 20, or 25 ppb on June 16, July 21, August 20, and October 15, 1990, with a 35-day contact time. Hydrilla was also exposed to the compound on August 9, 1991 at concentrations of 10, 20, 30, 40, or 50 ppb. In 1990, the August 20 exposure resulted in the greatest inhibition of tuber production for a single application. Exposure in June caused hydrilla to produce at least twice as many tubers as unexposed plants by April 10, 1991. Exposure in October arrested tuber production, which had already begun. Exposure in June and August delayed tuber formation until after February 9, 1991. Exposure in June, July, August, and October inhibited tuber formation for the entire growing season. Hydrilla treated with all concentrations of bensulfuron methyl on August 9, 1991 produced tubers only sporadically through March 16, 1992, compared to unexposed hydrilla, which produced an average of 48 tubers/531 sq cm by January 4, 1992. With the onset of warmer weather after March 16, tubers produced by unexposed hydrilla more than doubled, and comparable numbers of new tubers were produced by plants that were exposed to 10 or 20 ppb. Tuber production was inhibited for the entire growing season by exposure to 50 ppb on August 9, 1991. In spite of the promise that bensulfuron methyl showed for use in aquatic plant management, the Experimental Use Permit was not renewed in 1992 and efforts to register the compound were discontinued.     

Crop responses to applications of both chlorsulfuron and monoammonium phosphate

Field studies were conducted at two locations of different soil pH in Saskatchewan using preemergence chlorsulfuron and granular monoammonium phosphate. In year one, barley was not adversely affected by chlorsulfuron applied in the spring at 11.25 and 22.5 g a.i. ha^–1 (Kernen location, soil pH 7.5) or 22.5 and 45.0 g a.i. ha^–1 (Scott location, soil pH 5.5). In year two, the residual chlorsulfuron did not affect flax seed yield on the acid soil, but caused substantial crop damage to flax and lentil on the neutral soil. In general, flax and lentil were more injured by chlorsulfuron residues where lower rates of MAP had been applied the previous year, suggesting that the presence of a MAP band in the soil at the time of herbicide application may have slowed the rate of chlorsulfuron degradation. A corn-root bioassay procedure was adapted to estimate residue levels in soil samples taken from the field trials.     

Use of two grasses for the phytoremediation of aqueous solutions polluted with terbuthylazine

The capacity of two grasses, tall fescue (Festuca arundinacea) and orchardgrass (Dactylis glomerata), to remove terbuthylazine (TBA) from polluted solutions has been assessed in hydroponic cultures. Different TBA concentrations (0.06, 0.31, 0.62, and 1.24 mg/L) were chosen to test the capacity of the two grasses to resist the chemical. Aerial biomass, effective concentrations (to cause reductions of 10, 50, and 90% of plant aerial biomass) and chlorophylls contents of orchardgrass were found to be more affected. Tall fescue was found to be more capable of removing the TBA from the growth media. Furthermore, enzymes involved both in the herbicide detoxification and in the response to herbicide-induced oxidative stress were investigated. Glutathione S-transferase (GST, EC. 2.5.1.18) and ascorbate peroxidase (APX, EC. 1.11.1.11) of tall fescue were found to be unaffected by the chemical. GST and APX levels of orchardgrass were decreased by the treatment. These negative modulations exerted by the TBA on the enzyme of orchardgrass explained its lower capacity to cope with the negative effects of the TBA.     

腈水解酶基因bxn的结构与功能研究

前期研究发现腈水解酶基因bxnRD127表达产物无功能,测序证明该基因结构中存在4处突变。本文对4处突变逐点进行结构与功能回复突变研究,发现其中2处突变能使基因表达产物丧失功能,1处使基因表达产物活性降低,1处对基因表达产物活性基本无影响。进一步将完全回复突变的bxn基因转化烟草,获得的转基因烟草具有抗溴苯腈除草剂特性,结果证明结构中的3处突变与活性中心功能有关。     

几种除草剂对薇甘菊的杀灭试验

在广东内伶仃岛保护区东湾和深圳福田红树林自然保护区两地,用2,4-D、草甘膦、25%森泰、森草净4种除草剂对薇甘菊进行杀灭试验,结果表明50～500倍液的2,4-D和草甘膦均只能杀灭薇甘菊的地上营养体,不能杀死根部;用25%森泰水剂注射薇甘菊主根,0.1～3mL/株的用量,可以在5～6个月内彻底杀灭薇甘菊;用森草净水溶性粉剂配制成的溶液,喷洒薇甘菊的茎叶,用药量在1～1000g/hm     

几种除草剂对薇甘菊的杀灭试验

在广东内伶仃岛保护区东湾和深圳福田红树林自然保护区两地,用2,4－D、草甘膦、25％森泰、森草净4种除草剂对薇甘菊进行杀灭试验,结果表明：50－500倍液的2,4－D和草甘膦均只能杀灭微甘菊地地上营养体,不能杀死根部;用25％森泰水剂注射微甘菊主根,0．1－3mL株的用量,可以在5－7个月内彻底杀灭微甘菊;用森草净水溶性粉剂配制成的溶液,喷洒薇甘菊的茎叶,用药量在1－1000g/hm^2时,可在2－3个月内彻底杀灭微甘菊。     

Role of calcium in the inhibition of nitrogenase activity by Methylparathion and Benthiocarb in the cyanobacterium Nostoc muscorum

Methylparathion and Benthiocarb inhibition of N₂ fixation in the cyanobacterium Nostoc muscorum was reversed by Ca²⁺ at 1 mm but not at 0.1 mm. The concentration of intracellular Ca²⁺ was relatively high in the presence of these pesticides when 1 mm Ca²⁺ was also present, indicating that intracellular Ca²⁺ may participate in protecting nitrogenase activity against Methylparathion and Benthiocarb.The authors are with the Department of Biochemistry, University College of Science, 35 Ballygunge Circular Road, Calcutta 700 019, India.     

VA-mycorrhizae and mycorrhiza stimulating isoflavonoid compounds reduce plant herbicide injury

Imazaquin, imazethapyr and pendimethalin showed high toxicity to sorghum plants grown in a greenhouse soil mix. However, mycorrhizal sorghum plants were less affected by herbicide toxicity than non-mycorrhizal ones, at low to moderate herbicide concentrations. VAM herbicide safening effects were more evident on imazaquin-treated plants, than for those treated with the other two herbicides. Applications of imazethapyr and pendimethalin at the two highest concentrations, but not imazaquin, reduced VAM colonization rates in sorghum. Applications of the VAM stimulating isoflavonoids, biochanin A and formononetin, at 5 ppm solutions to a field soil sample containing toxic levels of imazaquin (13 ppb) and indigenous VAM fungi, reduced herbicide-induced injury in corn and sorghum under growth chamber conditions. The benefits of isoflavonoids were reduced when additional propagules of Glomus intraradix were added into field-soil samples, and were eliminated when VAM fungi were inactivated by autoclaving. This indicates that herbicide safening effects of biochanin A, and formononetin are VAM-mediated and also suggests the potential use of these isoflavonoids as herbicide safeners.     

农作物抗除草剂遗传工程研究进展

控制杂草提高农作物产量是农业生产中共同面临的问题,发展抗除草剂农作物将是最经济最方便控制杂草的技术。由于对除草剂的作用模式和除草剂代谢途径的了解,弄清了除草剂的关键靶酶及其基因,因此分离除草剂靶酶基因,克隆能解毒除草剂的酶基因,通过转化技术可获得抗除草剂农作物,大量的抗除草剂转基因农作物大田试验表明,将最有希望在２０００年进入市场。     

Effect of the Herbicide Atrazine on the Bean Leaf Lipids

Eight-day-old bean plants, grown in a nutrient solution, were sprayed with 10⁻⁵, 10⁻⁴, 10⁻³, and 10⁻² M atrazine. The lipid changes in primary and trifoliate leaves were studied 6 d after the herbicide application. The atrazine treatment inhibited the growth of the trifoliate leaves, and decreased photosynthetic and transpiration rates, the stomatal conductance, and the total lipid content. Atrazine treatment increased 16:0 and 18:3 acids and decreased trans-3-hexadecenoic and 18:2 acids in the phospholipids. The herbicide also increased 16:0 and 18:1 acids in glycolipids and decreased 18:3 acid in monogalactosyl diacylglycerols and digalactosyl diacylglycerols. In most cases the marked changes in fatty acid composition of the main lipid classes were observed at 10⁻⁴ and 10⁻³ M atrazine. This revised version was published online in June 2006 with corrections to the Cover Date.     

Contrasting effects of simazine on the photosynthetic physiology and leaf morphology of two Populus clones

Differential effects of simazine (2-chloro-4,6-bis(ethylamino)- s -triazine) on the physiology of two Populus clones were investigated in a greenhouse study. Additions of 5 mg/pot simazine to young plants had no deleterious morphological or physiological effects on clone NC 5328 ( P. x euramericana cv. I 45/51; Section Aigeiros), but reduced the rate of CO₂ fixation, increased CO₂ compensation concentrations and lowered the specific leaf weight of clone NE 388 ( P. maximowiczü x P. trichocarpa cv. Kingston; section Tacamahaca). Abaxial leaf conductance to water vapor was not affected in NE 388. Deleterious effects of simazine on NE 388 were detected ca 48 h after exposure of plants to simazine and generally became more pronounced thereafter. Visual symptoms of injury were evident at ca 2 weeks after simazine application.
Toxic responses to simazine in clone NE 388 varied in different portions of the crown. Inhibition of photosynthesis and increased CO₂ compensation concentrations were more pronounced in the region of recently matured leaves, but were somewhat less in the region of expanding leaves. Older mature leaves in the lower crown region showed no visual symptoms of injury and the rate of photosynthesis and CO₂ compensation concentrations were largely unaffected.     

A Comparative Risk Assessment of Genetically Engineered,Mutagenic, and Conventional Wheat Production Systems

Wheat (Triticum aestivum L.) varieties produced using modern biotechnologies, such as genetic engineering and mutagenic techniques, have lagged behind other crop species, but are now being developed and, in the case of mutagenic wheat, commercially grown around the world. Because these wheat varieties have emerged recently, there is a unique opportunity to assess comparatively the potential environmental risks (human health, ecological, and livestock risks) associated with genetically engineered, mutagenic, and conventional wheat production systems. Replacement of traditional herbicides with glyphosate in a glyphosate-tolerant (genetically engineered) wheat system or imazamox in an imidazolinone-tolerant (mutagenic) wheat system may alter environmental risks associated with weed management. Additionally, because both systems rely on plants that express novel proteins, the proteins and plants themselves may impose risks. The purpose of our study was to examine comparatively the multiple aspects of risk associated with different wheat production systems in the US and Canada using the risk assessment paradigm. Specifically, we used tier 1 quantitative and qualitative risk assessment methods to compare specific environmental risks associated with the different wheat production systems. Both glyphosate and imazamox present lower human health and ecological risks than many other herbicides associated with conventional wheat production systems evaluated in this study. The differences in risks were most pronounced when comparing glyphosate and imazamox to herbicides currently with substantial market share. Current weight-of-evidence suggests that the transgenic CP4 EPSPS protein present in glyphosate-tolerant wheat poses negligible risk to humans, livestock, and wildlife. Risk for mutated AHAS protein in imidazolinone-tolerant wheat most likely would be low, but there are not sufficient effect and exposure data to adequately characterize risk. Environmental risks for herbicides were more amenable to quantitative assessments than for the transgenic CP4 EPSPS protein and the mutated AHAS protein.     

Degradation of the herbicide diclofop-methyl in soil and influence of pesticide mixtures on its persistence

The degradation of the herbicide [¹⁴C]-diclofopmethyl was investigated in moist parabrown podzol soil at 22°C. Radiochemical procedures were used to monitor the herbicide breakdown. The mineralization of the uniformly labelled aromatic ring was pursued by trapping the¹⁴CO₂ generated for 96 days. Diclofop-methyl was rapidly degraded in the soil with a half-life of about 8 days. The major breakdown product was the corresponding acid-diclofop, formed by a very rapid hydrolysis of the esterbond. With time the acid appeared to undergo strong binding or complexing to the soil. An intermediate 4-(2,4-dichlorophenoxy) phenol was recovered from the treated soil. Concentration of the phenoxyphenol increased upto 6 days followed by quick decline. Insecticide combination of parathion + Demeton-Smethylsulphoxide partially inhibited diclofop degradation in the soil     

盐藻对除草剂草丁膦的抗性研究

对分离出的单藻落盐藻进行除草剂草丁膦的抗性实验,液体培养结果显示,野生盐藻对除草剂草丁膦敏感,3.0mg/L剂量的草丁膦能完全抑制盐藻的生长和增殖。     

Management of Microstegium vimineum Invasions and Recovery of Resident Plant Communities

Restoration of communities invaded by exotic plants requires effective eradication of the invader and reestablishment of the resident plant community. Despite the commonly cited need for techniques to accomplish such goals, studies that test strategies for removing invasive plants, monitor effects on resident communities, and incorporate replicate sites are generally lacking. Microstegium vimineum is an exotic annual grass that is rapidly invading forests in the eastern United States and threatening to reduce biodiversity and inhibit forest regeneration. I conducted a field experiment at eight sites over two growing seasons in southern Indiana to evaluate hand-weeding (HW), a postemergent grass-specific herbicide (POST), and the postemergent herbicide plus a preemergent herbicide (POST + PRE) for removing Microstegium . Compared to reference plots (REF), the three treatments each reduced Microstegium biomass at the end of the growing seasons to relatively low levels. However, after the second year of the experiment, POST and POST + PRE resulted in very little spring cover of Microstegium , but HW plots were significantly reinvaded. HW and POST, but not POST + PRE, increased resident plant community productivity and spring resident community cover compared to reference plots. The amount of light at the research sites did not alter the effectiveness of treatments, but the recovery of resident communities was positively correlated with light availability under HW and POST + PRE. These results indicate that natural systems invaded by Microstegium can be restored using the POST or HW treatments, which will effectively remove the invasion and allow the resident plant community to recover when used over multiple growing seasons.