HPPD抑制剂的研究进展

本从五个方面综述了对羟基苯基丙酮酸双氧化酶（HPPD）抑制剂的研制进展；1HPPD的特性和它的抑制剂的除草基本原理，2HPPD抑制剂品种，3HPPD抑制剂的结构特性和活性关系，4HPPD抑制剂的合成方法，5近期HPPD抑制剂的研制状况。     

对羟基苯基丙酮酸双氧化酶抑制剂筛选方法研究进展

对羟基苯基丙酮酸双氧化酶(HPPD)是植物体正常生长所必需的质体醌和生育酚生物合成路径中的关键酶,已成为当前最重要的除草剂作用靶标之一。发展快速、可靠的HPPD抑制剂筛选方法对研究小分子化合物与HPPD之间的相互作用,以及开展基于靶酶结构的新型HPPD抑制剂设计均具有重要意义。结合HPPD的结构和功能,文章从生物分析的角度分别就高效液相色谱、同位素标记、偶联法、电化学及简易筛选模型等方法在HPPD抑制剂筛选中的运用进行了总结,并对发展HPPD抑制剂的高通量筛选方法进行了展望。     

HPPD抑制剂的研究进展

对新型除草剂对羟苯基丙酮酸双氧化酶 (HPPD)抑制剂的发现过程、作用机制、结构特征、构效关系以及合成方法作了较为详细的综述     

新型吡唑-喹唑啉二酮类对羟基苯丙酮酸双加氧酶抑制剂的设计、合成及生物活性

对羟基苯丙酮酸双加氧酶 (HPPD) 抑制剂近年来因其高活性和低抗性风险成为了除草剂领域的研究热点。喹唑啉二酮被证明是一类具有潜力的骨架结构,为了继续发挥该骨架的优势,在前期工作的基础上,结合已有的构效关系研究结果,设计并合成了30个新型含喹唑啉二酮结构的吡唑类HPPD抑制剂,其结构均经过了高分辨质谱 (HRMS)、核磁共振氢谱 (1H NMR) 和碳谱 (13C NMR) 的表征。酶水平和活体活性测试结果表明,大部分目标化合物展现出了与对照药剂喹草酮相当甚至更优异的酶抑制活性。温室除草活性测试结果表明,目标化合物对6种供试杂草均有一定的生长抑制作用,特别是化合物 9-28 ,在有效成分150 g/hm2剂量下,对6种杂草的防治效果均在80%以上,其中对稗草和马唐的防治效果达100%。最后,培养了代表性化合物 9-28 与拟南芥HPPD的复合物晶体结构,从分子层面说明了该类抑制剂与靶标的结合模式,也为后续该类抑制剂的开发提供了一定的分子基础和设计思路。     

环吡氟草酮

正中文通用名称:环吡氟草酮环吡氟草酮是青岛清原化合物有限公司自主研发的新型HPPD抑制剂类除草剂,由江苏清原农冠杂草防治有限公司独家登记的新农药品种。环吡氟草酮通过抑制HPPD的活性,使对羟基苯基丙酮酸转化为尿黑酸的过程受阻,从而导致生育酚及质体醌无法正常合成,影响靶标体内类胡萝卜素合成,导致叶片发白。环吡氟     

农药中色素合成抑制剂的研究和应用进展

光合作用在植物的生长过程中起着至关重要的作用,以光合色素生物合成过程中的酶作为靶标,是研发除草剂的一个重要方向和热点。其中原卟啉原氧化酶(PPO),八氢番茄红素去饱和酶(PDS),ζ-胡萝卜素去饱和酶(ZDS),对羟苯基丙酮酸双氧化酶(HPPD)等作为除草剂靶酶非常成功。本文综述了近年来农药中色素合成抑制剂的作用机制及最新应用进展,并展望未来的发展趋势。     

农药学学报第3卷(2001年)总目次

综述光活化农药的研究与应用蒋志胜　尚稚珍　万树青　徐汉虹　赵善欢 ( 1 ) 1…………………植物病原菌对杀菌剂抗性风险评估王文桥　马志强　张小风　张文吉 ( 1 ) 6…………………QSAR及其在农药设计中的应用和进展钟国华　胡美英 ( 2 ) 1…………………………………植物白粉病的化学防治进展周益林　段霞瑜　盛宝钦 ( 2 ) 1 2……………………………………HPPD抑制剂的研究进展吴彦超　胡方中　杨华铮 ( 3) 1………………………………………细胞色素 P4 50 s的纯化与鉴定郑明奇　张文吉　邱星辉 ( 3) 1 1………………………………分…     

双环磺草酮在我国稻田应用的现状、存在问题及对策建议

双环磺草酮是登记用于水稻移栽和直播田防除一年生杂草的对羟基苯基丙酮酸双氧化酶(HPPD)抑制剂类除草剂。本文简述了双环磺草酮的研发概况及其产品特性,并结合笔者近年来对双环磺草酮的研究以及在推广实践中了解掌握的情况,全面分析了双环磺草酮在我国稻区的应用现状,指出了存在的问题,提出了解决问题的对策建议,以期为双环磺草酮的科学使用提供参考。     

含咪唑啉酮结构的吡唑类对羟基苯基丙酮酸双加氧酶抑制剂的除草活性及选择性研究

对羟基苯基丙酮酸双加氧酶（HPPD）是一种重要的除草剂作用靶标。为了发现具有高活性和高选择性的新型HPPD抑制型除草剂,对前期合成的23个含咪唑啉酮结构单元的吡唑类衍生物（2A~2W）进行了深入的生物活性评价和构效关系研究,比较了它们对拟南芥HPPD（AtHPPD）和人源HPPD（hHPPD）抑制活性的差异,从酶水平上总结了该类化合物的结构-活性关系和种属选择性规律,从活体植株水平研究了它们的除草活性。结果表明:部分化合物表现出良好的除草活性和作物安全性,其中化合物2E和2G在150 g/hm2剂量下对荠菜、繁缕、小藜和棒头草抑制活性达到80%以上,且其对作物的安全性也明显优于商品化除草剂硝磺草酮。此外,化合物2P在酶水平上的选择性倍数高达93倍,展示出良好的应用潜力。     

美国发现新型抗性杂草

在美国内布拉斯加州一块种子用玉米生产土地上,发现一簇水萱麻杂草对苗后HPPD抑制类除草剂（如：Callisto,Laudis和Impact）产生抗性。这里已经连续5年反复使用这些HPPD抑制类除草剂。除了那些已经在该地发现的对草甘膦产生抗性的杂草如杉叶藻（marestail）、三裂豚草（giant ragweed）、地肤（kochia）,这是该州出现的新杂草抗性类型。     

广东省稻菜轮作区中牛筋草对十种除草剂的抗性水平及靶基因序列分析

为明确广东省稻菜轮作区中牛筋草对10种常用除草剂的抗性水平及抗性分子机制,采用整株生物测定法测定广东省稻菜轮作区内8个牛筋草种群P1～P8对草甘膦、草铵膦和乙酰辅酶A羧化酶(acetyl-CoA carboxylase,ACCase)抑制剂类等10种除草剂的抗性水平,并进一步分析P1和P8种群相关靶标酶基因5-烯醇丙酮酰莽草酸-3-磷酸合酶(5-enolpyruvyl-shikimate-3-phosphate synthase,EPSPS)、谷氨酰胺合成酶(glutamine synthetase,GS)和ACCase的部分功能区序列特征。结果显示,牛筋草P1～P8种群对草甘膦抗性指数为敏感种群的5.9倍～17.7倍,其中P8种群对草甘膦的抗性水平最高;8个种群对草铵膦也产生了不同程度的抗性,抗性指数为敏感种群的2.3倍～14.2倍,其中P1种群抗性最高。牛筋草P1和P8种群均对ACCase抑制剂类除草剂精喹禾灵、氰氟草酯和噁唑酰草胺产生了交互抗性;P1种群ACCase基因在第2 041位氨基酸处发生突变,该突变在牛筋草种群中首次发现;而P8种群ACCase基因则在第2 027位氨基...     

Resistance to HPPD-inhibiting herbicides in a population of waterhemp (Amaranthus tuberculatus) from Illinois, United States

BACKGROUND: A population of waterhemp in a seed maize production field in central Illinois, United States, was not adequately controlled after post‐emergence applications of herbicides that inhibit 4‐hydroxyphenylpyruvate dioxygenase (HPPD). RESULTS: Progeny from the field population survived following treatment with mesotrione, tembotrione or topramezone applied to the foliage either alone or in combination with atrazine in greenhouse experiments. Dose–response experiments indicated that the level of resistance to the HPPD inhibitor mesotrione is at least tenfold relative to sensitive biotypes. CONCLUSION: These studies confirm that waterhemp has evolved resistance to HPPD‐inhibiting herbicides. Copyright © 2011 Society of Chemical Industry     

Herbicide safety relative to common targets in plants and mammals

Most modern herbicides have low mammalian toxicity. One of the reasons for this safety is that the target site for the herbicides is not often present in mammals. There are approximately 20 mechanisms of action that have been elucidated for herbicides. Of these, some do share common target sites with mammals. The mechanisms include formation of free radicals, protoporphyrinogen oxidase (PROTOX), glutamine synthetase (GS) and 4-hydroxyphenylpyruvate dioxygenase (HPPD). PROTOX, HPPD and GS inhibitors have been shown to inhibit these enzymes in both plants and mammals and there are measurable effects in mammalian systems. However, the consequences of inhibiting a common target site in plants can be quite different than in animals. What may be a lethal event in plants, eg inhibition of HPPD, can have a beneficial effect in mammals, eg treatment for tyrosinemia type I. These chemicals also have low mammalian toxicity due to rapid metabolism and/or excretion of the herbicide from mammalian systems.     

Purification and Characterization of 4-Hydroxyphenylpyruvate Dioxygenase from Maize

The proposed target enzyme for benzoylcyclohexanedione herbicides, 4-hydroxyphenylpyruvate dioxygenase (HPPD) was purified from etiolated maize seedlings with a purification factor of 105. Enzyme activity was measured by detection of carbon dioxide formed from radiolabelled substrate. The enzyme has a pH optimum of 7·3 and an apparent molecular mass of 43 kDa, similar to that of the mammalian liver enzyme. Activity needs the presence of a reducing system glutathione/dichlorophenol indophenol or ascorbate and catalase. Surprisingly, a commercial catalase preparation of low specific activity—generally used for the enzyme assay—showed HPPD activity which was separable from the catalase activity on a gel filtration column. According to kinetic studies with purified maize HPPD, experimental herbicides from the family mentioned were strong competitive inhibitors of the plant enzyme in nanomolar range withK_i values of 5 and 15 nM for 2-(2-nitro-4-chlorobenzoyl)-5-(2-methoxyethyl) cyclohexane- 1,3-dione and 2-(2-chloro-4-methanesulfonylbenzoyl)- cyclohexane-1,3-dione (SC-0051; sulcotrione), respectively.     

p-Hydroxyphenylpyruvate dioxygenase inhibitor-resistant plants

The enzyme p-hydroxyphenylpyruvate dioxygenase (HPPD) catalyzes the formation of homogentisic acid, the aromatic precursor of plastoquinone and vitamin E. HPPD is the specific target of several herbicide families: isoxazoles, triketones and pyroxazoles. Its inhibition results in the depletion of the plant plastoquinone and vitamin E pools, leading to bleaching symptoms. These herbicides are very potent for the selective pre- and in some cases post-emergence control of a wide range of broadleaf and grass weeds in maize and rice. Their herbicidal potential raised interest in the development of highly resistant transgenic crops. This goal was first achieved by over-expression of a bacterial HPPD in crop plants, and an increased level of resistance was obtained by using a mutant enzyme. A second strategy based on bypassing HPPD in the production of homogentisate was then developed. Recently, a third strategy of resistance based on the increase of p-hydroxyphenylpyruvate substrate flux has been developed. This was achieved by the introduction of the yeast prephenate dehydrogenase gene (PDH) into transgenic plants already overexpressing HPPD. In addition to a high level of herbicide resistance, a massive accumulation of vitamin E, mainly tocotrienols, was observed in leaves of the transgenic HPPD-PDH plants.     

Weed control and selectivity in maize (Zea mays L.) with tembotrione mixtures

Field and pot trials were conducted in two sites of northern Greece (Thermi and Lepti) to study selectivity and weed control in maize (Zea mays L.) with herbicide mixtures based on tembotrione. Treatments included tembotrione (plus isoxadifen-ethyl safener) applied alone at 100 g ai/ha and three mixtures of tembotrione with: (i) rimsulfuron at 10 g ai/ha, (ii) nicosulfuron at 40 g ai/ha and (iii) foramsulfuron at 60 g ai/ha (label rates for weed control in maize). Herbicides were applied at the 7- to 8-leaf growth stage of maize in Thermi and at the 6- to 7-leaf growth stage in Lepti. Six weeks after treatment, control of rhizomatous Sorghum halepense with tembotrione alone was 63% in Thermi and 60% in Lepti, with plants showing the typical bleaching symptoms of the HPPD-inhibiting herbicides. Control of rhizomatous S. halepense with the mixtures was improved from 63% to 86% in Thermi and from 60% to 82% in Lepti compared with the single treatment of tembotrione, with plants showing only the symptoms of reddish/purplish color of the ALS-inhibiting herbicides. Control of common broadleaf weeds of maize, such as Xanthium strumarium, Amaranthus retroflexus, Datura stramonium, and Chenopodium album was excellent (92%–100%) either with tembotrione alone or with the mixtures, with plants showing the typical bleaching symptoms of the HPPD-inhibiting herbicides. None of the mixtures affected the ear length and grain yield of maize. All chemical treatments gave higher grain yield than that of the non-treated control and comparable yield to that of the weed-free control. Grass control (Echinochloa crus-galli, E. phyllopogon, and Setaria viridis) with tembotrione alone was above 90% in the pot experiments which was similar to that achieved with the mixtures. Overall, there was an adequate margin of safety in the use of tembotrione in mixture with ALS-inhibiting herbicides for improved weed control in maize, particularly where rhizomatous S. halepense is problematic.     

Activity of mesotrione on resistant weeds in maize

Mesotrione is a new callistemone herbicide that inhibits the HPPD enzyme (p-hydroxyphenylpyruvate dioxygenase) and introduces a new naturally selective tool into weed-management programmes for use in maize. Mesotrione provides control of the major broad-leaved weeds, and it can be used in integrated weed-management programmes depending on the grower's preferred weed-control strategy. At post-emergence rates of 150 g AI ha-1 or less, mesotrione provides naturally selective control of key species that may show triazine resistance (TR), e.g. Chenopodium album L, Amaranthus species, Solanum nigrum L, as well as species of weed that show resistance to acetolactase synthase (ALS) inhibitors e.g. Xanthium strumarium L, Amaranthus spp and Sonchus spp. The data presented show that resistant and susceptible biotypes of these species with resistance to triazine herbicides, such as atrazine, simazine, terbutylazine and metribuzin, or ALS-inhibitor herbicides, such as imazethepyr, remain susceptible to mesotrione. These results confirm that there is no cross-resistance in biotypes with target site resistance to triazine or ALS-inhibiting herbicides. It is important that herbicide choice and rotation becomes an integral part of planning weed management, so as to minimise the risks of crop losses from weed competition, build-up of weed seed in the soil and the further development of weed resistance across a range of herbicide modes of action.     

Sensitivity of recently naturalised Digitaria spp. populations to 4‐hydroxyphenyl pyruvate dioxygenase‐ and acetolactate synthase‐inhibiting herbicides in maize

Until recently, Digitaria aequiglumis var. aequiglumis, native to South America, and Digitaria ciliaris subsp. nubica, native to Northeast Africa, were completely overlooked in Belgium due to their close morphological resemblance to Digitaria sanguinalis and Digitaria ischaemum. One of the possible reasons for their expansion in maize fields, besides for example the lack of crop rotation, might be a lower sensitivity to post‐emergence herbicides acting against panicoid grasses. Dose – response pot experiments were conducted in the glasshouse to evaluate the effectiveness of four foliar‐applied HPPD‐inhibiting herbicides (mesotrione, sulcotrione, tembotrione, topramezone) and two foliar‐applied ALS‐inhibiting herbicides (foramsulfuron, nicosulfuron) for controlling Belgian populations of D. aequiglumis and D. ciliaris subsp. Nubica, as well as local D. sanguinalis and D. ischaemum populations. In another dose–response pot experiment, the influence of growth stage at time of herbicide application on efficacy of topramezone and nicosulfuron for Digitaria spp. control was evaluated. In general, D. aequiglumis and D. ciliaris subsp. nubica populations were less sensitive to HPPD inhibitors than D. ischaemum and D. sanguinalis populations, except for D. aequiglumis treated with topramezone. Contrary to other herbicides tested, topramezone adequately controlled all D. aequiglumis populations at doses well below maximum authorised field dose. All species tested showed a progressive decrease in sensitivity to topramezone and nicosulfuron with seedling age. A satisfactory post‐emergence control of Digitaria species in the field will require appropriate choice of herbicide and dose, as well as more timely application.