油菜乙酰乳酸合成酶抑制剂类除草剂抗性突变体M9的遗传和基因克隆

【目的】分离和鉴定自然突变的油菜乙酰乳酸合成酶抑制剂类除草剂抗性突变体M9抗性基因,阐明抗性产生的分子基础。【方法】通过配置杂交组合研究其遗传规律,应用同源序列法克隆油菜ALS家族基因,采用杂交转育和小孢子培养技术将抗性基因转育到陆奥-五十铃细胞质雄性不育（MICMS）恢复系中,进行PCR鉴定。【结果】该突变体抗性由1个显性核基因控制。克隆获得BnALS1-BnALS3,核酸序列比对发现,M9抗性是由BnALS1的点突变使蛋白序列的第638位丝氨酸（AGT）残基被天冬酰胺酸（AAT）替代所致,将此抗性基因命名为BnALS1R。抗性基因BnALS1R转育到MICMS恢复系中的PCR检测表明,所有抗除草剂的恢复系都携带有BnALS1R。【结论】突变体M9抗性由1个显性核基因控制,BnALS1第638位丝氨酸突变成天冬酰胺酸是抗性产生的分子基础。     

Cross-resistance pattern to four AHAS-inhibiting herbicides of tribenuron-methyl-resistant flixweed(Descurainia sophia) conferred by Asp-376-Glu mutation in AHAS

Flixweed(Descurainia sophia L.) is a problematic and widespread weed in winter wheat fields and has been controlled by tribenuron-methyl for more than twenty years in China. In this study, a flixweed accession(Hebei 25, HB25) with an Asp-376-Glu mutation in acetohydroxy acid synthase(AHAS) was identified and purified. The purified HB25 accession(p HB25) developed 758.1-fold resistance to tribenuron-methyl and exhibited obvious cross-resistance to four AHAS-inhibiting herbicides. The resistant/susceptible(R/S) ratios of 50% plant growth reduction(GR_(50)) to herbicides of halosulfuron-methyl, flumetsulam, imazethapyr and pyribenzoxim were 346.1, 15.7, 8.1 and 7.1, respectively. The reduced AHAS sensitivities to four different AHAS-inhibiting herbicides, which were caused by the Asp-376-Glu mutation, were responsible for the resistance and cross-resistance to AHAS-inhibiting herbicides. The R/S ratios of 50% inhibition of AHAS activity(I50) to tribenuron-methyl, halosulfuron-methyl, flumetsulam, imazethapyr and pyribenzoxim were 844.5, 532.9, 74.5, 13.3 and 5.5, respectively. The results of AHAS activity in vitro were highly correlated with that of whole-plant response experiments.     

抗药性杂草产生原因及治理策略

分析了长时间或多次使用一类或一种除草剂及使用作用靶标相同的除草剂致使杂草抗药性产生的原因,同时指出除草剂和杂草的特性也对杂草的抗性产生影响。提出了治理杂草抗性的措施：以预防为主,多种手段相结合的方式延缓和治理杂草抗药性的产生。     

抗药性杂草种群的发展及其防治对策

叙述了全球抗药性杂草种群的分布及其发展动态,杂草抗药性的原因和抗性杂草种群对作物的危害,提出了以作物轮作,栽培,来茬等农业措施为主,结合生态防治,生物防治和合理使用除草剂来治理抗性杂草,延缓杂草产生抗性的综合防治对策。     

Development of Sequence Characterized Amplified Region (SCAR) Primers for the Detection of Resistance to Sporisorium reiliana in Maize

Head smut of maize (Zea mays L.), which was caused by Sporisorium reiliana, occurred in most of the maize growing areas of the world. The purpose of this study was to develop SCAR markers for map-based cloning of resistance genes and MAS. Two sets of BC3 progenies, one (BC3Q) derived from the cross Qi319 (resistance)x Huangzao 4 (susceptible),the other (BC3M) from Mol 7 (resistance)x Huangzao 4 (susceptible), were generated. Huangzao 4 was the recurrent parent in both progenies. A combination of BSA (bulked segregant analysis) with AFLP (amplified fragment length polymorphism) method was applied to map the genes involving the resistance to S. Reiliana, and corresponding resistant and susceptible bulks and their parental lines were used for screening polymorphic AFLP primer pairs. One fragment of P13M61-152 was converted into SCAR (sequence charactered amplified fragment) marker S130. The marker was mapped at chromosome bin 2.09, the interval of a major QTL region previously reported to contribute to S. Reiliana resistance.Furthermore, S130 was highly associated with resistance to S. Reiliana, and could be useful for marker-assisted selection and facilitate map-based cloning of resistance genes.     

Alleles contributing to acetyl coenzyme A carboxylase(ACCase)resistance in keng stiffgrass(Pseudosclerochloa kengiana)populations from China

Keng stiffgrass is a grass weed that affects wheat-rice cropping systems in China.The extensive reliance on acetyl coenzyme Acarboxylase(ACCase)-inhibiting herbicides has resulted in keng stiffgrass developing resistance to these herbicides.The objective of this research was to evaluate the resistance level of the putative resistant keng stiffgrass populations to ACCase-inhibiting herbicides and to identify their molecular resistance mechanism.Whole-plant dose-response experiments demonstrated that SD-4(R),SD-11(R),and JS-25(R) populations were highly resistant to fenoxaprop,clodinafop,and fluazifop,moderately resistant to diclofop,had low resistance to sethoxydim and pinoxaden,but were sensitive to clethodim.Partial chloroplastic ACCase sequences showed that there were two copies of ACCase gene in keng stiffgrass,and all homoeologous genes were expressed.The results of sequence analyses of the ACCase CT domain revealed an isoleucine-to-asparagine substitution at position 2041 in SD-4(R) and SD-11(R) populations,and a tryptophan-to-cysteine substitution at position 2027 in the JS-25(R) population.To our knowledge,this is the first report of lle-2041-Asn and Trp-2027-Cys mutations in ACCase-resistant keng stiffgrass.In addition,three robust(derived) cleaved amplified polymorphic sequence((d)CAPS) markers have been developed to rapidly identify these mutations in the ACCase gene of keng stiffgrass.     

影响抗药性杂草发生的因素

文章主要讨论了影响杂草抗药性快速发展的一些主要因素。影响杂草抗药性发展的主要因素是由于一类或一种除草剂以及使用作用靶标相同的不同类型的除草剂的大面积和长期连续使用。另外除草剂和杂草的一些特性也影响杂草的抗药性的快速发展。     

抗除草剂转基因玉米研究

文中对抗除草剂转基因玉米的研究进行了综述。抗除草剂转基因玉米的转化方法主要有基因枪法和农杆菌介导法,其中基因枪法是转基因技术中经常使用的方法;也是在玉米中应用最成功的方法;农杆菌介导法因其具有费用低、拷贝数少、基因沉默现象少、能转化较大片段、遗传稳定性较高、再生植株可育性较好等优点也越来越受到重视。通常以Bar基因作为标记基因,采用喷施除草剂和PCR检测相结合的方法对抗除草剂转基因玉米进行检测和筛选。抗除草剂转基因玉米的安全性问题主要涉及演化为田间杂草的可能、基因漂移和田间杂草的抗药性3个方面。     

Crop/Weed Outcomes from Site-Specific and Uniform Soil-Applied Herbicide Applications

While uniform broadcast application (BC) of soil-applied herbicides is the norm, advances in geospatial technologies have enabled the deployment of site-specific herbicide application (SS). Since there are several, largely-untested decision rules for SS herbicide use, the objective of this work was to evaluate crop/weed outcomes and isoxaflutole use for a number of BC and SS strategies. Field experiments were established where isoxaflutole dose response functions were quantified from early-season Zea mays (L.), Sorghum bicolor (L.) Moench., and Abutilon theophrasti Medic. response data at multiple plots (64 in 1999; 62 in 2000) throughout a variable field. These data were then used to infer crop and weed outcomes from simulated application strategies. While BC and SS strategies that relied on the product label were similar in terms of total herbicide use, extent of crop tolerance, and A. theophrasti efficacy, the SS strategy resulted in a significant redistribution of herbicide. Using biologically effective doses (ED) resulted in the highest efficacy and herbicide use was highly species-dependent. By using a historical weed map of the field to guide herbicide application, herbicide use could be reduced some 20–48% when following the product label where seedlings were present, or >90% when using the ED for A. theophrasti. In order to maximize the potential agronomic benefits of SS, biological and spatial information of both the weed and soil should be taken into account.An erratum to this article can be found at     

Beyond patch spraying: site-specific weed management with several herbicides

Site-specific weed management can include both limiting herbicide application to areas of the field where weed pressure is above the economic threshold (patch spraying) and varying the choice of herbicide for most cost-effective weed control of local populations. The benefits of patch spraying with several, postemergence herbicides in irrigated corn were evaluated in simulation studies using weed counts from 16 fields. Patch spraying with one, two or the number of herbicides that maximized net return for a field was simulated. With patch spraying of one herbicide, the average area of a field left untreated is 34.5%. Net return increases by $3.09 ha⁻¹ compared to a uniform application without decreasing crop yield. Additional herbicides increase the average benefits with just 4% more of the field treated. With two herbicides, the increase in net return is almost tripled and herbicide use is reduced nearly 10-fold compared to patch spraying with one herbicide, and weed control is better than the uniform application in 10 fields. Using more than two herbicides for patch spraying further reduces weed escapes, but herbicide use is greater than a uniform application in 10 fields. Growers might be more willing to adopt patch spraying if more than one herbicide is used in a field.     

Development of Sequence Characterized Amplified Region （SCAR） Primers for the Detection of Resistance to Sporisorium reiliana in Maize

Head smut of maize （Zea mays L.）, which was caused by Sporisorium reiliana, occurred in most of the maize growing areas of the world. The purpose of this study was to develop SCAR markers for map-based cloning of resistance genes and MAS. Two sets of BC3 progenies, one （BC3Q） derived from the cross Qi319 （resistance）×Huangzao 4 （susceptible）, the other （BC3M） from Mol7 （resistance）× Huangzao 4 （susceptible）, were generated. Huangzao 4 was the recurrent parent in both progenies. A combination of BSA （bulked segregant analysis） with AFLP （amplified fragment length polymorphism） method was applied to map the genes involving the resistance to S. reiliana, and corresponding resistant and susceptible bulks and their parental lines were used for screening polymorphic AFLP primer pairs. One fragment of PI3M61-152 was converted into SCAR （sequence charactered amplified fragment） marker S130. The marker was mapped at chromosome bin 2.09, the interval of a major QTL region previously reported to contribute to S. reiliana resistance. Furthermore, S130 was highly and facilitate map-based cloni associated with resistance to S. reiliana, and could be useful for marker-assisted selection ng of resistance genes.     

假俭草草坪的芽后除草剂筛选研究

通过比较MCPA（2甲4氯）、Flur（使它隆）、Rim（砜嘧磺隆）3个单剂和CF（复配剂Ⅰ）、MS（复配剂Ⅱ） 2个复配除草剂对14种禾本科杂草、60种阔叶杂草和3种莎草科杂草的株防效以及对假俭草草坪中杂草的田间小区防除效果,旨在筛选出假俭草草坪的杂草综合防除效果最好的芽后除草剂。结果表明：不同除草剂对杂草的防除效果存在显著差异,除草剂对禾本科杂草的株防效从高到低为MS>CF>Rim=MCPA>Flur;对阔叶杂草的株防效排序为：MS=CF>MCPA=Flur>Rim;对莎草科防除效果排序为MS=CF>Rim>MCPA>Flur。田间小区试验结果也证明,2个复配除草剂（MS和CF）的干质量防效和株防效均显著高于3个单剂处理（MCPA、Flur、Rim）。上述结果表明,2个复配除草剂MS和CF对假俭草草坪中发生的杂草综合防除效果最好,可以推荐作为假俭草草坪芽后除草剂应用。     

杂草抗药性研究进展

 目前全球已有188种杂草（112种双子叶,76种单子叶）的324个生物型对19类化学除草剂产生了抗药性,几乎涵盖各类重要的除草剂。抗药性杂草已成为杂草治理和农业生产的严重威胁,由其引发的严重经济和安全问题倍受全球关注。研究证实,杂草体内除草剂作用位点发生改变、杂草代谢解毒能力增强、杂草的屏蔽作用或与作用位点的隔离作用是杂草对除草剂产生抗性的三大主要机制。杂草的抗药性机制相当复杂,同种杂草不仅对不同除草剂的抗药性机制不同,而且能以完全不同的抗药性机制对多种作用机制不同的除草剂产生抗药性,即交互抗性和多抗性。本文重点描述杂草对8大类重要除草剂的抗药性机制,杂草抗药性研究方法,以期为中国抗药性杂草研究和农田杂草科学治理提供参考和借鉴,并提出在杂草治理中必须汲取他人的经验教训,阐明了中国杂草抗药性研究应关注问题。     

水稻田杂草群落演化原因及趋势浅析

水稻不同的栽培制度、方式以及除草剂的选择等原因引起了水稻田杂草群落的不断演化,同时使得演化向抗药性、多年生、普遍性方向发展.从农业生产实际出发,对水稻田杂草群落演化原因及其趋势进行了综述.     

Dicamba resistance: enlarging and preserving biotechnology-based weed management strategies

The advent of biotechnology-derived, herbicide-resistant crops has revolutionized farming practices in many countries. Facile, highly effective, environmentally sound, and profitable weed control methods have been rapidly adopted by crop producers who value the benefits associated with biotechnology-derived weed management traits. But a rapid rise in the populations of several troublesome weeds that are tolerant or resistant to herbicides currently used in conjunction with herbicide-resistant crops may signify that the useful lifetime of these economically important weed management traits will be cut short. We describe the development of soybean and other broadleaf plant species resistant to dicamba, a widely used, inexpensive, and environmentally safe herbicide. The dicamba resistance technology will augment current herbicide resistance technologies and extend their effective lifetime. Attributes of both nuclear- and chloroplast-encoded dicamba resistance genes that affect the potency and expected durability of the herbicide resistance trait are examined.     

广东农田杂草防控的问题与对策

广东光温水热资源丰富,作物四季种植,杂草周年为害,已成为影响农作物生产的重要生物因子。目前,广东农田杂草防控主要依赖化学除草剂,在控制农田杂草危害和提高作物产量方面发挥了重要作用。但随着农业现代化进程的推进,杂草防控面临诸多挑战：化学除草剂的过量使用,导致杂草种群演替及抗药性问题突出,除草剂药害问题频发,防治成本增加的同时给农田生态环境带来巨大的压力;绿色防控技术储备不足,适应杂草防控的智能化和机械化手段不多,人才队伍总量不足等。针对上述问题,提出了以下相应对策：推进杂草群落动态和抗性监测预警工作;加强杂草抗性机理研究;研发推广除草剂药害诊断及防控技术;加强杂草绿色防控技术和精准除草技术研究;组建产学研联盟,增强协同创新能力;推进杂草统防统治及基层从业者培训。     

热除草技术现状和展望

除草剂的广泛使用引发了环境污染、生物多样性减少、杂草群落变迁和杂草抗药性等诸多问题。热除草方法是替代化学除草的潜在方法,热除草方法有火焰除草、热水除草、蒸汽除草、红外辐射除草、激光除草等。热除草是快速加热植物体内的水分使植物细胞破裂失去活性、死亡的方法。各种热除草技术的传热机制不同。重点介绍了各种热除草技术的机理和在国内外的发展现状及所适用的环境和发展方向。随着有机农作的发展,热除草技术将发展成熟并得到广泛应用。     

油-棉连作棉田油菜秸秆覆盖对棉田杂草发生及土壤杂草种子库的动态影响

在安徽省沿江棉区开展了油菜秸秆覆盖对棉田杂草发生、棉花生长及土壤杂草种子库影响的研究。结果表明,随着油菜秸秆覆盖量的增加,对棉田杂草的抑制效果增强。与未覆盖秸秆且不除草处理相比较,7000Kg/hm²秸秆覆盖量处理棉花单株铃数和子棉产量显著提高。7000Kg/hm²秸秆覆盖量处理在覆盖后30d、60d和120d逐步减少0-20cm土层杂草种子库密度,与全程除草剂处理较一致;随着覆盖量减少,对0-20cm土层杂草种子库密度降低幅度减小。全程除草剂处理降低0-5cm土层杂草种子库多样性,而油菜秸秆覆盖则可能增加0-5cm土层的杂草种子库多样性。3500Kg/hm²覆盖量+秸秆覆盖30d后喷施除草剂处理的抑草效果和增产效果与全程除草剂处理一致。因此,在安徽省沿江棉区油-棉连作棉田推荐使用3500Kg/hm²油菜秸秆覆盖量+秸秆覆盖30d后喷施除草剂。     

细胞色素P450酶系介导的除草剂抗性研究进展

细胞色素P450酶系在生物界中广泛分布,并且是大多数除草剂代谢第一阶段的重要酶系,对除草剂抗性的形成发挥着重要作用。结合国内外细胞色素P450酶系在除草剂代谢中的研究进展,对细胞色素P450酶系参与的杂草抗性形成及其在抗除草剂转基因作物培育中的应用等方面的研究进行了综述,旨为该领域的相关研究提供参考。     

The Economic Impact of Site-Specific Weed Control

In order to evaluate economic and ecological benefits, site-specific weed control was realised in a 4-year experiment on five fields with a GPS-guided sprayer. An average of 54% of the herbicides could be saved. Savings were strongly dependent on crop and year. For grass weed herbicides, the savings were 90% in winter cereals, 78% in maize, and 36% in sugar beet. For herbicides against broadleaf weeds, 60% were saved in winter cereals, 11% in maize, and 41% in sugar beet. The monetary savings resulting from the reduction in herbicide use varied between the crops, depending on the amount of herbicides saved and the price of the herbicides. In maize, savings of 42euro/ha were realised, in winter wheat of 32euro/ha, in winter barley of 27euro/ha and in sugar beet of 20euro/ha. Large sections of the fields needed herbicide treatment significantly less frequently. In those areas where weed density remained below the weed control threshold, flora and fauna were allowed to establish largely without disturbance.