Identifying proteins with insecticidal activity: use of encoding genes to produce insect-resistant transgenic crops

Crops resistant to insect attack offer an alternative strategy of pest control to a total reliance upon chemical pesticides. Transgenic plant technology can be a useful tool in producing resistant crops, by introducing novel resistance genes into a plant species. This technology is seen very much as forming an integral component of a crop management programme. Several different classes of plant proteins have been shown to be insecticidal towards a range of economically important insect pests from different orders; in some cases a role in the defence of specific plant species against phytophagous insects has been demonstrated. Genes encoding insecticidal proteins have been isolated from various plant species and transferred to crops by genetic engineering. Amongst these genes are those that encode inhibitors of proteases (serine and cysteine) and α-amylase, lectins, and enzymes such as chitinases and lipoxygenases. Examples of genetically engineered crops expressing insecticidal plant proteins from different plant species, with enhanced resistance to one or more insect pests from the orders Lepidoptera, Homoptera and Coleoptera are presented. The possibility of ‘pyramiding’ different resistance genes to improve the effectiveness of protection and durability is discussed and exemplified. The number of different crop species expressing such genes is very diverse and ever-increasing. The viability of this approach to crop protection is considered. © 1998 SCI.     

昆虫线虫抑制植物线虫的机理研究进展

斯氏科(Steinernematidae)、异小杆科(Heterorhabditidae)、索科(Mermithidae)和小杆科(Rhabditidae)线虫为几类较有生物防治前景的昆虫线虫。一些昆虫线虫除了能够有效地防治有害昆虫,还具有抑制植物线虫的作用,本课题组研究发现小杆科、小杆属Rhabditis(Oscheius)的某些昆虫线虫品系也有抑制植物线虫的作用。国内在这方面报道很少,国外许多线虫研究者自20世纪80年代以来有过一些研究。本文着重报道昆虫线虫抑制植物线虫的作用机理研究进展、植物线虫的生物防治途径和昆虫线虫的应用范围。     

‘99昆明世博会参展植物寄生线虫的检疫初报

本次博览会参展植物种类多,来源广,带土普遍疫情复杂。在严处国内外的参展植物中截获矛线目和垫刃目的１３个科的５６个属,其中在参展植物中截获我国对外检疫潜在危险性线。     

植物寄生线虫乙酰胆碱酯酶结构与功能研究进展

乙酰胆碱酯酶通过水解神经递质乙酰胆碱使得神经信号能够在生物体内进行正常的传递,从而有效调节正常的肌肉活动,在生物体神经传导的过程中发挥着重要作用。植物寄生线虫乙酰胆碱酯酶具有由多种基因编码的不同分子型,在线虫化感、运动、取食、抗药性等多个方面承担着不同的功能。本文简要综述了植物寄生线虫乙酰胆碱酯酶分子型、基因结构、基因功能、组织表达及其在线虫抗药性中的作用等方面的研究进展。     

小杆线虫(Rhabditis sp.)对温室黄瓜根际植物寄生线虫的抑制作用

近年来，随着我国温室黄瓜复种指数的增大，线虫病害日趋严重。植物线虫不仅本身直接危害植物根系，导致黄瓜减产20％～50％，还常诱发一些病菌的侵害。根结线虫（Meloidogyne spp．）是温室黄瓜发生较严重的一种植物线虫，在常规栽培管理条件下，矮化线虫（Tylenchorhynchus spp．）也具有相当高的种群密度。     

Root-lesion nematodes of potato: Current status of diagnostics,pathogenicity and management

Root-lesion nematodes of the genus Pratylenchus are migratory endoparasites with worldwide economic impact on several important crops including potato, where certain species like P. penetrans, P. neglectus, and P. scribneri reduce the yield and quality of potato tubers. Morphological identification of Pratylenchus spp. is challenging, and recent advancements in molecular techniques provide robust and rapid diagnostics to differentiate species without the need of specialist skills. However, the fact that molecular diagnostics are not available for all Pratylenchus species means that there are limitations in worldwide application. In general, root-lesion nematodes are difficult to manage once introduced into agricultural land and damage can be related to pathogenicity and population densities. In addition, root-lesion nematodes interact with fungi such as Verticillium dahliae, resulting in disease complexes that enhance the damage inflicted on the potato crop. Management interventions are often focused on limiting nematode reproduction before planting crops and include the application of nematicides, and cultural practices such as crop rotation, cover crops, biofumigation, and biological control. Understanding the limitations of the available crop protection strategies is important and there are many gaps for further study. This review discusses the status of the diagnosis, distribution, pathogenicity, and management of the main species of root-lesion nematodes, reported to infect potatoes worldwide, and highlights areas for potential future research.     

Subsurface drip application of alternative fumigants to methyl bromide for controlling nematodes in replanted grapevines

BACKGROUND: Many California grape growers use preplant fumigation to ensure uniform and healthy grapevine establishment in replant situations. A field study was conducted to evaluate the performance of subsurface drip‐applied chemical alternatives to methyl bromide on plant‐parasitic nematodes, plant vigor and fruit yield during the 6 year period following replanting. RESULTS: Subsurface drip fumigation with 1,3‐dichloropropene plus chloropicrin and with iodomethane plus chloropicrin had generally similar nematicide activity as methyl bromide in three grape types, while sodium azide was less effective. The combination of 1,3‐dichloropropene plus chloropicrin enhanced vine vigor similarly to methyl bromide. However, all plots treated with alternative fumigants produced less fruit yield than methyl bromide over the 4 years of evaluation. CONCLUSION: Subsurface drip fumigation with alternative chemicals to methyl bromide generally provided adequate management of plant‐parasitic nematodes during the vine establishment period. However, further research is required to increase the performance of alternative chemicals against other components of the replant problem, as grape yield in vines grown in the alternative treatments was lower than in methyl bromide. Copyright © 2011 Society of Chemical Industry     

Phenylalanine ammonia lyase activity in chilli CM-334 infected by Phytophthora capsici and Nacobbus aberrans

We tested the hypothesis that PAL activity in chilli plants CM-334 inoculated with Nacobbus aberrans (Na) alone or in combination with Phytophthora capsici (Pc), is lower than in those inoculated only with Pc. At 21 days after nematode inoculation, inoculated plants showed a significant (P < 0.01) reduction of 48% in PAL activity compared to those non-inoculated in two separate experiments. In two other tests, where plants were inoculated with the oomycete 21 days after inoculation with the nematode, PAL activity at 2, 4, 6, 8 and 24 h after inoculation with Pc was significantly higher (Tukey, P < 0.01) in plants inoculated only with Pc than in plants inoculated only with Na or both pathogens (Na+Pc).     

植物抗线虫基因与抗性机理研究进展

植物寄生线虫是严重危害农业生产的一类重要病原生物,对全球作物产量造成重大损失.抗线虫基因在植物抗线虫反应中发挥重要作用,发掘抗线虫基因并培育抗线虫品种是防治线虫病害的一条有效途径.抗线虫基因的定位与克隆对解析植物抗线虫性的分子机理做出了巨大贡献,明确线虫与寄主植物之间的互作关系及抗线虫机制,可以为制定和采取更加有效的防控策略提供借鉴.     

Insecticide toxicity and synergism by enzyme inhibitors in 18 species of pest insect and natural enemies in crucifer vegetable crops

The toxicities of three enzyme inhibitors and their synergistic effects on four insecticides were studied by using the dry film method on field populations of 18 species of insects collected in Jianxin and Shanjie, China, from 2003 to 2005. Meanwhile, the inhibitory effects of these enzyme inhibitors on the activities of acetylcholinesterases (AChE), carboxyesterases (CarE) and glutathione-S-transferases (GST), in vivo, were also studied. In general, triphenyl phosphate (TPP) and diethyl maleate (DEM) showed low toxicities to six herbivorous pest insects, four ladybirds and eight parasitoids. Piperonyl butoxide (PB) exhibited low toxicities to the herbivorous pest insects and ladybirds, but high toxicities to the eight parasitoids. The tolerance to the insecticides in 11 pest insects and natural enemies was mainly associated with the tolerance to PB. PB showed the highest synergism on methamidophos, fenvalerate, fipronil and avermectin in nine species of pest insects and natural enemies. In general, TPP and DEM showed significant synergisms to these four insecticides in four parasitoid species. However, in contrast to their effects on the parasitoids, the synergistic effects of TPP and DEM on the four insecticides by TPP and DEM against four pest insects and one ladybird varied depending on the insect species and enzyme inhibitor. Activity of AChE, CarE or GST could be strongly inhibited, in vivo, by PB, TPP or DEM, depending on the insect species and enzyme inhibitors. From the results obtained in this study, mixed-function oxidase (MFO) was thought to play the most critical role in insect tolerances to the tested insecticides in the field. Low competition existed in the evolution of insecticide resistance in the field populations of parasitoids, as compared with herbivorous pest insects and ladybirds. Possible causes of the high synergistic effects of PB on the four classes of insecticides, based on multiattack on the activity of CarE, GST or AChE in the insect species, are also discussed.     

近年水稻主要线虫病害的研究进展

目前,以拟禾谷根结线虫Meloidogyne graminicola、水稻干尖线虫Aphelenchoide besseyi、水稻潜根线虫Hirschmanniella oryzae、水稻茎线虫Ditylenchus angustus和旱稻孢囊线虫Heterodera elachista为主的水稻寄生线虫侵染水稻以及其它禾本科作物引致的线虫病害,已造成了严重的经济损失。近年来,对拟禾谷根结线虫的致病机制和寄主的防御机制等方面的研究已取得了显著进展,但对其它水稻寄生线虫与寄主互作机制的研究还存在较大空白。水稻寄生线虫的防治多依赖化学农药,抗性品种和生物防治等措施的实施存在一定的局限性。本文对上述5种主要水稻寄生线虫病的发生为害、致病机制以及防治方法等进行了综述,并对水稻寄生线虫致病机理的研究以及抗性品种、生物防治和诱导化合物的应用进行了探讨。     

南海口岸进境林木种苗线虫检疫情况分析

本文汇总了2008～2010年南海口岸进境林木种苗的线虫截获情况,共截获15科26属超过40种植物寄生线虫,其中检疫性线虫有7种,分别为短体线虫(非中国种)Pratylenchus Filipjev(non-Chinese species)、毛刺线虫属(传毒种类)Trichodorus Cobb(The species transmit viruses)、根结线虫(非中国种)Meloidogyne Goeldi(non-Chinese species)等;根据携带线虫状况分析了检疫性线虫的截获情况,提出了防止危险性线虫入境、扩散措施,为口岸防止线虫入境提供借鉴。     

植物寄生线虫滞育机制研究进展

植物寄生线虫防治困难与其在土壤中存活时间和抗逆能力有直接关系。休眠或滞育是其度过逆境条件或缺乏寄主时的主要策略。根据植物寄生线虫休眠或滞育的特点和规律可确定防治的关键时期。本文就植物寄生线虫休眠或滞育的影响因素和不同寄生线虫的滞育特点, 以及一些植物寄生线虫休眠或滞育生理生化机制的研究进展进行综述, 以期为植物寄生线虫的综合防治提供新的思路。     

RNA interference technology in crop protection against arthropod pests,pathogens and nematodes

Scientists have made significant progress in understanding and unraveling several aspects of double‐stranded RNA (dsRNA)‐mediated gene silencing during the last two decades. Now that the RNA interference (RNAi) mechanism is well understood, it is time to consider how to apply the acquired knowledge to agriculture and crop protection. Some RNAi‐based products are already available for farmers and more are expected to reach the market soon. Tailor‐made dsRNA as an active ingredient for biopesticide formulations is considered a raw material that can be used for diverse purposes, from pest control and bee protection against viruses to pesticide resistance management. The RNAi mechanism works at the messenger RNA (mRNA) level, exploiting a sequence‐dependent mode of action, which makes it unique in potency and selectivity compared with conventional agrochemicals. Furthermore, the use of RNAi in crop protection can be achieved by employing plant‐incorporated protectants through plant transformation, but also by non‐transformative strategies such as the use of formulations of sprayable RNAs as direct control agents, resistance factor repressors or developmental disruptors. In this review, RNAi is presented in an agricultural context (discussing products that have been launched on the market or will soon be available), and we go beyond the classical presentation of successful examples of RNAi in pest‐insect control and comprehensively explore its potential for the control of plant pathogens, nematodes and mites, and to fight against diseases and parasites in beneficial insects. Moreover, we also discuss its use as a repressor for the management of pesticide‐resistant weeds and insects. Finally, this review reports on the advances in non‐transformative dsRNA delivery and the production costs of dsRNA, and discusses environmental considerations. © 2017 Society of Chemical Industry     

TuMV management for brassica crops through host resistance: retrospect and prospects

Turnip mosaic virus (TuMV), the only potyvirus known to infect brassicas, is a devastating virus threatening many economically important brassica crops, including cabbage, Chinese cabbage, oilseed rape and mustard. TuMV disease, which was first discovered in the United States, is now found worldwide, especially in Europe, Asia and North America. TuMV results in a yield loss of up to 70% and has a wide host range, infecting most cruciferous plants, as well as many non-cruciferous species. This virus is also characterized by high pathotype diversity because of its highly variable genome structure and has been divided into 12 pathotypes. These characteristics, as well as its nonpersistent transmission mode by as many as 89 aphid species, mean the disease is difficult to prevent through traditional methods such as the application of chemicals, prompting researchers to seek host resistance for effective control. During the last decade, extensive studies have been conducted to investigate inheritance, mapping and cloning of the TuMV resistance genes, and several NB-LRR- or eIF-encoding loci with divergent molecular mechanisms have been uncovered. These studies have greatly facilitated resistance breeding for brassica crops and have advanced our understanding of virus−host interactions.     

拮抗南方根结线虫PKS基因的筛选及活性评价

土壤环境中蕴藏着巨大的微生物资源,而99%的土壤微生物不能用传统的纯培养。本研究基于西藏米拉山高寒草甸土壤微生物宏基因组Fosmid文库,采用PCR方法从文库中筛选PKS基因,以南方根结线虫(Meloido-gyne incognita)为靶标测定其杀线活性,并验证温室盆栽防效。从Fosmid文库中筛选得到2个含PKS基因的克隆K99和K82。K99属于Ⅰ型PKS,K82属于杂合PKS/NRPS;K99发酵液、热处理发酵液浸泡南方根结线虫J2 12 h后,杀线活性为100%。温室盆栽防效结果表明,K99的发酵菌液对南方根结线虫的防效为89%。K99能产生一种外分泌有杀线活性的物质,其热稳定性好,具有重要的开发利用价值。     

江苏进出口园林植物寄生线虫调查及检疫处理技术

1997－1998年，对南京口岸进口的荷兰、加拿大、美国、韩国、台湾和香港等国家和地区41批（次）园林植物检疫中，1次截获鳞球茎类线虫、19交截获咖啡短体线虫、草莓芽叶线虫、克鲁克剑线虫、毛刺尾线虫、南方根结线虫、花生根结线虫、北方根结线虫和其它根结线虫等，进口的园林植物中危险性疫情发生率高达34％。对江苏5个主要出口园林植物生长基地进行的植物寄生线虫种类调查，在59种经常性出口的园林植物中，发现27属植物寄生线虫，其中已鉴定出13种。对红花木莲苗上的南方根结线虫、牡丹苗上的腐烂茎线虫进行的检疫处理试验表明，用几种农药混配的处理液加热后处理植物的根部，可以有效地杀死这两种植物寄生线虫，保证货物安全出口。     

Diclofop‐resistant Lolium rigidum from northern Greece with cross‐resistance to ACCase inhibitors and multiple resistance to chlorsulfuron

Repeated use of ACCase‐ and ALS‐inhibiting herbicides in northern Greece has resulted in the evolution of a population of Lolium rigidum resistant to diclofop and chlorsulfuron. The biotype from Athos was highly resistant to diclofop and also exhibited differential cross‐resistance to clodinafop, fluazifop, tralkoxydim and sethoxydim. Assay of ACCase activity confirmed that the resistant biotype was tenfold more resistant to diclofop than the susceptible biotype, suggesting that the resistance mechanism could involve an altered target site. The diclofop‐resistant biotype has also exhibited multiple resistance to chlorsulfuron and the mechanism for this is unknown. Seed‐bioassay was found to be a rapid, cheap and reliable method to identify populations of L rigidum resistant to ACCase inhibitors and chlorsulfuron. Moreover, root elongation in the seed bioassay was more sensitive to ACCase inhibitors and chlorsulfuron than shoot elongation. © 2000 Society of Chemical Industry