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.     

Phylogeny of five predominant pospiviroid species in Belgium

In Belgium pospiviroids are routinely detected in various hosts. The most frequently found pospiviroids are: Citrus exocortis viroid (CEVd), Chrysanthemum stunt viroid (CSVd), Potato spindle tuber viroid (PSTVd), Tomato apical stunt viroid (TASVd) and Tomato chlorotic dwarf viroid (TCDVd). Apart from the high incidence of pospiviroids in latently-infected ornamentals, viroids have also been found in plants where they cause disease: PSTVd and TCDVd in tomatoes and CSVd in chrysanthemum. In order to gain more epidemiological data on these infections, this study has conducted phylogenetic analyses of Belgian isolates for each of these five pospiviroid species. PSTVd and CEVd-isolates show a clustering depending on host plant identity. This was not observed for TCDVd and TASVd. A very high degree of sequence similarity was noticeable for CSVd-isolates from various hosts. During the past decade, PSTVd and CSVd-infected mother plants have been systematically eradicated in Belgium after positive detection results, also when found in symptomless plants, leading to a decreased trend of these quarantine pests in the past few years. However, other non-quarantine pospiviroid species are still ubiquitously present in many ornamentals. Since these pospiviroids can be equally harmful to crops as the two quarantine pests PSTVd and CSVd, there is still a risk that transmission occurs from symptomless-infected ornamental plants to economically important crops in Belgium such as tomato, pepper and chrysanthemum.     

Identification of pests and pathogens recorded in Europe with relation to fruit imports

Within the framework of the EU project DROPSA (‘Strategies to develop effective, innovative and practical approaches to protect major European fruit crops from pests and pathogens’), a review on pests (including pathogens) that have fruit species as their host plants was conducted. The focus was on pests that have been introduced into Europe or were found in the fruit trade during the last 10–15 years. Among the 387 recorded pests, the following groups were identified: 1. fruit and vinegar flies, 2. scale insects, 3. fungi, 4. plant viruses, 5. bacteria, 6. pests of unknown risk, 7. tropical fruit pests, 8. pests that had an unexpected change of hosts, 9. fruit pests not likely to be transported on fruit and 10. ‘hitchhikers’ (non‐fruit pests intercepted on fruit/fruit plants, fruit pests intercepted on other commodities than fruit). The large number of pests identified, from different taxonomic groups and origins, shows that fruit are an important pathway for pests, threatening fruit production in Europe.     

Host-based life cycle traits and detoxification enzymes of major looper pests (Lepidoptera: Geometridae) of tea from Darjeeling Terai,India

Three polyphagous looper caterpillars, Buzura suppressaria Guenée, Hyposidra talaca Walker and Hyposidra infixaria Walker, have established themselves as severe pests of tea [Camellia sinensis (L.) Kuntze] in the plantations of sub Himalayan West Bengal (Terai region). Schima wallichii (DC.) Korth. is a naturally occurring alternative host of all these looper species. To gain an insight into looper and host plant relationships, the present work contemplates studies on host preference, host-based life cycle traits and levels of detoxification enzymes, such as general esterases (GEs) and glutathione S-transferases (GSTs). From the study, host-induction of feeding preference was evident in all the three looper species. Hyposidra spp. exhibited similar post-embryonic development periods both on tea and S. wallichii, whereas B. suppressaria reared on tea needed a longer development period than on S. wallichii. Tea-reared caterpillars of Hyposidra spp. were significantly heavier, having higher quantities of GEs and GSTs than S. wallichii-reared ones. B. suppressaria, however, exhibited similar body weights on tea and S. wallichii. While GST level was higher in tea-reared B. suppressaria, its GE quantity was higher on S. wallichii. Although tea was found to be a more suitable host for Hyposidra spp., the host S. wallichii proved marginally better than tea for supporting B. suppressaria. However, all the three looper species could utilize the foliage of tea and S. wallichii successfully. So S. wallichii trees can act as a sylvan reservoir of the looper species, prompting their possible invasion of tea plantations and thus making management of looper pests in tea more difficult.     

The role of weeds in the epidemiology of pospiviroids

Many weeds that are closely associated with horticultural activities are known as natural reservoirs of plant viruses. However, whether these weeds can also serve as hosts of pospiviroids is not well known. Pospiviroids are naked, non‐coding RNA pathogens that cause severe economic damage in many solanaceous crops. In this study, we examined the overall risk of pospiviroid spreading from weeds to economically important crops, by combining the results from previous inoculation studies with new results coming from a survey, a contact experiment and an inoculation experiment. A survey of commercial ornamental glasshouses revealed that ornamental plants mainly belonging to the Solanaceae harbour pospiviroids, in contrast to weed species sampled in the same places. No new weed hosts could be identified after testing weeds that grew in contact with Tomato apical stunt viroid (TASVd)‐infected plants of tomato (Solanum lycopersicum) and jasmine nightshade (Solanum jasminoides) in an experimental glasshouse. Finally, in mechanical inoculation experiments with TASVd, none of the six tested weed species were determined to be a host at 6 weeks after inoculation. Commonly occurring weed species therefore do not appear to play a significant role as reservoir hosts for pospiviroids. This does not rule out other potential weed hosts that have not yet been tested. Inoculation studies should include rigorous experimental protocols with a sufficient number of replicated as well as adequate positive controls. The information gained through this study may prove useful in future risk assessments for the pospiviroid group.     

Factors in the Success and Failure of Microbial Controlagents for Soil Dwelling Pests

The soil, being rich in microbes, should be an ideal environment in which to practise microbial control of insect pests, but relatively few organisms have been developed into successful microbial control agents for soil pests. Difficulty arises in implementation of microbial control, as the soil is a highly complex, competitive environment where survival of applied microbes is by no means assured. In addition, soil dwelling pests appear to show a high level of resistance to broad spectrum pathogens and microbial toxins. However, some significant successes have been achieved with bacteria, fungi and nematodes for control of a range of soil inhabiting pest species. The successful microbial controls are based on highly specific microbial/insect interactions where co-evolved pathogens can overcome the host's defences. These agents also have the ability to persist in the soil, either through the inherent stability of the organism or from recycling through the host population. While strain selection for high levels of pathogenicity and environmental competence is the essential first step in microbial control of soil dwelling pests, it must be followed by development of efficient production systems and quality control measures to ensure application of standardised, high quality products. Success in microbial control of soil dwelling pests depends on an appropriate combination of these factors, with failure likely to occur if any one of these essential components is overlooked.     

新疆有毒植物资源及其植物源杀虫剂开发应用前景

植物源杀虫剂(botanical insecticides)就是利用植物的某些部位或提取成分,加工成有杀虫作用的杀虫剂。在新疆,有毒植物资源种类繁多。通过介绍新疆有毒植物的资源概况,讨论有毒植物作为农业杀虫剂的应用现状和存在问题,提出其种质资源作为植物源杀虫剂的开发利用前景。     

Integrated pest management of coffee for small-scale farmers in East Africa: needs and limitations

Coffee in East Africa (Kenya, Tanzania and Uganda) is an important cash and export crop for small-scale farmers. The crop suffers heavy yield losses due to damage caused by a wide range of indigenous pests (insects, diseases, nematodes and weeds). Current recommended pest control measures include a combination of cultural, resistant/tolerant cultivars and the use of broad spectrum chemical pesticides. Chemical pesticides are far more popular at the farm level than any of the other recommended pest control measures. Coffee pest control strategies are often aimed at individual pests with little consideration of the implications for the total coffee pest complex and its agro-ecosystem. This unilateral approach has resulted in increased pest pressure on coffee and some of its companion crops, outbreak of new pests of coffee, development of pest strains resistant to the cheap and commonly available chemical pesticides, increased environmental problems, increased health risks to man and livestock and an overall increase in the costs of coffee production, thus forcing many farmers to neglect their coffee plantations. Measures to alleviate the above problems, particularly the high production costs, are needed to improve coffee production and increase the cash return to the small-scale farmer. Integrated pest management (IPM) offers the best prospects for solving the above problems. However, lack of national IPM policies, poor extension systems, inefficient research-extension-farmer linkage and the lack of a holistic approach will delay the development and implementation of appropriate, acceptable and sustainable IPM practices.     

The transmission of geminiviruses by Bemisia tabaci

The cosmopolitan whitefly species, Bemisia tabaci (Gennadius) and Trialeurodes taporariorum (Westwood) have always been regarded as pests to a large range of worldwide crops. Both species are capable of transmitting plant viruses, with T. vaporariorum being the vector of only a few ‘clostero’-like viruses and B. tabaci the vector of viruses in several groups. The largest group of viruses transmitted by B. tabaci are the geminiviruses and B. tabaci is known to transmit around 60 members. Until recently, B. tabaci had been associated with only a limited range of host plants within any one region, although its total potential host range was large. Virus transmission was confined within the plant host range of each regional population of B. tabaci. The emergence of the polyphagous ‘B’ biotype of B. tabaci with its increased host range of more than 600 plant species, has resulted in geminiviruses infecting previously unaffected crops. As the ‘B’ biotype spreads further into Europe, European field and glasshouse crops have been shown to be susceptible to whitefly-transmitted viruses already endemic to other parts of the world. More than 20 colonies of B. tabaci, including both ‘B’ and non-‘B’ biotypes from disparate global locations have been compared for their ability to transmit more than 20 geminiviruses. All but two highly host-specific colonies were capable of transmitting most geminiviruses tested. However, some viruses were transmitted more efficiently than others. The virus coat protein or capsid is essential for vector recognition and transmission. By comparing transmissible viruses at the molecular level to viruses that are no longer whitefly-transmissible, the active epitope on the virus coat protein could be identified for designing future virus control strategies.     

Host matrix has major impact on the morphology of <Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis>

Oomycetes contain some of the economically most important pathogens of flowering plants. Most have a rather narrow host range, often being restricted to single host species. In downy mildews and other obligate biotrophic plant parasites, like powdery mildews and rusts, delimitating species on grounds of morphological characteristics is often hardly possible and thus often based on only subtle differences. This has led to the widespread application of a broad species concept for these organisms. Consequently, despite the fact that morphological differences were reported for Pseudoperonospora cubensis from different host species, the corresponding new pathogen species were not accepted as being independent, and the host range of Pseudoperonospora cubensis is reported to encompass more than 50 host species in the Cucurbitaceae in temperate to tropical climates. However, recent studies have reported narrow host ranges for other downy mildew genera and advocated a narrow species concept. Here, we report successful colonisation of five different tribes of the Cucurbitaceae by a strain of Pseudoperonospora cubensis and demonstrate that the host matrix has a major impact on the morphology of the pathogen. On the basis of five morphological criteria significant differences could be found for all hosts. These differences were more pronounced in phylogenetically unrelated than in related hosts. Our results provide evidence for a broad host range of Pseudoperonospora cubensis and demonstrate that species delimitation based on morphological characters is not feasible in Pseudoperonospora on Cucurbitaceae. Also in other biotrophic plant pathogens, the situation could be similar, thus necessitating thorough morphological, molecular phylogenetic and cross inoculation experiments for species recognition.     

Biopesticide based sustainable pest management for safer production of vegetable legumes and brassicas in Asia and Africa

Vegetables are one of the important crops which could alleviate poverty and malnutrition among the smallholder farmers in tropical Asia and Africa. However, a plethora of pests limit the productivity of these crops, leading to economic losses. Vegetable producers overwhelmingly rely on chemical pesticides in order to reduce pest‐caused economic losses. However, over‐reliance on chemical pesticides poses serious threats to human and environmental health. Hence, biopesticides offer a viable alternative to chemical pesticides in sustainable pest management programs. Baculoviruses such as nucleopolyhedrovirus (NPV) and granulovirus (GV) have been exploited as successful biological pesticides in agriculture, horticulture and forestry. Maruca vitrata multiple nucleocapsid NPV (MaviMNPV) was found to be a unique baculovirus specifically infecting pod borer on food legumes, and it has been successfully developed as a biopesticide in Asia and Africa. Entomopathogenic fungi also offer sustainable pest management options. Several strains of Metarhizium anisopliae and Beauveria bassiana have been tested and developed as biopesticides in Asia and Africa. This review specifically focuses on the discovery and development of entomopathogenic virus and fungi‐based biopesticides against major pests of vegetable legumes and brassicas in Asia and Africa. © 2019 Society of Chemical Industry     

Zucchini yellow mosaic virus

Zucchini yellow mosaic potyvirus (ZYMV), first isolated in Italy in 1973, described in 1981, and then identified in all continents within a decade, is one of the most economically important viruses of cucurbit crops. It is efficiently aphid-transmitted in a nonpersistent manner and it is also seed-borne in zucchini squash, which could have contributed to its rapid spread worldwide. Biological variability has been observed among ZYMV isolates, concerning host range, symptomatology and aphid transmissibility. More recent studies also revealed a serological and molecular variability. The survival of ZYMV in areas where cucurbits are not grown throughout the year remains to be elucidated, because very few natural over-wintering hosts have been identified so far. Partial control of ZYMV can be achieved by limiting transmission of the virus to the crops by aphids, using adapted cultural practices. Cross-protection with a mild strain has been shown to be effective against most ZYMV isolates. Resistance genes found in cucurbit germplasms are currently being introduced into cultivars with good agronomical characteristics. Pathogen-derived resistance strategies using the expression of ZYMV genes in transgenic plants have also been developed and appear promising. Nevertheless, the high biological variability of ZYMV justifies a careful evaluation of the deployment of genetic control strategies in order to increase their durability.     

Molecular characterization,comparison of screening methods,and evaluation of cross‐pathogenicity of black rot (Xanthomonas campestris pv. campestris) strains from cabbage,choy sum,leafy mustard and pak choi from Taiwan

Choy sum (Brassica rapa var. parachinensis), leafy mustard (Brassica juncea) and pak choi (B. rapa var. chinensis) are highly nutritious components of diets in Taiwan and other Asian countries, and bacterial black rot caused by Xanthomonas campestris pv. campestris (Xcc) is a major biotic constraint in these crops. As very little was known about the Xcc strains from these crops in these regions, including their cross‐pathogenicity and aggressiveness on different hosts, Xcc strains were obtained from cabbage (Brassica oleracea var. capitata), choy sum, leafy mustard and pak choi crops in Taiwan. Two previously published PCR‐based assays reliably distinguished the Xcc strains from other Xanthomonas species and subspecies. Phylogenetic analysis based on repetitive sequence‐based PCR assays placed the Xcc strains in a clade distinct from other Xanthomonas species, and also showed host specificity. Although all of the Xcc strains from the different host species were pathogenic on all five Brassica test species in both a detached leaf assay and an intact plant assay, in the intact plant assay they showed differences in virulence or aggression on the different test hosts. The Xcc strains from leafy mustard and pak choi were consistently highly aggressive on all the test host genotypes, but the strains from choy sum and cabbage were less aggressive on leafy mustard and choy sum. The intact plant assay proved more discriminating and reliable than the detached leaf assay for comparing the aggressiveness of Xcc strains on different host genotypes, and so, with the new Xcc strains isolated in this study, will be useful for screening leafy brassica germplasm accessions for resistance to black rot.     

韭菜迟眼蕈蚊研究进展

韭菜迟眼蕈蚊是葱韭蒜类蔬菜重要的地下害虫,尤喜食韭菜,其幼虫俗称韭蛆。因该虫发生量大且为害严重,近几年有些菜农大量使用化学农药,造成韭菜农药残留超标,"毒韭菜"事件频发,已引起社会各界的广泛关注。有关韭菜迟眼蕈蚊的研究主要集中在生物学特性和防治方法两方面,如生活习性、发生规律、化学杀虫剂室内毒力测定和田间药效比较、筛选等。有关组织学、解剖学、生理学的研究甚少,而分子生物学方面的研究尚未见报道。本文对韭菜迟眼蕈蚊的研究成果进行了综述,分析了当前防治方法存在的问题,为今后的研究和防治提供新的思路。     

应用昆虫毒素基因构建重组病毒杀虫剂研究进展

由于基因工程技术的发展和安全性研究的深入,以重组杆状病毒为主的重组昆虫病毒杀虫剂的应用正面临着突破。文章综述了通过插入昆虫专一性毒素基因构建重组昆虫病毒杀虫剂的技术路线,展望重组病毒杀虫剂的发展趋势。     

Most Colletotrichum species associated with tree tomato (Solanum betaceum) and mango (Mangifera indica) crops are not host‐specific

An important constraint for crop production in Colombia is the high incidence of anthracnose caused by Colletotrichum species. Although several studies have focused on these fungi, the relationship between the different fungal species within the genus and their hosts and whether they display any host preference or host specificity has yet to be examined. In Colombia, diseases caused by Colletotrichum species are particularly severe in mango (Mangifera indica) and tree tomato (Solanum betaceum) crops. In a previous investigation, the Colletotrichum phylogenetic species attacking these crops were identified. The present study aimed to determine whether isolates collected from tree tomato and mango showed host preference or host specificity by assessing aggressiveness, spore density, latent period, and fitness of each strain on the two hosts. In the departments of Cundinamarca and Tolima, Colombia, isolates were collected from plants that presented typical anthracnose symptoms and were identified as C. acutatum, C. asianum, C. boninense, C. gloeosporioides, C. tamarilloi and C. theobromicola. Inoculation of conidia of each isolate onto both hosts showed isolates had no host preference and only the C. gloeosporioides isolate showed host specificity. However, in general, isolates produced a higher spore density when inoculated on the alternate host, which may indicate a difference in the degree of adaptation to each host. Statistical analyses of the assessed parameter values revealed that isolates use different infection strategies when infecting each host. In light of these results, the implications of using quantitative estimations of fitness when studying fungal pathogens are discussed.     

Effect of host plants on developmental and population parameters of invasive leafminer, <Emphasis Type="Italic">Tuta absoluta</Emphasis> (Meyrick) (Lepidoptera: Gelechiidae)

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is one of the most serious pests of tomato, capable of causing 100% crop losses under favourable conditions. We studied the effect of different host plants on developmental and population parameters of this pest at 25?±?0.5 °C temperature, 70?±?5% relative humidity and 12 L: 12D photoperiod. Host plant had significant effect on the developmental biology and the population growth parameters of the pest. The leafminer developed fastest on tomato leaves and slowest on potato tubers. Population growth parameters like intrinsic rate of increase, net reproductive rate, finite rate of increase, doubling time and weekly multiplication rate of T. absoluta were highest on tomato leaves and lowest on potato tubers. Mean generation time was minimum on tomato leaves and maximum on potato tubers. Females developed on tomato leaves were more fecund than other hosts. Though, tomato was found to be the most suitable host plant of T. absoluta, yet, the pest developed and grew successfully on other alternate hosts like potato (Solanum tuberosum L.), brinjal (Solanum melongena L.) and pepino (Solanum muricatum Aiton). These hosts can, therefore, play an important role in the survival, population build up and overwintering of the miner. Further, under favourable conditions the miner can become a serious pest on these crops and need to be monitored on these crops as well.     

人工繁殖中华甲虫蒲螨的替代寄主研究

室内条件下,观察了中华甲虫蒲螨Pyemotes zhonghuajia对替代寄主和自然寄主的寄生选择性,比较了不同寄主繁殖的蒲螨发育情况,蒲螨后代的生物学差异.结果表明,中华甲虫蒲螨最喜好的是替代寄主麦蛾Sitotroga cerealella和自然寄主台湾狭天牛Stenhomalus taiwanus,选择系数分别为0.6743和0.6493,差异不显著;其次为多毛小蠹虫Scolytus seulensis和玉米象Sitophilus zeamais,选择系数分别为0.5006和0.4619,差异不显著,但前一组的选择系数显著高于后一组.通过对单头蒲螨发育和生物学特性观察,替代寄主麦蛾、玉米象与自然寄主台湾狭天牛、多毛小蠹虫繁殖蒲螨从接螨至开始产后代历期分别为6.8、7.9、7.4、7.8d;最大膨腹体直径分别为0.9871、0.9243、0.9662、0.9423mm;每雌产后代雌螨数分别为109.6、105.5、130.3、115.9头,雌雄性比分别为24.6:1、22.7:1、27.4:l、22.8:1;后代雌螨爬行速度分别为5.51、5.20、5.89、5.34cm·min-1;单头蒲螨杀死供试寄主多毛小蠹虫幼虫数量分别为9.3、9.3、9.2、9.6头.差异显著性分析表明,除寄主麦蛾繁殖蒲螨完成1代所需时间显著短于其它寄主外,用替代寄主与自然寄主多毛小蠹虫繁殖的蒲螨在其他方面差异均不显著.因此麦蛾幼虫和玉米象幼虫作为中华甲虫蒲螨替代寄主是可行的.     

Survey of mealybug (Hemiptera: Pseudococcidae) vectors of <Emphasis Type="Italic">Ampelovirus</Emphasis> and <Emphasis Type="Italic">Vitivirus</Emphasis> in vineyards of northwestern Italy

Mealybugs (Hemiptera: Pseudococcidae) are sap-sucking insects which infest a broad range of crops worldwide. They represent an important threat to viticulture as they are vectors of viruses associated with leafroll and rugose wood complex diseases. In this study, we surveyed the presence of mealybugs and their associated viruses in vineyards of the Piemonte and Liguria regions, northwestern Italy. In order to determine the collected specimens correctly, we added a species-specific marker for Heliococcus bohemicus to an existing molecular identification key. The only species collected in Piemonte was H. bohemicus, whereas in Liguria, H. bohemicus, Planococcus ficus and Pseudococcus longispinus were found; Ps. longispinus has never before been reported in Italian vineyards. Several specimens of all three species were infected by the ampeloviruses GLRaV-1 and GLRaV-3 and the vitivirus GVA. Both nymphs and adult females tested positive for the viruses and mixed infections were commonly found within the same insect. Population levels and virus incidence were higher in Liguria than in Piemonte, suggesting a greater risk of disease spread. We conclude that the mild, Mediterranean climate of Liguria favors the development of a diverse mealybug fauna while only H. bohemicus, known to be tolerant to the severe continental winter temperatures, colonize grapevines in colder viticultural areas.