Bacillus thuringiensis isolates from northeastern Brazil and their activities against Plutella xylostella (Lepidoptera: Plutellidae) and Spodoptera frugiperda (Lepidoptera: Noctuidae)

Northeastern Brazil has been little explored in the search for Bacillus thuringiensis (Berliner) variants for the control of agricultural pests such as Plutella xylostella (L.) and Spodoptera frugiperda (J.E. Smith). The aim of this study was to isolate B. thuringiensis from soil and insect samples collected from the northeastern states and to evaluate their lethal and sublethal activities against the neonate larvae of P. xylostella and S. frugiperda. One hundred and four isolates were bioassayed and visualised for the presence of crystals. Bipyramidal crystals were present in 31% of the isolates. In pathogenicity tests, 13 and 19 isolates caused >30% mortality in P. xylostella and S. frugiperda, respectively. The Laboratory of Insect-Toxic Interactions (LIIT)-4311 isolate was the most toxic for P. xylostella, with toxicity similar to B. thuringiensis var. kurstaki (Dipel® WP) and B. thuringiensis var. aizawai (Xentari® WDG). For S. frugiperda, the LIIT-4311, LIIT-4306 and LIIT-4406 isolates were more active than B. thuringiensis var. aizawai. The LIIT-4311 and LIIT-4306 isolates caused high rates of larval growth inhibition in both P. xylostella and S. frugiperda. These results suggest a broad distribution of B. thuringiensis variants in areas of northeastern Brazil. Because LIIT-4306 and LIIT-4311 provided the highest levels of toxicity and larval growth inhibition for both P. xylostella and S. frugiperda, these isolates can be exploited to develop new technologies for pest management.     

小菜蛾抗药性及其治理对策的研究进展

小菜蛾（Plutella xylostella L.）的抗药性研究是一项全球性的重要课题。本从其抗药性发展、抗药性机理研究进展、抗药性检测及研究手段的状况、抗药性治理对策等方面,对小菜蛾抗药性的研究进展进行综述。目前,对小菜蛾抗药性的分子水平上的探讨甚少。     

Detection of insecticide resistance in Plutella xylostella (L.) (Lepidoptera: Plutellidae) populations in South Australian crucifer crops

In response to reports of failures of sprays to control Plutella xylostella infestations in vegetable-growing areas around Adelaide in 1991, the extent of resistance to six insecticides in several South Australian populations of P. xylostella was determined. Populations sampled from vegetable crops in the Adelaide region were five- to 200-fold resistant to organophosphates (chlorpyrifos, methamidophos and mevinphos), eight- to 400-fold resistant to pyrethroids (esfenvalerate and permethrin) and six-fold resistant to the carbamate methomyl compared with a susceptible strain. Mevinphos and esfenvalerate were the most effective of the tested insecticides. These, together with Bacillus thuringiensis products, have continued until 1998 to be used almost exclusively for the control of P. xylostella in South Australian vegetable crops in the absence of any resistance management strategy.     

Silicon dioxide in the defence induction of soybean against Spodoptera frugiperda

Silicon can increase the natural defence of plants against stresses including herbivorous insects. Silicon dioxide (SiO₂) is one of the forms of silicon, and despite its wide use in the industrial sector, its use in agriculture is still poorly adopted. The aim of this study was to evaluate the effectiveness of foliar application of SiO₂ in inducing defence against Spodoptera frugiperda in soybean. The experiments were conducted in a completely randomised design with four treatments (0%, 1%, 2.5%, and 5% of SiO₂). The effect on cannibalism, biological parameters (mortality, duration of the larval stage, duration of the pupal stage and pupal weight) and wear on the mandible of S. frugiperda were evaluated. The foliar silicon content was also determined. The supply of SiO₂ prolonged the duration of the larval and pupal stages of the S. frugiperda by 0.56 and 0.17 days for each 1% of SiO₂ applied, respectively. The use of SiO₂ at 5% increased the mortality rate of caterpillars in the larval stage by approximately 25%. There was no effect of SiO₂ application on cannibalism and weight of S. frugiperda pupae. There was wear on the caterpillars' jaws in the third and fourth instar at the highest SiO₂ concentration. The application of SiO₂ promoted greater accumulation of silicon in soybean leaves. It is concluded that the foliar application of SiO₂ affects the biological performance of S. frugiperda through the induction of defence in the soybean crop and presents itself as a promising strategy in integrated pest management programmes.     

转双价基因抗虫棉及其杂交F1代对棉铃虫幼虫的抗性研究

利用室内生物测定法和田间调查法研究双价抗虫棉及其杂交F1代不同生育期主茎功能叶和花铃期不同组织器官的抗虫性。结果表明:双价抗虫棉及其杂交F1代的抗虫性变化规律与B t基因棉的抗虫性变化规律相近,为生育前期(苗期、蕾期)抗性强,生育后期(花铃期、吐絮期)抗性弱,在花铃期为:棉铃>棉蕾>花>棉叶,且差异极显著。与B t基因棉相比,生育前期双价抗虫棉及其杂交F1代的抗虫性略弱,但差异不显著;生育后期双价抗虫棉及其杂交F1代的抗虫性显著强于B t基因棉。     

小菜蛾在温带地区越冬研究进展

小菜蛾是世界性重要害虫。在热带和亚热带地区冬季的十字花科植物上能正常发育繁殖,可见到各种虫态。但在温带冬季十字花科植物不能生长的地区,小菜蛾的越冬成为一个重要生态学问题。综述了亚洲、北美洲和欧洲小菜蛾越冬的研究进展。小菜蛾在日本的北海道、本州岛的北陆和东北大部分地区不能越冬,越冬北限相当于冬季积雪覆盖时间为60d的区域;在中国,小菜蛾不能在寒冷的东北地区越冬,在长江中下游以南地区冬季可见各虫态,但越冬北限尚不清楚;小菜蛾在冬天气候温和的韩国以及澳大利亚东南部继续发生;北美洲的加拿大西部和安大略地区大量的试验证明小菜蛾不能成功越冬,在美国南部小菜蛾冬季可正常发生,北部小菜蛾的越冬尚未见系统研究报道,但确认春季从南部运输的受小菜蛾感染的甘蓝等种苗是美国北部的重要虫源。小菜蛾在欧洲各地越冬的系统研究未见报道,没有证据表明在英国小菜蛾会发生有显著意义的越冬。目前小菜蛾越冬研究主要采用冬季直接试验观察和基于耐寒性试验的越冬预测两种方法。冬季直接试验观察法包括:(1)利用人工饲养的小菜蛾在田间各种潜在的越冬场所的越冬试验;(2)在秋播、野生或残留的十字花科植物上进行冬季种群的系统抽样调查;(3)越冬前后在前茬为十字花科植物的田块广泛搜寻普查小菜蛾的存活个体。基于耐寒性试验的越冬预测法:在获取小菜蛾越冬场所温度的基础上设计低温处理模式,试验低温处理后小菜蛾的存活率及后续发育和生殖。将试验数据和各地气温或小气候相结合,对小菜蛾在的越冬可能性进行推断。     

Equivalent intensity but differential dominance of sodium channel blocker insecticide resistance conferred by F1845Y and V1848I mutations of the voltage-gated sodium channel in Plutella xylostella

Two point mutations (F1845Y and V1848I) in the voltage-gated sodium channel gene of Plutella xylostella are involved in the target-site resistance to sodium channel blocker insecticides (SCBIs). The contribution of the individual mutations to the SCBI resistance and the associated inheritance modes is as yet unclear. Through 2 rounds of single-pair crossing and marker-assisted selection, 2 P. xylostella strains (1845Y and 1848I) bearing homozygous F1845Y or V1848I mutant alleles were successfully established from a field-collected population, and the contribution of each mutation to SCBI resistance, as well as associated inheritance patterns, was determined. When compared with the susceptible SZPS strain, each of the mutations individually conferred equally high-level resistance to indoxacarb (378 and 313 fold) and metaflumizone (734 and 674 fold), respectively. However, dominance levels of resistance to SCBIs were significantly different between the 2 resistant strains. Resistance of the 1845Y strain to indoxacarb and metaflumizone was inherited as an autosomal and incompletely dominant trait (D values ranged from 0.43 to 0.76). In contrast, that of the 1848I strain followed an autosomal but incompletely recessive to semidominant mode (D values: −0.24 to 0.09). Our findings enriched the current understanding of inheritance and mechanisms of SCBI resistance in P. xylostella, and will help develop resistance management programs for P. xylostella and other economic pests.     

Selection for resistance to acetamiprid and various other insecticides in the diamondback moth, Plutella xylostella (L.) (Lep., Plutellidae)

Abstract:  A diamondback moth (DBM) colony (KOBII) originally collected from cabbage fields of Iwaoka-cho in Kobe City, Japan, where acetamiprid is the primary insecticide for DBM control was selected for studies on resistance to acetamiprid, phenthoate, esfenvalerate and cartap, using the leaf-dipping method. Resistance to acetamiprid was slow to develop in KOBII compared with the other insecticides. Three selection experiments each with phenthoate and esfenvalerate yielded a resistance ratio (RR) of 140 and 222, respectively, in the selected strains compared with a part of KOBII that was not exposed to any insecticide (KOBII-NS). Three selection experiments with cartap also gave an RR of 15. However, six selection experiments with acetamiprid yielded an RR of just 9.5 in the selected strain. When cross-resistance to acetamiprid was evaluated, the cartap-resistant strain showed a moderate level of cross-resistance to acetamiprid. In the case of the phenthoate-resistant strain, a very low cross-resistance to acetamiprid was observed. There was however no discernible cross-resistance to acetamiprid in the esfenvalerate-resistant strain.     

小菜蛾抗性治理及可持续防控技术研究与示范——公益性行业(农业)科研专项“小菜蛾可持续防控技术研究与示范”进展

针对我国小菜蛾Plutella xylostella(L.)严重为害及高抗药性的现状,在华南、华中、华东、华北和西南等具代表性地区开展小菜蛾灾变规律、抗性监测及治理、越冬迁飞、抗性机理及可持续防控技术研究集成与示范。灾变规律研究结果表明,我国各地小菜蛾的年发生世代从北至南呈逐渐增加趋势,各地小菜蛾发生起始峰时间从南至北逐渐向后推移,每年不同区域有一至两个发生高峰,年度间受温度和降水等气候因素、天敌等生物因子以及耕作制度等人为操作等多种因素的影响。抗性监测及治理研究显示,全国5个十字花科蔬菜主产区小菜蛾对11种代表性杀虫剂都有较强的抗药性,不同药剂在全国的抗药性水平有很大差异,在华南、西南和华东十字花科蔬菜主产区抗性水平相对较高,华中和华北呈现抗性上升趋势,并根据抗性监测结果制定区域性抗性治理策略。越冬和迁飞研究证明,武汉至驻马店区域为小菜蛾的越冬北限,小菜蛾存在远距离迁移的特性,并确定小菜蛾有迁入迁出和迁入定殖两种迁飞模式。抗性机理研究结果表明,小菜蛾对Bt制剂、阿维菌素、氟虫腈、茚虫威和丁醚脲的抗性遗传方式不同,各药剂交互抗性普也存在差异。可持续防控技术方面,制定了适合各区域的小菜蛾可持续防控技术体系,并在华南、华东、华北、华中和西南等地区建立了45个示范基地,依区域不同分别示范推广具区域特色的以农业措施、生物防治、生态调控和合理用药为主的可持续防控等技术,累计印刷技术宣传手册16万份,培训各类农民和基层农技人员5.19万人次,累计示范面积近4万公顷,带动技术示范近20万公顷,取得了显著的经济、社会和生态效益。     

Conserving the efficacy of insecticides against Plutella xylostella (L.) (Lep., Plutellidae)

Abstract:  The diamondback moth (DBM), Plutella xylostella (L.) (Lep., Plutellidae), is one of the most destructive insect pests of crucifers worldwide. It was the first crop insect reported to be resistant to DDT and now in many crucifer-producing regions it has shown significant resistance to almost every insecticide applied in field including biopesticides such as crystal toxins from Bacillus thuringiensis and spinosyns from Saccharopolyspora spinosa . In certain parts of the world, economical production of crucifers has become almost impossible because of its resistance to insecticides and resulting control failure. A coordinated resistance management program needs to be implemented with the involvement of pesticide industry, local pesticide regulatory authorities, scientists and farmers. The judicious use of chemicals in conjunction with other control measures (e.g. biological control agents, resistant varieties, proper fertilization rates) is the best way to manage DBM and other pests of cruciferous crops. Introduction of glucosinolate-sulphatase inhibitors as plant-incorporated-products or sprayable material may also lead to a novel pest management strategy.     

cDNA representational difference analysis of the deltamelthrin-resistant and -susceptible populations in diamondback moth (Plutella xylostella L.)

Abstract:  The improved cDNA representational difference analysis was preliminarily used to analyse the genetic differences between the deltamethrin-susceptible and -resistant strain of the diamondback moth. The driver amplicon is the cDNA from the susceptible strain while the tester amplicon is from the resistant strain. After four rounds of subtractive hybridization, we obtained one different product, the size of which was about 200 bp. Comparisons between our sequencing results and data in GenBank show that there is one sequence which has a relatively high homology to the ubiquitin gene, but there are no reports indicating that its upregulated expression is correlated with insecticide resistance. Hence further studies are warranted.     

Resistance of the fall armyworm,Spodoptera frugiperda,to transgenic Bacillus thuringiensis Cry1F corn in the Americas: lessons and implications for Bt corn IRM in China

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a major target pest of Bt crops (e.g., corn, cotton, and soybean) in North and South America. This pest has recently invaded Africa and Asia including China and the invasion has placed a great threat to the food security in many countries of these two continents. Due to the extensive use of Bt crops, practical resistance of S. frugiperda to Cry1F corn (TC 1507) with field control problems has widely occurred in Puerto Rico, Brazil, Argentina, and the mainland United States. Analyzing data generated from decade-long studies showed that several factors might have contributed to the wide development of the resistance. These factors include (1) limited modes of action of Bt proteins used in Bt crops; (2) cross-resistance among Cry1 proteins; (3) use of nonhigh dose Bt crop traits; (4) that the resistance is complete on Bt corn plants; (5) abundant in initial Cry1F resistance alleles; and (6) lack of fitness costs/recessive fitness costs of the resistance. The long-term use of Bt crop technology in the Americas suggests that Bt corn can be an effective tool for controlling S. frugiperda in China. IRM programs for Bt corn in China should be as simple as possible to be easily adopted by small-scale growers. The following aspects may be considered in its Bt corn IRM programs: (1) use of only “high dose” traits for both S. frugiperda and stalk borers; (2) developing and implementing a combined resistance monitoring program; (3) use “gene pyramiding” as a primary IRM strategy; and (4) if possible, Bt corn may not be planted in the areas where S. frugiperda overwinters. Lessons and experience gained from the global long-term use of Bt crops should have values in improving IRM programs in the Americas, as well as for a sustainable use of Bt corn technology in China.     

Interactions of Bacillus thuringiensis bioinsecticides and the predatory stink bug Podisus nigrispinus to control Plutella xylostella

The diamondback moth (DBM), Plutella xylostella (L.), is a major pest of brassica crops worldwide. Control of this pest is difficult because it rapidly develops resistance to synthetic and biological insecticides and because of the effects of insecticides on its natural enemies. Podisus nigrispinus (Dallas) is a predator that feeds on its prey, as well as on the host plants of its prey, and is an important biological control agent of DBMs. The aim of this study was to determine the susceptibility of P. xylostella larvae to two bioinsecticides: the HD1 strain of Bacillus thuringiensis (B. thuringiensis var. kurstaki) and the commercial product Agree^® (B. thuringiensis var. aizawai CG 91). In addition, the impact of these bioinsecticides on the P. nigrispinus consumption of DBM larvae and phytophagy was evaluated. Both the HD1 strain and Agree^® caused 100% mortality in P. xylostella larvae. P. nigrispinus nymphs fed only with kale leaves (Brassica oleracea var. acephala) sprayed with water, the HD1 strain, or Agree^® did not complete their nymphal development. When prey was also available, P. nigiripinus fed on kale leaves to obtain water. Both nymphs and adults of P. nigrispinus consumed greater numbers of DBM larvae, and fed less on kale leaves, when sprayed with the HD1 strain or Agree^®. These results suggest a positive interaction of B. thuringiensis‐based products and the predator P. nigrispinus in the control of P. xylostella larvae.     

Fitness consequences of cannibalism in the fall armyworm, Spodoptera frugiperda

We investigated the consequences of cannibalism for some correlatesof fitness in the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae).The benefits gained by cannibals were ascertained by comparing survival,development rate, and pupal weight of larvae that had the opportunityto cannibalize with those that did not, at two levels of food availability.Larvae in the cannibalism treatments were provided with a conspecificone instar younger than themselves on five dates throughoutlarval development. Cannibalism was frequent; given the opportunityall larvae predated at least one younger conspecific. The frequencyof cannibalism was not affected by the sex of the cannibal orby the availability of alternative food. However, cannibalssuffered a significant reduction in survival at both high andlow food availability. Furthermore, cannibals had a lower pupal weightin the high food treatment and a reduced development rate inthe low food treatment. The only detectable fitness benefitassociated with cannibalism was a positive correlation betweenthe number of victims consumed and development rate under conditionsof low food availability. On balance, cannibalism appears tobe costly; alternative explanations for its occurrence in thisspecies are discussed.     

Complex inheritance of larval adaptation in Plutella xylostella to a novel host plant

Studying the genetics of host shifts and range expansions in phytophagous insects contributes to our understanding of the evolution of host plant adaptation. We investigated the recent host range expansion to pea, in the pea-adapted strain (P-strain) of the crucifer-specialist diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae). Larval survivorship on the novel host plant pea and a typical crucifer host (kale) was measured in reciprocal F(1), F(2) and backcrosses between the P-strain and a strain reared only on crucifers (C-strain). Reciprocal F(1) hybrids differed: offspring from P-strain mothers survived better on pea, indicating a maternal effect. However, no evidence for sex-linkage was found. Backcrosses to the P-strain produced higher survivorship on pea than C-strain backcrosses, suggesting recessive inheritance. In a linkage analysis with amplified fragment length polymorphism markers using P-strain backcrosses, two, four and five linkage groups contributing to survival on pea were identified in three different families respectively, indicating oligogenic inheritance. Thus, the newly evolved ability to survive on pea has a complex genetic basis, and the P-strain is still genetically heterogeneous and not yet fixed for all the alleles enabling it to survive on pea. Survivorship on kale was variable, but not related to survivorship on pea. This pattern may characterize the genetic inheritance of early host plant adaptation in oligophagous insect species.     

Chitosan nanoparticles help double-stranded RNA escape from endosomes and improve RNA interference in the fall armyworm,Spodoptera frugiperda

RNA interference (RNAi) is a promising technology for the development of next-generation insect pest control products. Though RNAi is efficient and systemic in coleopteran insects, it is inefficient and variable in lepidopteron insects. In this study, we explored the possibility of improving RNAi in the fall armyworm (FAW), Spodoptera frugiperda by conjugating double-stranded RNA (dsRNA) with biodegradable chitosan (Chi). dsRNA conjugated with chitosan was protected from degradation by endonucleases present in Sf9 cell-conditioned medium, hemolymph, and midgut lumen contents collected from the FAW larvae. Chi–dsRNA complexes showed reduced accumulation in the endosomes of Sf9 cells and FAW tissues. Exposing chitosan formulated dsRNA in Sf9 cells and the tissues induced a significant knockdown of endogenous genes. Chi–dsIAP fed to FAW larvae induced knockdown of iap gene, growth retardation, and mortality. Processing of dsRNA into small interfering RNA was detected with chitosan-conjugated ³²P-UTP-labeled ds green fluorescent protein in Sf9 cells and FAW larval tissues. Overall, these data suggest that dsRNA conjugated with chitosan helps dsRNA escape from the endosomes and improves RNAi efficiency in FAW cells and tissues.     

The ability of synthetic sex pheromone traps to forecast Plutella xylostella infestations depends on survival of immature stages

The potential for using synthetic sex pheromone traps as a simple and practical method of monitoring population densities of insect pests has been investigated in many crop systems. Yet, factors enabling the forecast of infestations based on pheromone trap catches are not fully understood. This study tested the prediction that high survival of immature stages of the target pest is a pre‐requisite for trap catches to correlate well with future infestations on the crop. The influence of parasitoids, as an important natural mortality factor of diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), larvae and pupae in South Africa, on the ability of pheromone traps to forecast infestations was investigated continuously at weekly intervals over 6 years on unsprayed cabbage. During late October–May, when parasitism levels were high (≥50%), infestations and trap catches were significantly lower than during low parasitism (<50%) observed during June–early October. Because infestations were negatively related to parasitism level, trap catches correlated with infestations better when observations were made in the same week during periods of high parasitism. Conversely, when survival of P. xylostella immature stages was high due to low parasitism levels, trap catches correlated with future infestations well for up to 2 weeks. Thus, trap catches can be used to forecast infestations during September–October, a period that coincides with high P. xylostella infestations as a consequence of low natural control by parasitoids. This is the first study to show that the ability of pheromone trap catches to forecast infestations depends on survival of the immature stages of the target pest.     

Helicoverpa armigera baseline susceptibility to Bacillus thuringiensis Cry toxins and resistance management for Bt cotton in India

Transgenic cotton that produces insecticidal proteins from Bacillus thuringiensis (Bt), often referred to as Bt cotton, is widely grown in many countries. Bt cotton with a single cry1A gene and stacked also with cry2A gene has provided satisfactory protection against the damage by the lepidopteran bollworms, especially the cotton bollworm, Helicoverpa armigera (Hübner) which is considered as a key pest. The baseline susceptibility of the larvae of H. armigera to Cry1Ac and other toxins carried out in many countries has provided a basis for monitoring resistance. There is no evidence of development of field-level resistance in H. armigera leading to the failure of Bt cotton crop anywhere in the world, despite the fact that Bt cotton was grown on the largest ever area of 12.1 million hectares in 2006 and its cumulative cultivation over the last 11 years has surpassed the annual cotton area in the world. Nevertheless, the Bt resistance management has become a necessity to sustain Bt cotton and other transgenic crops in view of potential of the target insects to evolve Cry toxin resistance.     

Patterns of genomic and allochronic strain divergence in the fall armyworm,Spodoptera frugiperda (J.E. Smith)

Speciation is the process through which reproductive isolation develops between distinct populations. Because this process takes time, speciation studies often necessarily examine populations within a species that are at various stages of divergence. The fall armyworm, Spodoptera frugiperda (J.E. Smith), is comprised of two strains (R = Rice & C = Corn) that serve as a novel system to explore population divergence in sympatry. Here, we use ddRADSeq data to show that fall armyworm strains in the field are largely genetically distinct, but some interstrain hybridization occurs. Although we detected F1 hybrids of both R‐ and C‐strain maternal origin, only hybrids with R‐strain mtDNA were found to contribute to subsequent generations, possibly indicating a unidirectional barrier to gene flow. Although these strains have been previously defined as “host plant‐associated,” we recovered an equal proportion of R‐ and C‐strain moths in fields dominated by C‐strain host plants. As an alternative to host‐associated divergence, we tested the hypothesis that differences in nightly activity patterns could account for reproductive isolation by genotyping temporally collected moths. Our data indicates that strains exhibit a significant shift in the timing of their nightly activities in the field. This divergence in phenology creates a prezygotic reproductive barrier that likely maintains the genetic isolation between strains. Thus, we conclude that it may be ecologically inaccurate to refer to the C‐ and R‐ strain as “host‐associated” and they should more appropriately be considered “allochronic strains.”