Differential toxicity of Bacillus thuringiensis strains and their crystal toxins against high-altitude Himalayan populations of diamondback moth, Plutella xylostella L

BACKGROUND: Diamondback moth, Plutella xylostella (L.), is a major insect pest of crucifers in the biodiversity‐rich north‐western Indian Himalayan hills. The present investigation was aimed at determining the susceptibility pattern of P. xylostella populations collected from different locations of this region to autochthonous and standard Bacillus thuringiensis strains. RESULTS: Among the reference as well as indigenous B. thuringiensis strains tested, sub spp. kurstaki HD‐1, kurstaki HD‐73, galleriae HD‐8, local galleriae/colmeri strain BtOa1 and some of their Cry1 class toxins were found to be highly toxic. Surprisingly, the sub sp. tolworthi HD‐125, local tolworthi strain BtHa1 and Cry9 class toxins were found to be non‐toxic. Midgut homogenate from fourth‐instar larvae was found to activate 130 kDa protoxin from the local tolworthi strain BtHa1 into 68 kDa toxin, but failed to exert any larval mortality, probably owing to lack of receptor binding. CONCLUSION: The present study provides valuable baseline susceptibility data for the deployment of B. thuringiensis‐based control methods, as well as for future monitoring of development of resistance in P. xylostella to B. thuringiensis in this ecologically sensitive region. Copyright © 2008 Society of Chemical Industry     

Characterisation and toxicity of Bacillus thuringiensis strains from hazelnut pests and fields

BACKGROUND: In order to find and identify more toxic insecticidal Bacillus thuringiensis Berliner (Bt) strains, a survey was carried out of B. thuringiensis isolate pests belonging to Coleoptera, Lepidoptera and Diptera and from soils in hazelnut fields. Of 16 isolates having Bacillus cereus‐B. thuringiensis morphology, eight were classified as B. thuringiensis because of the production of parasporal δ‐endotoxin crystals. RESULTS: In this study, eight isolates of B. thuringiensis from hazelnut pests (isolates Bn1, Mm2, Mnd and Xd3) and from hazelnut soils (isolates 6, 27, 40 and 46) have been characterised in detail. These isolates were compared with reference strains by electron microscopy, SDS‐PAGE analysis, cry gene content, serological test and insecticidal activity. CONCLUSION: Results indicate that Bn1 and MnD are B. thuringiensis subsp. kurstaki, and Mm2 and Xd3 are B. thuringiensis subsp. tenebrionis. In addition, isolate 6 is B. thuringiensis subsp. israelensis, isolates 27 and 46 are B. thuringiensis subsp. kumamotoensis and isolate 40 is B. thuringiensis subsp. indiana. The four B. thuringiensis isolates from hazelnut pests may be valuable as biological control agents against coleopteran and lepidopteran insects. Copyright © 2009 Society of Chemical Industry     

Insect pests of pearl millet in West Africa: A review

Abstract

Pearl millet (Pennisetum americanum (L.) K. Schum), a major food crop in the sahelian and savanna regions of West Africa, is attacked by about 100 species of insects. Of these, the only regular pests are stem borers (Acigona ignefusalis Hmps. and Sesamia spp.) and earhead caterpillars (Masalia spp., Raghuva spp.). Sporadic pests such as hairy caterpillars (Amsacta moloneyi Druce), armyworms (Spodoptera spp. and Mythimna spp.) and grasshoppers (Acrididae) may cause severe losses to crops during prolonged droughts early in the season. A grain midge (Geromyia penniseti (Felt)) attacks late millets and causes a considerable loss in yield. Information is presented on the biology, distribution, hosts and natural enemies of these pests, and some methods of control are discussed.     

An effective strain ofBacillus thuringiensis against fourth-instar caterpillars ofBoarmia selenaria

A strain ofBacillus thuringiensis Berliner effective against fourth-instarBoarmk selenaria Schiffermüller (Lepidoptera: Geometridae) was found. Strain HD-251 of the varietykurstaki H-serotype 3a, 3b was more effective than the standard HD-1-S-1980 in laboratory bioassays. The potency of HD-251 is two to four times higher than that of the standard, and varies from 19,821 to 68,645 IU/mg. Two other strains of the varietyaizawai H-serotype 7,viz., HD-249 and T63L4, were inferior to HD-251, with potencies of 6,982 and 13,714 IU/mg, respectively, against first-instar larvae.     

Characterization of field‐evolved resistance to Bacillus thuringiensis‐derived Cry1F δ‐endotoxin in Spodoptera frugiperda populations from Argentina

BACKGROUND

Transgenic maize (Zea mays L.) event TC1507 (Herculex^® I insect protection), expressing Cry1F δ‐endotoxin derived from Bacillus thuringiensis var. aizawai, was commercialized in 2003 in the Americas. Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) susceptibility to Cry1F was monitored annually across several regions in Argentina using diagnostic concentration bioassays. Reduced performance of TC1507 maize against S. frugiperda was reported in 2013. A resistant population was established in the laboratory and the dominance of Cry1F resistance was characterized.

RESULTS

During 2012–2015, high‐survivorship of several populations was observed in the resistance monitoring program. Reciprocal crosses of a Cry1F‐resistant population with a Cry1F‐susceptible population were evaluated to calculate effective dominance (D_ML) based on mortality levels observed at 100 µg/ml Cry1F. Two additional dominance levels (D_LC and D_EC) were calculated using lethal (LC₅₀) or effective concentration (EC₅₀) derived from concentration–response bioassays. Estimates indicated that Cry1F resistance in S. frugiperda in Argentina was either highly recessive (D_ML = 0.005) or incompletely recessive (D_LC < 0.26 and D_EC < 0.19).

CONCLUSION

This study is the first documented confirmation and characterization of S. frugiperda Cry1F field‐evolved resistance in Argentina. The resistance to Cry1F in S. frugiperda populations collected in Argentina, is autosomal and incompletely recessive similar to the resistance reported in Brazil. © 2017 The Authors. Pest Management Science published by John Wiley © Sons Ltd on behalf of Society of Chemical Industry.     

苏云金芽胞杆菌一新菌株的鉴定及其杀虫活性

为分离并鉴定对亚洲玉米螟具有高毒力的苏云金芽胞杆菌菌种,通过棋盘式采集法从吉林省农业科学院试验田土壤中分离获得野生菌株,进行了形态、生化特性、伴孢晶体观察及基因型鉴定,并测定了其对3种鳞翅目害虫的室内生物活性。结果显示:该菌株在LB培养基上为圆形、暗白色菌落,边缘不整齐,革兰氏染色阳性,产生的伴孢晶体形状多为不规则形。16S r DNA序列与苏云金芽胞杆菌属的NBRC 13865同源性达99%;该菌株为cry2+cry9复合基因型,编码氨基酸序列与Cry2Ab和Cry9Ea蛋白同源性分别为94%和99%,鉴定为苏云金芽胞杆菌,命名为JN001。该菌株对亚洲玉米螟具有较高毒性,以1.0×109个/m L菌体浓度接种3龄幼虫72 h后,其校正死亡率为95.06%,致死中浓度(LC50)为4.12×103个/m L菌体,而对斜纹夜蛾和粘虫毒性较弱,接种72 h后校正死亡率分别为12.88%和7.34%。表明该菌株是1株对亚洲玉米螟具有较强毒力的苏云金芽胞杆菌新菌株,具有较好的开发利用潜力。     

Application of some bacterial varieties ofBacillus thuringiensis and its bioproduct Delfin onSynanthedon myopaeformis Borkh. (Lep. Aegeriidae) in apple orchards

Field experiments were conducted using the biological agentBacillus thuringiensis to control the clearwing moth,Synanthedon myopaeformis (Lep. Aegeriidae) in apple orchards at Alexandria governorate during 1995, 1996 and 1997. Three bacterial varieties ofB. thuringiensis showed significant influence on the reduction of borer infestation. Four applications with the sporic concentration 12×10⁷ spores/ml of each ofB. th. var.kurstaki, B. th. var.alesti andB. th. var.thuringiensis induced 82.5, 51.1 and 42.1% reduction of infestation, respectively. Methods of application showed highly significant effect on infestation level. Sparying after scraping was the best method (80.0%, reduction) followed by painting after scraping (71.5%, reduction) then spraying alone (59.7%, reduction) or painting alone (43.0%, reduction). A control programme was carried out using three applications of the biomaterial, Delfin at the rate of 200 g/100 I.w. during mid-March, early June and mid-August resulted in 88.5–91.4% reduction of infestation.     

Cross‐resistance,inheritance and biochemical mechanisms of imidacloprid resistance in B‐biotype Bemisia tabaci

BACKGROUND: The B‐type Bemisia tabaci (Gennadius) has become established in many regions in China, and neonicotinoids are extensively used to control this pest. Imidacloprid resistance in a laboratory‐selected strain of B‐type B. tabaci was characterised in order to provide the basis for recommending resistance management tactics. RESULTS: The NJ‐Imi strain of B‐type B. tabaci was selected from the NJ strain with imidacloprid for 30 generations. The NJ‐Imi strain exhibited 490‐fold resistance to imidacloprid, high levels of cross‐resistance to three other neonicotinoids, low levels of cross‐resistance to monosultap, cartap and spinosad, but no cross‐resistance to abamectin and cypermethrin. Imidacloprid resistance in the NJ‐Imi strain was autosomal and semi‐dominant. It is shown that enhanced detoxification mediated by cytochrome‐P450‐dependent monooxygenases contributes to imidacloprid resistance to some extent in the NJ‐Imi strain. Results from synergist bioassays and cross‐resistance patterns indicated that target‐site insensitivity may be involved in imidacloprid resistance in the NJ‐Imi strain of B. tabaci. CONCLUSION: Although oxidative detoxification mediated by P450 monooxygenases is involved in imidacloprid resistance in the NJ‐Imi strain of B‐type B. tabaci, target‐site modification as an additional resistance mechanism cannot be ruled out. Considering the high risk of cross‐resistance, neonicotinoids should be regarded as a single group when implementing an insecticide rotation scheme in B. tabaci control. Copyright © 2009 Society of Chemical Industry     

Mode of inheritance and stability of resistance to Bacillus thuringiensis var kurstaki in a diamondback moth (Plutella xylostella) population from Malaysia

Genetic inheritance of resistance to Bacillus thuringiensis var kurstaki (BTK) was examined in a diamondback moth (Plutella xylostella) population collected from the Melaka region of Malaysia. A BTK‐selected sub‐population (BTK‐SEL) which was more than 100‐fold resistant to BTK compared with a susceptible (ROTH) population of P xylostella was used with standard reciprocal crosses and back‐crosses between ROTH and BTK‐SEL. Logit regression analysis of F ₁ reciprocal crosses indicated that BTK resistance was inherited as an incompletely recessive autosomal trait and controlled by a single locus. In contrast, other studies have shown that resistance to Cry1Ac is inherited as an incompletely dominant autosomal trait in a Cry1Ac‐selected sub‐population of the same Melaka population. The frequency of the allele responsible for resistance decreased without exposure to insecticide in the laboratory. © 2000 Society of Chemical Industry     

Susceptibility pattern and development of resistance in the diamondback moth,Plutella xylostella L,to Bacillus thuringiensis Berl var kurstaki in India

Base‐line susceptibility for six‐day‐old larvae of the diamondback moth, Plutella xylostella, against Bacillus thuringiensis var kurstaki (Biobit^®) was studied by a cabbage leaf disc dip bioassay technique. Diamondback moth from 13 locations in seven different states spread over a distance of about 3000 km longitudinally was used for these studies. Forty‐eight‐hour LC₅₀ values varied from 1.0 to 10.97 mg AI litre⁻¹. Further investigations on the development of resistance under laboratory conditions showed an increase in LC₅₀ from 2.76 (for unselected F₁ generation) to 5.28 mg AI litre⁻¹ (for selected F₉ generation), using a selection concentration of 6.4 mg AI litre⁻¹. This suggested a possibility of the development of resistance under field conditions if there were to be extensive and indiscriminate use of B thuringiensis. These findings are discussed in relation to integrated pest management and the mechanisms of resistance in resistance management tactics. © 2000 Society of Chemical Industry     

Biological means of plant protection in glasshouses in Belarus1

Biological control methods introduced and developed for Belarussian glasshouse crops are described. The agents used against arthropod pests include the natural enemies Phytoseiulus persimilis, Encarsia formosa and Amblyseius spp., the fungi Verticillium lecanii, Beauveria bassiana and Aschersonio, and the preparation Bitoxibacillin (based on Bacillus thuringiensis var. thuringiensis). Against fungal pathogens, Trichodermin (based on Trichuderma Iignorum) and peat oxyhumates are used.     

Breeding systems and seed production for six weedy taxa of Bidens

Successful reproduction is considered to be the major determining factor for a weedy population to establish, colonize and infest. The breeding systems and seed production of six weedy taxa of Bidens were studied at an experimental garden in Jiangsu province in China. The results from four treatments (open pollination, bagged capitulum, decapitated capitulum, and pollen supplement) indicated that, with the exception of Bidens pilosa var. radiata, all the other five Bidens taxa; namely, B. pilosa var. pilosa, B. pilosa var. minor, B. frondosa, B.bipinnata, and B. biternata, were self‐fertile with a high seed‐set in bagged capitula, and a high autofertility index. Including the trait of vegetative propagation in self‐incompatible B. pilosa var. radiata, all the six weedy taxa of Bidens had the capacity of uniparental reproduction. The pollen limitation values for all six Bidens taxa were low in the present study. However, the seed production of B. pilosa var. radiata is likely to decrease in the absence of sufficient pollinators, related to its high pollen contribution and potential pollen limitation indices. The seed production per plant and per capitulum of the three varieties of B. pilosa was the highest among the six taxa and might contribute to their serious weediness. The breeding system type and seed number per capitulum could be used as a classification reference for Bidens taxa.     

Stability and controlled release properties of carboxymethylcellulose-encapsulated Bacillus thuringiensis var. Israelensis

The mosquito and blackfly pathogen, Bacillus thuringiensis Berl. var. israelensis (B.t.i.). containing spores and toxin crystal, was successfully encapsulated in an insolubilized carboxymethylcellulose (CMC)-aluminum matrix creating a controlled-release bead formulation. The release rate of B.t.i. from this formulation was sufficient to kill Culex sp. larvae during bioassays performed in the laboratory. The CMC concentration significantly affected the release rate of the bacterium. All types of beads tested exhibited a high initial release of B.t.i. in the first week of experiments, followed by a slower rate later. Levels of 10 g kg^?1 CMC solution and 0–05 m aluminium sulfate were found to be optimal in the formulation mixture for polysaccharide and gellant, respectively. This type of bead gave the fastest release of the bacterium and caused 100% larval mortality as from the second day of treatment. While acidic pH, high temperature and UV exposure drastically decreased spore viability of B.t.i., the CMC-encapsulated formulation was more stable to these conditions. Larvicidal activity of encapsulated B.t.i. was more resistant to high temperature (50°C) than the unformulated B.t.i. The high response of Culex larvae to the encapsulated B.t.i. suggests that the formulation could be employed effectively in the field.     

A comparative review of the mammalian toxicity of Bacillus thuringiensis-based pesticides

The extensive mammalian toxicity studies performed to support the safety of Bacillus thuringiensis-containing pesticides clearly demonstrate that the tested isolates are not toxic or pathogenic. Toxicity studies submitted to the US Environmental Protection Agency to support the registration of B. thuringiensis subspecies have failed to show any significant adverse effects in body weight gain, clinical observations, or upon necropsy. Infectivity/pathogenicity studies have shown that the intact rodent immune system responds as expected to eliminate B. thuringiensis gradually from the body after oral, pulmonary or intravenous challenge. Similar clearance patterns were observed with B. subtilis and B. sphaericus, which also were non-toxic to test animals. The results also indicate that the currently used protocols for toxicity/pathogenicity evaluations of micro-organisms in laboratory animals have provided useful and necessary information for risk assessment. The study results raise the issue, however, of whether some reduced set of toxicily/pathogenicity studies could be required for new B. thuringiensis isolates of already registered subspecies, and whether this reduced data set would also apply to subspecies not yet registered as pesticidal active ingredients.     

Control of tomato leafminer (Scrobipalpula absoluta) in Bolivia

Abstract

The leafminer Scrobipalpula absoluta Meyr. is a serious pest of tomatoes in the Los Valles region of the eastern foothills of the Bolivian Andes. An initial insecticide screening trial showed that the local population of S. absoluta had developed a high resistance to organophosphorous insecticides. Soil‐applied carbamates and Bacillus thuringiensis Berliner were also ineffective. Further trials were carried out to establish the optimum rates and timing of applications of two synthetic pyrethroids, fenvalerate and permethrin. The results showed that organophosphate‐resistant S. absoluta can be satisfactorily controlled under commercial growing conditions with tow doses of pyrethroid insecticides. Permethrin had greater toxicity and persistence than fenvalerate, and could be applied at longer intervals. It is recommended that permethrin, either as Ambush 50 e.c. at 100 ml/ha or as Pounce 30 e.c. at 170 ml/ha, be applied weekly or at 10–14‐day intervals at the beginning of the season or when infestations and damage are light.     

Field‐evolved resistance to Bt maize in sugarcane borer (Diatraea saccharalis) in Argentina

BACKGROUND

Maize technologies expressing Bacillus thuringiensis (Bt) insecticidal proteins are widely used in Argentina to control sugarcane borer (Diatraea saccharalis Fabricius). Unexpected D. saccharalis damage was observed to Bt maize events TC1507 (expressing Cry1F) and MON 89034 × MON 88017 (expressing Cry1A.105 and Cry2Ab2) in an isolated area of San Luis Province. Diatraea saccharalis larvae were sampled from MON 89034 × MON 88017 fields in the area to generate a resistant strain (RR), which was subsequently characterized in plant and diet bioassays.

RESULTS

Survivorship of the RR strain was high on TC1507 leaf tissue, intermediate on MON 89034 × MON 88017, and low on MON 810 (expressing Cry1Ab). The RR strain had high resistance to Cry1A.105 (186.74‐fold) and no resistance to Cry2Ab2 in diet bioassays. These results indicate resistance to Cry1F and Cry1A.105 (and likely cross‐resistance between them) but not to Cry1Ab or Cry2Ab2. Resistance to MON 89034 × MON 88017 was functionally recessive. Reviews of grower records suggest that resistance initially evolved to Cry1F, conferring cross‐resistance to Cry1A.105, with low refuge compliance as the primary cause. A mitigation plan was implemented in San Luis that included technology rotation, field monitoring, and grower education on best management practices (BMPs) including refuges.

CONCLUSION

In the affected area, the resistance to Cry1F and Cry1A.105 is being managed effectively through use of MON 89034 × MON 88017 and MON 810 in combination with BMPs, and no spread of resistance to other regions has been observed. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Conference Reports

Abstract

In an insect pests survey of soybean during 1977–1980, in the rabi and kharif seasons, 14 species were more, and 33 species less abundant, representing 19 families under seven orders. The occurrence of Aphis spp., Atractomorpha spp., Spilosoma obliqua W., Monolepta signata Olv., Nezara viridula L., Myllocerus spp., Riptortus spp., and Jassids were most notable.     

Putative new species of the genus Dickeya as major soft rot pathogens in Phalaenopsis orchid production

Bacterial soft rots are a serious limitation to the production of orchids and other horticultural plants. Here, the characterization of causative bacteria isolated from Phalaenopsis orchids showing symptoms, from a commercial production site, is reported. The most commonly isolated bacteria were identified as Dickeya spp. Partial sequencing of 16S rDNA, fliC and dnaX showed diversity among the isolates and divided the isolates into two groups, with greatest similarity to previously reported undefined Dickeya lineages from orchids (UDL‐3 and UDL‐4). Two isolates (B16, S1) were sequenced using next‐generation sequencing, which has provided draft genomes of these two isolates for further studies (Ali? et al., 2015 ). Newly developed fliC‐based lineage‐specific quantitative real‐time PCR assays were used to distinguish among the lineages and to assess their relative abundances in diseased tissues. Virulence and aggressiveness comparison tests in vivo on Phalaenopsis orchids, potato plants and witloof chicory leaves indicated high virulence and extreme maceration potential of these novel Dickeya isolates, compared to a reference panel of other Dickeya spp. Pantoea cypripedii (formerly Pectobacterium cypripedii), which has previously been reported as a soft rot pathogen of orchids, was not detected, and isolates obtained from culture collections did not cause symptoms on artificially infected Phalaenopsis orchids.     

Evaluation of powdered fruits of four peppers (Capsicum spp.) for the control of Callosobruchus maculatus (F) on stored cowpea seed

Abstract

The efficacy of two application rates of powdered fruits of four types of peppers (Capsicum spp.) was evaluated for the control of Callosobruchus maculatus (F). The various pepper powders at 2.5 g‐5.0 g per 250 g of seed were effective in reducing oviposition and damage to cowpea seeds. The pepper type Capsicum frutescens var. yarmunchi was the most effective in discouraging oviposition and minimizing damage to the seed, as indicated by the significantly lower number of emergence holes. The seed quality and viability were not affected by the powdered pepper fruit treatment. The result is indicative of the potential for the use of powdered fruits of some peppertypes in protecting stored cowpea seeds against C. maculatus.     

Resistance of Bemisia tabaci (Homoptera: Aleyrodidae) to insecticides in southern Spain with special reference to neonicotinoids

The tobacco whitefly, Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) which occurs in various parts of the world, has developed a high degree of resistance against several chemical classes of insecticide, including organophosphates, carbamates, pyrethroids, insect growth regulators and chlorinated hydrocarbons. The present studies were done in order to monitor the susceptibility of whitefly populations in southern Spain to insecticides commonly used there. Systemic bioassays using Spanish field populations of B tabaci collected in 1994, 1996 and 1998 indicated an increase, albeit a slow one, in resistance to imidacloprid over this period. Comparative studies of other neonicotinoids using the same bioassay revealed a high degree of cross‐resistance to acetamiprid and thiamethoxam. Leaf‐dip bioassays with adult females from these populations revealed a high level of resistance to cyfluthrin, endosulfan, monocrotophos, methamidophos, and pymetrozine, each at 200 mg litre⁻¹. Buprofezin and pyriproxyfen were tested against second‐instar nymphs and eggs, respectively. Buprofezin also showed a lower efficacy against ESP‐98, a strain of B tabaci received from Almeria in 1998, but pyriproxyfen resistance was not obvious when tested against eggs of strain ESP‐98. Field trials in 1998 revealed good efficacy of imidacloprid in one farm in the Almeria region and two greenhouses in Murcia and Sevilla, but a loss of activity by imidacloprid in another farm in the Almeria region. Cross‐resistance between imidacloprid and thiamethoxam was also confirmed under field conditions. © 2000 Society of Chemical Industry