scholarly journals THE EFFECT OF Bacillus thuringiensis TOXIN Cry1A.105 AND Cry2Ab2 ON THE SURVIVAL OF THE NON-TARGET PEST, Spodoptera litura (PENGARUH TOKSIN Bacillus thuringiensis Cry1A.105 DAN Cry2Ab2 TERHADAP KELANGSUNGAN HIDUP HAMA BUKAN SASARAN, Spodoptera litura)

2017 ◽  
Vol 20 (1) ◽  
pp. 7
Author(s):  
Kurnia Pratiwi ◽  
Y. Andi Trisyono ◽  
Edhi Martono

ABSTRACTSpodoptera litura is one of the important insect pest of maize besides the notoriously damaging corn borer, Ostrinia furnacalis. S. litura has been the target of various controls including the use of Bacillus thuringiensis (Bt) toxin Cry1A.105 and Cry2Ab2. This study was conducted to evaluate the acute effect of Bt toxin Cry1A.105 and Cry2Ab2 on the growth and development of S. litura from larval to adult stages. Two sublethal concentrations were used; 0.1875 and 0.0469 ppm for Cry1A.105, and 0.0008 and 0.0003 ppm for Cry2Ab2. The bioassay using diet dipping was carried out on a CRD with three experiments and five repetitions. The observation was carried out on the mortality and survival rates of S. litura. The mortality reached 28% when the larvae were treated with 0.1875 ppm and 20% with 0.0469 ppm of Cry 1A.105. The exposed larvae and pupae were smaller than control. Larval and pupal weight were 117.0 and 165.6 g with 0.1875 ppm, while control were 212.9 and 211.2 g. Cry1A.105 also longer the larval stage, larval stage with higher and lower concentration were 24.5 and 22.3 day, while control was 20.5 day. The resulted pupae from larve which exposed by Cry1A.105 were less than control; there were 40% at concentration 0.1875 ppm and control 61%. The two concentration of Cry2Ab2 produced similar mortality of 20%. Similarly, Cry2Ab2 affected pupal to adult stages development. The longevity of pupal stage with concentration 0.0003 ppm was 9.5 days, followed by 0.0008 ppm (9.1 days) and control (10.1 days). The adult emerge on the highest concentration was 47.4% while control only 34.6%. There results showed that both Cry1A.105 and Cry2Ab2 were detrimental to the survival of S. litura which is the non-target insect of transgenic Bt maize.Keywords: Cry1A.105, Cry2Ab2, maize, non-target pest, Spodoptera litura, survival INTISARISpodoptera litura merupakan salah satu hama penting yang menyerang tanaman jagung, selain Ostrinia furnacalis. Belakangan ini O. furnacalis diketahui telah menjadi target dari berbagai macam cara pengendalian termasuk penggunaan toksin Bacillus thuringiensis (Bt) Cry1A.105 dan Cry2Ab2. Penelitian ini dilakukan untuk mengetahui efek akut toksin Bt Cry1A.105 dan Cry2Ab2 terhadap pertumbuhan dan perkembangan S. litura dari larva sampai imago. Dua konsentrasi subletal yang akan digunakan adalah; 0,1875 dan 0,0469 ppm untuk Cry1A.105, dan 0,0008 dan 0,0003 ppm untuk Cry2Ab2. Pengujian dilakukan dengan menggunakan metode celup pakan dan Rancangan Acak Legkap dengan tiga perlakuan dan lima ulangan. Pengamatan dilakukan terhadap mortalitas dan kelangsungan hidup S. litura. Mortalitas mencapai 28% pada larva yang dipaparkan dengan 0,1875 ppm dan 20% dengan 0,0469 ppm Cry 1A.105. Larva dan pupa yang terkena toksin berukuran lebih kecil. Berat larva dan pupa yang terpapar toksin dengan konsentrasi 0,1875 ppm, masing – masing 117,0 dan 165,6 g, sedangkan kontrol masing – masing 212,9 dan 211,2 g. Cry1A.105 juga dapat memperpanjang stadia larva. Lama stadia larva dengan konsentrasi tertinggi dan terendah adalah 24,5 dan 22,3 hari, sedangkan kontrol 20,5 hari. Jumlah pupa yang berhasil terbentuk dari larva yang terpapar toxin Cry1A.105 lebih sedikit dibandingkan dengan kontrol; pada konsentrasi 0,1875 ppm sebesar 40%, sedangkan kontrol sebesar 61%. Kedua konsentrasi dari toksin Bt Cry2Ab2 menyebabkan mortalitas yang sama yaitu 20%. Cry2Ab2 juga berpengaruh terhadap lama stadia pupa dan tingkat keberhasilan pembentukan imago. Lama stadia pupa dengan konsentrasi 0,0003 ppm adalah 9,5 hari, diikuti dengan konsentrasi 0,0008 ppm (9,1 hari) dan kontrol (10,1 hari). Jumlah imago terbanyak terdapat pada perlakuan dengan konsentrasi 0,0008 ppm sebesar 47,4% sedangkan pada kontrol hanya 34,6%. Hal ini menunjukkan bahwa toksin Bt Cry1A. 105 dan Cry2Ab2 juga berpengaruh terhadap kelangsungan hidup S. litura yang merupakan serangga bukan sasaran dari tanaman jagung transgenik Bt.Kata kunci: Cry1A.105, Cry2Ab2, hama bukan sasaran, jagung, kelangsungan hidup, Spodoptera litura

Toxins ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 658
Author(s):  
Wenzhong Jin ◽  
Yuqian Zhai ◽  
Yihua Yang ◽  
Yidong Wu ◽  
Xingliang Wang

Transgenic crops expressing Bacillus thuringiensis (Bt) insecticidal proteins have been extensively planted for insect pest control, but the evolution of Bt resistance in target pests threatens the sustainability of this approach. Mutations of cadherin in the midgut brush border membrane was associated with Cry1Ac resistance in several lepidoptera species, including the Asian corn borer, Ostrinia furnacalis, a major pest of maize in Asian–Western Pacific countries. However, the causality of O. furnacalis cadherin (OfCad) with Cry1Ac resistance remains to be clarified. In this study, in vitro and in vivo approaches were employed to examine the involvement of OfCad in mediating Cry1Ac toxicity. Sf9 cells transfected with OfCad showed significant immunofluorescent binding with Cry1Ac toxin and exhibited a concentration-dependent mortality effect when exposed to Cry1Ac. The OfCad knockout strain OfCad-KO, bearing homozygous 15.4 kb deletion of the OfCad gene generated by CRISPR/Cas9 mutagenesis, exhibited moderate-level resistance to Cry1Ac (14-fold) and low-level resistance to Cry1Aa (4.6-fold), but no significant changes in susceptibility to Cry1Ab and Cry1Fa, compared with the original NJ-S strain. The Cry1Ac resistance phenotype was inherited as autosomal, recessive mode, and significantly linked with the OfCad knockout in the OfCad-KO strain. These results demonstrate that the OfCad protein is a functional receptor for Cry1Ac, and disruption of OfCad confers a moderate Cry1Ac resistance in O. furnacalis. This study provides new insights into the mode of action of the Cry1Ac toxin and useful information for designing resistance monitoring and management strategies for O. furnacalis.


2017 ◽  
Vol 108 (5) ◽  
pp. 658-666 ◽  
Author(s):  
X. Lin ◽  
Y. Jiang ◽  
L. Zhang ◽  
Y. Cai

AbstractSpodoptera litura is a widespread polyphagous insect pest that can develop resistance and cross-resistance to insecticides, making it difficult to control. Insecticide exposure has previously been linked with induction of specific olfactory-related proteins, including some chemosensory proteins (CSPs) and odorant-binding proteins (OPBs), which may disrupt detection of environmental factors and reduce fitness. However, functional evidence supporting insecticide and OBPs/CSPs mediation remains unknown. Here we fed male S. litura moths with sucrose water containing one of three insecticides, chlorpyrifos, emamectin benzoate or fipronil, and used real-time quantitative polymerase chain reaction and RNAi to investigate OBPs and CSPs expression and their correlations with survival. Chlorpyrifos and emamectin benzoate increased expression of 78% of OBPs, plus 63 and 56% of CSP genes, respectively, indicating a major impact on these gene families. RNAi knockdown of SlituCSP18, followed by feeding with chlorpyrifos or fipronil, decreased survival rates of male moths significantly compared with controls. Survival rate also decreased significantly with the downregulation of SlituOBP9 followed by feeding with chlorpyrifos. Thus, although these three insecticides had different effects on OBP and CSP gene expression, we hypothesize that SlituOBPs and SlituCSPs might mediate their effects by increasing their expression levels to improve survival. Moreover, the differential response of S. litura male moths to the three insecticides indicated the potential specificity of chlorpyrifos affect SlituCSP18 and SlituOBP9 expression.


Toxins ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 246 ◽  
Author(s):  
Xingliang Wang ◽  
Yanjun Xu ◽  
Jianlei Huang ◽  
Wenzhong Jin ◽  
Yihua Yang ◽  
...  

The adoption of transgenic crops expressing Bacillus thuringiensis (Bt) insecticidal crystalline (Cry) proteins has reduced insecticide application, increased yields, and contributed to food safety worldwide. However, the efficacy of transgenic Bt crops is put at risk by the adaptive resistance evolution of target pests. Previous studies indicate that resistance to Bacillus thuringiensis Cry1A and Cry1F toxins was genetically linked with mutations of ATP-binding cassette (ABC) transporter subfamily C gene ABCC2 in at least seven lepidopteran insects. Several strains selected in the laboratory of the Asian corn borer, Ostrinia furnacalis, a destructive pest of corn in Asian Western Pacific countries, developed high levels of resistance to Cry1A and Cry1F toxins. The causality between the O. furnacalis ABCC2 (OfABCC2) gene and resistance to Cry1A and Cry1F toxins remains unknown. Here, we successfully generated a homozygous strain (OfC2-KO) of O. furnacalis with an 8-bp deletion mutation of ABCC2 by the CRISPR/Cas9 approach. The 8-bp deletion mutation results in a frame shift in the open reading frame of transcripts, which produced a predicted protein truncated in the TM4-TM5 loop region. The knockout strain OfC2-KO showed much more than a 300-fold resistance to Cry1Fa, and low levels of resistance to Cry1Ab and Cry1Ac (<10-fold), but no significant effects on the toxicities of Cry1Aa and two chemical insecticides (abamectin and chlorantraniliprole), compared to the background NJ-S strain. Furthermore, we found that the Cry1Fa resistance was autosomal, recessive, and significantly linked with the 8-bp deletion mutation of OfABCC2 in the OfC2-KO strain. In conclusion, in vivo functional investigation demonstrates the causality of the OfABCC2 truncating mutation with high-level resistance to the Cry1Fa toxin in O. furnacalis. Our results suggest that the OfABCC2 protein might be a functional receptor for Cry1Fa and reinforces the association of this gene to the mode of action of the Cry1Fa toxin.


2005 ◽  
Vol 40 (3) ◽  
pp. 307-315 ◽  
Author(s):  
Kerry C. Allen ◽  
Henry N. Pitre

Laboratory studies were conducted to compare the response of Diatraea grandiosella (Dyar) larvae to lyophilized transgenic corn tissue expressing the Cry1Ab endotoxin protein of Bacillus thuringiensis (Berliner) (Bt) with conventional corn tissue incorporated into a BioServ™ artificial diet. Whorl leaf, stalk, shank, husk, silk or kernel tissues were tested independently in the diet. Larvae fed diet containing conventional corn weighed more and were longer in length than larvae fed diet containing Bt corn for all tissue types included in the study. The number of larvae that survived depended on the tissue type and age of the tissue. Larvae fed diet with kernel tissue expressing Bt toxin had greater weight and body length than larvae fed the other Bt tissue types. The negative effects of Bt corn tissues expressing the Cry1Ab endotoxin protein on growth of D. grandiosella was observed, even at the diluted concentrations of toxic tissues incorporated into the diet in this study.


2006 ◽  
Vol 73 (3) ◽  
pp. 956-961 ◽  
Author(s):  
Jun Fang ◽  
Xiaoli Xu ◽  
Ping Wang ◽  
Jian-Zhou Zhao ◽  
Anthony M. Shelton ◽  
...  

ABSTRACT Bacillus thuringiensis vegetative insecticidal proteins (Vip) are potential alternatives for B. thuringiensis endotoxins that are currently utilized in commercial transgenic insect-resistant crops. Screening a large number of B. thuringiensis isolates resulted in the cloning of vip3Ac1. Vip3Ac1 showed high insecticidal activity against the fall armyworm Spodoptera frugiperda and the cotton bollworm Helicoverpa zea but very low activity against the silkworm Bombyx mori. The host specificity of this Vip3 toxin was altered by sequence swapping with a previously identified toxin, Vip3Aa1. While both Vip3Aa1 and Vip3Ac1 showed no detectable toxicity against the European corn borer Ostrinia nubilalis, the chimeric protein Vip3AcAa, consisting of the N-terminal region of Vip3Ac1 and the C-terminal region of Vip3Aa1, became insecticidal to the European corn borer. In addition, the chimeric Vip3AcAa had increased toxicity to the fall armyworm. Furthermore, both Vip3Ac1 and Vip3AcAa are highly insecticidal to a strain of cabbage looper (Trichoplusia ni) that is highly resistant to the B. thuringiensis endotoxin Cry1Ac, thus experimentally showing for the first time the lack of cross-resistance between B. thuringiensis Cry1A proteins and Vip3A toxins. The results in this study demonstrated that vip3Ac1 and its chimeric vip3 genes can be excellent candidates for engineering a new generation of transgenic plants for insect pest control.


2019 ◽  
Vol 113 (1) ◽  
pp. 194-202
Author(s):  
Xiujing Shen ◽  
Xiaowei Fu ◽  
Yunxin Huang ◽  
Jianglong Guo ◽  
Qiulin Wu ◽  
...  

Abstract The Asian corn borer (ACB), Ostrinia furnacalis (Guenée), is a notorious polyphagous insect pest in China and other regions of East Asia. The long-distance flight behavior of the ACB moths, however, is not well understood, especially in the field conditions. In this study, nightly monitoring data for multiple years (2003–2017) on the long-distance flight of adults crossing the Bohai Strait in northern China, showed that a large number of the adults frequently flew across the Bohai Strait from May to September with the peak migrations usually occurred, followed in order by September, June, July, and May, and the number of southward migrants was larger than that of northward migrants. From May to September in 2010, 2011, and 2017, a subsample of trapped ACB females was dissected (879 individuals in 2010, 197 individuals in 2011, and 247 individuals in 2017), and the results showed that the sex ratio of the trapped ACB moths was unbiased each month from May to September. While the proportion of mated females in northward populations (May to July = 92.85 ± 4.86%) was significantly higher than that in southward population (August and September = 74.53 ± 8.55%) . The seasonal pattern in the proportion of sexually mature females was similar to the above proportion of mated females. We conclude that the ACBs have a strong propensity and ability for long-distance migration, although local dispersal is generally considered to be the primary movement of the ACB. These findings may be helpful to improve the forecasting systems and the pest management schemes for the ACBs.


Insects ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 718
Author(s):  
Carmen López ◽  
Pilar Muñoz ◽  
Daniela Zanga ◽  
Patricia Sarai Girón-Calva ◽  
Matilde Eizaguirre

Serious malnutrition problems occur in developing countries where people’s diets are mainly based on staple crops. To alleviate this, high-production crops are being developed that are better adapted to climate change, enriched in micronutrients and vitamins, or resistant to pests. In some cases, new varieties have been developed with several of the characteristics mentioned above, such as biofortified and pest-resistant crops. The development of biofortified Bacillus thuringiensis (Bt) crops raises the question of whether vitamin enrichment of Bt crops can in any way favor those pests that are not very susceptible to the Bt toxin that feed on these crops, such as Helicoverpa armigera (Hübner) or Mythimna unipuncta (Haworth) (Lepidoptera: Noctuidae). In this study, the response to a Bt diet enriched with vitamins A (β-carotene) and C (ascorbic acid) was somewhat different between the two species. M. unipuncta was less sensitive to the toxin than H. armigera, although the ingestion of the Bt diet resulted in oxidative stress (longer larval development and lower pupal weight) which was not mitigated by the vitamins. However, the two vitamins reduced the mortality of H. armigera larvae fed on a Bt-enriched diet; in addition, ß-carotene reduced the activity of the antioxidant glutathione S-transferase (GST) of both species, suggesting it has an antioxidant role. The results obtained here indicate that biofortified Bt crops will not favor the development of H. armigera very much and will not affect M. unipuncta’s development at all, although the effect of the increase in vitamins may be very variable and should be studied for each specific phytophagous.


2015 ◽  
Vol 84 (1) ◽  
pp. 47-53 ◽  
Author(s):  
Vladimír Ostrý ◽  
František Malíř ◽  
Annie Pfohl-Leszkowicz

Transgenic Bt maize is a potentially important tool against insect pest in the EU and other countries. Bt maize (e.g. MON 810, Bt 11) which carries the Bt gene is highly resistant to larval feeding of European corn borer, stalk borer, and Southwestern corn borer, depending on Bt toxin (δ toxin) production. Effective measures used to fight pests may often have positive side-effects in that they may also contribute to reducing mycotoxin concentrations. A systematic review has been used for the purposes of evaluating the studies on the reduction of aflatoxins in Bt maize. According to five studies, Bt maize has significantly lower concentrations of aflatoxins than non-Bt maize hybrids, only one study has shown no significant effect of Bt maize. Other studies have shown mixed results (four studies). The results of these studies were influenced by the year of sampling or by using maize breeding lines selected for resistance to aflatoxin accumulation.


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Karine da Silva Carvalho ◽  
Duschinka Ribeiro Duarte Guedes ◽  
Mônica Maria Crespo ◽  
Maria Alice Varjal de Melo-Santos ◽  
Maria Helena Neves Lobo Silva-Filha

Abstract Background Aedes aegypti can transmit arboviruses worldwide, and Bacillus thuringiensis svar. israelensis (Bti)-based larvicides represent an effective tool for controlling this species. The safety of Bti and lack of resistance have been widely reported; however, little is known regarding the impact of the extensive use of these larvicides on the life traits of mosquitoes. Therefore, this study investigated biological parameters, including susceptibility to arbovirus, of an Ae. aegypti strain (RecBti) subjected to 29 generations of exposure to Bti compared with the RecL reference strain. Methods The biological parameters of individuals reared under controlled conditions were compared. Also, the viral susceptibility of females not exposed to Bti during their larval stage was analysed by oral infection and followed until 14 or 21 days post-infection (dpi). Results RecBti individuals did not display alterations in the traits that were assessed (fecundity, fertility, pupal weight, developmental time, emergence rate, sex ratio and haematophagic capacity) compared to RecL individuals. Females from both strains were susceptible to dengue serotype 2 (DENV-2) and Zika virus (ZIKV). However, RecBti females showed significantly higher rates of ZIKV infection compared with RecL females at 7 (90% versus 68%, Chi-square: χ2 = 7.27, df = 1, P = 0.006) and 14 dpi (100% versus 87%, Chi-square: χ2 = 7.69, df = 1, P = 0.005) and for dissemination at 7 dpi (83.3% versus 36%, Fisher’s exact test: P < 0.0001, OR = 0.11, 95% CI 0.03–0.32). Quantification of DENV-2 and ZIKV viral particles produced statistically similar results for females from both strains. Conclusions Prolonged exposure of Ae. aegypti larvae to Bti did not alter most of the evaluated biological parameters, except that RecBti females exhibited a higher vector susceptibility for ZIKV. This finding is related to a background of Bti exposure for several generations but not to a previous exposure of the tested females during the larval stage. This study highlights mosquito responses that could be associated with the chronic exposure to Bti in addition to the primary larvicidal effect elicited by this control agent. Graphical abstract


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