scholarly journals Current Insights on Vegetative Insecticidal Proteins (Vip) as Next Generation Pest Killers

Toxins ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 522
Author(s):  
Tahira Syed ◽  
Muhammad Askari ◽  
Zhigang Meng ◽  
Yanyan Li ◽  
Muhammad Ali Abid ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Mechanism Of Action ◽  
Insecticidal Activity ◽  
Binding Sites ◽  
Growth Phases ◽  
Cry Proteins ◽  
Insecticidal Proteins ◽  
Gram Negative ◽  
Vip Proteins ◽  
Evolution Of Resistance

Bacillus thuringiensis (Bt) is a Gram negative soil bacterium. This bacterium secretes various proteins during different growth phases with an insecticidal potential against many economically important crop pests. One of the important families of Bt proteins is vegetative insecticidal proteins (Vip), which are secreted into the growth medium during vegetative growth. There are three subfamilies of Vip proteins. Vip1 and Vip2 heterodimer toxins have an insecticidal activity against many Coleopteran and Hemipteran pests. Vip3, the most extensively studied family of Vip toxins, is effective against Lepidopteron. Vip proteins do not share homology in sequence and binding sites with Cry proteins, but share similarities at some points in their mechanism of action. Vip3 proteins are expressed as pyramids alongside Cry proteins in crops like maize and cotton, so as to control resistant pests and delay the evolution of resistance. Biotechnological- and in silico-based analyses are promising for the generation of mutant Vip proteins with an enhanced insecticidal activity and broader spectrum of target insects.

scholarly journals Insecticidal Activity of Bacillus thuringiensis Proteins against Coleopteran Pests

Toxins ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 430 ◽  
Author(s):  
Mikel Domínguez-Arrizabalaga ◽  
Maite Villanueva ◽  
Baltasar Escriche ◽  
Carmen Ancín-Azpilicueta ◽  
Primitivo Caballero
Keyword(s):  
Bacillus Thuringiensis ◽  
Insecticidal Activity ◽  
Mode Of Action ◽  
Insect Species ◽  
Specific Information ◽  
Cry Proteins ◽  
Microbial Insecticide ◽  
Bt Toxins ◽  
Vip Proteins ◽  
Agro Forestry

Bacillus thuringiensis is the most successful microbial insecticide agent and its proteins have been studied for many years due to its toxicity against insects mainly belonging to the orders Lepidoptera, Diptera and Coleoptera, which are pests of agro-forestry and medical-veterinary interest. However, studies on the interactions between this bacterium and the insect species classified in the order Coleoptera are more limited when compared to other insect orders. To date, 45 Cry proteins, 2 Cyt proteins, 11 Vip proteins, and 2 Sip proteins have been reported with activity against coleopteran species. A number of these proteins have been successfully used in some insecticidal formulations and in the construction of transgenic crops to provide protection against main beetle pests. In this review, we provide an update on the activity of Bt toxins against coleopteran insects, as well as specific information about the structure and mode of action of coleopteran Bt proteins.

scholarly journals Bacillus thuringiensis Vip1 Functions as a Receptor of Vip2 Toxin for Binary Insecticidal Activity against Holotrichia parallela

Toxins ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 440 ◽  
Author(s):  
Jianxun Geng ◽  
Jian Jiang ◽  
Changlong Shu ◽  
Zeyu Wang ◽  
Fuping Song ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Affinity Chromatography ◽  
Insecticidal Activity ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Lethal Concentration ◽  
Insecticidal Proteins ◽  
Vegetative Phase ◽  
Holotrichia Parallela ◽  
Weak Binding

Bacillus thuringiensis is a well-known entomopathogenic bacterium that produces vegetative insecticidal proteins (Vips, including Vip1, Vip2, Vip3, and Vip4) during the vegetative phase. Here, we purified Vip1 and Vip2 from B. thuringiensis and characterized the insecticidal effects of these protoxins. Bioassay results showed that a 1:1 mixture of Vip1Ad and Vip2Ag, purified by ion-affinity chromatography independently, exhibited insecticidal activity against Holotrichia parallela larvae, with a 50% lethal concentration value of 2.33 μg/g soil. The brush border membrane (BBM) in the midgut of H. parallela larvae was destroyed after feeding the Vip1Ad and Vip2Ag mixture. Vacuolization of the cytoplasm and slight destruction of BBM were detected with Vip2Ag alone, but not with Vip1Ad alone. Notably, Vip1Ad bound to BBM vesicles (BBMVs) strongly, whereas Vip2Ag showed weak binding; however, binding of Vip2Ag to BBMV was increased when Vip1Ad was added. Ligand blotting showed that Vip2Ag did not bind to Vip1Ad but bound to Vip1Ad-t (Vip1Ad was activated by trypsin), suggesting the activation of Vip1Ad was important for their binary toxicity. Thus, our findings suggested that Vip1Ad may facilitate the binding of Vip2Ag to BBMVs, providing a basis for studies of the insecticidal mechanisms of Vip1Ad and Vip2Ag.

scholarly journals Oligomer Formation and Insecticidal Activity of Bacillus thuringiensis Vip3Aa Toxin

Toxins ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 274 ◽  
Author(s):  
Ensi Shao ◽  
Aishan Zhang ◽  
Yaqi Yan ◽  
Yaomin Wang ◽  
Xinyi Jia ◽  
...  
Keyword(s):  
Amino Acid ◽  
Bacillus Thuringiensis ◽  
Complex Formation ◽  
Insecticidal Activity ◽  
Spodoptera Litura ◽  
Insecticidal Proteins ◽  
Proteolytic Activation ◽  
Oligomer Formation ◽  
Lepidopteran Insects ◽  
Substitution Mutations

Bacillus thuringiensis (Bt) Vip3A proteins are important insecticidal proteins used for control of lepidopteran insects. However, the mode of action of Vip3A toxin is still unclear. In this study, the amino acid residue S164 in Vip3Aa was identified to be critical for the toxicity in Spodoptera litura. Results from substitution mutations of the S164 indicate that the insecticidal activity of Vip3Aa correlated with the formation of a >240 kDa complex of the toxin upon proteolytic activation. The >240 kDa complex was found to be composed of the 19 kDa and the 65 kDa fragments of Vip3Aa. Substitution of the S164 in Vip3Aa protein with Ala or Pro resulted in loss of the >240 kDa complex and loss of toxicity in Spodoptera litura. In contrast, substitution of S164 with Thr did not affect the >240 kDa complex formation, and the toxicity of the mutant was only reduced by 35%. Therefore, the results from this study indicated that formation of the >240 kDa complex correlates with the toxicity of Vip3Aa in insects and the residue S164 is important for the formation of the complex.

scholarly journals Use of Bacillus thuringiensis Toxins for Control of the Cotton Pest Earias insulana (Boisd.) (Lepidoptera: Noctuidae)

2006 ◽  
Vol 72 (1) ◽  
pp. 437-442 ◽  
Author(s):  
María A. Ibargutxi ◽  
Anna Estela ◽  
Juan Ferré ◽  
Primitivo Caballero
Keyword(s):  
Bacillus Thuringiensis ◽  
Binding Sites ◽  
Specific Binding ◽  
Significant Inhibition ◽  
The Other ◽  
Cross Resistance ◽  
Earias Insulana ◽  
Cry Proteins ◽  
Cry Toxins ◽  
Laboratory Colony

ABSTRACT Thirteen of the most common lepidopteran-specific Cry proteins of Bacillus thuringiensis have been tested for their efficacy against newly hatched larvae of two populations of the spiny bollworm, Earias insulana. At a concentration of 100 μg of toxin per milliliter of artificial diet, six Cry toxins (Cry1Ca, Cry1Ea, Cry1Fa, Cry1Ja, Cry2Aa, and Cry2Ab) were not toxic at all. Cry1Aa, Cry1Ja, and Cry2Aa did not cause mortality but caused significant inhibition of growth. The other Cry toxins (Cry1Ab, Cry1Ac, Cry1Ba, Cry1Da, Cry1Ia, and Cry9Ca) were toxic to E. insulana larvae. The 50% lethal concentration values of these toxins ranged from 0.39 to 21.13 μg/ml (for Cry9Ca and Cry1Ia, respectively) for an E. insulana laboratory colony originating from Egypt and from 0.20 to 4.25 μg/ml (for Cry9Ca and Cry1Da, respectively) for a laboratory colony originating from Spain. The relative potencies of the toxins in the population from Egypt were highest for Cry9Ca and Cry1Ab, and they were both significantly more toxic than Cry1Ac and Cry1Ba, followed by Cry1Da and finally Cry1Ia. In the population from Spain, Cry9Ca was the most toxic, followed in decreasing order by Cry1Ac and Cry1Ba, and the least toxic was Cry1Da. Binding experiments were performed to test whether the toxic Cry proteins shared binding sites in this insect. 125I-labeled Cry1Ac and Cry1Ab and biotinylated Cry1Ba, Cry1Ia, and Cry9Ca showed specific binding to the brush border membrane vesicles from E. insulana. Competition binding experiments among these toxins showed that only Cry1Ab and Cry1Ac competed for the same binding sites, indicating a high possibility that this insect may develop cross-resistance to Cry1Ab upon exposure to Cry1Ac transgenic cotton but not to the other toxins tested.

scholarly journals Fatores de virulência de Bacillus thuringiensis: o que existe além das proteínas Cry

2012 ◽  
Vol 5 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Gislayne Trindade Vilas-Bôas ◽  
Rita C. Alvarez ◽  
Clelton A. Dos Santos ◽  
Laurival A. Vilas-Boas
Keyword(s):  
Biological Control ◽  
Bacillus Thuringiensis ◽  
Virulence Factors ◽  
Insect Pests ◽  
Cry Proteins ◽  
Control Programs ◽  
Wide Range ◽  
Vip Proteins ◽  
Bacterium Bacillus ◽  
And Control

As proteínas Cry produzidas pela bactéria entomopatogênica Bacillus thuringiensis Berliner são bem conhecidas devido a alta citotoxicidade que exibem a uma variedade de insetos-alvo. O modo de ação destas proteínas é específico e torna os produtos à base de B. thuringiensis os mais amplamente utilizados em programas de controle biológico de pragas na agricultura e de importantes vetores de doenças humanas. Contudo, embora as proteínas Cry sejam os fatores de virulência inseto-específico mais conhecidos, linhagens de B. thuringiensis apresentam também uma ampla gama de fatores de virulência, os quais permitem à bactéria atingir a hemolinfa e colonizar eficientemente o inseto hospedeiro. Dentre estes fatores, destacam-se as proteínas Vip, Cyt, enterotoxinas, hemolisinas, fosfolipases, proteases, enzimas de degradação, além das recentemente descritas parasporinas. Essa revisão aborda a ação desses fatores de virulência, bem como a caracterização e o controle da expressão de seus genes. Adicionalmente, são discutidos aspectos relacionados ao nicho ecológico da bactéria com ênfase nas características envolvidas com a biossegurança da utilização dos produtos à base de B. thuringiensis para o controle biológico de insetos-alvo. Virulence Factors of Bacillus thuringiensis Berliner: Something Beyond of Cry Proteins? Abstract. The Cry proteins produced by the entomopathogenic bacterium Bacillus thuringiensis Berliner are widely known due to its high toxicity against a variety of insects. The mode of action of these proteins is specific and becomes B. thuringiensis-based products the most used in biological control programs of insect pests in agriculture and of important human disease vectors. However, while the Cry proteins are the best-known insect-specific virulence factor, strains of B. thuringiensis show also a wide range of other virulence factors, which allow the bacteria to achieve the hemolymph and colonize efficiently the insect host. Among these factors, we highlight the Vip proteins, Cyt, enterotoxins, hemolysins, phospholipases, proteases and enzymes of degradation, in addition to the recently described parasporin. This review explores the action of these virulence factors, as well as, the characterization and control of expression of their genes. Additionally, we discuss aspects related to the ecological niche of the bacteria with emphasis on the characteristics involved in the biosafety of the use of B. thuringiensis-based products for biological control of target insects.

scholarly journals Vegetative Insecticidal Protein (Vip): A Potential Contender From Bacillus thuringiensis for Efficient Management of Various Detrimental Agricultural Pests

2021 ◽  
Vol 12 ◽  
Author(s):  
Mamta Gupta ◽  
Harish Kumar ◽  
Sarvjeet Kaur
Keyword(s):  
Bacillus Thuringiensis ◽  
Insecticidal Protein ◽  
Current Status ◽  
Specific Class ◽  
Agricultural Pests ◽  
Cry Proteins ◽  
Vegetative Insecticidal Protein ◽  
Receptor Sites ◽  
Vip Proteins ◽  
The Difference

Bacillus thuringiensis (Bt) bacterium is found in various ecological habitats, and has natural entomo-pesticidal properties, due to the production of crystalline and soluble proteins during different growth phases. In addition to Cry and Cyt proteins, this bacterium also produces Vegetative insecticidal protein (Vip) during its vegetative growth phase, which is considered an excellent toxic candidate because of the difference in sequence homology and receptor sites from Cry proteins. Vip proteins are referred as second-generation insecticidal proteins, which can be used either alone or in complementarity with Cry proteins for the management of various detrimental pests. Among these Vip proteins, Vip1 and Vip2 act as binary toxins and have toxicity toward pests belonging to Hemiptera and Coleoptera orders, whereas the most important Vip3 proteins have insecticidal activity against Lepidopteran pests. These Vip3 proteins are similar to Cry proteins in terms of toxicity potential against susceptible insects. They are reported to be toxic toward pests, which can’t be controlled with Cry proteins. The Vip3 proteins have been successfully pyramided along with Cry proteins in transgenic rice, corn, and cotton to combat resistant pest populations. This review provides detailed information about the history and importance of Vip proteins, their types, structure, newly identified specific receptors, and action mechanism of this specific class of proteins. Various studies conducted on Vip proteins all over the world and the current status have been discussed. This review will give insights into the significance of Vip proteins as alternative promising candidate toxic proteins from Bt for the management of pests in most sustainable manner.

scholarly journals Characterization of Two Novel Bacillus thuringiensis Cry8 Toxins Reveal Differential Specificity of Protoxins or Activated Toxins against Chrysomeloidea Coleopteran Superfamily

Toxins ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 642
Author(s):  
Changlong Shu ◽  
Guixin Yan ◽  
Shizhi Huang ◽  
Yongxin Geng ◽  
Mario Soberón ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Genome Sequencing ◽  
Insecticidal Activity ◽  
Insect Pest ◽  
Anoplophora Glabripennis ◽  
Insecticidal Proteins ◽  
Holotrichia Parallela ◽  
Larval Stages ◽  
Differential Toxicity

Scarabaeoidea and Chrysomeloidea insects are agriculture-destructive coleopteran pests. Few effective Bacillus thuringiensis (Bt) insecticidal proteins against these species have been described. Bt isolate BtSU4 was found to be active against coleopteran insects. Genome sequencing revealed two new cry8 genes in BtSU4, designated as cry8Ha1 and cry8Ia1. Both genes expressed a 135 kDa protoxin forming irregular shape crystals. Bioassays performed with Cry8Ha1 protoxin showed that it was toxic to both larvae and adult stages of Holotrichia parallela, also to Holotrichia oblita adults and to Anoplophora glabripennis larvae, but was not toxic to larval stages of H. oblita or Colaphellus bowringi. The Cry8Ia1 protoxin only showed toxicity against H. parallela larvae. After activation with chymotrypsin, the Cry8Ha1 activated toxin lost its insecticidal activity against H. oblita adults and reduced its activity on H. parallela adults, but gained toxicity against C. bowringi larvae, a Chrysomeloidea insect pest that feeds on crucifer crops. The chymotrypsin activated Cry8Ia1 toxin did not show toxicity to any one of these insects. These data show that Cry8Ha1 and Cry8Ia1 protoxin and activated toxin proteins have differential toxicity to diverse coleopteran species, and that protoxin is a more robust protein for the control of coleopteran insects.

scholarly journals Interaction of Bacillus thuringiensis Toxins with Larval Midgut Binding Sites of Helicoverpa armigera (Lepidoptera: Noctuidae)

2004 ◽  
Vol 70 (3) ◽  
pp. 1378-1384 ◽  
Author(s):  
Anna Estela ◽  
Baltasar Escriche ◽  
Juan Ferr�
Keyword(s):  
Bacillus Thuringiensis ◽  
Concanavalin A ◽  
Helicoverpa Armigera ◽  
Binding Sites ◽  
Inhibitory Effect ◽  
Toxin Gene ◽  
Bt Cotton ◽  
Cry1ab Protein ◽  
Cry Proteins ◽  
Helicoverpa Armígera

ABSTRACT In 1996, Bt-cotton (cotton expressing a Bacillus thuringiensis toxin gene) expressing the Cry1Ac protein was commercially introduced to control cotton pests. A threat to this first generation of transgenic cotton is the evolution of resistance by the insects. Second-generation Bt-cotton has been developed with either new B. thuringiensis genes or with a combination of cry genes. However, one requirement for the “stacked” gene strategy to work is that the stacked toxins bind to different binding sites. In the present study, the binding of 125I-labeled Cry1Ab protein (125I-Cry1Ab) and 125I-Cry1Ac to brush border membrane vesicles (BBMV) of Helicoverpa armigera was analyzed in competition experiments with 11 nonlabeled Cry proteins. The results indicate that Cry1Aa, Cry1Ab, and Cry1Ac competed for common binding sites. No other Cry proteins tested competed for either 125I-Cry1Ab or 125I-Cry1Ac binding, except Cry1Ja, which competed only at the highest concentrations used. Furthermore, BBMV from four H. armigera populations were also tested with 125I-Cry1Ac and Cry1Ab to check the influence of the insect population on the binding results. Finally, the inhibitory effect of selected sugars and lectins was also determined. 125I-Cry1Ac binding was strongly inhibited by N-acetylgalactosamine, sialic acid, and concanavalin A and moderately inhibited by soybean agglutinin. In contrast, 125I-Cry1Ab binding was only significantly inhibited by concanavalin A. These results show that Cry1Ac and Cry1Ab use different epitopes for binding to BBMV.

scholarly journals A Bacillus thuringiensis S-Layer Protein Involved in Toxicity against Epilachna varivestis (Coleoptera: Coccinellidae)

2006 ◽  
Vol 72 (1) ◽  
pp. 353-360 ◽  
Author(s):  
Guadalupe Peña ◽  
Juan Miranda-Rios ◽  
Gustavo de la Riva ◽  
Liliana Pardo-López ◽  
Mario Soberón ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Insecticidal Activity ◽  
Pathogenic Bacteria ◽  
Bacillus Sphaericus ◽  
Epilachna Varivestis ◽  
Gene Encoding ◽  
Parasporal Crystal ◽  
Cry Proteins ◽  
Bean Beetle ◽  
Paracrystalline Array

ABSTRACT The use of Bacillus thuringiensis as a biopesticide is a viable alternative for insect control since the insecticidal Cry proteins produced by these bacteria are highly specific; harmless to humans, vertebrates, and plants; and completely biodegradable. In addition to Cry proteins, B. thuringiensis produces a number of extracellular compounds, including S-layer proteins (SLP), that contribute to virulence. The S layer is an ordered structure representing a proteinaceous paracrystalline array which completely covers the surfaces of many pathogenic bacteria. In this work, we report the identification of an S-layer protein by the screening of B. thuringiensis strains for activity against the coleopteran pest Epilachna varivestis (Mexican bean beetle; Coleoptera: Coccinellidae). We screened two B. thuringiensis strain collections containing unidentified Cry proteins and also strains isolated from dead insects. Some of the B. thuringiensis strains assayed against E. varivestis showed moderate toxicity. However, a B. thuringiensis strain (GP1) that was isolated from a dead insect showed a remarkably high insecticidal activity. The parasporal crystal produced by the GP1 strain was purified and shown to have insecticidal activity against E. varivestis but not against the lepidopteran Manduca sexta or Spodoptera frugiperda or against the dipteran Aedes aegypti. The gene encoding this protein was cloned and sequenced. It corresponded to an S-layer protein highly similar to previously described SLP in Bacillus anthracis (EA1) and Bacillus licheniformis (OlpA). The phylogenetic relationships among SLP from different bacteria showed that these proteins from Bacillus cereus, Bacillus sphaericus, B. anthracis, B. licheniformis, and B. thuringiensis are arranged in the same main group, suggesting similar origins. This is the first report that demonstrates that an S-layer protein is directly involved in toxicity to a coleopteran pest.

scholarly journals Cyt1A of Bacillus thuringiensis Delays Evolution of Resistance to Cry11A in the Mosquito Culex quinquefasciatus

2005 ◽  
Vol 71 (1) ◽  
pp. 185-189 ◽  
Author(s):  
Margaret C. Wirth ◽  
Hyun-Woo Park ◽  
William E. Walton ◽  
Brian A. Federici
Keyword(s):  
Bacillus Thuringiensis ◽  
Culex Quinquefasciatus ◽  
Laboratory Experiments ◽  
Principal Factor ◽  
Resistance Ratio ◽  
Cry Proteins ◽  
Lepidopteran Larvae ◽  
Evolution Of Resistance ◽  
Low Levels ◽  
Selection Of

ABSTRACT Insecticides based on Bacillus thuringiensis subsp. israelensis have been used for mosquito and blackfly control for more than 20 years, yet no resistance to this bacterium has been reported. Moreover, in contrast to B. thuringiensis subspecies toxic to coleopteran or lepidopteran larvae, only low levels of resistance to B. thuringiensis subsp. israelensis have been obtained in laboratory experiments where mosquito larvae were placed under heavy selection pressure for more than 30 generations. Selection of Culex quinquefasciatus with mutants of B. thuringiensis subsp. israelensis that contained different combinations of its Cry proteins and Cyt1Aa suggested that the latter protein delayed resistance. This hypothesis, however, has not been tested experimentally. Here we report experiments in which separate C. quinquefasciatus populations were selected for 20 generations to recombinant strains of B. thuringiensis that produced either Cyt1Aa, Cry11Aa, or a 1:3 mixture of these strains. At the end of selection, the resistance ratio was 1,237 in the Cry11Aa-selected population and 242 in the Cyt1Aa-selected population. The resistance ratio, however, was only 8 in the population selected with the 1:3 ratio of Cyt1Aa and Cry11Aa strains. When the resistant mosquito strain developed by selection to the Cyt1Aa-Cry11Aa combination was assayed against Cry11Aa after 48 generations, resistance to this protein was 9.3-fold. This indicates that in the presence of Cyt1Aa, resistance to Cry11Aa evolved, but at a much lower rate than when Cyt1Aa was absent. These results indicate that Cyt1Aa is the principal factor responsible for delaying the evolution and expression of resistance to mosquitocidal Cry proteins.

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