Vegetative Insecticidal Proteins (VIP): Bt toxin to protect agricultural crops

from insect pest

Naveenarani .M, Appunu .C
Bacillus thuringiensis (Bt) is well known for its ability to produce insecticidal crystal
proteins, called -endotoxins during its sporulation phase. The pathogenicity of these bacteria
against insects is connected with the action of various virulence factors of which the most
important are crystalline proteins Cry and Cyt, vegetative insecticidal proteins (Vips) and
Secreted insecticidal protein (Sip) (Palma et al., 2014). Insecticidal crystal (Cry) toxins produced
by B. thuringiensis during sporulation are used as biopesticides for the control of target pests and
many of them have narrow-spectrum activity (Sanahuja et al., 2011). In recent years, a number
of insecticidal proteins called vegetative insecticidal proteins (Vip) secreted during the vegetative
growth phase of Bt have been identified. Vip have received considerable attention since their
discovery and they have a broad insecticidal spectrum that includes a wide variety of
lepidopteran and coleopteran species (Palma et al., 2014). Vegetative insecticidal protein (Vip) is
now being used for transgenic expression in crops plants; conferring resistance against
lepidopteron pests. This toxin exhibits different mode of action to pests compared with that of
crystal proteins (Yu et al., 1997; Lee et al., 2003), However, extensively used in bio-pesticide
industry to control crop pests and will become a potentially useful tool in the prevention or
management of pest resistance to ICPs. Vip genes are not much exploited for control of insect
pests compared to any genes.
Vip protein diversity reports and an alternative source to control insects pests
So far, more than 100 Vip protein sequences have been recognized and their classification
has been established on a basis of amino acid sequence similarity resulting in four main groups:
Vip1, Vip2, Vip3, and Vip4 (Crickmore et al., 2014) (Table: 1). Vip1 and Vip2 are proteins of
100 and 52 kDa, respectively and the proteins are binary toxins with activity against Coleoptera
(Palma et al., 2014) and Hemiptera (Sattar and Maiti, 2011). Vip3 proteins shows insecticidal
activity against Lepidoptera pests and the member of the new family Vip4, no target insects have
been found until now (Sameh Sellami et al., 2016). Among the four types of Vip, Vip3 is well
studied and reported to have insecticidal activity against economically important Lepidoptera
pests: Spodoptera exigua Spodoptera frugiperda (Estruch et al., 1996; Figueiredo et al., 2013),
Helicoverpa zea, Chilo partellus, Phthorimaea opercullela, Heliothis virescens, Manduca sexta
(Lee et al., 2003), Ephestia kuehniella (Abdelkefi-Mesrati et al., 2005), Spodoptera litura
(Bhalla et al., 2005) and Trichoplusia ni (Fang et al., 2007), Chrysodeixis chalcites, Lobesia
botrana, Mamestra brassicae, Spodoptera littoralis, Plutella xylostella (Palma et al., 2013),
Ectomyelois ceratoniae (Iigo Ruiz de Escudero et al., 2014; Boukedi et al., 2015).
The Vip3 proteins are divided into three classes (Vip3A, Vip3B and Vip3C) and thirteen
subclasses (Vip3Aa to Vip3Aj, Vip3Ba, Vip3Bb and Vip3Ca (Crickmore et al., 2014).Vip3 toxins
display one of the most important features that it shares no sequence homology with known Bt -

endotoxins or other toxin genes. Vip3A proteins have been shown to target receptors in the insect
midgut, the ingested toxin get activated by the trypsin like proteases in the midgut is different to
those used by Cry proteins (Ruiz de Escudero et al., 2014), which makes them a good alternative
or complement to Cry proteins for insect pests management. Plants expressing the Vip3Aa gene
along with Cry genes have been developed and transgenic corn and cotton expressing Vip3Aa
have been commercialized for the control of lepidopteran pests (Raybould and Quemada, 2010).
The expression of Vip3Aa alone and in combination with Cry1A have been achieved in cotton,
maize and rice (Chen et al., 2010), and this new generation of Bt-crops display very valuable
features in pest control. However, differences in insecticidal spectrum of activity of particular
Vip toxins were noted for Vip3Aa1, Vip3Aa2, Vip3Aa10, Vip3Ab and Vip3Ae (Ruiz de
Escudero et al., 2014) display activity against Agrotis ipsilon, whereas Vip3Aa9 (Selvapandiyan
et al., 2001) and Vip3Ad (Hernandez-Martinez et al., 2013) are not active toward this insect.
High insecticidal activity of Vip3 proteins has opened the possibility of using them in
pest management in order to broaden the host spectrum activity of biopesticides based on B.
thuringiensis toxins and to be a part of the strategy aiming for preventing insect resistance to
those proteins (Abdelkefi-Mesrati et al., 2011). Furthermore, Vip3Aa has no effect on non
targeted organisms, which are not pests in the agricultural system (Raybould and Vlachos, 2011).
Vip3Aa58 and Vip3Aa59, insecticidal activity of these toxins was estimated against both insect
orders, Lepidoptera economically important pests of woodlands (Dendrolimus pini), orchards
(Cydia pomonella) and crop fields (S. exigua) and also tested against mealworm beetle Tenebrio
molitor (Coleoptera), a pest of stored grains (Baranek et al., 2015). So far, the toxicity of any
Vip3A protein against D. pini and C. pomonella has not been determined. Vip proteins seem to
bind to different target sites than Cry proteins in midgut epithelium cells of the species tested
(Lee et al., 2003, 2006), so these proteins are suitable candidates to be used together with Cry
proteins for efficient control of insect pest in agricultural crops.
Table 1: List of Vip toxin grouped according to their primary ranking
within the nomenclature (Crickmore et al., 2014)
VIP 1
Vip2Ac
Vip3Af
Vip1Aa
Vip2Ad
Vip3Ag
Vip1Ab
Vip2Ae
Vip3Ah
Vip1Ac
Vip2Af
Vip3Ai
Vip1Ad
Vip2Ag
Vip3Af
Vip1Ba
Vip2Ba
Vip3Ag
Vip1Bb
Vip2Bb
Vip3Ah
Vip1Bc
VIP3
Vip3Ai
Vip1Ca
Vip3Aa
Vip1Da
Vip3Ab
VIP 4
VIP2
Vip3Ac
Vip4Aa
Vip2Aa
Vip3Ad

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