Occurrence of Spodoptera frugiperda (J. E. Smith) (Insecta: Lepidoptera: Noctuidae), commonly known as fall armyworm, in southern India is reported along with associated natural enemies. Severe damage was noticed in Chikkaballapur, Hassan, Shivamogga, Davanagere and Chitradurga during July-August 2018. The incidence ranged from 9.0 to 62.5 percent at various locations, maximum incidence was recorded in Hassan district followed by Chikkaballapur, Davanagere, Shivamogga and Chitradurga. Morphology and molecular based taxonomic tools were used for the identification of this pest. The GenBank accession number MH704433 of Chikkaballapur population was released on 1 st August, 2018 and Barcode obtained from BOLD System-ID: AGIMP054-18. The survey also revealed natural parasitism by egg parasitoids viz., Telenomus sp. (Hymenoptera: Platygastridae) and Trichogramma sp. (Hymenoptera: Trichogrammatidae), gregarious larval parasitoid, Glyptapanteles creatonoti (Viereck) (Hymenoptera: Braconidae) solitary larval parasitoid, Campoletis chlorideae Uchida (Hymenoptera: Ichneumonidae), and a solitary indeterminate larval-pupal (Hymenoptera: Ichneumonidae: Ichneumoninae) parasitoid. Spodoptera frugiperda is the first host record for G. creatonoti across the globe. Glyptapanteles creatonoti, being a well established parasitoid of various noctuids in India and Malaysia, was capable of parasitizing S. frugiperda. Besides these, other commonly found bioagents viz., Forficula sp. (Dermaptera: Forficulidae) and entomopathogenic fungus Nomuraea rileyi (Farl.) Samson was also collected in large numbers. We report the natural enemy complex of S. frugiperda for the first time from India. The electro physiological response of Indian population of S. frugiperda male adults to pheromone was established. The studies to manage this pest by any/all means are in progress.

A vegetative insecticidal protein (VIP)-encoding gene from a local isolate of Bacillus thuringiensis has been cloned, sequenced, and expressed in Escherichia coli. The expressed protein shows insecticidal activity against several lepidopteran pests but is ineffective against Agrotis ipsilon. Comparison of the amino acid sequence with those of reported VIPs revealed a few differences. Analysis of insecticidal activity with N-and C-terminus deletion mutants suggests a differential mode of action of VIP against different pests.The gram-positive bacterium Bacillus thuringiensis is known to produce parasporal crystalline inclusions during the late exponential phase of growth (8). These crystals consist of several polypeptides, some of which are insecticidal or nematocidal. Upon ingestion by insects, these toxins are proteolytically activated, and after interaction with specific receptors at the mid-gut, they cause larval death (5). Since these toxins are highly specific, they are extremely useful in controlling targeted agricultural pests. Over the past several years, more than 100 different polypeptides have been identified, and several of them have been employed in insect management programs (8). The diversity, specificity, and usefulness of these insecticidal polypeptides have encouraged searches among diverse niches for new strains displaying novel insecticidal polypeptides. In addition to the crystal-associated toxic polypeptides, some insecticidal proteins produced during vegetative growth of the bacteria have also been identified. These proteins, called vegetative insecticidal proteins (VIPs), were reported from about 15% of the B. thuringiensis strains analyzed (2). We have screened several strains of B. thuringiensis obtained from soil samples collected from different parts of India for the presence of homologues of the VIP. Based on the reported gene sequences, we designed PCR DNA primers for the detection of the vip gene in strains held in our collection. As a result of the screening program, we have cloned, sequenced, and expressed a vegetative insecticidal toxin-coding gene from one of the isolates in our collection. The toxicity spectrum of the Escherichia coli-expressed recombinant protein has been evaluated against five lepidopteran pests. By deletion analysis, we have characterized the minimal toxic polypeptide segment that retains insecticidal activity. The toxicity of deleted VIP against lepidopteran pests suggested a differential mode of action against different pests.Bacterial strains and plasmids. Different isolates of B. thuringiensis were enriched from soil samples collected from different geographical locations within India. For routine use in the laboratory, the isolates were maintained in nutrient medium (Difco), and for long-term storage, the isolates were stored as glycerol stocks at Ϫ70°C. E. coli strain M15 was obtained from Qiagen (Braunschweig, Germany) and, when required, was grown in Luria-Bertani (LB) medium at 37°C with shaking at 200 rpm.Oligonucleotide PCR primers. Primers to sc...

Diamondback moth (DBM), Plutella xylostella (Linnaeus), is a notorious pest of brassica crops worldwide and is resistant to all groups of insecticides. The insect system harbors diverse groups of microbiota, which in turn helps in enzymatic degradation of xenobiotic-like insecticides. The present study aimed to determine the diversity of gut microflora in DBM, quantify esterase activity and elucidate their possible role in degradation of indoxacarb. We screened 11 geographic populations of DBM in India and analyzed them for bacterial diversity. The culturable gut bacterial flora underwent molecular characterization with 16S rRNA. We obtained 25 bacterial isolates from larvae (n = 13) and adults (n = 12) of DBM. In larval gut isolates, gammaproteobacteria was the most abundant (76%), followed by bacilli (15.4%). Molecular characterization placed adult gut bacterial strains into three major classes based on abundance: gammaproteobacteria (66%), bacilli (16.7%) and flavobacteria (16.7%). Esterase activity from 19 gut bacterial isolates ranged from 0.072 to 2.32 μmol/min/mg protein. Esterase bands were observed in 15 bacterial strains and the banding pattern differed in Bacillus cereus – KC985225 and Pantoea agglomerans – KC985229. The bands were characterized as carboxylesterase with profenofos used as an inhibitor. Minimal media study showed that B. cereus degraded indoxacarb up to 20%, so it could use indoxacarb for metabolism and growth. Furthermore, esterase activity was greater with minimal media than control media: 1.87 versus 0.26 μmol/min/mg protein. Apart from the insect esterases, bacterial carboxylesterase may aid in the degradation of insecticides in DBM.

Molecular characterization and DNA barcoding is a taxonomic method that uses a short genetic marker in an insect DNA to identify a species, including an unknown species. The DNA barcode method of identification includes, for example, identifying insect species from any developing stage and part; otherwise, identifying insects morphologically generally depends on adult stage and male genitalia. The utility of DNA barcoding for these purposes is subject to debate; however, in insects at least, it is approximately 650 bp of the mitochondrial gene, cytochrome oxidase I ( COX I). The approximate number of described insect species in India is 59,000; however, the number of barcodes generated from India is 4.6 % of known species, while the corresponding global scenario is about 16 % of described species; a lot of emphasis is required to catch up with the world scenario. In order to speed up taxonomic identification, DNA barcoding is now been considered as an alternative tool for insect biodiversity identification in India and the world. The present chapter deals with the use of barcode in the identification of insects belonging to different orders and families, using the neighbour-joining approach with the bootstrapping method and the Kimura-2 parameter to obtain a clear phylogenetic signal. In a neighbour-joining tree for all sequences, two clades were obtained: the first cluster consisting of hymenopteran insects and the other consisting of other orders. This phylogeny also agrees with the traditional phylogeny of insects. The present results, thus, favour DNA barcoding as a decisive tool in quick and reliable identification of insects.

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