Double-stranded RNA mediated knockdown of sucrase gene induced mortality and reduced offspring production in Aphis gossypii.

The importance of gut sucrase in maintaining osmotic equilibrium and utilizing phloem contents as a carbon source has been widely investigated and proven in sap-sucking insects. In the present study, silencing of Aphis gossypii sucrase1 (Agsuc1) was carried out by double-stranded RNA (dsRNA), which would be lethal to it due to disruption of osmotic balance. The dsRNA corresponding to Agsuc1 was synthesized by two methods, i.e., in vitro synthesis using T7/SP6 RNA polymerase and in vivo synthesis by bacterial expression, i.e., Escherichia coli strain HT115 transformed with the L4440 vector system. Oral delivery of double-stranded Agsuc1 synthesized in vitro (dsAgsuc1) and in vivo (HT115Agsuc1) induced around 50% mortality in nymphs of A. gossypii. Moreover, the number of offspring produced by the survived aphids decreased by 39-43%. Parthenogenetic reproduction of the aphids is the critical factor for their fast population build-up, leading to yield losses of economic significance. Thus, the present study demonstrated that the silencing of the Agsuc1 gene reduced the aphid population by killing it and inhibited the population buildup by reducing the number of offspring produced by the survived aphids, likely to result in a significant reduction in crop damage. The production of dsRNA by bacterial expression is a cost-effective method. It has the potential to be used as a biopesticide. The sucrase gene is an excellent putative target gene for RNAi against A. gossypii. It could be used to develop a transgenic plant that produces dsAgsuc1 to keep A. gossypii populations below the economic threshold level.

Diversity of transgenes in sustainable management of insect pests.

Insecticidal transgenes, when incorporated and expressed in plants, confer resistance against insects by producing several products having insecticidal properties. Protease inhibitors, lectins, amylase inhibitors, and chitinase genes are associated with the natural defenses developed by plants to counter insect attacks. Several toxin genes are also derived from spiders and scorpions for protection against insects. Bacillus thuringiensis Berliner is a microbial source of insecticidal toxins. Several methods have facilitated the large-scale production of transgenic plants. Bt-derived cry, cyt, vip, and sip genes, plant-derived genes such as lectins, protease inhibitors, and alpha-amylase inhibitors, insect cell wall-degrading enzymes like chitinase and some proteins like arcelins, plant defensins, and ribosome-inactivating proteins have been successfully utilized to impart resistance to insects. Besides, transgenic plants expressing double-stranded RNA have been developed with enhanced resistance. However, the long-term effects of transgenes on insect resistance, the environment, and human health must be thoroughly investigated before they are made available for commercial planting. In this chapter, the present status, prospects, and future scope of transgenes for insect pest management have been summarized and discussed.

Leaf Damage Based Phenotyping Technique and Its Validation Against Fall Armyworm, Spodoptera frugiperda (J. E. Smith), in Maize.

Globally, maize is an important cereal food crop with the highest production and productivity. Among the biotic constraints that limit the productivity of maize, the recent invasion of fall armyworm (FAW) in India is a concern. The first line of strategy available for FAW management is to evaluate and exploit resistant genotypes for inclusion in an IPM schedule. Screening for resistant maize genotypes against FAW is in its infancy in India, considering its recent occurrence in the country. The present work attempts to optimize screening techniques suited to Indian conditions, which involve the description of leaf damage rating (LDR) by comparing injury levels among maize genotypes and to validate the result obtained from the optimized screening technique by identification of lines potentially resistant to FAW under artificial infestation. Exposure to 20 neonate FAW larvae at the V5 phenological stage coupled with the adoption of LDR on a 1-9 scale aided in preliminary characterize maize genotypes as potentially resistant, moderately resistant, and susceptible. The LDR varies with genotype, neonate counts, and days after infestation. The genotypes, viz., DMRE 63, DML-163-1, CML 71, CML 141, CML 337, CML 346, and wild ancestor Zea mays ssp. parviglumis recorded lower LDR ratings against FAW and can be exploited for resistance breeding in maize.

Sex Biased Variance in the Structural and Functional Diversity of the Midgut Bacterial Community of Last Instar Larvae of Pectinophora gossypiella (Lepidoptera: Gelechiidae).

Elucidating the midgut bacterial diversity in an important cotton bollworm Pectinophora gossypiella can be a stepping stone in understanding the possible role of midgut bacteria in field evolved resistance against Bt cotton as well as to commonly used insecticides. Present study targeted metagenomics of 16S rRNA V3-V4 region to understand the influence of sex, if exists, in community diversity of gut microbes vis a vis their function in pink bollworm larvae. The results of the present study revealed that Proteobacteria, Firmicutes, and Actinobacteria were the predominant phyla in the midgut of pink bollworm. Distinctive differences were found in the Shannon and Simpson diversity indices, ChaoI and ACE richness estimates in male and female larvae. The alpha diversity analysis showed that the gut bacteria of male were diverse and rich as compared to that of female. Further, beta diversity analysis indicated that the gut bacterial communities of both larval groups were unique from each other. These findings are the maiden report on sex-based variation in gut bacteria in P. gossypiella larvae. Role of candidate phyla OD1 (Parcubacteria) and TM7 (Saccharibacteria) in the living organisms needs to be studied, and their fairly significant composition in male and negligible composition in female larva raises question on their obvious role. Taxonomic to phenotypic mapping revealed that these gut bacteria play vital role in many metabolic and physiological activities of pink bollworm. Difference in potential functions of gut bacteria also varied with the sex.