Pest management based on CRISPR/Cas9-mediated site-specific mutations is an effective and environmentally safer strategy to suppress the pest population. However, the potential of this approach is yet to be tested on many important agricultural pests such as Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), a fit candidate for area-wide pest management. Therefore, in the present study, 2 spermatogenesis-related genes viz. Testis-specific zinc finger protein (topi) and Testis-specific serine protein kinase 1 (Tssk1) of B. dorsalis were edited to impart male sterility and its impact on further progeny. In this regard, topi and Tssk1 mutant populations deposited significantly fewer eggs per day (6.12 +/- 0.36 and 3.60 +/- 0.24, respectively) as compared to the control (11.16 +/- 0.58 eggs per day). About the hatching rate, the above trend was observed, topi (44.51) and Tssk1 (30.04) as compared to the control (73.96). Furthermore, the total number of viable offspring for topi and Tssk1 populations decreased as a result of the cumulative progeny production ten days after the post-mating phase. It suggests that topi and Tssk1 from B. dorsalis could be potential targets for imparting male sterility in B. dorsalis. Graphical Abstract

Insects have a rich diversity of RNA viruses that can either cause acute infections or persist in host populations without visible symptoms. The melon fly, Zeugodacus cucurbitae (Tephritidae) causes substantial economic losses through infestation of diverse cucurbit and other crops. Of Indomalayan origin, it is now established in many tropical regions of the world. The virome diversity of Z. cucurbitae is largely unknown across large parts of its distribution, including the Indian subcontinent. We have analysed three transcriptomes each of one field-collected and one laboratory-reared Z. cucurbitae population from Bangalore (India) and discovered genomes of ten putative RNA viruses: two sigmaviruses, one chimbavirus, one cripavirus, one noda-like virus, one nora virus, one orbivirus, one partiti-like virus, one sobemovirus and one toti-like virus. Analysis of the only available host genome of a Hawaiian Z. cucurbitae population did not detect host genome integration of the detected viruses. While all ten viruses were found in the Bangalore field population only seven were detected in the laboratory population, indicating that these seven may cause persistent covert infections. Using virus-specific RNA-dependent RNA polymerase gene primers, we detected nine of the RNA viruses with an overall low variant diversity in some but not all individual flies from four out of five Indian regions. We then screened 39 transcriptomes of Z. cucurbitae laboratory populations from eastern Asia (Guangdong, Hainan, Taiwan) and the Pacific region (Hawaii), and detected seven of the ten virus genomes. We found additional genomes of a picorna-like virus and a negev-like virus. Hawaii as the only tested population from the fly's invasive range only had one virus. Our study provides evidence of new and high RNA virus diversity in Indian populations within the original range of Z. cucurbitae, as well as the presence of persistent covert infections in laboratory populations. It builds the basis for future research of tephritid-associated RNA viruses, including their host effects, epidemiology and application potential in biological control.

The Fall armyworm, Spodoptera frugiperda , is a globally important invasive pest, primarily on corn, causing severe yield loss. Overuse of synthetic chemicals has caused significant ecological harm, and in many instances control has failed. Therefore, developing efficient, environmentally friendly substitutes for sustainable management of this pest is of high priority. CRISPR/Cas9-mediated gene editing causes site-specific mutations that typically result in loss-of-function of the target gene. In this regard, identifying key genes that govern the reproduction of S. frugiperda and finding ways to introduce mutations in the key genes is very important for successfully managing this pest. In this study, the pheromone biosynthesis activator neuropeptide ( PBAN ) gene of S. frugiperda was cloned and tested for its function via a loss-of-function approach using CRISPR/Cas9. Ribonucleoprotein (RNP) complex (single guide RNA (sgRNA) targeting the PBAN gene + Cas9 protein) was validated through in vitro restriction assay followed by embryonic microinjection into the G0 stage for in vivo editing of the target gene. Specific suppression of PBAN by CRISPR/Cas9 in females significantly affected mating. Mating studies between wild males and mutant females resulted in no fecundity. This was in contrast to when mutant males were crossed with wild females, which resulted in reduced fecundity. These results suggest that mating disruption is more robust where PBAN is edited in females. The behavioural bioassay using an olfactometer revealed that mutant females were less attractive to wild males compared to wild females. This study is the first of its kind, supporting CRISPR/Cas9 mediating editing of the PBAN gene disrupting mating in S. frugiperda . Understanding the potential use of these molecular techniques may help develop novel management strategies that target other key functional genes.

The Oriental fruit fly, Bactrocera dorsalis (Hendel), is a highly destructive polyphagous pest of horticultural crops. Owing to the magnitude of destruction it causes, this species is an attractive target for control through genetic biocontrol techniques such as pgSIT and CRISPR/Cas9 based gene drive. In the present communication, we are presenting detailed information on the types of edits (inter and inter individual) in B. dorsalis contributing to white eye generated through the delivery of ribonucleoproteins specific to white locus in G0 embryos. A total of 457 embryos were microinjected, and a partial white eye phenotype was evident in 14 adult flies in G0 generation. Further, these edited flies were batch crossed to check the heritability and types of the edits in G1 generation from randomly selected 79 flies that exhibited complete white eye phenotype. Three flies (partial and complete white eye phenotype) were randomly chosen from each generation and sequenced closer to the edit site to decipher the type of edits obtained. Edit characterization revealed the predominant deletions of different lengths 2, 4, 6, 11 and 14 bp. Additionally, multiple edits, pointing towards genetic mosaicism, were obtained from individual flies with complete and partial white eye phenotypes (G0M3 and G0M4, respectively). Hence, this study provides detailed documentation of genetic mosaicism in G0 CRISPR/Cas edited individuals.

Biology of the burrowing cockroach Pycnoscelus surinamensis (L.) revealed that there are four instars and took 134.8±4.94 days to complete lifecycle. Morphometric studies showed increase in antennal body length, head width, pronotum length and width from first to fourth instar with peak at adult and other developmental stages well fit into Brooks-Dyar rule. All body parameters showed hypoallometry except pronotum length which showed hyperallometry. In substrate preference studies, the survival was lowest for the treatment without substrate and against the treatments with substrates like leaf litter, cocopeat and vermiculite. The mean number of offspring produced and survived was found to be lower in the treatments without substrate and with leaf litter against the treatments with cocopeat and vermiculite.

The melon fly, Zeugodacus cucurbitae (Diptera: Tephritidae) is an important pest of cucurbit crops in the Oriental and warm regions of the world. We conducted an age-stage, two-sex life table study of Z. cucurbitae on four different hosts, viz cucumber (Cucumis sativus), ridge gourd (Luffa acutangula), bitter gourd (Momordica charantia) and pumpkin (Cucurbita pepo) under laboratory conditions at 25 +/- 1 degrees C, 65 +/- 5% relative humidity and a 14:10 h (L:D) photoperiod. The developmental time from egg to pupa was shortest when reared on cucumber (15.4 days), and longest when reared on pumpkin (17 days). The intrinsic rate of increase (r) of the Z. cucurbitae population on cucumber, ridge gourd, bitter gourd and pumpkin was 0.1143, 0.1062, 0.0071 and 0.0089 per day, respectively. The net reproduction rate (R0) was highest on cucumber (77 offspring per female) and lowest on bitter gourd (15.9 offspring per female). The mean generation time (T ) ranged from 35.9 days when reared on ridge gourd to 38.7 days when reared on bitter gourd. Larval hatching rate, pupation rate and adult eclosion rate were highest when reared on cucumber compared to other hosts. In host preference experiments (choice and no-choice tests), the flies were most attracted to cucumber and oviposited most on cucumber when compared to other hosts. We found that cucumber is a suitable host for Z. cucurbitae growth and development and can be used for mass rearing of this fly species under controlled conditions for research purposes.

Overuse of synthetic chemicals over a long period of time has not only resulted in control failures but also enormous ecological damage. This necessitates developing eco-friendly, effective alternatives for sustainable pest management. In this regard, potential of CRISPR/Cas9 mediated genome editing to introduce site specific mutations that mostly resulting in loss of function has been successfully demonstrated in wide varieties of organisms including insects. This has opened a new avenue to design and implement futuristic pest management strategies like precision guided sterile insect technique (pgSIT) for an area wide suppression. This requires validation of target genes employing Ribonucleoprotein (RNP) complex containing sgRNA and Cas9 protein before undertaking transgenesis for achieving pgSIT. In the present study, we have for the first time, supporting CRISPR/Cas9 mediated editing of the eye colour gene, Tryptophan 2, 3-dioxygenase of the eggplant shoot and fruit borer, Leucinodes orbonalis using RNP complex. The mutant moths of both sexes exhibited reddish brown eyed phenotype. Having established an RNP mediated editing system in L. orbonalis, the key genes involved in sex determination and spermatogenesis will be validated for developing a pgSIT system for L. orbonalis.

Most chemically produced fertilisers emit methane and nitrous oxide into the atmosphere, which are toxic to humans, composting is a great alternative to artificial fertilisers. Half of the organic waste generated by mankind is compostable. Blatticomposting is an innovative method for managing organic waste using cockroaches as bio converters in sustainable waste management. It is an eco-friendly strategy and an efficient method for nutrient recycling and organic waste decomposition. Overall blatticomposting presents a unique opportunity to transform waste management practices and contribute to a more sustainable future. This review paper aims to provide an overview of blatticomposting and its process, its benefits, challenges and prospects. Furthermore, we explore the potential applications of this technique and its contribution to waste reduction, resource recovery, and sustainable agriculture which leads to the upliftment of mankind leading to a healthy life.

Genome editing mediated by programmable CRISPR/Cas system is a new, emerging technology that holds greater potential in developing genetic pest management strategies via precision guided sterile insect technique; genetic improvement of pollinators, natural enemies etc. The seemingly innocuous prokaryotic immunity has now been translated into a mammoth site specific editing technology for its use in medical, agricultural, diagnostic fields etc. Until recently most of the genome editing work was dependent on RNA directed site specific cleavage by Cas9 enzyme followed by error prone, non-homologous end joining (NHEJ) resulting in random mutations. Later development of improved editing systems like base editor and prime editor have enabled obtaining site-specific, pre determined mutations without a double stranded break. Similarly many engineered Cas9 variants and other Cas proteins belonging to different class and types such as Cas3, Cas12a, Cas13a have improved the existing editing tool box. Currently efforts are being made to design and implement suitable gene drive methods for various pests such as spotted wing Drosophila, Drosophila suzukii Matsumura, Diamondback moth, Plutella xylostella (Linnaeus) , many species mosquito, Med fly, Ceratitis capitata Wiedem etc..

Rugose spiralling whitefly (RSW), Aleurodicus rugioperculatus Martin (Hemiptera: Aleyrodidae), an invasive pest to the Oriental region was found infesting several plantation crops like coconut, banana, guava, Indian almond, sapota, oil palm, mango, cashew, custard apple and several ornamental plants in southern states of India as well as Assam, West Bengal, Maharashtra, Meghalaya, Odisha, Chhattisgarh and Gujarat. Intensive surveys across different agroclimatic zones of Karnataka revealed that infestation of the pest was severe in coastal and southern region of Karnataka compared to northern region. Molecular characterization of partial mitochondrial cytochrome c oxidase–I gene (658 bp) and morphological characters of the pest and its natural enemies confirms its identity. During the study, 35 host plants were recorded, and several natural enemies viz., parasitoids, Encarsia guadeloupae, Encarsia dispersa (Hymenoptera: Aphelinidae), predators, Pseudomallada astur (Neuroptera: Chrysopidae), Jauravia pallidula, Cheilomenes sexmaculata (Coleoptera: Coccinellidae) and Cybocephalus indicus (Coleoptera: Cybocephalidae) were also observed feeding on RSW. The parasitoid E. guadeloupae was the major parasitoid associated with the pest. Natural parasitism by the parasitoids was ranging from 7–80% in the study area.

A field experiment on the efficacy of some insecticides in combination with neem against rice green leaf hoppers (GLH), Nephotettix virescens Distant, N. nigropictus Stal, and gundhi bug Leptocorisaacuta Thunberg was conducted during kharif 2017 at the Central Research Farm, Department of Entomology, Odisha University of Agriculture and Technology (OUAT), Bhubaneswar. Seven treatments viz., flonicamid 50%WG @ 75g a.i./ha, buprofezin 25%SC @ 200 ml a.i/ha, imidacloprid17.8% SL@35g a.i/ha, flonicamid 50%WG @ 37.5 g a.i./ha + multineem (300ppm) @ 1.25l/ha, buprofezin 25%SC @ 100 ml a.i./ha+ multineem (300ppm) @ 2.5l/ha, imidacloprid 17.8%SL @ 17.5 ml a.i./ha + multineem (300ppm) @ 1.25l/ ha, and multineem (300ppm) @ 2.5l/ha were evaluated. Results revealed that foliar spray of imidacloprid 17.8%SL @ 17.5 ml a.i./ha+ multineem (300 ppm) @ 1.25l/ha at 55 days after spraying (DAS) and 70 DAS were the most effective with 87.77% and 88.91% reduction of GLH and gundhi bug, respectively followed by imidacloprid 17.8%SL@35g a.i/ha with 85.73% and 85.14% reduction, respectively.

Comparative biology of invasive rugose spiralling whitefly (RSW) Aleurodicus rugioperculatus Martin (Hemiptera: Aleyrodidae) was studied on three host plants viz., coconut (Cocos nucifera L), banana (Musa spp. L) and Indian shot (Canna indica) under caged as well as field conditions. The RSW life cycle was shorter on coconut as compared to that of banana and Indian shot plant. Thus, coconut is the preferred host for its growth and development with a fecundity of 49.50± 4.09 eggs and total duration of the nymphal instars being of 23.2± 1.38 days. Duration to lifestages on banana was less under field conditions as compared to that under caged conditions. These results on the comparative biology and growth will help to develop suitable IPM strategies for this invasive pest.

Laboratory and field evaluation of some insecticides (buprofezin 25 SC, pymetrozine 50 WG, flonicamid 50 WG, dinotefuran 20 SG, thiamethoxam 25 WG, profenophos 50 EC), botanical (azadirachtin 10000 ppm) and entomopathogenic fungus (Isaria fumosorosea Wize) were carried out against the invasive rugose spiralling whitefly Aleurodicus rugioperculatus Martin on banana. The results revealed that azadirachtin 10000 ppm (86.49%), dinotefuran 20 SG (82.64%), pymetrozine 50 WG (72.72%) and thiamethoxam 25 WG (79.59%) caused significant mortality of first and second instar nymphs at 2 days after treatment (DAT). Profenophos 50 EC (77.61%) and azadirachtin 10000 ppm (71.53%) were significantly effective and on par with mortality at 2 DAT of third and fourth instar nymphs. Flonicamid 50 WG was the most toxic with the least LC50 value (507.8 and 646.1) and buprofezin 25 SC was the least toxic with maximum value (1245.1 and 1752.0) for early and late instar nymphs, respectively. Profenophos 50 EC and LC50 buprofezin 25 SC also showed maximum ovicidal effect. Under field conditions, I. fumosorosea with 100% mortality was significantly superior compared to dinotefuran 20 SG (97.23%), thiamethoxam 25 WG (95.02%) and azadirachtin 10000 ppm (94.98%) at 6 DAT.