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Oogenesis in insects is a carefully orchestrated process, facilitating the formation of female gametes, which is regulated by multiple extrinsic and intrinsic factors, including ovary serine protease (Osp). As a member of the serine protease family, Osp is a homolog of Nudel, a maternally required protease defining embryonic dorsoventral polarity in Drosophila. In this study, we used CRISPR/Cas9-mediated mutagenesis to functionally characterize Osp in the Asian corn borer, Ostrinia furnacalis, a devastating maize pest throughout Asia and Australia. Building on previous knowledge, we hypothesized that knockout of Osp would disrupt embryonic development in O. furnacalis females. To examine this overarching hypothesis, we (1) cloned and characterized Osp from O. furnacalis, (2) designed target sites on exons 1 and 4 to construct a CRISPR/Cas9 mutagenesis system, and (3) documented phenotypic impacts among O. furnacalis Osp mutants. As a result, we (1) examined the temporal-spatial expression profiles of OfOsp, which has an open reading frame of 5648 bp in length and encodes a protein of 1873 amino acids; (2) established O. furnacalis Osp mutants; and (3) documented recessive, female-specific sterility among OfOspF mutants, including absent or deformed oviducts and reduced fertility in female but not male mutants. Overall, the combined results support our initial hypothesis that Osp is required for embryonic development, specifically ovarian maturation, in O. furnacalis females. Given its substantial impacts on female sterility, Osp provides a potential target for the Sterile Insect Technique (SIT) to manage Lepidoptera pests in general and the species complex Ostrinia in particular. [ABSTRACT FROM AUTHOR]

Simple Summary: Delaying or preventing the evolution of resistance to Bacillus thuringiensis (Bt) toxins produced by transgenic crops in insect pests is a major challenge for agriculture. A simulation model suggested that the rotation of different single-gene crops planted in subsequent seasons has the potential to delay resistance. To test this prediction, we set out laboratory selection experiments under the alternation of multi-toxins by mixing individual toxins (Cry1Ab, Cry1F, and Cry1Ie) in an artificial diet to emulate single-gene Bt maize plants. Two (Cry1Ab-Cry1F, Cry1Ab-Cry1Ie) or three (Cry1Ab–Cry1F–Cry1Ie) toxin alternation regimes were tested to imitate two or three single-gene crops in a rotation fashion. The species tested was the Asian corn borer, Ostrinia furnacalis (Guenée), the most economically important species of maize pest in Asia. The present study suggested that rotation of multiple toxins did not slow the evolution of resistance to Cry1F or Cry1Ie. Data generated from the study will assist in the development of sustainable resistance-management strategies. A common strategy for delaying the evolution of resistance to transgenic crops that produce insecticidal proteins from Bacillus thuringiensis is to ensure that insect pests are exposed to multiple toxins with different mechanisms of action (MoAs). This can take the form of planting crops in a rotation pattern when different crops expressing single toxins are available on the market. The efficacy of a rotation strategy is reliant on mathematical models based on biological assumptions. Here, we designed laboratory evolution experiments to test whether Bt-based insecticidal proteins with different MoAs used in rotation could delay resistance from developing in Asian corn borer (ACB), Ostrinia furnacalis. We investigated the proteins Cry1Ab, Cry1F, and Cry1Ie, which are widely utilized for commercial insect control. We found that rotation of multiple toxins did not slow the evolution of resistance to Cry1F or Cry1Ie. Furthermore, the evolution of ACB to the Cry1Ab toxin develops faster when Cry1F or Cry1Ie is present, as compared to Cry1Ab exposure only. Our results suggest that toxins used in a rotation fashion do not work as an effective strategy in delaying ACB resistance evolution to Cry toxins over one-toxin exposure. Our result highlights the need to better understand the biological factors leading to insecticidal protein resistance and to develop IRM strategies against target insects. [ABSTRACT FROM AUTHOR]

Insect brains play important roles in the regulation of sex-biased behaviors such as mating and oviposition. The neural structure and function of brain differences between males and females have been identified, in which the antenna lobes (AL) showed the most discrepancy, however, the whole repertoire of the genes expressed in the brains and the molecular mechanism of neural signaling and structural development are still unclear. In this study, high-throughput transcriptome analysis of male and female brains was carried on in the Asia corn borer, Ostrinia furnacalis, and a total of 39.23 Gb data and 34,092 unigenes were obtained. Among them, 276 genes displayed sex-biased expression by DEG analysis, of which 125 genes were highly expressed in the males and 151 genes were highly expressed in the females. Besides, by homology analysis against genes that have been confirmed to be related to brain neurodevelopment, a total of 24 candidate genes were identified in O. furnacalis. In addition, to further screen the core genes that may be important for sex-biased nerve signaling and neurodevelopment, proteinprotein interaction networks were constructed for the sex-biased genes and neurodevelopment genes. We identified 10 (Mhc, Mlc1, Mlc2, Prm, Mf, wupA, TpnC25D, fln, l(2)efl, and Act5C), 11 (PPO2, GNBP3, Spn77Ba, Ppn, yellow-d2, PGRP-LB, PGRP-SD, PGRP-SC2, Hml, Cg25C, and vkg) and 8 (dac, wg, hh, ci, run, Lim1, Rbp9, and Bx) core hub genes that may be related to brain neural development from male-biased, female-biased, and neurodevelopment gene groups. Our results provide a reference for further analysis of the dimorphism of male and female brain structures in agricultural pests. [ABSTRACT FROM AUTHOR]

Asian corn borer, Ostrinia furnacalis (Guenée), is an important insect pest of maize throughout most of Asia. The rind of a maize stalk is a key barrier against corn borer larvae boring into the plant. There is a need to better understand the relationship between stalk strength and O. furnacalis larval injury, particularly for elite maize genotypes. To determine whether stalk strength is involved in maize resistance to O. furnacalis larval injury, 39 maize lines were evaluated in 2012 and 2013. Rind penetration strength (RPS) was measured at tassel (VT) and milk (R3) stages as a possible stalk resistance trait for O. furnacalis. RPS of primary ear internode at VT and R3 accounted for 37 and 38% of the variance in O. furnacalis injury (measured as number of holes) for simulated (artificially infested) first and second generation O. furnacalis, respectively. Relationships between stalk RPS values and tunnel length were weak. Results suggest that harder stalks have enhanced resistance to stalk boring but not to pith feeding or tunneling of O. furnacalis larvae. The RPS measures could provide classical maize breeders an important tool for evaluating stalk strength and corn borer resistance in maize. The assessments should focus on the internodes primary ear or above/below primary ear during both VT stage for first generation and R3 stage for second generation O. furnacalis resistance. [ABSTRACT FROM AUTHOR]

Ostrinia furnacalis (Lepidoptera: Pyralidae) is one of the most destructive agricultural pests in Asia. Traditional pest‐management methods include sex pheromone capture, transgenic crops that produce Bacillus thuringiensis toxin, and pesticides. Although these strategies control pest populations effectively, they also cause negative side effects, including dramatically increased pesticide resistance, severe pollution, and hazards for human health. Recently developed genome editing tools provide new prospects for pest management and have been successfully used in several species. However, few examples have been reported in the agricultural pest O. furnacalis due to a lack in genomic information. In this report, we identified only one transcript of O. furnacalis Argonaute 1 (OfAgo1) gene from the genome and cloned the open reading frame. OfAgo1 presented the maximum expression at the embryo stage or in the fat body during the larval stages. To understand its function, an OfAgo1 mutant was constructed using the Clustered Regularly Interspaced Short Palindromic Repeat/RNA‐guided Cas9 nuclease (CRISPR/Cas9). Mutagenesis of OfAgo1 disrupted cuticle pigmentation by down‐regulating micro RNAs and pigmentation‐related genes. This is the first report for the cloning and functional analysis of OfAgo1, revealing a role of OfAgo1 in cuticle pigmentation. The current report also established a CRISPR/Cas9 system in O. furnacalis, providing a new insight for pest management. [ABSTRACT FROM AUTHOR]

The Asian corn borer, Ostrinia furnacalis (Guenée), is one of the most destructive lepidopteran pests affecting maize in Asia. Previous research on the feeding behavior of O. furnacalis has mostly focused on larvae, but the adult feeding preference has been unclear hitherto. In this study, we investigated the ultramorphology of the proboscis and sensilla of O. furnacalis using scanning electron microscopy and made morphological comparisons between the sexes, attempting to make predictions on the food preference and feeding behavior of the adults. The proboscis of O. furnacalis is divided into zones 1 and 2 with three types of sensilla, namely chaetica, basiconica, and styloconica. The entire proboscis and zone 2 of the females are significantly longer than those of the males. The main sexual difference in the sensilla is shown in the number of sensilla styloconica, which are eight in females but nine in males. The feeding preference of adult O. furnacalis is discussed based on the morphology of the proboscis and sensilla. • The proboscis of adult Ostrinia furnacalis is divided into zones 1 and 2. • Sexual differences exist in the lengths of the entire proboscis and zone 2. • The proboscis contains three types of sensilla: chaetica, basiconica, and styloconica. • Sensilla styloconica per galea are eight in the females and nine in the males, respectively. [ABSTRACT FROM AUTHOR]

Two cornborer species, Ostrinia furnacalis (Lepidoptera: Crambidae) and O. nubilalis, are major corn pests in Asia and Europe, respectively. In both continents, the larval endoparasitoid Macrocentrus cingulum (Hymenoptera: Braconidae) develops on another, closely related stemborer, O. scapulalis, which feeds on mugwort and other dicotyledons. M. cingulum also emerges from O. furnacalis in Asia and O. nubilalis in North America, but not from O. nubilalis in Europe. We assessed the ability of three populations of each of the three Ostrinia species to encapsulate foreign bodies of a size similar to that of a M. cingulum egg. We conclude that variations in encapsulation ability alone cannot account for the differences observed in the field between parasite emergence rates in these different host species and geographic areas. [ABSTRACT FROM AUTHOR]

Simple Summary: The performance of two egg parasitoids T. dendrolimi and T. ostriniae were compared on Asian corn borer (ACB) Ostrinia furnacalis eggs of different ages under choice and no-choice conditions. Both parasitoid species showed significant preferences in parasitizing ACB eggs of different ages. Younger ACB eggs (0–8-h-old) showed high suitability while eggs older than 8 h were not suitable for T. dendrolimi. The age of ACB eggs did not affect the biological parameters of T. ostriniae. Overall, our results highlighted the T. ostriniae species as the best candidate biocontrol agent for managing ACB populations. The Asian corn borer (ACB), Ostrinia furnicalis, is a serious corn pest in south-east Asia, causing huge economic losses every year. Trichogramma dendrolimi and Trichogramma ostriniae, two egg parasitoids, have previously been identified as key biological control agents. To determine the age impact of ACB eggs on their effective biocontrol potential, herein we compared the biological parameters (i.e., number of parasitized eggs, emergence, developmental time, and sex ratio) of both parasitoids on ACB eggs of various ages (i.e., 0–4, 4–8, 8–12, 12–16, 16–24, 24–36, and 36–48 h old), respectively. Our results showed that the age of ACB eggs had a significant impact on the parasitization activity of T. dendrolimi in both choice and no-choice conditions. Trichogramma dendrolimi preferred to parasitize 0–8-h-old ACB eggs, and its parasitization dramatically declined on ACB eggs older than 8 h under choice and no-choice conditions. On the other hand, T. ostriniae showed high preference to parasitize all tested ACB egg ages. The age of ACB eggs had no significant impact on the parasitization of T. ostriniae under choice and no-choice conditions. Furthermore, the female progeny of T. dendrolimi decreased as the age of ACB increased, while no differences were found in female progeny of T. ostriniae. Trichogramma ostriniae also developed faster on each ACB egg age group in comparison with T. dendrolimi. Overall, the age of ACB eggs had a significant impact on T. dendrolimi performance, leading us to conclude that T. ostriniae is more effective than T. dendrolimi as a biocontrol agent of the ACB. [ABSTRACT FROM AUTHOR]

The Asian corn borer, Ostrinia furnacalis (Guenée, 1854), is a highly damaging pest in Asia and the Pacific islands, and larvae feed mainly from corn crops. To determine the suitability of Bt-corn technology for the future control of this pest, understanding the potential to develop resistance to Cry1Ab and the basis of cross-resistance to other Cry1 proteins is of great interest. Here, we have explored the binding of Cry1A proteins to brush border membrane vesicles from two O. furnacalis colonies, one susceptible (ACB-BtS) and one laboratory-selected with Cry1Ab (ACB-AbR). The insects developed resistance to Cry1Ab and showed cross-resistance to Cry1Aa, Cry1Ac, and Cry1F. Binding assays with radiolabeled Cry1Ab and brush border membrane vesicles from susceptible insects showed that Cry1A proteins shared binding sites, though the results were not conclusive for Cry1F. The results were confirmed using radiolabeled Cry1Aa. The resistant insects showed a reduction of the specific binding of both Cry1Ab and Cry1Aa, suggesting that part of the binding sites were lost or altered. Competition binding assays showed full competition between Cry1Ab and Cry1Aa proteins in the susceptible colony but only partial competition in resistant insects, confirming the alteration of some, but not all, binding sites for these two proteins. The binding site model for Cry1A proteins in O. furnacalis is in agreement with the occurrence of multiple membrane receptors for these proteins. [ABSTRACT FROM AUTHOR]

In this study, the complete mitogenome sequence of Ostrinia furnacalis was described. The assembled mitogenome is 15,241 bp in length with an extreme bias of high AT content (80.9%) (GenBank accession no. MN747041). The mitochondrial genome contains 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (12S rRNA and 16S rRNA) genes, and a control region (D-loop region). The mitochondrial gene order was identical to that observed in most lepidopteran genomes, nine PCGs were located on the H-strand, others were located on the L-strand. 12 PCGs were initiated by typical ATN codons, except for COI with CGA instead. 21 tRNAs had the typical cloverleaf structure, while the DHU arm of the trnS1 gene did not form a stable stem-loop structure. The 'ATAGT(A)'-like motif and a 19 bp poly-T stretch at the down-stream of the rrnS gene were observed in the A + T-rich region. The phylogenetic analysis showed that the relationship of O. furnacalis is very close to the three species in the subfamily Pyraustinae: O. nubilalis, O. penitalis and Loxostege sticticalis, and all the subfamilies of Spilomelinae, Pyraustinae, Crambinae and Nymphulinae within Crambidae formed monophyletic groups with the highest bootstrap value support. [ABSTRACT FROM AUTHOR]

The Asian corn borer Ostrinia furnacalis is one of the most destructive pests of maize throughout eastern Asia and the South Pacific. In the present study the fine structure of the compound eyes of adult O. furnacalis was investigated under light/dark adaptation using light and electron microscopy. The compound eyes of male and female O. furnacalis are superposition eyes with electron-lucent clear zones. The sexual differences of the compound eyes of O. furnacalis are mainly reflected in eye size rather than ommatidial ultrastructure. Each ommatidium of both sexes contains 12 retinula cells, one of which is the basal retinula cell. All the retinula cells form a centrally-fused, two-tiered rhabdom, whose distal layer passes through the clear zone and distally connects with the crystalline cone. The ultrastructural changes under light/dark conditions mainly involve the rhabdom occupation ratio to retinula cell volume in the proximal layer of the rhabdom as well as the dimensions of the subcorneal zone and the crystalline tract. Pigment movements occur within the retinula cells and primary pigment cells, but are undetectable within the secondary pigment cells. Regardless of light or dark adaptation, in other words, the pigments never migrate into the clear zone. • The compound eyes of O strinia furnacalis are superposition eyes. • Each ommatidium contains 12 retinula cells forming a two-tired rhabdom. • Rhabdom occupation ratio (ROR) of the proximal rhabdom decreases from dark to light adaptation. • Pigment migrations are not detectable in the clear zone. [ABSTRACT FROM AUTHOR]