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Simple Summary: The colonization of new areas by invasive insects has been a prominent concern in entomology. Bactrocera dorsalis serves as a typical invasive insect, which has expanded its range from southern to northern regions of China in recent years. Host suitability plays a crucial role in the successful establishment of B. dorsalis populations. In this study, we have observed a decline in the fitness of B. dorsalis when feeding on cucumber, primarily due to significant changes in the gut microbiota composition of the insect. Among them, Empedobacter brevis and Enterococcus faecalis were identified as key factors leading to the reduced fitness of B. dorsalis. These findings confirm the close association between insect fitness and symbiotic bacteria within the insect's body. Furthermore, the source of food and gut bacteria have significant impacts on the invasion of exotic insects. How alien pests invade new areas has always been a hot topic in invasion biology. The spread of the Bactrocera dorsalis from southern to northern China involved changes in food sources. In this paper, in controlled conditions, we take Bactrocera dorsalis as an example to study how plant host transformation affects gut bacteria by feeding it its favorite host oranges in the south, its favorite host peaches and apples in the north, and feeding it cucumbers as a non-favorite host plant, thereby further affecting their fitness during invasion. The result showed that, after three generations of feeding on cucumbers, Bactrocera dorsalis took longer to develop as a larva while its longevity and fecundity decreased and pre-adult mortality increased. Feeding it cucumbers significantly reduced the overall diversity of gut microbiota of Bactrocera dorsalis. The relative abundance of Enterobacter necessary for survival decreased, while the Empedobacter and Enterococcus increased, resulting in decreased carbohydrate transport and metabolism and increased lipid transport and metabolism. Feeding Bactrocera dorsalis Empedobacter brevis and Enterococcus faecalis resulted in a 26% increase in pre-adult mortality and a 2–3 d increase in adult preoviposition period (APOP). Additionally, Enterococcus faecalis decreased the longevity of female and male adults by 17 and 12 d, respectively, and decreased fecundity by 11%. We inferred that the shifted plant hosts played an important role in posing serious harm to Bactrocera dorsalis invading from the south to the north. Therefore, after an invasion of Bactrocera dorsalis into northern China, it is difficult to colonize cucumbers for a long time, but there is still a risk of short-term harm. The findings of this study have established that the interactions between an insect's food source and gut bacteria may have an important effect on insect invasions. [ABSTRACT FROM AUTHOR]

Simple Summary: Cas9 is limited by protospacer adjacent motif (PAM) sequences during the application process, which is particularly evident in insect genome editing. Researchers have discovered Cas9 variants with a wider range of recognition due to the development of genome-editing technology. However, the application of these variants in insects has been ineffective and time-consuming. To tackle this issue, this study established a high-throughput assay system capable of rapidly detecting recognizable PAM sequences of different Cas9 variants in Bombyx mori and the corresponding editing efficiency of the sites. We utilized this system to evaluate the editing ability of two protein variants, xCas9 and Cas9-NG, in Bombyx mori cells. Based on the editing ability of these two proteins, the editable range of the Bombyx mori genome can be greatly expanded, accelerating research on the genome's function. This system is also applicable to other insects for detecting the editing ability of Cas9 variants, thus accelerating the study of silkworm genome function. Genome editing provides novel opportunities for the precise genome engineering of diverse organisms. Significant progress has been made in the development of genome-editing tools for Bombyx mori (B. mori) in recent years. Among these, CRISPR/Cas9, which is currently the most commonly used system in lepidopteran insects, recognizes NGG protospacer adjacent motif (PAM) sequences within the target locus. However, Cas9 lacks the ability to target all gene loci in B. mori, indicating the need for Cas9 variants with a larger editing range. In this study, we developed a high-throughput screening platform to validate Cas9 variants at all possible recognizable and editable PAM sites for target sequences in B. mori. This platform enabled us to identify PAM sites that can be recognized by both xCas9 3.7 and SpCas9-NG variants in B. mori and to assess their editing efficiency. Cas9 shows PAM sites every 13 base pairs in the genome, whereas xCas9 3.7 and SpCas9-NG have an average distance of 3.4 and 3.6 base pairs, respectively, between two specific targeting sites. Combining the two Cas9 variants could significantly expand the targeting range of the genome, accelerate research on the B. mori genome, and extend the high-throughput rapid screening platform to other insects, particularly those lacking suitable NGG PAM sequences. [ABSTRACT FROM AUTHOR]

Simple Summary: The fibroin heavy chain (FibH) is the core protein of silkworm silk, and a systematic analysis of its sequence (primary structure) is essential for understanding silkworm silk fibers. However, only a few FibH sequences have been identified. The recently published high-resolution silkworm pan-genome was useful in analyzing the FibH gene systematically. In this study, we extracted the complete FibH sequences of wild silkworms, local and improved strains from the silkworm pan-genome, bringing the total number of known silkworm FibH genes to 286. We characterized in detail the FibH variation during domestication and breeding and established a silkworm pedigree based on the FibH gene sequence. Additionally, we investigated the relationship between the cis-regulatory elements of FibH gene and silk yield in wild and domesticated silkworms. Our study laid the foundation for artificial silk as well as provided guidance for silkworm breeding. The highly repetitive and variable fibroin heavy chain (FibH) gene can be used as a silkworm identification; however, only a few complete FibH sequences are known. In this study, we extracted and examined 264 FibH gene complete sequences (FibHome) from a high-resolution silkworm pan-genome. The average FibH lengths of the wild silkworm, local, and improved strains were 19,698 bp, 16,427 bp, and 15,795 bp, respectively. All FibH sequences had a conserved 5′ and 3′ terminal non-repetitive (5′ and 3′ TNR, 99.74% and 99.99% identity, respectively) sequence and a variable repetitive core (RC). The RCs differed greatly, but they all shared the same motif. During domestication or breeding, the FibH gene mutated with hexanucleotide (GGTGCT) as the core unit. Numerous variations existed that were not unique to wild and domesticated silkworms. However, the transcriptional factor binding sites, such as fibroin modulator-binding protein, were highly conserved and had 100% identity in the FibH gene's intron and upstream sequences. The local and improved strains with the same FibH gene were divided into four families using this gene as a marker. Family I contained a maximum of 62 strains with the optional FibH (Opti-FibH, 15,960 bp) gene. This study provides new insights into FibH variations and silkworm breeding. [ABSTRACT FROM AUTHOR]

Simple Summary: Interactions between insects and microorganisms facilitate the spread of the microorganisms, which in turn affect insect behaviors. In the current paper, we reviewed how fungi, bacteria, and viruses act on insect olfaction, ranging from volatiles of microorganism, microorganism-induced changes in host volatiles, and symbiotic microorganisms that directly affect insect olfaction. Finally, we highlighted the importance of olfaction in insect-microorganism interactions and provided insights into future research directions. Olfaction plays an essential role in various insect behaviors, including habitat selection, access to food, avoidance of predators, inter-species communication, aggregation, and reproduction. The olfactory process involves integrating multiple signals from external conditions and internal physiological states, including living environments, age, physiological conditions, and circadian rhythms. As microorganisms and insects form tight interactions, the behaviors of insects are constantly challenged by versatile microorganisms via olfactory cues. To better understand the microbial influences on insect behaviors via olfactory cues, this paper summarizes three different ways in which microorganisms modulate insect behaviors. Here, we deciphered three interesting aspects of microorganisms-contributed olfaction: (1) How do volatiles emitted by microorganisms affect the behaviors of insects? (2) How do microorganisms reshape the behaviors of insects by inducing changes in the synthesis of host volatiles? (3) How do symbiotic microorganisms act on insects by modulating behaviors? [ABSTRACT FROM AUTHOR]

Simple Summary: Influenced by global climate, trade, and transportation factors, Bactrocera dorsalis (Hendel) has gradually spread from southern regions to Beijing, Hebei, and other northern areas in China. In order to evaluate the risk of damage of B. dorsalis to the dominant northern fruits, an age-stage two-sex life table was used to study the fitness of B. dorsalis for peaches and apples, with oranges as the control. Our results showed that the pest can cause continuous damage on northern fruit hosts, with the damage degree being basically the same on peaches as on the southern fruit of oranges. The B. dorsalis population on peaches increased by 12,112.1 times 90 days after oviposition, easily causing great damage. Additionally, the population increased by 4311 times on apples, which may also become a potentially new host of B. dorsalis in northern China, though with relatively lower fitness. This research lays a foundation for monitoring and the formulation of efficient control strategies for B. dorsalis. Bactrocera dorsalis (Hendel), as a quarantine pest in many countries and regions, has shown a trend of northward diffusion in the past century in China. In order to determine whether B. dorsalis will cause great harm to the dominant northern fruits, the age-stage two-sex life tables of peaches and apples were constructed, with oranges as the control. The results showed that the developmental rate, intrinsic rate of increase (r), and finite rate of increase (λ) on oranges and peaches were significantly greater than on apples. Additionally, the prediction of population growth 90 days after oviposition revealed that the whole population on oranges and peaches increased by 13,667.3 and 12,112.1 times, respectively, indicating that B. dorsalis is very likely to endanger peach orchards. The population increased on apples by 4311 times, though this is lower than that on oranges and peaches. Overall, peaches with high fitness similar to oranges are very suitable as a host for B. dorsalis and are likely to become a new favorable host, while apples may also become a potentially new host, though with lower fitness. Therefore, the most pressing solutions to take are population monitoring, comprehensive prevention, and control in the case of any potential large-scale outbreak of B. dorsalis in northern China. [ABSTRACT FROM AUTHOR]

Simple Summary: Bombyx mori (B. mori), an important economic insect, is famous for its silk. B. mori silk is mainly composed of silk fibroin coated with sericin. Among them, the silk fibroin heavy chain protein has the highest content and the largest molecular weight, which is encoded by the silk fibroin heavy chain (FibH) gene. At present, apart from the complete sequence of the FibH of the B. mori strain p50T, there are no other reports regarding this protein. This is mainly because the special structure formed by the GC-rich repetitive sequence in FibH hinders the amplification of polymerase and the application of Sanger sequencing. Here, the FibH sequence of Dazao, which has 99.98% similarity to that of p50T, was obtained by means of CEO. As far as we know, this is the first complete FibH sequence of the Chinese B. mori strain. Additionally, the methylated CG sites in the FibH repeat unit were identified. To study the evolution of gene function and a species, it is essential to characterize the tandem repetitive sequences distributed across the genome. Cas9-based enrichment combined with nanopore sequencing is an important technique for targeting repetitive sequences. Cpf1 has low molecular weight, low off-target efficiency, and the same editing efficiency as Cas9. There are numerous studies on enrichment sequencing using Cas9 combined with nanopore, while there are only a few studies on the enrichment sequencing of long and highly repetitive genes using Cpf1. We developed Cpf1-based enrichment combined with ONT sequencing (CEO) to characterize the B. mori FibH gene, which is composed of many repeat units with a long and GC-rich sequence up to 17 kb and is not easily amplified by means of a polymerase chain reaction (PCR). CEO has four steps: the dephosphorylation of genomic DNA, the Cpf1 targeted cleavage of FibH, adapter ligation, and ONT sequencing. Using CEO, we determined the fine structure of B. moriFibH, which is 16,845 bp long and includes 12 repetitive domains separated by amorphous regions. Except for the difference of three bases in the intron from the reference gene, the other sequences are identical. Surprisingly, many methylated CG sites were found and distributed unevenly on the FibH repeat unit. The CEO we established is an available means to depict highly repetitive genes, but also a supplement to the enrichment method based on Cas9. [ABSTRACT FROM AUTHOR]