ME_2024v15n1

Molecular Entomology 2024, Vol.15 http://emtoscipublisher.com/index.php/me © 2024 EmtoSci Publisher, registered at the publishing platform that is operated by Sophia Publishing Group, founded in British Columbia of Canada. All Rights Reserved.

Molecular Entomology 2024, Vol.15 http://emtoscipublisher.com/index.php/me © 2024 EmtoSci Publisher, registered at the publishing platform that is operated by Sophia Publishing Group, founded in British Columbia of Canada. All Rights Reserved. Publisher EmtoSci Publisher Editedby Editorial Team of Molecular Entomology Email: edit@me.emtoscipublisher.com Website: http://emtoscipublisher.com/index.php/me Address: 11388 Stevenston Hwy, PO Box 96016, Richmond, V7A 5J5, British Columbia Canada Molecular Entomology (ISSN 1925-198X) is an open access, peer reviewed journal published online by EmtoSci Publisher. The journal is committed to publishing and disseminating all the latest and outstanding research articles, letters and reviews in all areas of molecular entomology. The range of topics including genome structure of insects, gene expression and their function analysis, molecular evolution, molecular ecology, molecular genetics, insect physiology and biochemistry and other topical advisory subjects. Meanwhile we also publish the articles related to basic research, such as anatomy, morphology and taxonomy, which are fundamental to molecular technique’s innovation and development. All the articles published in Molecular Entomology are Open Access, and are distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. EmtoSci Publisher uses CrossCheck service to identify academic plagiarism through the world’s leading plagiarism prevention tool, iParadigms, and to protect the original authors’ copyrights. EmtoSci Publisher is an international Open Access publisher specializing in insect science, and entomology-related research registered at the publishing platform that is operated by Sophia Publishing Group (SPG), founded in British Columbia of Canada.

Molecular Entomology (online), 2024, Vol.15, No.1 ISSN 1925-198X http://emtoscipublisher.com/index.php/me © 2024 EmtoSci Publisher, registered at the publishing platform that is operated by Sophia Publishing Group, founded in British Columbia of Canada. All Rights Reserved. Latest Content Evolution of Butterfly Wing Patterns and Their Ecological Functions BaohuaXu Molecular Entomology, 2024, Vol. 15, No. 1, 1-7 Research on Insect Pathogen Resistance Based on GWAS: Methods, Challenges, and Prospects Fangqi Xu International Journal of Horticulture, 2024, Vol. 14, No. 1, 8-17 Analysis of the Mechanism of Action of Trametinib: Extending the Lifespan of Female Fruit Flies in Intestinal Stem Cells Henry Smith International Journal of Horticulture, 2024, Vol. 14, No. 1, 18-22 Advances in Biological Control Methods for Managing Sugarcane Insects YulinZhou International Journal of Horticulture, 2024, Vol. 14, No. 1, 23-31 Rice Varietal Resistance to Insect Pests: Genetic Mechanisms and Breeding Approaches JiaChen International Journal of Horticulture, 2024, Vol. 14, No. 1, 32-42

Molecular Entomology 2024, Vol.15, No.1, 1-7 http://emtoscipublisher.com/index.php/me 1 Review and Progress Open Access Evolution of Butterfly Wing Patterns and Their Ecological Functions BaohuaXu Chengxi Elementary School in Zhuji City, Zhuji, 311800, Zhejiang, China Corresponding author email: 1073985433@qq.com Molecular Entomology, 2024, Vol.15, No.1 doi: 10.5376/me.2024.15.0001 Received: 03 Nov., 2023 Accepted: 12 Dec., 2023 Published: 01 Jan., 2024 Copyright © 2024 Xu, This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Xu B.H., 2024, Evolution of butterfly wing patterns and their ecological functions, 15(1): 1-7 (doi: 10.5376/me.2024.15.0001) Abstract This study delves into the evolutionary mechanisms of butterfly wing patterns and their ecological functions. Through a comprehensive analysis of classical theories and new models in evolutionary biology concerning the evolution of butterfly wing patterns, the study reveals the relationship between diversity and evolution. Factors influencing evolution are examined, with a particular focus on the potential impact of survival advantages, reproductive success, and external environmental pressures on pattern formation. The study compares the diversity of butterfly wing patterns, including differences among different species and regions. Attention is given to the ecological functions of wing patterns, especially protective colors and evolutionary adaptability, with an in-depth exploration of their roles in ecosystems. Through an in-depth analysis of relevant theories, models, and empirical studies, the study provides valuable guidance for future research and ecological conservation efforts. This research expands our understanding of biological evolution and contributes scientific support to the protection and sustainable development of ecosystems. Keywords Butterfly wings; Evolutionary mechanisms; Ecological functions; Diversity; Evolutionary adaptability Butterflies, as unique creatures among insects, have attracted widespread interest with their beautiful wing patterns (Figure 1). The diversity of these patterns encompasses a rich array of colors, shapes, and patterns, ranging from brilliant colors to unique spots and textures, demonstrating nature's infinite creativity in biomorphic design, and providing a rich source of material for bioaesthetics (Li et al., 2023). Figure 1 Butterfly The diversity and aesthetic appeal of butterfly wings have made them the subject of much attention in the fields of biology and ecology. These patterns are not only a landscape in nature, but also an enigma in evolutionary and ecological processes (Wang et al., 2019). Ecologists and evolutionary biologists have long shown interest in

Molecular Entomology 2024, Vol.15, No.1, 1-7 http://emtoscipublisher.com/index.php/me 2 butterfly wing patterns in an attempt to unravel the evolutionary story behind them and their ecological function in the ecosystem. By tracing changes in wing patterns in butterfly populations, researchers have endeavored to uncover evolutionary traces and decipher the effects of natural selection and genetic drift on pattern formation. Scientists have come to realize that wing patterns may be more than just aesthetic expressions, but may be the product of evolutionary adaptations. This has led to in-depth studies of the evolutionary mechanisms behind butterfly wing patterns. The aim of this study is to thoroughly investigate the evolutionary mechanism of butterfly wing patterns, and to explore the variation of patterns among different species and individuals and the genetic basis behind them (Quan et al., 2023). Through comprehensive analysis of related theories, models and empirical studies, this study will reveal the evolutionary laws and driving forces behind butterfly wing patterns. In addition to aesthetic appeal, butterfly wing patterns may also play important functions in ecosystems. This study aims to promote a deeper understanding of the mysteries behind butterfly wing patterns and provide useful guidance for future research and ecological conservation. 1 The Evolution of Butterfly Wing Patterns The evolution of butterfly wing patterns is an integrative process, subject to a combination of factors. The classical theory provides the basic framework of evolution, while the new model provides a more in-depth analysis at the molecular and population levels. Meanwhile, the relationship between survival advantages and external environmental pressures has enabled butterfly populations to maintain a dynamic balance in evolution. 1.1 Evolutionary theory and models The evolution of butterfly wing patterns has been a topic of great interest in evolutionary biology. Classical theories include Darwin's theory of natural selection, which states that an individual's adaptive wing pattern may be passed on over a long evolutionary process. The theory of natural selection suggests that individuals with wing patterns that are more compatible with their environment are more likely to stand out in the struggle for survival and increase their reproductive success. In addition, sexual selection theory also proposes that certain wing patterns may be better able to attract the attention of the opposite sex, improving the chances of reproduction (Figure 2). Figure 2 Butterfly wing pattern In recent years, with advances in molecular biology and genetics techniques, new models have played an important role in resolving the evolution of butterfly wing patterns. Factors at the molecular level, including gene mutation, gene flow and gene recombination, play a crucial role in evolution. In addition, new models of population genetics emphasize selection and evolution at the population level.

Molecular Entomology 2024, Vol.15, No.1, 1-7 http://emtoscipublisher.com/index.php/me 3 1.2 Factors affecting evolution There is a close correlation between the evolution of butterfly wing patterns and survival advantage and reproductive success. Through long-term field observations and experimental studies, ecologists and geneticists have attempted to investigate the relationship between different wing patterns and an individual's survivability and reproductive success in the natural environment. Certain wing patterns may provide better protective coloration, making it more difficult for butterflies to be detected by predators and thus improving their survivability (Zhang et al., 2022). At the same time, some wing patterns may be associated with reproductive behavior as a signal of opposite-sex attraction. External environmental pressures are another key factor influencing the evolution of butterfly wing patterns. Changes in the environment may lead to changes in survival pressures, thus driving the evolution of butterfly populations towards adaptation to new environments. Global climate change and anthropogenic disturbances may have far-reaching impacts on butterfly ecosystems, thus giving rise to new wing patterns (Han et al., 2023). 2 Diversity of Butterfly Wing Patterns The diversity of butterfly wing patterns is a rich phenomenon at both the species and geographic levels. Variation in wing patterns across species and geographic regions is constrained by both a genetic basis and shaped by environmental factors. 2.1 Species differences The diversity of butterfly wing patterns shows striking contrasts between species. By observing and studying butterfly species globally, researchers have found significant differences in their wing patterns in terms of color, shape and texture. These differences may be caused by species-specific evolutionary paths, ecological environments, and genetic variation (Duan et al., 2023). The wing patterns of different species reflect their roles and adaptive strategies in the ecosystem. Some species may display more cryptic and protective color wing patterns to avoid the attention of natural enemies. Conversely, other species may employ more vibrant colors and distinctive spots to attract the opposite sex during the breeding season or as a sign of territory. Differences in wing patterns between individuals within the same species are also an important aspect to study. Even if they belong to the same species, individuals may exhibit certain differences in wing patterns. These differences may be caused by a variety of factors such as genetic inheritance, environmental factors, or special experiences during individual development. Studying differences in wing patterns of individuals within the same species not only provides insights into genetic variation and phenotypic shaping, but also helps to reveal the role of the environment in shaping individual differences within the same species (Maheshwari et al., 2021). Such differences may play a key role in natural selection, sexual selection and population dynamics, influencing individual survival and reproductive success. 2.2 Regional differences The regional differences in butterfly wing patterns are another noteworthy research direction. In different geographical regions, the same or similar butterfly species may exhibit completely different wing patterns. This regional difference may be due to limited gene flow caused by geographical isolation, or it may be the result of adapting to specific local environments. The study of regional differences helps to understand the adaptive evolution of butterfly populations in different ecosystems. The climate, vegetation, and other ecological factors in different geographical regions may shape the unique characteristics of butterfly wing patterns, making them better adapted to the local environment. Climate and environment are one of the main factors leading to regional differences. The climate differences in different geographical regions can affect vegetation types, temperature ranges, and seasonal changes, thereby

Molecular Entomology 2024, Vol.15, No.1, 1-7 http://emtoscipublisher.com/index.php/me 4 having a profound impact on the ecological environment of butterflies. This influence may be shaped by the power of natural selection, shaping butterfly wing patterns to better adapt to the local environment. 3 Ecological Functions and Adaptability The ecological functions and adaptations of butterfly wing patterns are key factors for their survival and reproduction in nature (Hao et al., 2019). The development of protective colors makes butterflies more hidden in the ecological environment and reduces the risk of predation (Figure 3). At the same time, evolutionary adaptive wing patterning has allowed butterflies to better adapt to different ecological pressures and maintain balance in the ecosystem. Figure 3 Protective colors of butterflies 3.1 Protective colors that integrate with the environment The protective coloration of butterfly wings is an important part of their survival strategy. By coloring in harmony with their surroundings, butterflies are better able to blend into their habitat and reduce the probability of detection by natural predators. The evolution of this protective coloration has made some butterflies almost imperceptible in leaves, flowers, or other natural backgrounds. The study found that butterflies blend in with their environment in an extremely subtle and complex way by mimicking the texture and color of plant surfaces. This natural selection has resulted in some excellent protective coloration, making the butterflies more secretive in their habitats and reducing the risk of predation. 3.2 Mimicry with toxic plants and food Some butterflies choose to mimic toxic plants or foods as a way to develop a mimetic protective coloration. This evolutionary strategy reduces the risk of predation by mimicking the color and texture of toxic substances, causing natural enemies to misidentify toxic organisms. Such mimicry not only gives butterflies a survival advantage in ecosystems, but also provides ecologists with a unique object of study to deepen their understanding of evolution and ecological adaptations. 3.3 Genetic diversity in relation to wing pattern The evolutionary adaptability of butterfly wing patterns is closely related to genetic diversity. The existence of genetic diversity in butterfly populations provides a basis for the variation of wing patterns (She, 2022). Through the diversity of genetic mechanisms, butterflies can form different phenotypes in populations to adapt to different ecological environments and natural enemy pressures.

Molecular Entomology 2024, Vol.15, No.1, 1-7 http://emtoscipublisher.com/index.php/me 5 Studies have shown that there is a strong correlation between the presence of some specific genes and certain wing pattern characteristics. This genetic relationship has driven the evolution of butterfly populations to exhibit a diversity of wing patterns, thereby enhancing their ability to survive in the ecosystem. 3.4 The evolutionary advantages of wing patterns in predation and predation The evolution of butterfly wing patterns is not only related to genetic diversity, but also influenced by the dynamic balance between predation and prey. During the long evolutionary process, butterfly populations have gained the evolutionary advantage of finding a balance between predation and prey through the continuous adjustment of wing patterns (Figure 4). Figure 4 Butterflies in predation Some studies have shown that butterflies with more subtle wing patterns are more likely to avoid the attention of predators, thus improving survival rates. Conversely, some butterflies with striking colors and spots may be more likely to attract the opposite sex, improving their chances of reproductive success. This evolutionary advantage of predation and prey has enabled butterfly populations to achieve ecological balance in complex and changing ecosystems. 4 The Impact of Human Activities on Butterfly Wing Patterns 4.1 The impact of urbanization and agricultural activities Butterfly habitat loss and destruction is a serious problem caused by human urbanization and agricultural activities (Xie et al., 2019). With the increase of urban expansion and agricultural land use, the native habitats of many butterflies have suffered serious threats. Urbanization has led to the covering of large areas of land, and natural environments originally used for butterfly reproduction and foraging have been gradually replaced by concrete. Agricultural activities are also one of the important factors affecting butterfly habitats. Large-scale reclamation of agricultural land, the widespread use of chemical pesticides and the monoculture of agriculture pose a threat to the survival and breeding environment of butterflies. These activities have not only led to a reduction in plant diversity, but have also negatively affected gene flow and the evolution of wing patterns in butterfly populations. 4.2 Impact of deforestation on species diversity and wing patterns Deforestation is another important factor affecting butterfly habitats. Large-scale logging activities have led to the destruction of many butterflies' natural habitats, depriving them of their ideal habitat. Deforestation has triggered a decline in species diversity, with far-reaching effects on the gene pools of butterfly populations and the formation of wing patterns.

Molecular Entomology 2024, Vol.15, No.1, 1-7 http://emtoscipublisher.com/index.php/me 6 The disappearance of forests poses new challenges to the genetic diversity and adaptability of butterfly populations. The original lush forest environment not only provided sufficient food resources for butterflies, but also ideal breeding sites. Deforestation has gradually weakened these conditions, posing a threat to the survival of butterflies. 4.3 Possible effects of global warming on butterfly distribution and wing patterns Global warming is an issue of great concern today, with potentially far-reaching effects on the distribution and wing patterns of butterfly populations. Warming has led to changes in temperature and humidity in many regions, which pose new challenges to butterfly habitats. On the one hand, rising temperatures may lead to a reduction in the habitat of some butterfly species as they are unable to adapt to the new climatic conditions. On the other hand, climate change may prompt some butterfly species to expand their ranges in search of more suitable habitats. This will trigger competition between different species, creating new pressures on the evolution of wing patterns (Mortazavi and Moloodpoor, 2021) 4.4 Potential impacts of extreme weather events Climate change is also accompanied by an increase in extreme weather events, such as droughts, floods and hurricanes. These extreme weather events may have a direct impact on the survival of butterfly populations and the formation of wing patterns. For example, droughts may cause herbaceous plants in butterfly habitats to die out, reducing the butterflies' food source and thus affecting their reproduction and survival. Meanwhile, floods may destroy butterfly eggs and larvae, reducing the birth rate of new generations. Overall, the impact of climate change on butterfly wing patterns is a comprehensive issue that requires a concerted effort by the global community to address in order to protect this unique and beautiful creature. 5 Summary and Outlook The evolution of butterfly wing patterns is a complex and fascinating process involving the interaction of multiple ecological, evolutionary, and genetic factors (Zhang and Fang, 2019). This study delves into the diversity of butterfly wing patterns, evolutionary mechanisms, and associations with ecological functions. In terms of evolutionary theories and models, this study reviewed the classical theories on the evolution of butterfly wing patterns in evolutionary biology and introduced new models that have emerged in recent years. Among the factors affecting evolution, this study explored the association between survival advantage and reproductive success, as well as the potential effects of external environmental stress on the evolution of wing patterns. For the diversity of butterfly wing patterns, this study compares the differences between species and between individuals within the same species, and also investigates the territorial differences in butterfly wing patterns in different geographical regions. Future butterfly wing pattern research can leverage advanced technological innovations and research methods to increase the depth and breadth of research. Among them, molecular biology techniques such as genomics and transcriptomics can help researchers gain a deeper understanding of the molecular mechanisms underlying the formation of butterfly wing patterns and reveal the association between genes and phenotypes. Meanwhile, the application of advanced imaging techniques and mathematical models will provide more detailed information on the three-dimensional structure and evolution of wing patterns. In order to conserve butterfly diversity, one needs to adopt a comprehensive strategy. First of all, the protection of butterfly habitats should be strengthened, especially by formulating sustainable development policies with regard to urbanization and agricultural expansion. In this process, an ecological compensation mechanism can be adopted to compensate for habitat damage. Secondly, monitoring and response to climate change should be strengthened. A global cooperation mechanism should be established to promote the reduction of greenhouse gas emissions and slow down the rate of climate change. At the same time, corresponding protection strategies should be formulated for butterfly groups in different regions to ensure that they can adapt to the new climate environment. In addition,

Molecular Entomology 2024, Vol.15, No.1, 1-7 http://emtoscipublisher.com/index.php/me 7 promote education and public participation to raise people's awareness of butterfly conservation. Through popularization of science activities, establishment of protected areas and promotion of regulations, the whole society can form a synergy for the conservation of butterfly diversity (Alweshah et al., 2020). In conclusion, the in-depth study of butterfly wing patterns not only expands people's understanding of biological evolution, but also provides scientific support for ecological conservation. Through technological innovation and comprehensive conservation strategies, it is expected that this unique and beautiful creature can be conserved and contribute to the sustainable development of biodiversity. References Alweshah M., Khalaileh S.A., Gupta B.B., Almomani A., Hammouri A.I., and Azmi Al-Betar M., 2020, The monarch butterfly optimization algorithm for solving feature selection problems, Neural Computing and Applications, 27: 1-15. https://doi.org/10.1007/s00521-020-05210-0 Duan M.Y., Zhu H., Qu Y.K., Wang W.H., Jiang S.Q. Yuan K., and Ren B.Z., 2023, Diversity of butterfly communities in different habitats in Songnen Plain and conservation suggestions, Shengtai Xuebao (Acta Ecologica Sinica), 43(18):7682-7692. Han D., Han C.H., Wang C., She J.Y., Bian Q., Han W.J., and Yin L.Q., 2023, Butterfly diversity and dominant species niche in different urbanization zones of beijing, Zhongguo Chengshi Linye (Journal of Chinese Urban Forestry), 21(5): 74-81. Hao S.L., Xue Q.Q., Feng D.D., Li X.F., Liu Y., Zhang Z.W., and Men L.N., 2019, Comparative study on butterfly diversity and niche difference in mountainous region of southern shanxi province, Shengtai yu Nongcun Huanjing Xuebao (Journal of Ecology and Rural Environment), 35(10): 1314-1321. Li F., Zhao K.X., Yan C.Y., Yan J.W., Xing J.C., and Xie B.L., 2023, Identification of butterfly species in the natural environment based on residual network, Kunchong Xuebao (Acta Entomologica Sinica), 66(3): 409-418. Maheshwari P., Sharma A.K., and Verma K., 2021, Energy efficient cluster based routing protocol for wsn using butterfly optimization algorithm and ant colony optimization, Ad Hoc Networks, 110: 102317. https://doi.org/10.1016/j.adhoc.2020.102317 Mortazavi A., and Moloodpoor M., 2021, Enhanced butterfly optimization algorithm with a new fuzzy regulator strategy and virtual butterfly concept, Knowledge-Based Systems, 228: 107291. https://doi.org/10.1016/j.knosys.2021.107291 Quan L.F., Yao Q., Dong Y.Z., Xu S., Chi Y.Y., and Chen B.X., 2023,Research progress and prospects of circadian clock in lepidoptera, Nongxue Xuebao (Journal of Agriculture), 13(9): 38-45. She J.Y., Han D., Wang C., Yin L.Q., Sun Z.K., and Han C.H., 2022, Butterfly diversity in pocket parks at urban core of beijing, 20(3): 1-6. Wang G.G., Deb S., and Cui Z., 2019, Monarch butterfly optimization, Neural computing and applications, 31: 1995-2014. https://doi.org/10.1007/s00521-015-1923-y Xie J.Y., Cao J.W., Ma L.B., Zhen W.Q., Chen Z.Y., Li X.D., Li H.H., and Xu S.Q., 2019, A dataset of butterfly ecological images for automatic species identification, Zhongguo Kexue Shuju Zhongyingwen Wangluoban (China Scientific Data), 4(3): 189-194. Zhang Y.H., Huang F., Tan X.Y., Xia Z.Q., He X.Y., and Wu G., 2022, Resource survey and diversity analysis of butterflies in huangmei county,hubei province, Zhiwu Baohu Xuebao (Journal of Plant Protection), 49(4): 1277-1278. Zhag Y.J., and Fang L.J., 2019, Evaluation on eco-environment in the danjiang river basin based on gis and butterfly diversity, Shijie Shengtaixue (International Journal of Ecology), 8(3): 223-232. https://doi.org/10.12677/IJE.2019.83030 Disclaimer/Publisher's Note The statements, opinions, and data contained in all publications are solely those of the individual authors and contributors and do not represent the views of the publishing house and/or its editors. The publisher and/or its editors disclaim all responsibility for any harm or damage to persons or property that may result from the application of ideas, methods, instructions, or products discussed in the content. Publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Molecular Entomology 2024, Vol.15, No.1, 8-17 http://emtoscipublisher.com/index.php/me 8 Research Article Open Access Research on Insect Pathogen Resistance Based on GWAS: Methods, Challenges, and Prospects Fangqi Xu Insect Breeding and Biotesting Laboratory, Cuixi Academy of Biotechnology, Zhuji, 311800, Zhejiang, China Corresponding author email: 1049879736@qq.com Molecular Entomology, 2024, Vol.15, No.1 doi: 10.5376/me.2024.15.0002 Received: 14 Nov., 2023 Accepted: 24 Dec., 2023 Published: 14 Jan., 2024 Copyright © 2024 Xu, This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Xu F.Q., 2024, Research on insect pathogen resistance based on gwas: methods, challenges, and prospects, Molecular Entomology, 15(1): 8-17 (doi: 10.5376/me.2024.15.0002) Abstract This study reviews the application of genome-wide association studies (GWAS) in the field of insect pathogen resistance, and discusses the main methods, challenges, and future development prospects of this research direction. This study introduces the basic principles of GWAS and its application in revealing the genetic basis of insect resistance to pathogens. By analyzing genetic variations in the insect genome, GWAS helps scientists identify key genes and functional regions related to resistance. This study discusses the main challenges encountered in conducting GWAS research, including sample size limitations, genetic diversity, environmental factors, and difficulties in detecting rare variations. It also explores issues such as data sharing and privacy protection. This study looks forward to the potential of utilizing GWAS results to improve insect resistance strategies, including the application of gene editing techniques such as CRISPR-Cas9 in insect resistance improvement, and emphasizes the importance of interdisciplinary collaboration in solving complex scientific problems. This study aims to provide a comprehensive perspective for the research and management of insect pathogen resistance, promoting scientific progress and technological innovation in related fields. Keywords Genome-wide association studies; Insect resistance; Gene editing technology; Interdisciplinary cooperation; Pathogen control Insects, as one of the most diverse and widely distributed biomes on Earth, are closely linked to human life. They play a variety of roles in natural ecosystems, both as beneficial and harmful insects, the latter being particularly known for their enormous damage to agricultural crops. Insect pathogens, including viruses, bacteria, fungi and parasites, pose a major threat to agriculture and forestry and can cause significant reductions in crop yields, thus challenging global food security. Some insects are also capable of transmitting deadly diseases to humans, such as mosquito-borne malaria and dengue fever, further highlighting the importance of controlling the spread of insect pathogens (Pan et al., 2023). In this context, understanding the mechanisms of insect resistance to these pathogens has become particularly important. This can not only help develop more effective pest management strategies and reduce the use of pesticides, but also provide new ideas for the prevention and control of infectious diseases. In recent years, genome-wide association studies (GWAS) have been widely used as a powerful genetic research tool in the study of insect pathogen resistance. By analyzing the association between genetic variants in the insect genome and resistance phenotypes, GWAS can help scientists to reveal the genetic basis of insect resistance and thus its complex genetic mechanisms. The application of GWAS has greatly contributed to the understanding of the genetic basis of insect resistance. Compared with traditional genetic analysis methods, GWAS is able to provide a rapid and comprehensive scan of the entire genome without relying on a priori genetic information. This means that GWAS can efficiently identify key genetic variants even for resistance traits that have complex genetic backgrounds and are associated with multiple genes. This ability is critical for resolving the complexity of insect pathogen resistance, especially in the current context of global change and rapidly changing agricultural practices.

Molecular Entomology 2024, Vol.15, No.1, 8-17 http://emtoscipublisher.com/index.php/me 9 Although GWAS shows great potential for insect resistance research, its application faces a number of challenges, including the huge amount of data, computational complexity, and difficulties in interpreting the results. How to translate the genetic information discovered by GWAS into practical pest management strategies is also an important direction of current research (Li et al., 2022). The aim of this study was to overview GWAS-based insect pathogen resistance research, analyze its methodology, challenges and future prospects, introduce the basic principles of GWAS technology and its application in insect resistance research, and discuss in detail the progress of research on the genetic basis of insect resistance, including the key genes and functional regions discovered through GWAS technology. This study explores the major challenges encountered in the implementation of GWAS research, such as the effects of sample size, genetic diversity and environmental factors, etc. It will also demonstrate the application and achievements of GWAS technology in revealing insect resistance mechanisms through several specific research cases. This study will discuss the potential applications and perspectives of GWAS technology in future entomopathogen resistance studies, and how complex problems in entomopathogen resistance studies can be solved through interdisciplinary collaborations. 1 Application of GWAS Technical Methods 1.1 The role of GWAS in insect pathogen resistance research Genome-wide association studies (GWAS) is a scientific method used to study the association between genetic variation and complex traits, and has been widely used in insect pathogen resistance research in recent years.The basic principle of GWAS is to search for genetic markers associated with specific traits by scanning genome-wide genetic variation in a large number of individuals. genetic markers associated with specific traits. This approach can reveal the genetic basis behind traits and provide new ideas for understanding the mechanisms of insect resistance to pathogens. The GWAS technique is based on the premise that differences in traits can to some extent be explained by genetic variation in the genome. These variations usually refer to single nucleotide polymorphisms (SNPs), which are the most common genetic markers in the genome. By comparing the distribution of SNPs in populations of individuals with different traits (e.g., disease resistance versus disease susceptibility), it is possible to identify genetic loci that are significantly associated with a particular trait.GWAS studies usually require a large number of samples in order to ensure the reliability of the statistical results (Liu et al., 2023). The basic steps in conducting a GWAS study include: sample collection, DNA extraction, genotyping, statistical analysis, and validation of candidate loci. Each step requires precise technical support and strict quality control to ensure the accuracy and reliability of the data. 1.2 Special considerations and applicability of GWAS technology in insect research Insects, as research objects, have their own special characteristics when applying GWAS technology. There are many kinds of insects, and the genetic backgrounds of different kinds vary greatly, which requires representativeness and comparability when choosing research objects. Insects have a short life cycle and fast reproduction rate, which facilitates the rapid acquisition of a large amount of genetic material, but also requires researchers to be able to effectively manage and maintain the experimental population. The small size of insects and the limited amount of DNA extracted require the use of highly sensitive genotyping techniques. The application of GWAS techniques in insect resistance research also requires consideration of the impact of environmental factors. Insect resistance traits are often affected by a combination of multiple genetic and environmental factors, which requires the use of appropriate models to control these confounding factors when performing GWAS analysis to ensure the accuracy of the research results.

Molecular Entomology 2024, Vol.15, No.1, 8-17 http://emtoscipublisher.com/index.php/me 10 1.3 Data processing and statistical analysis methods In genome-wide association studies (GWAS), data processing and statistical analysis are crucial steps that ensure the accuracy and reliability of the study results. This process involves a complex transformation from quality control of raw data to final association analysis aimed at identifying statistically significant correlations between insect pathogen resistance and specific genetic markers. Overall, data processing and statistical analysis play a central role in GWAS, involving not only the initial stages of study design, but also every detail of data analysis and interpretation of results. Through these fine steps, researchers are able to mine meaningful information from massive amounts of genetic data to provide a scientific basis for understanding pathogen resistance in insects. 2 Genetic Basis of Insect Resistance 2.1 Genetic mechanisms of insect resistance to major pathogens As the most diverse group of organisms on earth, the diversity and complexity of resistance mechanisms in insects have been the focus of scientific research. Insects are capable of resisting a wide range of pathogens, including viruses, bacteria, fungi and parasites, and this ability is based on their complex genetic and molecular mechanisms. In recent years, genome-wide association studies (GWAS) have provided powerful tools for resolving these resistance mechanisms, revealing many key genes and functional regions, and these findings have opened up new horizons for understanding the genetic basis of insect resistance (Wang et al., 2022). The ability of insects to defend themselves against pathogen attacks is largely dependent on the effective response of their immune system. For different types of pathogens, insects exhibit different resistance strategies. For example, against viruses, the RNA interference (RNAi) machinery in insects is activated to prevent viral replication by specifically degrading viral RNA. In contrast, when confronted with bacteria and fungi, insects initiate an immune response that produces antimicrobial peptides to directly kill or inhibit the growth of these microorganisms. Insects are also able to limit parasite infestation through, for example, physical isolation. 2.2 Key genes and functional regions identified in GWAS studies Through GWAS analysis, scientists have successfully identified several key genes and functional regions associated with insect pathogen resistance. For example, in studies against certain viruses, GWAS has helped identify several key genes associated with the RNAi pathway, such as Dicer-2 and Argonaute-2, which play important roles in the recognition and cleavage of viral RNA. In terms of viral resistance, GWAS studies have revealed several genes associated with the RNA interference (RNAi) pathway, such as Dicer-2 and Argonaute-2, which play key roles in the recognition and degradation of viral RNA, one of the major defense mechanisms against viral infestation in insects. This mechanism protects insects from viral infection by specifically cleaving the genetic material of viruses and blocking their replication and spread. For bacterial and fungal resistance, GWAS studies have similarly identified key components of insect immune signaling pathways, particularly some components of the Toll and Imd signaling pathways. These pathways play a central role in activating the insect's natural immune response, including defense mechanisms such as recognition of pathogens, activation of immune cells, and production of antimicrobial peptides. Through these mechanisms, insects are able to effectively defend themselves against bacterial and fungal attacks. In the face of bacterial attack, some key components of signaling pathways, such as those of the Toll and Imd signaling pathways, have also been pointed out by the GWAS study to be closely related to the anti-bacterial capacity of insects. These findings not only deepen the understanding of insect immune mechanisms, but also offer the possibility of developing novel biopesticides or improving insect resistance.

Molecular Entomology 2024, Vol.15, No.1, 8-17 http://emtoscipublisher.com/index.php/me 11 2.3 Association of resistance with physiological and behavioral characteristics of insects Pathogen resistance in insects is not only limited to its genetic basis, but is also strongly influenced by its physiological and behavioral characteristics. These traits are closely related to insect resistance, and together they constitute a complex set of defense mechanisms that enable insects to display amazing adaptability and resistance in the face of pathogen attack. From a physiological point of view, an insect's immune system is its first line of defense against pathogens. Immune cells and antimicrobial peptides in insects are able to respond rapidly to pathogen invasion and directly kill or inhibit pathogen growth. The metabolic pathways of insects are also closely linked to their resistance capabilities. For example, insects are able to resist or eliminate toxins from their bodies by metabolizing specific compounds that may be inhibitory to certain pathogens. Detoxification enzyme systems in insects, such as cytochrome P450 enzymes, are also able to help insects detoxify, thereby enhancing their resistance to various pathogens in the environment (Figure 1). Figure 1 Correlation between resistance and insect physiological and behavioral characteristics Insect resistance is a complex trait with multiple levels and dimensions, in which physiological and behavioral traits are adapted and optimized to provide insects with additional means of defense. These adaptive behaviors and physiological responses are not only part of an insect's survival strategy, but also important mechanisms developed during its evolution to counteract environmental stresses, especially pathogen threats. An in-depth understanding of the physiological and behavioral characteristics of insects and their association with resistance is important for unraveling the mechanisms of insect disease resistance and even for developing novel pest management strategies. 3 Challenges and Limitations 3.1 Impact of sample size, genetic diversity and environmental factors on GWAS accuracy Genome-wide association studies (GWAS), as a powerful genetic research tool, have made a series of breakthroughs in the field of insect pathogen resistance. However, this research method still faces a series of

Molecular Entomology 2024, Vol.15, No.1, 8-17 http://emtoscipublisher.com/index.php/me 12 challenges and limitations, including the effects of sample size, genetic diversity, environmental factors, and the difficulties of detecting rare variants and complex traits, as well as issues of data sharing and privacy protection. First, sample size is one of the important determinants of GWAS accuracy. A sufficiently large sample size can improve the statistical efficacy of a study, allowing even small genetic effects to be detected. However, in entomopathogen resistance studies, obtaining large numbers of high-quality samples is often difficult, especially for rare or geographically diverse insect species. Genetic diversity also affects the results of GWAS. Highly diverse genetic backgrounds may mask or obscure important genetic signals, making it difficult to analyze the genetic basis of traits. The influence of environmental factors should not be ignored. Insects live in extremely complex environments, and environmental variables such as temperature, humidity, and food sources may affect their resistance to pathogens. Even under the same genetic background, different environmental conditions may lead to significant differences in insect resistance. Failure to adequately control these environmental factors in GWAS studies may lead to an increase in false positive results and affect the accuracy of the study. 3.2 Difficulties in detecting rare variants and complex traits A major challenge in genome-wide association studies (GWAS) is the detection of rare variants and complex traits. These two problems center on the fact that, on the one hand, rare variants are difficult to be effectively detected in conventional GWAS sample sizes due to their extremely low frequency in populations; on the other hand, complex traits are usually the result of multiple genes as well as environmental factors, which makes it particularly difficult to accurately identify all the relevant genetic factors (Du et al., 2021). Rare variants, despite their low frequency, may in some cases have a decisive impact on pathogen resistance in insects. For example, a rare variant may make an insect highly resistant to a specific pathogen, but because the frequency of such variants in a population is extremely low, it is difficult for conventional GWAS designs to be statistically efficacious enough to detect these rare variants that are significantly correlated with traits. Detection of rare variants is also affected by sample selection and genotype quality control criteria, which further increases the difficulty of detection. For the detection of complex traits, the complexity of the polygenic genetic mechanisms and gene-environment interactions underlying the trait are involved. Insect resistance traits are often not simple genetic traits determined by a single gene, but are the result of the interaction of multiple genes under specific environmental conditions. In this case, even if GWAS are able to identify some genetic markers associated with the trait, it is difficult to fully explain the genetic variation in the trait, especially when the trait is strongly influenced by the environment. Interactions between genes (phenotypically non-additive effects) and between genes and the environment also pose challenges in identifying relevant genetic factors. 3.3 Challenges of data sharing and privacy protection Data sharing is an important aspect of accelerating scientific discovery and technological advancement when conducting genome-wide association studies (GWAS). By sharing data, researchers can validate the results of other studies, discover new research directions, or improve statistical validity through meta-analysis. However, the process of data sharing also faces the challenge of privacy protection, and although individual privacy issues may not be as prominent in insect research as in human research, a range of privacy and sensitive information protection issues are still involved. In order to address these challenges, it is crucial to develop sound data management and sharing policies. For example, the legality and legitimacy of the purpose of data use can be ensured through the establishment of a data access committee (DAC) to scrutinize data access requests. De-identify or anonymize data through technical means to reduce the risk of exposing sensitive information. The establishment of clear data use agreements and

Molecular Entomology 2024, Vol.15, No.1, 8-17 http://emtoscipublisher.com/index.php/me 13 copyright notices can protect the interests of data providers to a certain extent and promote the construction of a healthy sharing ecology for scientific data. Data sharing and privacy protection is also an issue that needs to be emphasized in insect GWAS research. Through the formulation of reasonable policies and the application of technical means, the value of data sharing can be maximized, while protecting the interests of all parties involved in the research, and promoting the healthy development of entomological research and related application fields. 4 Research Case Studies 4.1 Insect resistance to agricultural pest pathogens The research team of Leeuwen et al. (2020) in 2020 carried out the interpretation of saliency and molecular diagnostics in the management of resistance to agricultural pests in the current agriculture, where insecticide options against agricultural pests are diminishing due to environmental and health concerns, and insecticide-resistant pests are becoming increasingly difficult to control. Rational decisions on insecticide use are needed to ensure effective resistance management. However, monitoring programs that can inform about pest susceptibility and resistance are currently not widely available in agriculture. Sparks et al. (2020) team used insecticides, biological agents, and nematicides: an update of the IRAC classification of modes of action-tools for resistance management. Insecticide resistance is an important issue and the IRAC mode of action classification update provides the latest information for implementing effective resistance management strategies. Pests and pathogens are common problems in agricultural production and they pose a significant threat to crop growth and yield. However, some insect populations exhibit resistance to these pest pathogens, which provides an important biological control method for agricultural production. By studying and analyzing these resistant insect populations, the principles of their resistance mechanisms can be revealed, thus providing a theoretical basis for the development of more effective control strategies. 4.2 Role of rare genetic variation in insect resistance The team of Nam et al. (2019) tested whether copy number variation was responsible for increased levels of insect resistance in two populations of fall stickleback (Spodoptera frugiperda) in different geographic locations and different host plants. Mckenzie (2000) conducted experiments on the nature or variation of resistance and genetic analysis of insecticide resistance phenotypes in insects, and discussed the genetic basis of resistance evolution as dependent on the manner in which phenotypes and their underlying genotypic variation are directed during selection responses. A polygenic response is favored if selection acts within the distribution of susceptible phenotypes, and a monogenic response is predicted if selection screens for rare mutations whose phenotypes lie outside that susceptibility distribution. Insect populations are rich in genetic variation, some of which may be closely associated with their resistance phenotypes. Rare genetic variation, an important form of genetic variation, plays an important role in insect resistance. Through the study of these rare genetic variants, a deeper understanding of the genetic basis of insect resistance and its evolutionary mechanism can be achieved. 4.3 Utilizing GWAS results to improve resistance strategies Chen et al. (2021) conducted cotton disease and insect resistance: this study identified loci significantly associated with yellow wilt resistance by GWAS and found that two non-specific lipid transfer protein genes (GhnsLTPsA10) were highly expressed under yellow wilt pathogen stress. The expression of these genes was significantly increased in roots against the yellow wilt pathogen, but significantly decreased in leaves under insect attack. Siddiqui et al. (2023) conducted Resistance mechanisms and control strategies: this study discussed resistance mechanisms in invasive species including behavioral, biochemical, physiological, genetic and metabolic

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