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Genome editing in insects: current status and challenges

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National science Review 2019年第3期6卷 399-401页

作者： Jun Xu Xia Xu Shuai Zhan Yongping Huang CAS Key Laboratory of insect Developmental and evolutionary biology CAS Center for Excellence in Molecular Plant Sciences Shanghai Institute of Plant Physiology and Ecology Chinese Academy of Sciences

insects constitute the largest group in the animal kingdom, and include many economically important species as well as pests. This abundance provides largescale genetic resources, and research on insects is beneficial from the perspectives of human health, agricultural production and basic science (Fig.1).The availability of many genetic tools has made Drosophila melanogaster a crucially important model organism

关键词： Genome editing in insects current status and challenges

MicroRNA-2738 regulates gene expression in the sex determination pathway in Bombyx mori

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insect science 2020年第4期27卷 646-654页

作者： Zu-Lian Liu Jun Xu Lin Ling De-Hong Yang Shu-Qing Chen Yong-Ping Huang CAS Key Laboratory of insect Developmental and evolutionary biology CAS Center for Excellence in Molecular Plant SciencesInstitute of Plant Physiology and EcologyCASShanghaiChina

MicroRNAs(miRNAs)are a class of short,non-coding transcripts that bind to B'-untranslated regions to trigger messenger RNA degradation or translational *** we explored how miRNAs regulate sex determination in Bombyx mori,a lepidopteran model *** known to be involved in sex determination,BmPSI,Bmdsx,and BmMasc\are predicted targets of the species-specific *** a dual luciferase reporter assay in HEK293T cells,we confirmed that miR-2738 suppressed transcription of BmPSl,Bmdsx,and *** levels of BniPSI and BmMasc were significantly down-regulated in *** miR-2738 *** contrast,the genetic disruption of miR-2738 using the clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 transgenic system increased the levels of BmPSI and BmMasc transcripts,whereas splicing of Bmdsx was unaltered by miR-2738 depletion or *** together,this study implicates miR-2738 as a minor regulator of sex determination genes in the silkworm.

关键词： CRISPR/Cas9 miRNA sex determination silkworm

Natural selection contributes to the myopia epidemic

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National science Review 2021年第6期8卷 128-135页

作者： Erping Long Jianzhi Zhang department of ecology and evolutionary biology University of Michigan State Key Laboratory of Ophthalmology Zhongshan Ophthalmic CenterSun Yat-sen University

The prevalence of myopia, or nearsightedness, has skyrocketed in the past few decades, creating a public health crisis that is commonly attributed to lifestyle changes. Here we report an overall increase in the frequencies of myopia-associated mutant alleles over 25 years among participants of the UK *** myopia itself appears to be selected against, many of the mutant alleles are associated with reproductive benefits, suggesting that reproduction-related selection inadvertently contributes to the myopia epidemic. We estimate that, in the UK alone, natural selection adds more than 100 000 myopia cases per generation, and argue that antagonistic pleiotropy be broadly considered in explaining the spreads of apparently disadvantageous phenotypes in humans and beyond.

关键词： allele frequency natural selection antagonistic pleiotropy evolution myopia reproduction

CRISPR/Cas9-based mutation reveals Argonaute 1 is essential for pigmentation in Ostrinia fumacalis

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insect science 2019年第6期26卷 1020-1028页

作者： Lang You Hong-Lun Bi Yao-Hui Wang Xiao-Wei Li Xi-En Chen Zhi-Qian Li CAS Key Laboratory of insect Developmental and evolutionary biology CAS Center for Excellence in Molecular Plant Sciences Shanghai Institute of Plant Physiology and Ecology Chinese Academy of Sciences Shanghai China

Ostrinia fumacalis (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. fumacalis due to a lack in genomic information. In this report, we identified only one transcript of O. fumacalis Argonaute 1 (OfAgo 1) gene from the genome and cloned the open reading frame. OfAgol presented the maximum expression at the embryo stage or in the fat body during the larval stages. To understand its function, an OfAgol mutant was constructed using the Clustered Regularly Interspaced Short Palindromic Repeat/RNA-guided Cas9 nuclease (CRISPR/Cas9). Mutagenesis of OfAgol disrupted cuticle pigmentation by dowregulating micro RNAs and pigmentation-related genes. This is the first report for the cloning and functional analysis of OfAgol, revealing a role of OfAgol in cuticle pigmentation. The current report also established a CRISPR/Cas9 system in O. fumacalisy providing a new insight for pest management.

关键词： Agol CRISPR/Cas9 cuticle pigmentation Ostrinia fumacalis pest management

Temperature Responses to Infrared-Loading and Water Table Manipulations in Peatland Mesocosms

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Journal of Integrative Plant biology 2008年第11期50卷 1484-1496页

作者： Jiquan Chen Scott Bridgham Jason Keller John Pastor Asko Noormets Jake F.Weltzin department of Environmental sciences University of Toledo center for ecology and evolutionary biology University of Oregon Smithsonian Environmental Research center department of Biological sciences University of Minnesota National Phenology Network

We initiated a multi-factor global change experiment to explore the effects of infrared heat loading （HT） and water table level （WL） treatment on soil temperature （T） in bog and fen peatland mesocosms. We found that the temperature varied highly by year, month, peatland type, soil depth, HT and WL manipulations. The highest effect of HT on the temperature at 25 cm depth was found in June for the bog mesocosms （3.34-4.27 ℃） but in May for the fen mesocosms （2.32-4.33 ℃） over the 2-year study period. The effects of WL in the bog mesocosms were only found between August and January, with the wet mesocosms warmer than the dry mesocosms by 0.48-2.03 ℃ over the 2-year study period. In contrast, wetter fen mesocosms were generally cooler by 0.16-3.87℃. Seasonal changes of temperatures elevated by the HT also varied by depth and ecosystem type, with temperature differences at 5 cm and 10 cm depth showing smaller seasonal fluctuations than those at 25 cm and 40 cm in the bog mesocosms. However, increased HT did not always lead to warmer soil, especially in the fen mesocosms. Both HT and WL manipulations have also changed the length of the non-frozen season.

关键词： global change infrared mesocosm peatlands warming experiment water table.

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Heterogeneous signals in plant–biotic interactions and their applications

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science China(Life sciences) 2019年第12期62卷 1707-1709页

作者： Fang-Yan Chen Xiao-Ya Chen Ying-Bo Mao Chinese Academy of sciences (CAS) Key Laboratory of insect Developmental and evolutionary biology CAS Center for Excellence in Molecular Plant Sciences/Shanghai Institute of Plant Physiology and Ecology University of CAS Chinese Academy of Sciences

Plants have to overcome different types of environment stress including various insect and pathogen attacks during their life *** long-term evolution,plants have developed sophisticated systems to recognize different biotic attacks and initiate an integrated defense network for *** the other hand,pathogens and insects have devised multiple strategies to adapt to their host *** the past three decades,substantial progress has been made toward improving the understanding of plant-biotic *** increasing number of active molecules have been identified that not only have functions within an organism but also act as heterogeneous regulators that can migrate across interacting individuals(Kan et al.,2017;Qian et al.,2017).The discovery of new heterogeneous regulators and their mechanisms of migration across interacting organisms is potentially helpful in the development of novel strategies to protect crop plants from biotic threats.

关键词： plant overcome migrate

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Identification of a germline-expression promoter for genome editing in Bombyx mori

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insect science 2019年第6期26卷 991-999页

作者： Jun Xu Rong-Mei Chen Shu-Qing Chen Kai Chen Lin-Meng Tang De-Hong Yang Xu Yang Yong Zhang Hong-Sheng Song Yong-Ping Huang CAS Key Laboratory of insect Developmental and evolutionary biology CAS Center for Excellence in Molecular Plant Sciences Shanghai Institute of Plant Physiology and Ecology Chinese Academy of Sciences ShanghaiChina College of Life sciences Shanghai University ShanghaiChina department of biology University of NevadaRenoNevadaUSA

Identification of stage- and tissue-specific cis-regulatory elements will enable more precise genomic editing. In previous studies of the silkworm Bombyx mori, we identified and characterized several tissue- and sex-specific cisregulatory elements using transgenic technology, including a female- and fat body-specific promoter, vitellogenin, testis-specific promoters, Radial spoke head 1 (BmRl) and beta-tubulin 4 (Bmp4). Here we report a c/5-regulatory element specific for a somatic and germ cell-expressed promoter, nanos (Bmnos). We investigated activities of three truncated promoter sequences upstream of the transcriptional initiation site sequences of Bmnos in vitro (nos-0.6kb, nos-1 kb and nos-2kb) and in vivo (nos-2kb). In BmN cultured cells, all three lengths drove expression of the gene encoding enhanced green fluorescence protein (EGFP), although nos-2kb had the highest fluorescence activity. In transgenic silkworms, nos-2kb drove EGFP expression at the early embryonic stage, and fluorescence was concentrated in the gonads at later embryonic stages. In addition, this cisregulatory element was not sex differentiated. The fluorescence intensity gradually weakened following the larval developmental stage in the gonads and were broadly expressed in the whole body. The nos-2kb promoter drove the Cas9 system with efficiency comparable to that of the broad-spectrum strong IE1 promoter. These results indicate that Bmnos is an effective endogenous cisregulatory element in the early embryo and in the gonad that can be used in applications involving the clustered, regularly interspaced, short palindromic repeats (CRISPR)/Cas9 system.

关键词： Bombyx mori nanos promoter transgenic CRISPR/Cas9

The Masc gene product controls masculinization in the black cutworm, Agrotis ipsilon

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insect science 2019年第6期26卷 1037-1044页

作者： Yao-Hui Wang Xi-En Chen Yang Yang Jun Xu Gang-Qi Fang Chang-Ying Niu Yong-Ping Huang Shuai Zhan CAS Key Laboratory of insect Developmental and evolutionary biology CAS Center for Excellence in Molecular Plant Sciences Institute of Plant Physiology and Ecology ShanghaiChina Hubei Key Laboratory of insect Resource Application and Sustainable Pest Control College of Plant Science & Technology Huazhong Agricultural University Wuhan China School of Life science East China Normal UniversityShanghaiChina

Sex determination has been studied in the model lepidopteran species Bombyx mori, but it remains poorly understood in lepidopteran pests. In the present study, we identified and characterized the Masculinizer (Masc) gene in a Noctuidae pest species, Agrotis ipsilon. Sequence analysis revealed that AiMasc encodes a protein of 658 amino acids that has two CCCH-type zinc finger domains and two conserved cysteine residues (Cys-277 and Cys-280). We assessed the masculinizing activity AiMasc in BmN cells and found that/z'Masc induced expression of the male-specific doublesex isoform. Disruption of Masc via clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) in A. ipsilon caused abnormalities in abdominal segments and external genitalia, resulting in male-specific sterility. These results suggest that Masc participates in the process of sex determination in A. ipsilon. Successful identification of sex-determination gene in a pest species may enable the development of novel genetic approaches for pest control.

关键词： Agrotis ipsilon CRISPR/Cas9 Masc sex determination

Foraging valor linked with aggression:selection against completely abandoning aggression in the high-elevation ant Tetramorium alpestre?

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insect science 2024年第3期31卷 953-970页

作者： Marie-Luise Contala Patrick Krapf Florian M.Steiner Birgit C.Schlick-Steiner Molecular ecology Group Department of EcologyUniversitat InnsbruckInnsbruckAustria rganismal&evolutionary biology Research Programme University of HelsinkiHelsinkiFinland

Aggression has multiple benefits and is often coupled with other behaviors("behavioral syndromes").The level of aggressiveness is influenced by an adaptive benefit-cost ratio suggesting that benefits should outweigh the costs of ***,we assess if several behaviors are coupled in two behaviorally different populations(aggressive,peaceful)of the high-elevation ant Tetramorium *** three weeks,we collected colony fragments and analyzed boldness,exploring,foraging,and risk-taking *** hypothesized that the aggressive population is bolder,more explorative and risk-prone,and forages more food than the peaceful *** test whether(a)the combination of experiments and parameters used yields a good setup,(b)populations differ behaviorally,and(c)populations display behavioral syndromes,we assessed(a)the frequency of repeatable behaviors of each experiment,(b)the behavioral means among populations,and(c)the behavioral repeatability,*** found that(a)boldness and exploring were most repeatable and represent a good experimental setup,(b)the aggressive population was bolder and more explorative and risk-prone than the peaceful population,(c)boldness and exploring behaviors were highly repeatable in both populations,thus corroborating our *** results suggest that boldness,exploring,and risk-taking but not foraging are presumably coupled with aggression and indicate the presence of behavioral syndromes in this *** specific ecological conditions,aggression may be coupled with other behaviors and important for finding *** is probably adaptive in ***,possibly indicating that selection favors aggression at least partially,which may counteract the complete loss of intraspecific aggression.

关键词： adaptive value behavioral syndromes boldness exploration repeatability trade-off