害虫遗传不育技术中致死基因的研究与应用

基于遗传修饰手段的昆虫不育技术(SIT)作为一类物种特异、环境友好、科学高效的新兴策略,在害虫防治中具有广阔的应用前景。释放携带显性致死基因昆虫的技术(RIDL)是改进传统SIT的重要手段之一,主要包括四环素调控系统、特异性启动子、性别特异剪接系统和特异性致死基因等重要元件,其中根据不同昆虫的特点选择合适的特异性致死基因对于构建遗传不育品系至关重要。这些致死基因或受到阻遏调控系统的控制、或特异的在雌虫中表达、亦或直接作用于X染色体,导致后代在特定发育阶段或特定性别中条件致死。本文综述了RHG家族(reapr、hid、grim、michelob_x)细胞凋亡基因、转录激活因子t TA及Nipp1Dm、归巢内切酶基因等在害虫遗传不育技术中的研究和应用,讨论了特定致死基因的效应机理和应用特点,并对其可能的发展方向进行了展望。由于不同效应基因的致死作用和调控机理尚未完全明晰,因此深入研究特异致死基因的凋亡机制和在不同物种中的兼容作用,将为害虫遗传防控提供更多的研究思路和手段。     

蚊虫微生物群系及其在蚊媒疾病防控中的研究进展

蚊虫体内外栖居着大量的微生物,构成多样且动态的微生物群系(microbiome).蚊虫微生物群系(mosquito microbiome)在蚊虫的营养、代谢、免疫等诸多生理功能上发挥着重要作用,是蚊虫生长发育不可或缺的重要组成部分.同时,蚊虫微生物群系显著影响蚊媒病原体感染和传播,在蚊媒疾病防控中具有重要的应用价值.因此,全面了解蚊虫生理以及蚊媒疾病传播不能忽视微生物群系的关键作用.本文系统概述了蚊虫微生物群系的组成及其影响因素、微生物群系对宿主生理功能的影响与互作机制,以及基于蚊虫微生物群系的蚊媒疾病阻断控制等方面的研究进展,并对利用微生物群系防控蚊媒传染病的应用前景提出展望.     

转基因在苹果蠹蛾不育昆虫释放技术中的应用

使用不育昆虫释放技术是一项新兴的苹果蠹蛾Cydia pomonella(L.)防治方法,使用转基因得到不育雄蛾具有比传统辐射方法更多的优点。转基因技术通过使用基于piggyBac等转座子的质粒载体,并插入显性条件致死基因以培育遗传性别品系,同时插入荧光蛋白等标记基因来显示转基因的效果;在人工控制的发育条件,建立成熟的稳定苹果蠹蛾品系,用以最终的田间释放以达到防治的目的。     

苹果蠹蛾的综合防控和遗传控制研究进展

苹果蠹蛾是仁果类水果的重要检疫害虫,在世界各地造成了巨大的经济损失。目前对其化学防治、化学生态调控、病毒等防治方法研究较多,但仍不能满足防控该害虫的需要,对新型防控技术的需求日益增强。不育昆虫释放技术(SIT)是一种可控制甚至根除靶标害虫的环境友好型防控技术,但传统SIT技术存在一定的局限性,如较难区分性别与筛选雌雄虫、辐射不育昆虫的交配竞争力和适合度降低等问题,这些缺陷随着昆虫遗传修饰技术的发展将得以解决,并将在害虫防控进程中起到积极作用。本文综述了苹果蠹蛾主要防控技术研究现状,介绍了通过遗传修饰技术改善SIT的技术策略,并综合分析了我国开展苹果蠹蛾遗传修饰研究情况和将其应用在苹果蠹蛾防控体系中的可行性及优势。     

遗传控制技术在实蝇类害虫中的研究进展

实蝇类害虫严重危害多种水果和蔬菜,是世界果蔬产业最重要的害虫类群之一,严重影响了发生地的果蔬生产和出口贸易活动。昆虫不育技术(SIT)是一种物种特异和环境友好型防治措施,在多种实蝇类害虫的防治、阻截和根除中起到了不可替代的重要作用。通过分子生物学技术对昆虫的基因组进行遗传修饰,可对SIT进行改进,提高其防控效果并扩大应用的物种范围,近年来相关方面的研究已取得重要进展,成为害虫遗传控制的研究热点。本文阐述了通过受四环素调控的tet-off基因表达系统来实现昆虫"不育"的基本原理和在果蝇及其他几种主要实蝇类害虫中建立的不同类型的遗传控制体系,以及类似体系在其他农业昆虫中的应用情况。简要介绍了在橘小实蝇遗传控制技术体系构建方面的工作进展,并对该技术的在害虫综合治理(IPM)尤其是实蝇类害虫防治中的应用前景进行了讨论和展望。     

苹果蠹蛾不育昆虫释放技术研究进展

不育昆虫释放技术(sterile insect technique,SIT)是一种环境友好、可作为大面积害虫综合治理(AW-IPM)的防治技术,是以压倒性比例释放不育昆虫来减少田间同种害虫繁殖量的害虫治理方法。苹果蠹蛾Cydia pomonella(L.)是世界重要的梨果类害虫,现已入侵世界5洲71国。本文综述了苹果蠹蛾大规模饲养技术、辐射不育技术、释放技术3个关键环节的研究与技术进展,主要包括:苹果蠹蛾人工饲料、实验种群建立、饲养设备与条件、收集和质量评估、长距离运输、辐射源与设备、辐射剂量与敏感性、释放方法、释放标记和释放量等,并介绍了各国采用SIT技术的应用效果。苹果蠹蛾在我国新疆、甘肃、宁夏、内蒙、黑龙江、吉林6个省区发现,对我国苹果产业安全生产构成严重威胁,我国很有必要引进并建立苹果蠹蛾SIT防治技术体系。     

冈比亚按蚊性别决定基因 doublesex 的克隆、序列分析及表达谱

【目的】doublesex 是控制昆虫性别分化的关键基因,决定了昆虫体细胞与生殖细胞的性别。本研究旨在克隆、鉴定重要疟疾媒介冈比亚按蚊 Anopheles gambiae 性别决定基因 doublesex（Angdsx）,分析其在雌雄个体内的剪切体及在不同发育时期的表达模式。【方法】基于冈比亚按蚊转录组数据库,比对到 Angdsx 相关片段,分别以雌雄成蚊cDNA为模板,采用RT-PCR与RACE方法克隆分别获得雌雄个体内 Angdsx 全长基因,利用生物信息软件对所得序列进行结构域预测、氨基酸序列比对和进化树分析。根据 Angdsx 特异性表达引物,利用RT-PCR方法研究其在冈比亚按蚊雌雄个体及不同发育时期的表达谱。【结果】分别从冈比亚按蚊雌雄成虫中克隆获得 Angdsx cDNA全长序列,分别命名为Angdsx^F（GenBank登录号：KM978937）和 Angdsx ^M（GenBank登录号：KM978938）。Angdsx 位于2号常染色体右臂,基因横跨接近80 kb基因组长度。Angdsx^F 长度为4 874 nt,编码长度为265 个氨基酸的雌性特异性蛋白DSX^F;Angdsx ^M 长度为3 183 nt,编码长度为633个氨基酸的雄性特异性蛋白DSX^M。结构域分析发现 Angdsx 包括 doublesex 保守的TRA/TRA-2结合位点、dsx 重复序列、富含精氨酸/丝氨酸双肽区、多聚嘌呤增强子序列和RNA结合蛋白结合序列,以及连续的双核苷酸GT为主的重复序列。与Angdsx^F 相比, Angdsx ^M具有一个雌性特异性的外显子。Angdsx ^M 在0-2 h卵中高表达,随后逐渐减少,在12-24 h卵中降至最低,之后再次升高;Angdsx^F 则在6-8 h卵中开始表达。【结论】本研究获得了冈比亚按蚊性别决定基因 Angdsx 在雌雄个体内的全长序列,Angdsx 具有保守的结构域与表达特征。本研究结果为蚊虫性别分化的分子机制及将其最终应用于显性致死昆虫施放技术进行蚊媒的防制提供了理论基础。     

小麦显性核不育基因的遗传效应 Genetic Effects of a Dominant Male Sterile Gene on F1 in Common Wheat

采用NCII设计研究显性核不育基因在40个杂种F1代的遗传效应。结果表明，显性核不育基因对F1代形态性状影响不大，而对产量、产量性状及品质性状的影响较大。F1形态性状主要表现加性效应，且不育株和可育株趋势一致；可育株单株产量及大部分产量性状表现显性效应，百粒重、每小穗粒数表现加性效应，而不育株这些性状的加、显性效应起作用。F1两种育性植株蛋白质含量的遗传以加性效应为主、蛋白质产量以显性效应为主，籽粒饱满度则是可育株以加性效应为主，不育株以显性效应为主。显性核不育基因在F1代有明显的母性效应，且在这种基因的细胞质基础上核质互作也很重要。     

植物基因转化技术在苹果遗传改良中的应用

近年来,苹果基因转化技术研究取得了较大进展,本文从转化体系,选择标记,报告基因及目的基因的遗传学行为等方面综述了苹果遗传转化研究发展现状,着重论述了其在苹果遗传改良中的应用,并对植物基因转化技术在莱果上的应用前景和应注意的几个问题进行了讨论。     

RAPD技术及其在昆虫学研究中的应用

本世纪７０年代以前,对昆虫进行遗传研究,主要是利用它的各种外部形态和生理缺陷（如翅长、体色、致死突变等）等特征。所需周期长,且易受环境条件的影响。随着生化技术的不断发展,蛋白质电泳技术在许多昆虫遗传变异研究中得到了广泛的应用。但是随着研究的深人,发现蛋白质受外界环境条件的影响也很大,同时,能检测出的变异频率不能满足继续深人研究的需要［１］根据现代遗传学的观点,物种的遗传性状是由基因决定的。外部形态的变异及体内蛋白质分子的变化最本质的原因是由于基因中ＤＮＡ分子碱基序列发生了改变［’］。因此从ＤＮＡ…     

Genetic elimination of field-cage populations of Mediterranean fruit flies

The Mediterranean fruit fly (medfly, Ceratitis capitata Wiedemann) is a pest of over 300 fruits, vegetables and nuts. The sterile insect technique (SIT) is a control measure used to reduce the reproductive potential of populations through the mass release of sterilized male insects that mate with wild females. However, SIT flies can display poor field performance, due to the effects of mass-rearing and of the irradiation process used for sterilization. The development of female-lethal RIDL (release of insects carrying a dominant lethal) strains for medfly can overcome many of the problems of SIT associated with irradiation. Here, we present life-history characterizations for two medfly RIDL strains, OX3864A and OX3647Q. Our results show (i) full functionality of RIDL, (ii) equivalency of RIDL and wild-type strains for life-history characteristics, and (iii) a high level of sexual competitiveness against both wild-type and wild-derived males. We also present the first proof-of-principle experiment on the use of RIDL to eliminate medfly populations. Weekly releases of OX3864A males into stable populations of wild-type medfly caused a successive decline in numbers, leading to eradication. The results show that genetic control can provide an effective alternative to SIT for the control of pest insects.     

Genetic pest management and the background genetics of release strains

Genetic pest management (GPM) methods involve releasing modified versions of a pest species to mate with wild pests in the target area. Proposed for a wide range of applications in public health, agriculture and conservation, most progress has been made with pest insects. Offspring of the released modified insects and wild pests carry the modification—which might be transgenes, artificially introduced Wolbachia or genetic damage from radiation, for example—but they also carry a complete haploid genome from their laboratory-reared parent, as well as one from their wild parent. Unless these F₁ hybrids are completely unable to reproduce, further mating will lead to introgression of DNA sequences from the release strain into the wild population. We discuss issues around strain selection and the potential consequences of such introgression. We conclude that such introgression is probably harmless in almost all circumstances, and could, in theory, provide specific additional benefits to the release programme. We outline population monitoring approaches that could be used, going forward, to determine how background genetics may affect GPM.This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases’.     

Novel control strategies for mosquito-borne diseases

Mosquito-borne diseases are an increasing global health challenge, threatening over 40% of the world''s population. Despite major advances in malaria control since 2000, recent progress has stalled. Additionally, the risk of Aedes-borne arboviruses is rapidly growing, with the unprecedented spread of dengue and chikungunya viruses, outbreaks of yellow fever and the 2015 epidemic of Zika virus in Latin America. To counteract this growing problem, diverse and innovative mosquito control technologies are currently under development. Conceptually, these span an impressive spectrum of approaches, from invasive transgene cassettes with the potential to crash mosquito populations or reduce the vectorial capacity of a population, to low-cost alterations in housing design that restrict mosquito entry. This themed issue will present articles providing insight into the breadth of mosquito control research, while demonstrating the requirement for an interdisciplinary approach. The issue will highlight mosquito control technologies at varying stages of development and includes both opinion pieces and research articles with laboratory and field-based data on control strategy development.This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases''.     

害虫遗传防治的研究历史与现状

作为防治或根除重大害虫最为有效的手段之一,害虫遗传防治在世界范围内被广泛采用并取得了巨大成功。本文综述了不育昆虫技术、雌性致死系统和昆虫显性致死技术等经典害虫遗传防治策略的发展历史、技术特点和应用情况。近年来,许多新的分子生物手段被不断提出并整合到害虫遗传防治策略中,包括归巢核酸内切酶基因、锌指核酸酶、转录激活因子样效应因子核酸酶、CRISPR/Cas9系统、Medea元件、Killer-Rescue系统、Wolbachia-细胞质不亲和性系统等。基于这些新的工具手段,许多国家已经在不同程度上启动了下一代害虫遗传防治项目。而我国在该领域的研究相对薄弱,需要在借鉴国外成功经验的同时,进一步加强害虫遗传防治的基础和应用研究,从而实现本地有害生物的可持续治理和外来入侵生物的有效狙击,确保我国未来的粮食和生态安全。     

Towards the genetic control of insect vectors: An overview

Insects are responsible for the transmission of major infectious diseases. Recent advances in insect genomics and transformation technology provide new strategies for the control of insect borne pathogen transmission and insect pest management. One such strategy is the genetic modification of insects with genes that block pathogen development. Another is to suppress insect populations by releasing either sterile males or males carrying female‐specific dominant lethal genes into the environment. Although significant progress has been made in the laboratory, further research is needed to extend these approaches to the field. These insect control strategies offer several advantages over conventional insecticide‐based strategies. However, the release of genetically modified insects into the environment should proceed with great caution, after ensuring its safety, and acceptance by the target populations.     

Genetics‐based methods for agricultural insect pest management

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昆虫种群遗传控制技术中启动子的研究

利用昆虫遗传转化技术对害虫进行遗传控制是害虫防治研究的新方向,该技术具有物种特异、防效高且对环境友好的特点。启动子是基因表达调控的重要元件,选择合适的启动子是外源基因高效、准确表达的关键,对获得高效、稳定的遗传修饰昆虫品系至关重要。本文简要介绍了昆虫基因启动子的结构特征,重点描述了昆虫种群遗传防治中组成型启动子、性别和组织特异型启动子、特定发育时期启动子和诱导型启动子的研究和应用概况,并对这几类启动子在害虫遗传控制中的应用前景进行了展望。     

Environmentally sustainable pest control options for Drosophila suzukii

The spotted‐wing drosophila or cherry vinegar fly (Drosophila suzukii) is native to Asia but has invaded other continents since 2008 and has spread throughout Europe. The females have a serrated ovipositor allowing them to penetrate the skins of intact ripening fruits to deposit their eggs, and the developing larvae rapidly destroy the fruits close to harvest. Drosophila suzukii has a rapid life cycle and the larvae develop well beneath the fruit surface. This means that the use of pesticides is problematic and often not effective, first due to their restricted use close to harvest to protect consumers, and second because the larvae are deep enough inside the fruit to avoid contact. There are currently no effective and environmentally sustainable pest control methods for this species, resulting in extensive damage to fruit crops. Here, we review the current status of D. suzukii as a fruit crop pest and discuss the feasibility of current pesticide‐free control methods. We also consider the potential of new technologies as a basis for the urgently needed specific and long‐term control of this species.     

Genetic sexing strains for the population suppression of the mosquito vector Aedes aegypti

Aedes aegypti is the primary vector of arthropod-borne viruses including dengue, chikungunya and Zika. Vector population control methods are reviving to impede disease transmission. An efficient sex separation for male-only releases is crucial for area-wide mosquito population suppression strategies. Here, we report on the construction of two genetic sexing strains using red- and white-eye colour mutations as selectable markers. Quality control analysis showed that the Red-eye genetic sexing strains (GSS) is better and more genetically stable than the White-eye GSS. The introduction of an irradiation-induced inversion (Inv35) increases genetic stability and reduces the probability of female contamination of the male release batches. Bi-weekly releases of irradiated males of both the Red-eye GSS and the Red-eye GSS/Inv35 fully suppressed target laboratory cage populations within six and nine weeks, respectively. An image analysis algorithm allowing sex determination based on eye colour identification at the pupal stage was developed. The next step is to automate the Red-eye-based genetic sexing and validate it in pilot trials prior to its integration in large-scale population suppression programmes.This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases’.