Identification of novel juvenile-hormone signaling activators via high-throughput screening with a chemical library

Juvenile hormone (JH) is an insect-specific hormone that regulates molting and metamorphosis. Hence, JH signaling inhibitors (JHSIs) and activators (JHSAs) can be used as effective insect growth regulators (IGRs) for pest management. In our previous study, we established a high-throughput screening (HTS) system for exploration of novel JHSIs and JHSAs using a Bombyx mori cell line (BmN_JF&AR cells) and succeeded in identifying novel JHSIs from a chemical library. Here, we searched for novel JHSAs using this system. The four-step HTS yielded 10 compounds as candidate JHSAs; some of these compounds showed novel basic structures, whereas the others were composed of a 4-phenoxyphenoxymethyl skeleton, the basic structure of several existing JH analogs (pyriproxyfen and fenoxycarb). Topical application of seven compounds to B. mori larvae significantly prolonged the larval period, suggesting that the identified JHSAs may be promising IGRs targeting the JH signaling pathway.     

草鱼肠系膜脂肪组织高质量总RNA提取方法的研究

基于传统的酸性酚—异硫氰酸胍—氯仿一步提取法,比较分析多种优化操作步骤,摸索出一种高质量提取体质量为80~150 g草鱼肠系膜脂肪组织总RNA的改良方法。试验结果显示,相较于肝脏、脾脏、肠道等脏器组织,草鱼肠系膜脂肪组织RNA丰度低,且极易在样品前处理阶段出现顽固性降解问题。探索发现,将取样量增至约30 mg,可提升RNA产量以满足常规试验需求。针对降解难题,改良常规的样品前处理技术流程,采用鲜样液氮速冻,冻样直接放入TRIzol试剂中裂解,并即刻进行长时间机械匀浆,时长约3 min等核心操作步骤,可显著降低脂肪组织样品RNA的降解。Agilent生物分析仪检测结果显示,改良方法提取的草鱼肠系膜脂肪组织RNA完整度高,关键RIN值为8.7~9.0。研究推测,长时间机械匀浆所形成的持续剪切冲击力或许有助于TRIzol试剂中的异硫氰酸胍等成分突破油滴阻碍而有效抑制内源性RNA酶。本方法提取的草鱼脂肪组织总RNA质量可满足高通量转录组测序要求。     

桉树不同成熟度叶片营养元素内循环特征

揭示桉树人工林不同成熟度的叶片器官所需营养元素变化以及土壤肥力对其影响,对于桉树营林和科学施肥具有重要的生产指导意义。选取广西隆安县小林镇和田东县思林镇的桉树2代萌芽林作为研究对象,测定两个人工林嫩叶、成熟叶、黄叶的C、N、P、K、B、Ca、Mg以及土壤C、N、P、K、Ca、Mg元素含量,并进行相关分析。结果表明,植物叶片的N、P、K、Mg营养元素转移规律是黄叶向成熟叶和嫩叶转移,但C、Ca、B营养元素则是随着叶片的生长发育逐渐积累,黄叶中含量最高,说明施肥时应关注Ca、B元素的持续和有效供给。两片人工林都表现出类似的规律,说明土壤肥力的差异不影响桉树人工林叶片营养元素的内循环规律。桉树嫩叶营养元素含量及其生态化学计量比学特征具有与成熟叶和黄叶不同的表征,在营养物质研究中应予以足够重视。     

New ways and new hopes for IGR development

Insect Growth Regulators (IGRs) represent advanced, bio-rational insecticides. This Special Issue reflects progress in IGR development that has been enabled by insight into the molecular principles of biosynthetic or hormone signaling pathways. The unifying principle is aiming at processes and molecular targets that are unique to arthropods and ideally to narrower insect taxa representing pests or disease vectors. While some strategies of obtaining the desired compounds for chemical intervention rely on rational, structure-based design or computational power, others exploit technologies allowing automated, high-throughput screening of large chemical libraries. All avenues leading to selective and environmentally safe pest control are valid as we face the imminent threat of the declining world insect population.     

Assessing the efficacy and impact of management of an invasive species in a protected area: Poa annua on sub‐Antarctic Macquarie Island

Plant eradication is difficult, particularly in remote, protected areas. The Southern Ocean Islands are very isolated and highly protected, but the flora contains many alien plants. Small restricted populations have been eradicated, but eradication of established species has proven difficult. A better understanding of the efficacy of control methods at sub‐Antarctic temperatures and their off‐target impacts may increase eradication success. With interest in controlling non‐native Poa annua in the region, we aimed to determine if physical and chemical methods can control P. annua (the sub‐Antarctic biotype) in sub‐Antarctic conditions and examined their impact on native plants. We quantified the effectiveness of physical control methods on P. annua in situ on sub‐Antarctic Macquarie Island through field‐based experiments and assessed their selectivity on P. annua compared with native grasses. We also quantified the effectiveness of several herbicides on P. annua at sub‐Antarctic temperatures and assessed their selectivity on native grasses. Of the four physical disturbance methods tested, none effectively reduced P. annua cover as one‐off treatments. Of the herbicide treatments, glyphosate and trifloxysulfuron sodium were effective and were less damaging to native grass species, indicating potential selectivity. Physical control was of limited effectiveness, but did not affect native species richness. An integrated weed management programme utilising the strategic use of selective herbicides with follow‐up chemical and physical intervention may balance control and biodiversity outcomes. This research highlights the importance of site‐specific testing of control methods and understanding off‐target impacts of control when managing alien plant species in protected areas.     

鱼类组蛋白核苷酸的多态性及其在杀鱼爱德华氏菌感染中的作用

组蛋白是染色体核小体的重要组分,在调控染色质结构、基因转录、个体发育等不同生物学过程中起着重要作用。为了研究鱼类组蛋白基因是否存在核苷酸的多态性以及组蛋白核苷酸多态性是否会影响鱼类的抗病力,本实验以斑马鱼和草鱼为研究对象,通过PCR扩增克隆了组蛋白H2A的全长开放阅读框;利用过表达技术、菌落平板计数、感染存活分析以及荧光定量PCR技术,研究了斑马鱼和草鱼组蛋白H2A核苷酸多态性不同变异体在杀鱼爱德华氏菌感染中的作用。在本研究中,实验发现鱼类组蛋白H2A存在着丰富的核苷酸多态性。序列分析结果显示斑马鱼和草鱼组蛋白H2A核苷酸多态性的变异体核苷酸序列相似性为90%～100%;而两两H2A核苷酸多态性变异体氨基酸序列之间最多只有3个位点存在差异。通过体内和体外抗菌实验可知,斑马鱼和草鱼组蛋白H2A核苷酸的多态性显著影响H2A的抗菌活性。此外,筛选出的抗菌组蛋白H2A核苷酸多态性的变异体在斑马鱼体内的过表达,不仅具有免疫增强的作用,还能显著增强斑马鱼对杀鱼爱德华氏菌感染的抗病力。本研究为筛选具有抗病作用的组蛋白H2A免疫保护原奠定了重要基础。     

牛磺酸对急性氨中毒的鲫和草鱼抗氧化及炎症相关基因表达影响的比较

为了比较牛磺酸对急性氨中毒的鲫和草鱼缓释作用的差异,实验分别构建了4个处理组,组1实验鱼通过腹腔注射生理盐水,组2注射醋酸铵(鲫7 mmol/g,草鱼9 mmol/g),组3注射醋酸铵和牛磺酸(100μg/g),组4注射牛磺酸。毒性实验持续96 h。结果显示,组2鲫肝脏中SOD、CuZnSOD和CAT基因mRNA表达量显著低于组1和组3;组2和组3鲫肝脏中GPx基因mRNA表达量显著低于组1;组1鲫大脑中SOD、CuZnSOD、CAT和GPx基因mRNA表达量最高;组2草鱼肝脏中SOD、CuZnSOD、CAT和GPx基因mRNA表达量显著高于其他组;组2和组4草鱼大脑中SOD和CuZnSOD基因mRNA表达量显著低于组1和组3;组3草鱼大脑中CAT和GPx基因mRNA表达量最高;此外,组2鲫和草鱼肝脏及大脑中TNF和IL基因mRNA表达量均显著高于其他组。研究表明,鱼类氨中毒会扰乱机体的抗氧化酶系统和免疫应答,引起氧化损伤和炎症反应;草鱼通过提高抗氧化相关基因表达以应对氨中毒;牛磺酸能够有效缓解氨中毒对鲫和草鱼造成的氧化伤害,但牛磺酸并不能降低氨中毒对鲫和草鱼造成的炎症反应。     

5种药物对异育银鲫"中科3号"受精卵孵化的影响

在水温21~23℃,pH 6.2~6.5条件下,采用半静水试验法,按等对数间距设置药物质量浓度梯度,日换水50%,补充药物50%,24 h观察记录1次,比较美婷Ⅱ、霉灵、纳他霉素、杀毒矾和高锰酸钾5种药物对异育银鲫"中科3号"受精卵水霉抑制作用及孵化率的影响。试验结果显示,试验药物对受精卵水霉生长均有一定抑制作用,其抑霉率为美婷Ⅱ>霉灵>高锰酸钾、纳他霉素和杀毒矾,但杀毒矾对异育银鲫"中科3号"受精卵的毒副作用较大,显著降低了受精卵孵化率,不宜使用;美婷Ⅱ、霉灵、纳他霉素和高锰酸钾对异育银鲫"中科3号"受精卵的适宜使用质量浓度分别为4.90~6.50、1.97~3.87、1.50、2.50 mg/L,使用方法为长时间浸浴,可有效抑制受精卵孵化过程中水霉的生长,显著提高受精卵孵化率,适合在异育银鲫"中科3号"受精卵孵化过程中使用。     

草鱼3个6-磷酸葡萄糖酶催化亚基的基因表达分析及高糖饲料对其表达的影响

为了探讨6-磷酸葡萄糖酶催化亚基(glucose-6-phosphatase catalytic subunit,G6PC)在草鱼(Ctenopharyngodon idellus)糖代谢中的作用,采用同源序列比对的方式,在草鱼基因组中获取了3个g6pc基因的序列,通过序列比对和进化树分析,将其分别命名为g6pca、g6pcb1和g6pcb2,其编码的氨基酸序列与斑马鱼(Danio rerio)、虹鳟(Oncorhynchus mykiss)和人(Homo sapiens)具有较高的同源性,相似度分别为85%～94%、64%～83%和54%～66%。同线性分析表明g6pc在草鱼染色体上的分布与斑马鱼等高度相似,表明草鱼g6pc基因在进化中具有较高的保守性。利用RT-PCR检测3个基因在鳃、脂肪、脑、心脏、肝、肾、前肠、中肠、后肠和肌肉10个组织中的表达,结果显示, g6pca在脑和肝中表达量较高,脂肪组织次之; g6pcb1在肝中表达量最高,中肠次之; g6pcb2在心脏表达量最高,其次为脂肪组织。同时探讨了高糖饲料对草鱼不同g6pc亚型转录水平的影响,以及不同浓度葡萄糖和胰岛素刺激草鱼肝细胞(L8824)后对g6pca转录水平的影响。结果显示,饲喂高糖饲料(7周)后,与对照组相比,草鱼肝脏g6pca mRNA水平显著升高, g6pcb1和g6pcb2 mRNA水平无显著变化。离体情况,与5 mmol/L葡萄糖组相比,15 mmol/L葡萄糖显著增加了L8824的g6pca mRNA水平,且1 mol/L胰岛素可以抑制这种作用; 30 mmol/L葡萄糖对L8824 g6pca mRNA水平无显著性影响。本研究表明,草鱼g6pc发生加倍后存在功能分化,高糖可以诱导g6pca mRNA的表达,而对g6pcb1和g6pcb2的转录水平无影响,其具体功能还需进一步研究。     

盐度对珍珠龙胆石斑鱼幼鱼渗透调节与耗氧率的影响

为研究盐度对珍珠龙胆石斑鱼（）渗透调节与耗氧率的影响，设计实验一，将（162.5±12.1）g珍珠龙胆石斑鱼置于不同盐度（6、12、18、24、30）下养殖10 d，测定血清渗透压及Na⁺、Cl^-、K⁺离子浓度。实验结果表明，随盐度的升高血清渗透压及Na⁺、Cl^-、K⁺离子浓度也随着升高，各组[Na⁺]：[Cl^-]比值无显著差异（>0.05）；经回归分析得到血清等渗点渗透压为365.95 mOsm/kg，所对应盐度为12.75。实验二，将（26.4±2.7）g幼鱼置于不同盐度（6、12、18、24、30）下养殖30 d，测定在开始暴露后0 h、3 h、24 h、72 h鳃Na⁺/K⁺-ATPase活性及表达和第30天耗氧率，结果表明鳃Na⁺/K⁺-ATPase活性随盐度的增大呈“U”形变化；鳃Na⁺/K⁺-ATPase α1基因表达量波动较大，在72 h后随盐度增大先降低后增加，变化趋势与酶活性一致；第30天耗氧率随盐度的增加先降低后增加又降低。综上所述，珍珠龙胆石斑鱼幼鱼10 d内能够完全适应6~30盐度急性变化，耗氧率除了受离子渗透调节的影响，还可能与其生活史阶段有关。