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.     

Screening and identification of soybean varieties with high phosphorus efficiency at seedling stage

In order to simplify the workload and find a reliable method for screening soybeans with different phosphorus(P)efficiencies,47 soybean varieties were screened from 90 varieties according to yield under normal phosphorus,and 10 indicators including root phenotype,phosphorus utilization efficiency at the seedling stage and yield were measured.Through single-index cluster analysis,the performance value and relative value of the abovementioned indexes under low-phosphorus conditions were analyzed,and then,the combined indexes were analyzed by principal components method.The membership function method and the cluster analysis method was used to calculate and analyze the comprehensive score value.Three indexes of root P utilization efficiency(X₁),relative value of root P utilization efficiency(X₂),and root volume(X₃)under low P at the seedling stage were selected as the most consistent with the yield screening results under low P condition,and the mathematical model of the comprehensive score value was obtained:D=1.218X₁t 0.320X₂t 0.007X₃-0.664(P=0.000,R²=1.000).The comprehensive score can screen and identify the P efficiency of soybeans with fewer indexes in the early growth stages,which provides a more rapid and reliable mathematical model for screening and identifying a larger number of germplasm resources for P efficiency.     

不同生育阶段燕麦对干旱胁迫的响应

设中度干旱(MD,土壤田间最大持水量的55±5%)、重度干旱(SD,土壤田间最大持水量的35±5%)两个处理,研究干旱胁迫对温室盆栽燕麦不同生育阶段(苗期、分蘖期、拔节期)的生长性状、叶绿素荧光参数以及根系形态的影响。结果表明,与正常供水(CK)相比,干旱胁迫处理均显著(P<0.05)降低了燕麦苗期、分蘖期、拔节期3个生育阶段地上部生物量、株高以及叶片光合荧光参数Fv/Fm、Fv/Fo。干旱胁迫下,燕麦根系表现出明显的形态可塑性,苗期通过增加根长、分支数和细根数(直径≤0.48mm)来响应干旱胁迫,分蘖期主要通过减少根长、分支数来响应干旱胁迫,拔节期主要通过减缓各级根系的生长并增加根的分级数来响应干旱胁迫。     

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.     

罗氏沼虾细胞色素P450家族CYP302a1基因克隆及其在蜕皮周期中的表达

蜕皮是甲壳动物重要的生理活动,与其蜕皮激素的合成密切相关,细胞色素P450(CYP)302a1是甲壳动物蜕皮激素合成通路中的关键酶之一。本研究克隆了罗氏沼虾CYP302a1基因(Mr-CYP302a1),cDNA全长1859 bp,开放阅读框(ORF)为1629 bp,编码543个氨基酸(aa),分子量大小为61.09 ku,等电点为8.42。氨基酸序列分析显示CYP302a1基因的保守结构域含有5个P450基因家族特征保守区域:heme-binding、helix-K、helixC、helix-I及PERF。系统进化分析结果显示Mr-CYP302a1首先与绿虾CYP302a1聚为一支,然后与凡纳滨对虾及三疣梭子蟹等十足目甲壳动物的CYP302a1聚为一支,与甲壳动物的亲缘关系最近。实时荧光定量PCR(qRT-PCR)检测表明Mr-CYP302a1在罗氏沼虾的多个组织中均有表达,其中在Y器官中的表达量最高,性腺中次之。同时研究发现,MrCYP302a1基因在罗氏沼虾的蜕皮后期(A期和B期)表达量很低,蜕皮间期(C期)表达量开始上升,在蜕皮前期D1亚期达到峰值。对Mr-CYP302a1进行蛋白表达及多克隆抗体制备,蛋白印迹法(Western blot,WB)检测表明Mr-CYP302a1蛋白在罗氏沼虾Y器官中的表达量最高,在蜕皮过程中的蜕皮前期D1亚期达到峰值。综上所述,该基因在罗氏沼虾的蜕皮过程中扮演着十分重要的角色。     

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。     

Met基因在黏虫生殖中的功能分析

保幼激素(juvenile hormone, JH)是由咽侧体分泌的倍半萜类化合物,可以调控昆虫的很多生理过程,如发育、变态、生殖等。作为bHLH-PAS(helix-loop-helix-Per-ARNT-Sim)转录因子家族成员之一的methoprene-tolerant (Met)是JH的受体,在JH的信号传导过程中有非常重要的作用。本研究通过实时荧光定量PCR结合RNAi技术测定了沉默Met基因后黏虫Mythimna separata的Vg基因表达、卵巢发育和生殖行为,旨在探究Met基因在黏虫生殖过程中的功能。结果表明当Met基因沉默后,与卵巢发育密切相关的卵黄原蛋白(vitellogenin, Vg)基因表达量下调了50%,从而显著抑制了卵巢发育,并导致产卵显著延迟、产卵历期显著缩短,产卵量显著下降。结果表明Met基因是黏虫生殖发育过程中的关键受体基因,它通过调节后续Vg的表达和沉积,来达到控制卵巢发育,从而调控生殖作用。     

甘蓝型油菜种子萌发期耐铝毒特性综合评价及其种质筛选

种子萌发是油菜植株形态建成的重要阶段,铝毒是酸性土壤中影响其种子萌发的主要因素之一,因此筛选出种子萌发过程中耐铝毒种质对油菜生产及研究具有重要意义。本研究利用5份甘蓝型油菜品种(系)筛选出油菜萌发期耐铝毒种质资源处理的适宜胁迫浓度为90μg mL~(-1)。并以该浓度处理148份甘蓝型油菜品种(系)种子,于萌发期测定其发芽势、发芽率、鲜重、干重、根长和芽长等指标,通过铝毒胁迫耐性综合评价值(A值)、平均隶属函数值(ASF值)、对铝毒加权耐性系数(WAC值)、相关性、频数、主成分、灰色关联度、聚类和逐步回归分析等鉴定萌发期耐铝性,建立萌发期对铝毒耐性综合评价模型并筛选出适宜的评价鉴定指标。结果表明, 148份甘蓝型油菜的萌发期各指标在品种(系)间存在显著差异;筛选出萌发期耐铝毒甘蓝型油菜品种(系) 01188、WH-20、A109、甲预31棚等。根据灰色关联度及回归分析结果认为,在油菜萌发期测定其根长、芽长、鲜重、发芽率和发芽势,通过回归方程估算其A值,可以初步判断甘蓝型油菜种质的耐铝毒特性。     

利用SOM聚类分析及主成分分析方法鉴定小麦品种萌发期耐盐性

为了研究不同小麦品种萌发期的耐盐性,为小麦萌发期耐盐性鉴定提供快速、准确的鉴定方法和理论依据。本研究以40个冬小麦品种为试验材料,通过主成分分析法(PCA)与神经网络自组织映射(SOM)聚类分析法,研究盐胁迫对40个小麦品种萌发期各项形态指标的影响。主成分分析法结果表明,发芽势、发芽率、最长根长、第一片叶长及胚芽鞘长在PCA下载荷量最大;根据SOM聚类分析结果,‘德抗961’等4个小麦品种属于高度耐盐品种,‘济麦22’等14个小麦品种属于耐盐性品种,‘济麦20’等16个品种属于中等耐盐性品种,‘鲁麦21’等6个小麦品种属于盐敏感性品种。发芽势、发芽率、最长根长、第一片叶长、胚芽鞘长能够作为萌发期小麦耐盐性筛选的主要评价指标,并且结合主成分、SOM聚类分析方法进行小麦萌发期耐盐性鉴定更为科学、方便。