Effects of Melia azedarach, (Meliaceae) fruit extracts on the leafminer Liriomyza huidobrensis, (Diptera, Agromyzidae): Assessment in laboratory and field experiments

Control of the widely distributed pest Liriomyza huidobrensis, is complicated due to the protected habit of the leafmining larvae, and their resistance to insecticides. The effects of Melia azedarach, (Meliaceae) fruit extracts against adults and larvae of L. huidobrensis, were investigated. In the laboratory, leaves of Cucurbita, sp. infested with first and third instar larvae were treated with different extract solutions. Larval and pupal survival, as well as wing‐spread of adults, were ssessed. Female adult behaviour towards the extract was also analysed in terms of number of feeding punctures and number of offspring left on treated/untreated leaves. In the field, an infested Vicia faba, crop was sprayed four times at weekly intervals with plant extract, water, and a blank solution. The number of adult leafminers and parasitoids emerging from sampled leaves from each treatment were compared. The laboratory tests showed translaminar action of the extracts, which negatively affected leafminer pupal survival, while body size was not affected. The extracts also deterred feeding by adult females and may also have caused reduction in oviposition rates. All solutions and concentrations tested had similar effects. In the field, extract effects were consistent with those from laboratory trials, number of pupae and pupal survival being lower on treated plants. Percentage parasitism was not affected by plant extract treatment, suggesting a selective activity.     

入侵害虫南美斑潜蝇在西藏首次发现及其寄生蜂调查

【目的】南美斑潜蝇Liriomyza huidobrensis为我国重要的入侵害虫,我们于2017年7月首次发现其已扩散至西藏。本研究旨在了解西藏地区南美斑潜蝇的发生现状及其寄生蜂种类。【方法】2017和2018年6月上旬-9月中旬对西藏拉萨、林芝、山南和日喀则4个地区的南美斑潜蝇的发生危害及其寄生蜂组成进行了调查采样。【结果】调查的西藏这4个地区均发现南美斑潜蝇分布和危害;温室发生时间为6月上旬-9月中旬,而露地的发生高峰在7月下旬-8月上旬。其寄主植物有9科21种,主要有豆科(Fabaceae)、茄科(Solanaceae)和伞形科(Umbelliferae)等,主要嗜好经济作物有豇豆Vigna unguiculata, 扁豆Lablab purpureus , 莴笋Lactuca sativa var. angustana和白菜Brassica rapa ssp. pekinensis。其寄生蜂包括姬小蜂科(Eulophidae)、茧蜂科(Braconidae)和瘿蜂科(Cynipidae)共计3科9属16种,分别为双斑潜蝇姬小蜂Diglyphus bimaculatus, 豌豆潜蝇姬小蜂Diglyphus isaea, 小斑潜蝇姬小蜂Diglyphus minoeus, 万氏潜蝇姬小蜂Diglyphus wani, Diglyphus sp., 纵纹瑟姬小蜂Cirrospilus vittatus, Hemiptarsenus sp., 芙新姬小蜂Neochrysocharis formosa, 底比斯姬小蜂Chrysocharis pentheus, 普金姬小蜂Chrysocharis pubicornis, Chrysocharis sp., 潜蝇柄腹姬小蜂Pediobius metallicus, Pnigalio sp., 西伯利亚离颚茧蜂Dacnusa sibirica, Dacnusa sp.和Gronotoma sp.;其中优势种为西伯利亚离颚茧蜂、双斑潜蝇姬小蜂和豌豆潜蝇姬小蜂,其个体数量占比分别为总采集个体数的48.7%, 22.1%和12.9%。【结论】西藏南美斑潜蝇的寄生蜂种类丰富,发挥着重要的自然控制作用。建议进一步加强优势寄生蜂生物学、控害潜力评价以及利用研究,为南美斑潜蝇的绿色防控提供指导。     

南美斑潜蝇寄主选择性与植物次生化合物及叶毛的关系

采用非自由选择法,以刺伤孔（包括取食孔和产卵孔）数为指标,在室内测定了南美斑潜蝇对6科、16种、22个蔬菜品种的寄主选择性。结果表明,南美斑潜蝇对供试蔬菜品种的选择性存在显著差异,选择性大小依次为豆科>菊科>葫芦科>茄科>伞形科>十字花科; 菜豆和茼蒿是南美斑潜蝇最喜欢产卵和取食的寄主,而甘蓝和番茄是其选择性最差的寄主。化学分析和解剖镜观察表明,叶片单宁酸、黄酮含量及叶毛密度在供试品种间存在显著差异： 苦瓜的单宁酸含量最高,其次为豌豆、番茄和青椒,含量最低的为金丝搅瓜、茼蒿、菜豆和美国西芹; 黄酮含量最高的为甘蓝和番茄,其次为苦瓜,最低为茼蒿、菜豆和美国西芹; 南瓜和小白菜的叶毛密度最高,其次为西葫芦、金丝搅瓜和甘蓝,最低为青椒、莴苣和苦瓜。相关分析表明,南美斑潜蝇寄主选择性与叶片单宁酸和黄酮含量呈显著的负相关关系,相关系数分别为-0.4425（ P=0.0392）和 -0.5728（ P=0.0053）,而与叶毛密度相关关系不显著（ R＝-0.1807,P=0.4211）。说明黄酮和单宁酸对南美斑潜蝇产卵和取食具有明显的抑制作用。     

南美斑潜蝇为害对黄瓜体内主要营养物质、 次生代谢物质及叶绿素含量的影响

南美斑潜蝇Liriomyza huidobrensis (Blanchard)是一种危害多种蔬菜和观赏植物的多食性害虫。本研究采用蒽酮比色法、 考马斯亮蓝法、 磷钼酸 磷钨酸比色法、 索氏回流法及丙酮法, 分别测定了南美斑潜蝇1~3龄幼虫不同为害程度（系统受害、 轻度受害和重度受害）对黄瓜叶片内可溶性糖、 可溶性蛋白质、 单宁、 黄酮及叶绿素含量的影响。结果表明： 南美斑潜蝇幼虫为害后, 黄瓜叶片中可溶性糖、 可溶性蛋白质和叶绿素的含量随着为害程度的增强而显著降低（P<0.01）, 最大降幅分别为62%, 35%和40%; 单宁和黄酮的含量随着为害程度的增强而显著上升（P<0.01）, 最高升幅分别为26%和53%; 并且具有系统性影响。结果提示, 南美斑潜蝇幼虫为害导致寄主植物营养物质含量下降、 光合作用降低, 而次生代谢物质含量上升, 从而对植食者产生诱导抗性。本研究结果为进一步揭示南美斑潜蝇与其寄主植物相互作用关系及其机理提供了有益的参考。     

Variation of density-dependence with spatial scale in the leaf-mining fly Liriomyza commelinae (Diptera: Agromyzidae)

Abstract. 1. Mortality imposed on the leaf mining fly, Liriomyza commelinae (Frost) was studied at three spatial levels: (1) the leaf, (2) the stem, and (3) a metre quadrat of the food plant.
2. Real mortality (R.M.) (death as a percentage of eggs laid) during development for five sites over 3 years (forty-seven generations) was 96.3 + 1.9% -2.5% (±95% c.l.), due mainly to parasitoids and predatory ants, with a small component due to competition and other causes.
3. Real mortality due to the parasitoid complex averaged 41.2%. It was not related to population density in leaves or stems but was positively dependent on density in one metre quadrats.
4. Predation by Crematogaster brevispinosa Mayr was 21.2% and was independent of population density at all spatial levels.
5. Mortality due to other causes (e.g. competition and exposure to high temperature resulting from insolation) was positively dependent on the number of mines per leaf at three out of five sites studied and the number of mines per stem at two of the three sites studied; however, such mortality was independent of density per metre quadrat.     

Molecular evidence of cryptic species within the Liriomyza huidobrensis (Diptera: Agromyzidae)

Phylogenetic relationships among populations of the polyphagous pea leafminer, Liriomyza huidobrensis (Blanchard), were investigated using DNA sequence data. Maximum parsimony analysis of 941 bp of mitochondrial cytochrome oxidase I and II genes showed that L. huidobrensis contains two well-defined monophyletic groups, one composed of specimens from California and Hawaii and one composed of specimens from South and Central America together with populations that have been recently introduced into other parts of the world. The differentiation between the two clades within L. huidobrensis is equivalent to that seen between other agromyzid species, suggesting that L. huidobrensis as currently defined contains two cryptic species. This finding is consistent with field observations of differences in pest status and insecticide resistance between L. huidobrensis populations. Until additional studies are complete, no changes in L. huidobrensis taxonomy are proposed. However, researchers and quarantine officials may wish to consider the findings of the current study in designing research, pest management, and quarantine programs for L. huidobrensis.     

福建省发现加藤斑潜蝇（双翅目：潜蝇科Diptera：Agromyzidae）

1995年在调查潜叶蝇种类时，在菊花上发现加藤斑潜蝇，属中国大陆首次报道，本文对其形态特征进行描述。     

南美斑潜蝇幼虫为害对黄瓜叶片中水杨酸和茉莉酸的诱导作用

【目的】为揭示南美斑潜蝇Liriomyza huidobrensis (Blanchard)与其寄主相互作用的机理, 为利用诱导抗性控制南美斑潜蝇的发生为害奠定必要的基础。【方法】本文采用高效液相色谱法（HPLC）和超高效液相色谱法-质谱联用法（UPLC MS）, 分别测定了南美斑潜蝇幼虫为害对黄瓜叶片中茉莉酸（jasmonic acid, JA）和水杨酸（salicylic acid, SA）的诱导作用。【结果】南美斑潜蝇幼虫持续为害1 d后, 受害黄瓜叶片内JA含量即显著高于健康对照, 轻度受害处理和重度受害处理分别在第3天和第5天上升幅度最大, 分别比健康对照增加2.01倍和1.62倍; 而SA含量在3 d后才显著高于健康对照, 轻度受害处理和重度受害处理在第9天上升幅度最大, 分别比健康对照增加4.66倍和1.67倍; 轻度受害对JA和SA的系统诱导作用不明显, 而重度受害对JA和SA具有明显的系统诱导作用。【结论】南美斑潜蝇幼虫为害对黄瓜叶片内JA和SA具有诱导作用。     

南美斑潜蝇寄主选择性与植物营养物质及叶绿素含量的关系

采用非自由选择法,在室内研究了南美斑潜蝇寄主选择性及其与植物叶片营养物质和叶绿素含量的关系.结果表明,南美斑潜蝇成虫对不同供试植物的选择性存在显著差异,选择性最强的为菜豆、茼蒿和西葫芦,最差的为番茄和甘蓝.相关分析表明,南美斑潜蝇成虫寄主选择性与叶片可溶性糖含量存在显著的正相关关系(R＝0.5014,P=0.0174,F＝6.717),而与叶片可溶性蛋白和叶绿素含量相关关系不显著.     

不同地理种群美洲斑潜蝇及近缘种的rDNA-ITS1序列分析和比较

【目的】通过对美洲斑潜蝇Liriomyza sativae Blanchard 不同地理种群及近缘种间的核糖体DNA第一内转录间隔区（rDNA-ITS1）进行比较,分析美洲斑潜蝇不同地理种群间的遗传分化情况,并为美洲斑潜蝇与近缘种间提供分子鉴别标记。【方法】用PCR产物直接测序法及克隆测序法对我国美洲斑潜蝇8个地理种群的rDNA-ITS1序列进行测序,并调用GenBank中3个近缘种的rDNA-ITS序列,运用软件MEGA3.1对美洲斑潜蝇不同地理种群及近缘种间的rDNA-ITS1序列进行分析。【结果】美洲斑潜蝇8个地理种群间的分化程度较低,只有8个变异位点,遗传距离都在0.02以下,但4个近缘种间的碱基差异显著,遗传距离为0.149～0.390,有126个变异位点,12个美洲斑潜蝇特异性识别位点。【结论】虽然基于rDNA-ITS1序列所显示的美洲斑潜蝇各地理种群之间的遗传分化很小,但是其分化趋势与地理分布基本相吻合;得到的12个特异性识别位点不仅可以作为美洲斑潜蝇与其近缘种间鉴别的分子标记,而且可为今后设计鉴别性PCR引物提供重要的参考依据。     

温度对美洲斑潜蝇及南美斑潜蝇种群增长的影响

在恒温条件下观察了美洲斑潜蝇和南美斑潜蝇实验种群的发育、存活及繁殖情况,拟合了两种斑潜蝇发育速率与温度之间的关系模型,求出了各发育阶段的发育起点温度及有效积温,组建了不同温度下两种斑潜蝇实验种群的生殖力表,计算了主要的生命表参数.结果表明,温度对两种斑潜蝇的发育、存活、繁殖均有显著的影响,且两种斑潜蝇对温度的适应特性有着明显差异.美洲斑潜蝇适应的温度范围较广,相对较高的温度有利于种群的发育、生存及繁殖;而南美斑潜蝇适应的温度范围相对较窄,且适温范围明显偏低,高温不利于种群的增长;在各自的适温条件下,两种斑潜蝇都有很强的种群增殖能力.     

A genome-wide phylogeny and the diversification of genus Liriomyza (Diptera: Agromyzidae) inferred from anchored phylogenomics

The genus Liriomyza Mik (Diptera: Agromyzidae) is a diverse and globally distributed group of acalyptrate flies. Phylogenetic relationships among Liriomyza species have remained incompletely investigated and have never been fully addressed using molecular data. Here, we reconstruct the phylogeny of the genus Liriomyza using various phylogenetic methods (maximum likelihood, Bayesian inference, and gene tree coalescence) on target-capture-based phylogenomic datasets (nucleotides and amino acids) obtained from anchored hybrid enrichment (AHE). We have recovered tree topologies that are nearly congruent across all data types and methods, and individual clade support is strong across all phylogenetic analyses. Moreover, defined morphological species groups and clades are well-supported in our best estimates of the molecular phylogeny. Liriomyza violivora (Spencer) is a sister group to all remaining sampled Liriomyza species, and the well-known polyphagous vegetable pests [L. huidobrensis (Blanchard), L. langei Frick, L. bryoniae. (Kaltenbach), L. trifolii (Burgess), L. sativae Blanchard, and L. brassicae (Riley)]. belong to multiple clades that are not particularly closely related on the trees. Often, closely related Liriomyza species feed on distantly related host plants. We reject the hypothesis that cophylogenetic processes between Liriomyza species and their host plants drive diversification in this genus. Instead, Liriomyza exhibits a widespread pattern of major host shifts across plant taxa. Our new phylogenetic estimate for Liriomyza species provides considerable new information on the evolution of host-use patterns in this genus. In addition, it provides a framework for further study of the morphology, ecology, and diversification of these important flies.     

The feasibility of using the sterile insect technique against Liriomyza trifolii (Diptera: Agromyzidae) infesting greenhouse chrysanthemum

Liriomyza trifolii is a serious pest of chrysanthemum greenhouses and other crops around the world. The larvae feed within the leaves of the host plants and create serpentine mines. The Sterile Insect Technique (SIT) has been successfully used against some Dipteran species in the field, and it is especially efficient against isolated populations like those on islands. Therefore, SIT against pest populations in confined environments such as greenhouses should have high potential for success. The objectives of this study were to determine the optimum gamma‐irradiation dosage required for the sterilisation of L. trifolii adults, and to determine the quality (emergence percentage, flight ability, longevity, copulatory success, and sperm transfer) of the irradiated males compared with that of non‐irradiated (normal) males. We found that sterility (< 0.7 mine per female) was achieved with the dose of 170 ± 5% Gy. The copulatory success and sperm transfer during copulation of the sterile males were not significantly different from those of normal males. Moreover, the longevity, emergence percentage and flight ability of irradiated males were also not significantly different from normal males. The SIT experiments suggest that the release of sterile L. trifolii can significantly reduce the reproductive capacity of a wild leafminer population. Our studies indicate that sterilisation of L. trifolii flies is feasible and that sterile males are of high quality and competitive with normal males. Based on these data, research on the use of SIT against L. trifolii populations in greenhouses is ongoing.     

Polyandry and Reproduction in the Serpentine Leaf Miner Liriomyza trifolii (Diptera: Agromyzidae)

We investigated the hypotheses that Liriomyza trifolii is a polyandrous species and that multiple mating is required to maximize female fecundity. In addition, the hypothesis that male copulating frequency reduces female fecundity was examined. Using sterile and fertile males, we found that L. trifolii females mate more than once during their life span, and that multiple mating is required to maximize female fecundity. A similar amount of sperm was transferred during five successive male copulations with an equal insemination probability. However, the probability of reproduction failure was strongly correlated with the male copulating sequence. Moreover, the failure rate of sperm transfer was found to be 6.8%. In light of these results, possible selection pressures driving for polyandry in L. trifolii are discussed.     

醋酸纤维素薄膜电泳实验成绩量化评定模式的设计

通过对实验教学量化评定的内容、类型、考核形式及各部分内容占总成绩的权重,各部分内容中的详细考核项目,各详细考察项目的项目系数等多方面的综合权衡,建立了醋酸纤维素薄膜电泳实验成绩量化评定模式,其使用简便、合理、易行,能科学正确的量化评定学生实验成绩,有效地改进实验教学效果。     

福建省潜蝇科昆虫属种检索表

本文根据作者于1997－2000年对福建省九个地、市的蔬菜和花卉等作物上潜蝇科昆虫的调查结果,编制了现行福建省潜蝇科昆虫属、种检索表,以供鉴定时参考。     

Avermectin类农药对美洲斑潜蝇的生物活性

Avermectin类农药对美洲斑潜蝇Liriomyzasattvae Blanchard生物活性的室内研究结果表明： 揭阳霉素对美洲斑潜蝇1、2、3龄幼虫的LC₅₀分别是1.54×10^-4 g/L、3.73×10^-4 g/L、1.99×10^-3g/L;害极灭对上述幼虫的LC₅₀分别是1.48×10^-4g/L、3.68x10^-4 g/L和1.97×10^-3 g/L。美洲斑潜蝇幼虫对Avermectln类农药以1龄最敏感,其中揭阳霉素对3龄幼虫的LC₅₀是1龄的12.9倍。揭阳霉素对雌成虫24 h、48 h的LC₅₀分别是3.12×10^-3 g/L和2.08×10^-3 g/L,其对美 洲斑潜蝇取食、产卵拒避持效期分别是4-8天和10天。使用浓度0.005 g/L揭阳霉素处理6天、8天和10天后接虫,幼虫的存活率分别是0、16.13%和28.07%。田间使用浓度0.005 g/L的揭 阳霉素和0.0045 g/L几的害极灭分别处理,6天后的校正虫口减退率分别为91.0%和90.9%,两者差异不显著,而使用浓度0.0067 g/L揭阳霉素处理,6天后校正虫口减退率为93.6%。     

美洲斑潜蝇幼虫空间分布型和田间抽样技术

通过田间调查和计算,明确了美洲斑潜蝇幼虫呈聚集分布,且以负二项分布为主,理论抽样数当t＝1．00,D＝0．2时,n＝26．08／X＋16．12,如果防治指标定为10头／株,则最大抽样数为19株,序贯抽样的累积幼虫数量界限为．田间随机取样以平行线和Z字形为最佳。     

Mass production of Diglyphus isaea (Hymenoptera: Eulophidae), a biological control agent of a Korean population of potato leaf miner Liriomyza huidobrensis (Blanchard) (Diptera: Agromyzidae)

The potato leaf miner, Liriomyza huidobrensis (Blanchard), is an important pest of potato throughout the world, including Korea. A method was developed for mass rearing the parasitoid Diglyphus isaea (Walker) using faba bean, Vicia faba L. (Fabaceae), as the host‐plant and L. huidobrensis as the insect host. Faba bean plants were planted in pots and maintained in a greenhouse for about 15 d. Pots were then exposed to adult leaf miners in oviposition cages for 4 h. Plants containing late second to early third instar larvae were exposed to adult D. isaea in parasitoid cages for 24 h. The leaf area per pot after 15 d was 597.9 cm², which produced 103.33 larvae per pot. The number of adult parasitoids emerging per pot was 72.5; about 41% of these were female. The daily cost of parasitoid production was USD20.95 per 1000 individual parasitoids. The methodology developed for D. isaea could be used to rear other ectoparasitoids such as Hemiptarsenus spp. and Pnigalio sp. with different insect hosts like L. trifolii. This is why this mass‐rearing information is important for securing test insect materials for ecological and biological study of Liriomyza species, and also for developing a biological control for Liriomyza species other than L. huidobrensis by mass production of associated natural enemies. We are facing rapid agro‐ecosystem changes including pest systems. Continuous monitoring of Liriomyza in solanaceous crops is needed.