苯并噻唑对不同虫态韭菜迟眼蕈蚊的生物活性

【目的】明确室内条件下挥发性化合物苯并噻唑对韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang的生物活性。【方法】采用三角瓶密闭熏蒸法测定了苯并噻唑对韭菜迟眼蕈蚊成虫、 卵、 幼虫和蛹的熏蒸活性; 利用Oxytherm氧电极研究了苯并噻唑对韭菜迟眼蕈蚊成虫呼吸速率的影响; 利用“Y”型嗅觉仪测定了韭菜迟眼蕈蚊成虫对苯并噻唑的行为反应。【结果】苯并噻唑对雌雄成虫处理0.5~2.0 h的LC50分别为0.186~0.052和0.163~0.039 μL/L; 在0.01~0.13 μL/L剂量下对卵熏蒸处理24 h, 第6 天卵孵化率为4.83%~82.39%, 而对照组孵化率为96.97%; 苯并噻唑熏蒸4龄幼虫6~72 h的LC50变化范围为1.247~0.248 μL/L; 0.01~0.09 μL/L剂量熏蒸蛹24 h, 第5天的羽化率为8.17%~69.63%, 对照组羽化率为96.23%。用0.052和0.039 μL/L浓度分别处理雌雄成虫测定2.5 h内呼吸速率的变化, 表现为处理组初始呼吸速率明显高于对照组, 然后逐渐降低至与对照组持平, 最后明显低于相同处理时间对照组呼吸速率。“Y”型嗅觉仪测定结果表明, 苯并噻唑对韭菜迟眼蕈蚊成虫有较强的引诱作用。在0.5 L/min空气流速条件下, 0.5 μL的苯并噻唑对雌雄成虫的引诱率分别为88.33%和78.33%。【结论】苯并噻唑对韭菜迟眼蕈蚊各虫态有很好的毒杀效果, 并对成虫有强烈的引诱作用。     

Effects of short-term heat stress on the growth and development of Bradysia cellarum and Bradysia impatiens

Bradysia cellarum Frey and Bradysia impatiens Johansen are major pests of vegetable crops, as well as edible mushrooms and ornamental plants, and damage to hosts resulting in economic losses. Temperatures above the optimum levels for these pests have been predicted to regulate their population growth during summer. The aim of the present study was to examine the effects of both heat stress and exposure time on the growth and development of eggs, larvae and pupae for two Bradysia species. The egg stage, egg hatching rate, 4th instar larval stage, pupation rate, pupal stage and adult emergence rate were observed after exposing at high temperatures of 34°C, 37°C and 40°C for 1, 2, 4 and 6 hr. The results showed that 34°C, 37°C and 40°C for 1-, 2-, 4- and 6-hr exposure treatments prolonged the developmental stage of egg, 4th instar larva and pupa, while decreasing the egg hatching rate, pupation rate and adult emergence rate. This suggests that increasing temperature or prolonging exposure time to the heat stress could significantly affect insect survival, growth and development. Our study could provide an ecological basis for pests’ management strategy by using short-term heat stress.     

百合科寄主植物对韭菜迟眼蕈蚊的生物效应

室内分别用百合科韭菜、大葱、大蒜和圆葱饲喂韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang幼虫,研究了4种寄主植物对其生长发育和繁殖的影响;用气-质谱联用分析技术,检测了人工合成大蒜素及大蒜、圆葱和韭菜的乙醇提取物的主要化学成分。结果表明,4种供试寄主植物对韭菜迟眼蕈蚊生长发育和繁殖的影响存在差异,幼虫取食韭菜最有利于其生长发育和繁殖;取食大蒜和圆葱对其生长发育和繁殖表现出明显的不利性,主要表现为幼虫死亡率增加,幼虫期延长,蛹重减轻,单雌产卵量降低;而取食大葱的影响介于取食韭菜与取食大蒜和圆葱之间。大蒜、圆葱的乙醇提取物和人工合成的大蒜素均对韭菜迟眼蕈蚊1龄幼虫有不同程度的杀虫活性,大蒜和圆葱的乙醇提取物（干粉,2 g/mL）稀释100和200倍,处理后48 h对1龄幼虫的校正死亡率分别达54.7%、28.0%和49.4%、22.7%;10%大蒜素稀释500和1 000倍,处理后48 h校正死亡率达100%和80.0%。成分分析表明硫醚类化合物可能是大蒜和圆葱中含有的杀虫活性物质之一。     

环境颜色对韭菜迟眼蕈蚊生物学特性的影响

【目的】明确环境颜色对韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang生长发育及繁殖能力等生物学特征的影响。【方法】在室内条件下,利用黑、棕、绿、红和橙5种颜色卡纸模拟自然环境颜色,比较研究了其对韭菜迟眼蕈蚊生长发育和繁殖的影响。【结果】韭菜迟眼蕈蚊发育历期在橙色环境下最长,为26.15 d,黑色环境下最短,为22.35 d;世代存活率在黑色环境下最高,为43.88%,橙色环境下最低,为28.88%;性比在各颜色环境下无显著差异;雌雄成虫寿命均在棕色环境下最长,分别为2.06 d和2.99 d,橙色环境下最短,分别为1.58 d和2.57 d;单雌产卵量在棕色环境下最多,为62.99粒,橙色环境下最少,为45.64粒;种群趋势指数由高到低依次为:黑色(15.90)棕色(15.07)透明色对照(12.93)红色(11.69)绿色(11.67)橙色(8.50)。【结论】环境颜色影响韭菜迟眼蕈蚊生物学特性,黑色及棕色环境有利于其生长发育及繁殖,橙色环境可抑制其生长发育及繁殖。     

不同波长LED光源对韭菜迟眼蕈蚊生殖行为的影响

【目的】明确不同波长的LED光源对韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang求偶、交配及繁殖等生殖行为的影响。【方法】采用红(625~630 nm)、橙(600~605 nm)、黄(590~595nm)、绿(525~530 nm)、蓝(455~460 nm)和白(6 000~6 500 k)6种LED光源在韭菜迟眼蕈蚊成虫交配期进行照光处理,观察统计其求偶和交配行为以及单雌产卵量、卵孵化情况和有效后代数量。【结果】韭菜迟眼蕈蚊成虫求偶前期时长在橙光下最长,为28.48 min。求偶率在蓝光下最高,为86%;橙光下最低,为48%。交配期时长在蓝光下最长,为4.59 min;橙光下较短,为4.23 min。单雌产卵量在各波长光源下与对照均无显著差异。卵孵化率在蓝光下最低,仅为43.41%。有效后代数量在蓝光下最低,仅为27.00头;橙光下次之,为43.40头。【结论】LED光源的波长可影响韭菜迟眼蕈蚊的生殖行为,其中橙光(600~605 nm)不利于其求偶、交配和繁殖;蓝光(455~460nm)虽有利于其求偶和交配,但明显抑制其繁殖。     

韭菜迟眼蕈蚊在不同温度下的实验种群生命表

组建了韭菜迟眼蕈蚊Bradysia odoriphaga 在15℃、20℃、25℃和30℃温度下的实验种群生命表。结果表明,韭菜迟眼蕈蚊各虫态的发育速率随温度的升高而加快,在15℃下卵-成虫的发育历期最长（72.4天）,而30℃时仅为21.2天,20℃和25℃时分别为27.3天和23.9天;韭菜迟眼蕈蚊卵、幼虫,蛹和卵-成虫的发育起点温度分别为5.9℃、8.7℃,3.3℃和7.8℃,有效积温分别为77.7、267.2、75.7和418.2日·度;韭菜迟眼蕈蚊成虫寿命随温度升高而逐渐缩短,雌虫寿命在20℃时最长,为11.7天,在30℃下最短,仅存活4.1天,除30℃外在其它温度下雌蚊寿命均长于雄蚊;单雌产卵量在20℃时最高,平均为159.9粒,30℃时最低,为114.7粒。20℃～25℃是韭菜迟眼蕈蚊的最适生长温度,成虫寿命长、繁殖力强、种群趋势指数大,温度过高或过低均不利于种群的繁衍。     

Pest species of a fungus gnat genus Bradysia Winnertz (Diptera: Sciaridae) injuring agricultural and forestry products in Japan,with a review on taxonomy of allied species

Agricultural and forestry pest species of the Bradysia tilicola group from Japan were revised taxonomically after examination of the type specimens. A sciarid pest of Welsh onion occurred from northern Saitama, Japan since 2014 and later from south‐eastern Gunma, Japan, was identified as B. odoriphaga Yang & Zhang, 1985, sp. rev. It was removed from synonymy of B. cellarum Frey, 1948, being the first record of the species from Japan. Bradysia agrestis Sasakawa, 1978, was assigned to be a junior synonym of B. impatiens (Johannsen, 1912). Bradysia paupera Tuomikoski, 1960, sp. rev. , was removed from synonymy of B. impatiens and was recognized as a distinct species. We also compiled their host plants and fungi hitherto reported from Japan. Bradysia impatiens was here recorded from an indoor facility of cloud ear fungus, Auricularia polytricha (Montagne) Sacc., for the first time. At least 19 cultural crops belonging to 12 plant and fungal families were reported as actual and potential hosts of B. impatiens. We suggested that B. impatiens and B. paupera might have been confused in studies of mushroom production in Japan. Further survey of distribution of B. paupera in Japan needs to be done.     

韭蛆人工饲料配方筛选及饲养效果比较

【目的】研究实验室条件下韭蛆 Bradysia odoriphaga Yang et Zhang大量饲养的方法以解决试虫不足的问题。【方法】本实验在25℃,相对湿度75%,光周期16L:8D条件下,通过控制变量法筛选出人工饲料配方,运用两性生命表软件,分析比较人工饲料与韭菜Allium tuberosum的饲喂效果。【结果】对不同成分和不同含量的8种韭蛆人工饲料配方筛选,结果确定最优的人工饲料配方成分和含量为：平菇粉6 g,韭菜粉5 g,琼脂粉1.25 g,山梨酸0.04 g,苯甲酸0.05 g,酵母粉0.5 g,维生素C 2.5 g和水50 mL。该配方和韭菜饲养效果相当,不存在显著差异（P>0.05）。同时,两性生命表结果表明,人工饲料饲喂的韭蛆平均世代周期23.6800 d,韭菜饲喂的韭蛆世代周期为25.9000 d,两者之间差异显著（P<0.05）。生殖生物学特性分析发现：韭菜和人工饲料饲喂的韭蛆雌成虫净生殖率分别为16.3200和41.1800/d,内禀增长率分别为0.1078和0.1570/d。【结论】筛选的人工饲料和韭菜均可以稳定地完成韭蛆的世代繁殖,且人工饲料饲喂效果略优于韭菜,另外人工饲料较韭菜不但具有低成本省人工的优点,而且可解决韭菜反季节供应和农药残留问题,保证稳定的虫源供给。     

韭菜养根期干旱处理对韭菜迟眼蕈蚊种群动态的影响

【目的】本研究旨在明确韭菜养根期干旱处理对韭菜迟眼蕈蚊Bradysia odoriphaga种群动态的影响。【方法】通过控制浇水调控土壤含水量,调查15, 20, 25和30 d干旱胁迫下砂土和壤土栽培的韭菜中韭菜迟眼蕈蚊F1代成虫的发生量,并通过两性生命表方法分析干旱胁迫30 d对韭菜迟眼蕈蚊F₁代种群参数的影响。【结果】研究结果表明,不论砂土还是壤土,干旱胁迫均抑制了韭菜迟眼蕈蚊F1代成虫的发生: 随干旱胁迫时间的增加,韭菜迟眼蕈蚊成虫的数量显著下降;其中干旱胁迫30 d时,韭菜迟眼蕈蚊成虫的发生量最低,壤土或砂土中的F₁代成虫羽化量仅为1头,与其他干旱胁迫时间(15, 20和25 d)相比发生量差异显著。经历30 d干旱胁迫的韭菜迟眼蕈蚊F₁代1, 2, 3和4龄幼虫发育历期、未成熟期、雌成虫产卵前期和总产卵前期较对照组显著延长,分别延长0.82, 253, 2.82, 0.68, 6.32, 1.36和8.69 d,而净生殖率(R₀)、内禀增长率(r_m)以及周限增长率(λ)则显著低于对照组,分别较对照组下降32.38%, 37.50%和5.13%。【结论】在韭菜养根期进行干旱处理可有效抑制韭菜迟眼蕈蚊的发生。     

营养物质和水分对韭菜迟眼蕈蚊成虫繁殖和寿命的影响

【目的】研究韭菜迟眼蕈蚊Bradysia odoriphaga成虫对营养物质的选择补充习性,找出对其产卵繁殖起重要作用的关键因子,为其有效防控提供依据。【方法】通过室内成虫单独配对试验,研究不同营养物质(花粉、蜂蜜、维生素、糖、醋、酒)和水分对韭菜迟眼蕈蚊成虫繁殖和寿命的影响。【结果】随着花粉、蜂蜜和维生素浓度的增加,成虫的繁殖力下降;糖醋酒液单一成分和混合液对成虫繁殖和寿命的影响与水分相近;补充供试营养物质和水分对韭菜迟眼蕈蚊的繁殖力均明显高于干燥对照。补充水分的成虫繁殖力最高,产卵率超过70%,单雌平均产卵量达70粒,雌雄成虫寿命最长均超过3 d。土壤湿度对产卵有显著影响,土壤相对湿度为40%和50%时,成虫的落卵率超过25%,落卵量大于480粒,显著高于其它湿度(P<0.05)。【结论】水分是影响韭菜迟眼蕈蚊成虫繁殖和寿命的关键因子,供试的营养物质对韭菜迟眼蕈蚊繁殖没有促进作用,因此在韭菜迟眼蕈蚊成虫盛发期可以通过控制田间土壤湿度来影响韭菜迟眼蕈蚊繁殖。     

臭氧水对韭蛆的防治效果及对韭菜生长的影响

【目的】为科学合理地浇灌臭氧(O₃)水防治韭菜迟眼蕈蚊Bradysia odoriphaga幼虫(俗称韭蛆),明确最佳浇灌时期和使用条件。【方法】在平地覆膜环境中浇灌不同浓度(5, 10, 20和30 mg/L)O₃水,调查对韭蛆的防治效果;在平地覆膜环境中不同日光强度、3种不同灌溉环境［平地覆膜、小拱棚和小拱棚+草垫］、2种不同灌溉环境［平地覆膜、小拱棚］的不同浇灌频率等条件下分别浇灌30 mg/L O₃水,调查对韭蛆的防治效果;调查平地覆膜环境中浇灌30 mg/L O₃水对韭菜产量和根系生长的影响;分析日光强度对不同灌溉环境中土壤温度及浇灌O₃水对韭蛆的防治效果的影响。【结果】平地覆膜条件下,O₃水防治韭蛆的最佳浓度为20~30 mg/L。在平地覆膜条件下浇灌30 mg/L O₃水处理中,对韭蛆的防治效果与浇灌当天的日光强度呈正相关;当日光强度超过60 000 lx时,第1天韭蛆的死亡率高达100%;日光强度低于10 000 lx时,对韭蛆无显著的防治效果;日光强度处于中间水平时,前期对韭蛆的防治效果较差,而后期防治效果逐渐增强。土壤升温效果与日光强度呈正相关,也与灌溉环境显著相关,不同灌溉环境下土壤5 cm深处的温度为：平地覆膜>小拱棚＞小拱棚+草垫。小拱棚内浇灌O₃水对韭蛆的防治效果显著高于平地覆膜组和小拱棚+草垫组。采取连续多次浇灌O₃水有利于提高O₃水对韭蛆的防治效果,而且小拱棚内韭蛆的防治效果更显著。浇灌O₃水时,进水口离田埂末端的距离不能超过40 m。平地覆膜环境中浇灌30 mg/L O₃水后,韭菜的株高、重量和须根数量显著高于对照组(浇灌清水);韭菜植株分蘖数与对照组差异不显著。【结论】若能科学合理地浇灌O₃水,既能达到防治韭蛆的理想效果,又能促进韭菜生长。     

山东不同地区韭菜迟眼蕈蚊共生菌Wolbachia 的检测及鉴定

【目的】为了揭示山东省韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang种群共生菌 Wolbachia 的感染率及其分类地位,探讨该共生菌对韭菜迟眼蕈蚊的潜在影响。【方法】利用线粒体细胞色素氧化酶I（mtCOI）基因引物（LCO1490/HCO2198）,通过扩增测序和序列比对对采自山东省12个地区的根蛆种群进行了分类鉴定。在上述基础上,利用 Wolbachia 的16S rDNA和 wsp 基因特异引物（分别为16S-F/16S-R和81F/691R）对鉴别出的11个韭菜迟眼蕈蚊种群体内Wolbachia 感染情况进行了PCR检测;对感染个体体内 Wolbachia 依据16S rDNA基因片段序列进行分类鉴定。【结果】山东省12个根蛆种群中,11个种群为韭菜迟眼蕈蚊种群。基于 Wolbachia 的16S rDNA基因特异引物检测结果发现,这些韭菜迟眼蕈蚊种群广泛感染 Wolbachia （感染率为6.67%~93.33%）,而利用wsp 基因特异引物检测的感染率（0.00%~40.00%）相对较低些。基于 Wolbachia 的16S rDNA基因构建系统发育树表明,这些韭菜迟眼蕈蚊种群感染的Wolbachia 全部属于A组。【结论】确定了 Wolbachia 在山东省韭菜迟眼蕈蚊体内的感染率及其分类地位,为研究 Wolbachia 对韭菜迟眼蕈蚊生物学及生态学的影响奠定了基础。     

Insecticidal activity of Brassica juncea seed meal to the fungus gnat Bradysia impatiens Johannsen (Diptera:Sciaridae)

Fungus gnats (Bradysia impatiens) can be a serious pest especially to plants grown in confined areas, and although various methods of control are available, safer and more effective control measures are desirable. Mustard seed meal, a by‐product remaining after oil removal for use as a biodiesel feedstock, contains compounds called glucosinolates that hydrolyse to insecticidal 2‐propenyl isothiocyanate. Our objective was to produce a dose‐response curve for making recommendations of Brassica juncea seed meal applications that will result in fungus gnat larvae control. Twenty colony‐raised fungus gnat larvae were added to 20 g (226 per cm³) of potting media, and adult emergence monitored during 2 weeks using yellow sticky cards. Treatments included without meal, detoxified meal and 19 doses ranging from 0.05 to 3.0 g seed meal. A logistic model was used to predict an LC50 of 0.18 and an LC90 of 0.38 g seed meal for the 20‐g pot. The amounts of seed meal required to produce the observed LC50 and LC90 were predicted to produce 0.08 and 0.17 μmol 2‐propenyl isothiocyanate per cm³ potting medium, respectively. B. juncea seed meal has potential utility for the control of B. impatiens, thus warranting additional studies to determine the seed meal's chronic impact on fungus gnats, phytotoxicity and plant fertility benefits.     

Mechanisms of programmed cell death in the midgut and salivary glands from Bradysia hygida (Diptera: Sciaridae) during pupal–adult metamorphosis

Programmed cell death is involved with the degeneration/remodeling of larval tissues and organs during holometabolous development. The midgut is a model to study the types of programmed cell death associated with metamorphosis because its structure while degenerating is a substrate for the formation of the adult organ. Another model is the salivary glands from dipteran because their elimination involves different cell death modes. This study aimed to investigate the models of programmed cell death operating during midgut replacement and salivary gland histolysis in Bradysia hygida. We carried out experiments of real‐time observations, morphological analysis, glycogen detection, filamentous‐actin localization, and nuclear acridine orange staining. Our findings allow us to establish that an intact actin cytoskeleton is required for midgut replacement in B. hygida and nuclear condensation and acridine orange staining precede the death of the larval cells. Salivary glands in histolysis present cytoplasmic blebbing, nuclear retraction, and acridine orange staining. This process can be partially reproduced in vitro. We propose that the larval midgut death involves autophagic and apoptotic features and apoptosis is a mechanism involved with salivary gland histolysis.     

The development of Steinernema feltiae (Nematoda Steinernematidae) in the sciarid fly Bradysia paupera (Diptera: Sciaridae)

There was only one generation of Steinernema feltiae in Bradysia paupera. Infection occurred after 3 h, adults developed 27 h after invasion and new infective juveniles (IJ) were produced after 48 h. Stunted females were produced in B. paupera larvae and in other small hosts and these stunted females produced small IJs. The small IJs were capable of infecting hosts and normal sized Us were produced in succeeding generations in Galleria mellonella.     

DNA barcoding of Bradysia (Diptera: Sciaridae) for detection of the immature stages on agricultural crops

We investigated the usefulness of mitochondrial cytochrome c oxidase (COI) DNA barcoding of the genus Bradysia for the detection of immature stages and cryptic species complex. Although the larvae of some species in this genus are agricultural pests, immature stages are rarely identified due to the lack of key morphological characteristics. We constructed partial sequences of the COI gene for 25 species of Bradysia as a first step towards a DNA barcode. Using these data, Bradysia impatiens, B. procera and B. peraffinis were identified from larval specimens collected, respectively, from paprika, ginseng and oak sawdust beds used for cultivating shiitake. Our findings reveal a complex of three species within the B. tilicola group. These species were all identified as important pest B. ocellaris based on the morphology of male genital structures; however, the interspecific genetic divergence of the COI region was significantly greater (16.1–19.4%) than the intraspecific variation in each species. Therefore, B. ocellaris may consist of at least three species. The results demonstrate that COI DNA barcodes are useful for Bradysia species identification.     

Isolation of polymorphic microsatellite loci in the clear lake gnat and two other phantom midge species of the genus Chaoborus (Diptera: Chaoboridae)

Chaoborus is of great interest to many freshwater ecologists. The adults can become pests in certain areas in North America and the larvae are an important food source for fish. In this preliminary study, we identified variable microsatellite loci in three species: Chaoborus astictopus (H_E = 0.52–0.76), Chaoborus americanus (H_E = 0.46–0.80) and Chaoborus punctipennis (H_E = 0.66–0.81). Using a biotin/streptavidin capture technique of repetitive sequences in a 96‐well format, we obtained microsatellite‐enriched genomic libraries for all three species and identified six polymorphic microsatellite markers for each species. None of the primers did yield a polymerase chain reaction fragment in a cross‐species test.     

Female resistance behaviour and progeny sex ratio in two Bradysia species (Diptera: Sciaridae) with paternal genome elimination

The relationship between female mating preferences and sex allocation has received considerable theoretical and empirical support. Typically, choosier females adjust their progeny sex ratio towards sons, who inherit the attractive traits of their father. However, in species with paternal genome elimination, where male sperm do not contain the paternal genome, predictions for the direction of progeny sex ratio biases and their relationship with female choosiness are atypical. Paternal genome elimination also creates a potential for male–female conflict over sex allocation, and any influence of female mate choice on sex ratio outcomes have interesting implications for sexually antagonistic coevolution. Within the Sciaridae (Diptera) are species that produce single‐sex progeny (monogenic species) and others in which progeny comprise both sexes (digenic species). Paternal genome elimination occurs in both species. We explore female mate resistance behaviour in a monogenic and digenic species of mushroom gnat from the genus Bradysia. Our experiments confirmed our theoretical predictions, revealing that in the monogenic and digenic species, females producing female‐biased progeny were more likely to have resisted at least one mating attempt.     

温度对六斑平颜蚜蝇种群生长发育及繁殖的影响

六斑平颜蚜蝇Eumerus seximaculatum Huo et Liu是一种对肉苁蓉造成毁灭性危害的蛀茎害虫,本研究旨在探索温度对六斑平颜蚜蝇生长发育及繁殖影响,为其预测预报和防治提供依据。在室内测定了六斑平颜蚜蝇在16、20、24、28和32±1℃ 5个恒温处理下各发育阶段的发育历期、存活率、繁殖力以及种群增长指数。结果表明,在16~32℃恒温范围内,六斑平颜蚜蝇均能完成整个世代,各虫态的发育历期随着温度升高逐渐缩短;而存活率、繁殖力、种群增长指数随着温度升高先增多后减少。28℃时世代存活率、每雌平均产卵量、预计下一代产虫数、种群趋势指数均最大,分别为66.67%、150.77粒、2 277.25头、34.68,说明28℃为六斑平颜蚜蝇种群增长的最适宜温度。