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。     

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。     

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.     

Effects of four constant temperatures on the development of two Bradysia (Diptera: Sciaridae) species

In this study, the effects of temperature on the growth, development, survival, fecundity and other population parameters of two local Bradysia species B. odoriphaga and B. impatiens were studied at four constant temperatures (25, 28, 31 and 34°C). The results show that 25°C is the optimum temperature for the growth and development of B. odoriphaga, while 28°C is more favourable for B. impatiens. The temperature of 31°C restricted the growth and development, while the temperature of 34°C inhibited the eggs hatching in both species, resulting in no egg survival and no subsequent development. High temperatures (>28°C) prolonged the 4th larval stage duration, mean generation time (T) and population doubling time (Dt) of both species. The high temperature of 31°C greatly shortened the female longevity, weakened the oviposition and reduced the survival of both species. Moreover, the life table parameters R₀, r_m and λ were also suppressed by this high temperature. However, the high temperature of 31°C had little impact on the egg survival, pupal weight and male longevity. In addition, at 31°C, the values of R₀, r_m and λ of B. odoriphaga were higher than those of B. impatiens, suggesting that B. odoriphaga is more tolerant to high temperature than B. impatiens. The differences between two Bradydsia species seem determined genetically. Our findings are important for better understanding their biological characteristics at a certain constant temperature and demonstrate the possibility to control and manage those two Bradysia species by increasing ambient temperature.     

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。     

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。     

Transmission of Pythium aphanidermatum to greenhouse cucumber by the fungus gnat Bradysia impatiens (Diptera: Sciaridae)

The fungus gnat (Bradysia impatiens) was examined for its ability to transmit Pythium aphanidermatum to cucumber plants. Larvae that had ingested oospores and mycelium, and then fed on the roots of young cucumber plants growing in rockwool readily introduced the fungus to them. Trans-stadial transmission of oospores from the larval to the adult stage of B. impatiens was demonstrated, although decreasing to a very low level (1.67%) in adults. However, external transmission of P. aphanidermatum on the surface of adults could not be shown. These results suggest that the larval stages of fungus gnats play a role in the dissemination of the fungus between cucumber plants but that adults probably play only a minor role.     

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.     

Unequal X Chromosomes in Bradysia Hygida (Diptera:Sciaridae) Females: Karyotype Assembly and Morphometric Analysis

The mitotic chromosomes of Bradysia hygida(Diptera:Sciaridae) neuroblast cells are described together with their morphometric data. Giemsa-stained neuroblast chromosomes from female and male larvae confirm the chromosome number of this species, 2n=8 (XX) and 2n=7 (XO), respectively. The karyotype assembly reveals two metacentric autosomic pairs, the A and B chromosome; a subtelocentric, the C chromosome, the smallest one; and a sexual unequal metacentric pair, X chromosome, in female karyotype and a one sexual metacentric X chromosome in male. The implications of the unequal X chromosome pair are discussed.     

Immunocharacterization of the DNA puff BhC4-1 protein of Bradysia hygida (Diptera: Sciaridae)

The DNA puff BhC4-1 gene is amplified and highly expressed in the salivary gland of Bradysia hygida late larvae. Using affinity-purified polyclonal antibodies we have identified the product of the BhC4-1 gene as a 43 kDa polypeptide which is present in extracts of salivary glands from late fourth instar larvae and in the corresponding gland secretion, but not in glands from earlier stages. We also demonstrate that this protein is produced mainly in the S1 and S3 regions of the salivary gland, where BhC4-1 amplification levels are more pronounced and larger amounts of mRNA are produced. By immunoelectron microscopy the BhC4-1 protein was detected in secretory granules of the S1 and S3 regions, and localized in fibrous structures present in the saliva.     

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.     

Ingestion of Pythium spp. by larvae of the fungus gnat Bradysia impatiens (Diptera:Sciaridae)

Mycelium, oospores and zoospore cysts of Pythium spp. were fed to larvae of the fungus gnat Bradysia impatiens. The fungal structures were all ingested and provided a complete nutritional source for the insect's development from egg to adult. Mycelium seemed the major source of food as only empty fragments were found either in the digestive tract or in larval faeces. Oospores appeared intact and were viable both during passage through the tract and when expelled. Germination of oospores was normal. Most encysted zoospores also survived passage through the gut, although some appeared damaged. After transferring recently-fed larvae to a new food source, oospores were still detected in the digestive tract 48 h later. These results show that the larval stage of B. impatiens may serve as an important vector for Pythium spp.     

Programmed cell death in Bradysia hygida (Diptera, Sciaridae) salivary glands presents apoptotic features

In this work, we present biochemical and morphological evidence that the final steps of programmed cell death (PCD) in the salivary glands of the inferior Diptera, Bradysia hygida, present apoptotic characteristics. In B. hygida, elimination of salivary glands is preceded by the establishment of a typical pattern of protein synthesis; increase in caspase activity; decrease in cell volume; nuclear pyknosis; nuclear DNA breakage; changes in the actin cytoskeleton; and most importantly, destruction of giant cells via formation of apoptotic bodies containing broken DNA or cytoplasm remains. Thus, elimination of B. hygida salivary glands by this process suggests that such mode of PCD is also involved in the destruction of entire organs in insects and, therefore, adds more complexity to the regulation of tissue elimination during development.     

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.     

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。     

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。     

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.     

Grazing rate determination of Corynoneura scutellata Winnertz (Chironomidae: Diptera)

Corynoneura scutellata Winnertz (Chironomidae: Diptera) grazing rates were determined radiometrically at different temperatures. The resulting temperature-grazing function was used to calculate C. scutellata grazing rates in the littoral zone of a Rhode Island lake. These grazing rates were compared to periphyton net accumulation rates, grazing rate: accumulation rate ratios ranged from 3–15% with a maximum of 70%.     

Behavioural response of the fungus gnat,Bradysia impatiens (Diptera: Sciaridae) towards certain edible mushrooms and saprophytic fungi

The fungus gnat Bradysia impatiens (Johannsen) is a worldwide pest inhabiting organic-rich environments, including mushroom-cultivation substrates, which leads to massive production losses of edible mushrooms. To promote a non-insecticidal pest control strategy, we evaluated adult behavioural response towards two saprophytic fungal strains Aspergillus flavus Link and Mucor circinelloides Tiegh., isolated from B. impatiens-inhabiting substrates and six edible mushrooms species. Our results indicated that (a) B. impatiens was most attracted to the oyster mushroom Pleurotus ostreatus among all six edible mushrooms; (b) males and females demonstrated a significant attraction for M. circinelloides over A. flavus and P. ostreatus; (c) adults demonstrated dynamic attraction tendencies that varied with the culture periods of A. flavus and M. circinelloides, wherein females represented most (mean = 78.6%) of the attracted individuals; and (d) larvae that fed on mycelial cultures of M. circinelloides could complete their entire life cycles, but larvae could not survive on a non-fungal or A. flavus diet. These results demonstrate the potential for the development of M. circinelloides in a push-pull strategy to biocontrol this pest in edible mushroom cultivation.     

Selective breeding of Steinernema feltiae (Filipjev) (Nematoda: Steinernematidae) for improved efficacy in control of a mushroom fly,Lycoriella solani Winnertz (Diptera: Sciaridae)

A method of selecting a Steinernema feltiae strain that is effective against a mushroom fly, Lycoriella solani, is described in detail. The pest control efficacy of the selected nematode strain was evaluated and compared with the efficacy of two unselected strains. The selection procedure was designed to give preference to nematode individuals with the greatest ability (1) to search effectively for the target insect larvae in their natural habitat, (2) to infect them shortly after application and (3) to reproduce in their haemocoel. Thirty‐four rounds of selection achieved a 4‐fold improvement in nematode ability to find and parasitize third‐ and fourth‐instar larvae of the pest in the mushroom substrate. In 24‐h laboratory experiments, mortality of the insect caused by nematode juveniles rose from 22.5%, recorded for the original unselected isolate, to 92.5% for the selected strain. In a 51‐day experiment conducted on a mixed age mushroom house population of L. solani, the enhanced pest control ability of the selected strain was detected shortly after nematode application and remained high throughout the experimental period. During the first 4 weeks of the trial the selected nematode strain was significantly better than both unselected strains and caused 91.1–92.7% reduction of the fly emergence from the mushroom substrate. No difference was observed between the efficacy of the selected nematodes applied at 1 × 10⁶ and 3 ×10⁶ infective juveniles per m², while the unselected strains performed significantly better at the higher concentration. All the nematodes examined showed good persistence in the mushroom casing apparently due to recycling in the insect host.