东方黏虫[Mythimna separata(Walker)]田间自然种群的性信息素受体MsepOR6功能鉴定

为进一步理解东方黏虫[Mythimna separata(Walker)]的求偶行为,探明雄蛾对种内和种间性信息素的嗅觉分子识别机制,本研究以顺11-十六碳烯乙酸酯(Z11-16:Ac)和顺11-十六碳烯醇(Z11-16:OH)为性信息素进行田间诱捕到的东方黏虫作为研究对象,根据文献报道的MsepOR6基因片段序列,采用cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术扩增出其全长,采用双电极电压钳技术鉴定MsepOR6对东方黏虫及其同作物生境害虫的性信息素的电流反应,并通过田间诱捕试验验证雄性东方黏虫对其中的活性化合物的行为反应。电生理测定结果表明:MsepOR6是东方黏虫雄蛾的性信息素受体基因,其cRNA对种内性信息素顺11-十六碳烯醛(Z11-16:Ald)、十六醛(16:Ald),以及种间性信息素顺9-十四碳烯乙酸酯(Z9-14:Ac)、顺7-十二碳烯乙酸酯(Z7-12:Ac)、顺9-十六碳烯乙酸酯(Z9-16:Ac)、顺9-十四碳烯醇(Z9-14:OH)有显著的电生理反应。田间生物测定结果表明,Z9-14:Ac可显著抑制东方黏虫性信息素的引诱活性,Z7-12:Ac则显著提高东方黏虫性信息素对雄蛾的引诱效果。本研究有助于理解东方黏虫的性信息素行为调控机制,为开发应用性信息素技术提供了理论支撑。

草地贪夜蛾(Spodoptera frugiperda)与东方粘虫(Mythimna separata)种间竞争的室内模拟研究

研究草地贪夜蛾(Spodoptera frugiperda)和东方粘虫(Mythimna separata)在室内以不同的初始种群密度(50头草地贪夜蛾初孵幼虫+50头东方粘虫初孵幼虫;25头草地贪夜蛾初孵幼虫+50头东方粘虫初孵幼虫;10头草地贪夜蛾初孵幼虫+50头东方粘虫初孵幼虫)取食相同寄主时二者的种群竞争行为并分析原因。结果表明,在以小麦苗为寄主的条件下,当初始种群为50头草地贪夜蛾+50头东方粘虫时,混合后13 d,东方粘虫存活率为0,而草地贪夜蛾的存活率为51.0%;随着初始混合种群中草地贪夜蛾比例的下降,混合后13 d,各处理中东方粘虫的存活率逐渐增加,2种昆虫种群达到共存。在以玉米苗为寄主的条件下,当初始种群为50头草地贪夜蛾+50头东方粘虫时,混合后10 d,东方粘虫存活率即为0,而草地贪夜蛾的存活率为81.0%;随着初始混合种群中草地贪夜蛾比例的下降,混合后13 d,各处理中东方粘虫均不能完成幼虫历期,2种昆虫种群不能达共存。草地贪夜蛾和东方粘虫在相同生态位的种群竞争中,草地贪夜蛾具有明显的优势;无论寄主为小麦还是玉米,东方粘虫在与草地贪夜蛾的混合种群中每日存活率通常低于草地贪夜蛾及单独饲养的东方粘虫种群,甚至不能完成幼虫发育,造成东方粘虫种群可能被草地贪夜蛾取代。

Relationship between the Occurrence of Mythimna separata on Rice and Climate Factors in Menghai County

The changes of climate factors and the number of Mythimna separata in Menghai County during 1982 -2007 were analyzed, and the relationship between them was discussed. The results showed that there was certain relationship between the outbreak of M. separata and the climate conditions. There was extremely significant correlation between temperature and the number of M. separata. The temperature affected the occurfence amount of M. separata and the regression equation was y=2.691 x-57.123（ P^2 =0.110）. There was no significant correlation between the relative humidity and the amount of M. separata. But the rainfall had significant relationship with the large occurrence of M. separata.

粘虫(Mythimna separata Walker)迁飞行为研究

粘虫成虫期取食、飞翔、求偶或性反应交配和产卵一系列行为是各具相对独立性和顺序性的阶段。飞翔在取食、飞翔、求偶或性反应交配、产卵等期内都有发生 ,并有不同的飞翔特征 ,在迁飞中以飞翔期的飞翔最为重要。雌雄蛾的飞翔期在羽化后 3～ 1 0日龄 ,因求偶、性反应前期变化而有变异。粘虫雌蛾求偶前期 2～ 1 1 d,平均为 5.93d,雄蛾性反应前期 3～ 1 1 d,平均为 7.1 4 d。求偶、性反应与迁飞潜力有密切关系。随求偶、性反应前期加长 ,求偶、性反应前飞翔量呈“S"形曲线增长 ;求偶或性反应迟的个体有飞翔期 ,且有 1～ 2个飞翔峰 (日 )。根据有无飞翔期和求偶、性反应与飞翔量曲线确定雌蛾求偶前期大于 5d,雄蛾性反应前期大于 4 d的个体为迁飞型个体。自然种群中雌蛾迁飞型个体占 52 .4 7% ,雄蛾占97.1 4 % ;不同地理种群江苏、吉林、山东和广东 1～ 4代粘虫 ,雌蛾迁飞个体依次占 4 7.64%、4 9.2 9%、4 7.2 2 %和68.2 2 % ,求偶前期平均依次为 5.75、5.64、5.58和 6.77d。影响求偶前期的生态因素主要是温度逐渐上升和食料不断老化 ,两者可能是引起粘虫迁飞的主要因素。讨论了粘虫成虫期行为。

Effects of temperatures on the development and reproduction of the armyworm, Mythimna roseilinea: Analysis using an age-stage, two-sex life table

The armyworm Mythimna roseilinea（Walker） is a major pest of grain crops in South China. So far little is known about its basic biology and ecology, making prediction of population dynamics difficult. This study examined the relationships of individual development and population growth with temperature based on an age-stage, two-sex life table of M. roseilinea reared on maize in the laboratory at 18, 21, 24, 27 and 30℃. The highest values of net reproductive rate（R_0） and fecundity were observed at 21 and 24℃, respectively. Both the intrinsic rate of increase（r） and finite rate of increase（λ） increased significantly and mean generation time（T） decreased significantly with increasing temperature. M. roseilinea was able to develop, survive and lay eggs at all temperature regimes tested. Development rates of the egg, larval, pupal, as well as the whole pre-oviposition stages had a positive linear relationship with temperature. The calculated development threshold temperatures of egg, larval, pupal, pre-oviposition and total pre-oviposition stages were 13.29, 8.39, 14.35, 7.42, and 12.24℃, respectively, and their effective accumulated temperatures were 63.59, 445.00, 211.11, 89.02, and 698.95 degree-days, respectively. These results provide insight into temperature-based phenology and population ecology of this insect pest and will allow population prediction and management available in the field.

Effects of Terpinen-4-ol on Four Metabolic Enzymes and Polyphenol Oxidase (PPO) in Mythimna separta Walker

To study insecticidal mechanism of terpinen-4-ol, a main insecticidal composition in the essential oil of Sabina vulgaris, the 5th instar larvae of Mythimna separta, were investigated with terpinen-4-ol by topical application. The activities of phosphatase, glutathione S-transferase （GSTs）, cytochrome P450 （P450）, and polyphenol oxidase （PPO） of tested insects were determined in all poisoning stages, including exciting stage, convulsing stage, paralysis stage, and recover stage. The result showed that the activities of both acid phosphatase （ACP） and alkaline phosphatase （AKP） in treated insects were induced by terpinen-4-ol, but ACP was inhibited in paralysis stage. The activities of GSTs were inhibited in exciting stage, convulsing stage, and paralysis stage, but gradually recovered in recover stage. O-demethylase activity of cytochrome P450 was inhibited by terpinen-4-ol, and the inhibition rate in all poisoning stages were 26.27, 46.03, 80.24, and 90.22%, respectively. PPO activities were strongly inhibited by terpinen-4-ol both in vitro and in vivo. In conclusion, the activities of P450, GSTs, and PPO could have relation with toxicity of terpinen-4-ol against larvae of the Mythimna separta, but recover stage of the poisoning insects might be related to GSTs induced. As a new insecticide or synergist, terpinen- 4-ol has a potential value in field of insecticide resistance management.

Effects of constant and stage-specific-alternating temperature on the survival, development and reproduction of the oriental armyworm, Mythimna separata(Walker)(Lepidoptera: Noctuidae)

Migratory insects make diverse adaptive strategies in response to changes in external environment. Temperature has an impact on the survival, development, reproduction, and migration initiation of insects. Previous research has primarily been focused on the effects of constant temperature on populations, but changing temperature has received less attention. Three constant temperature treatments（20, 25 and 30℃） and three pupal-alternating temperature treatments（20–25, 25–20 and 25–30℃） were set up to study the relationship between temperature and population development by age-stage, two-sex life table analysis in the oriental armyworm, Mythimna separata Walker, a notorious migratory pest in grain crops. The 25℃ treatment was considered optimal with 20 and 30℃ as low suitable temperature and high temperatures, respectively. The survival rate was relatively low before third instar larvae at 20℃（63.0%） and 20–25℃（70.1%）, and extreme low after pupal stage at 30℃（20.6%）. Developmental duration of each stage was negatively correlated with temperature. The adult preoviposition period, when most migratory insects initiate migration, was the shortest at 25℃（2.69d） but was lengthened at both low suitable（7.48d for 20℃, 6.91d for 25–20℃ and 4.57d for 20–25℃） and high temperatures（3.74d for 25–30℃ and 5.00d for 30℃）. Both low suitable and high temperature decreased lifetime fecundity, net reproductive rate and the intrinsic rate of increase, with variability observed across developmental duration and stage during non-optimal temperature. The results expand knowledge of the relationship between changing temperature and armyworm population development, and adaptive strategies in complex ambient environment.

Insecticide resistance of the field populations of oriental armyworm,Mythimna separata(Walker) in Shaanxi and Shanxi provinces of China

Resistance of five field populations of Mythimna separata（Walker） collected from Shaanxi and Shanxi provinces of China to six different insecticides was evaluated by leaf dip method in the laboratory. The results showed that all populations were relatively sensitive to emamectin benzoate with a resistance ratio（RR） of 0.583–1.583 folds. All populations showed susceptible or low level resistance to chlorantraniliprole and beta-cypermethrin. Compared with a relatively susceptible strain of M. separata, the resistance level of the whole populations ranged from susceptible to moderate to chlorpyrifos and lambda-cyhalothrin, moderate to high to phoxim（RR=19.367–70.100） except for population from Sanyuan County（RR=2.567）. Pair-wise correlation analysis among different insecticides indicated that chlorpyrifos has a significantly positive and significant correlation with emamectin benzoate. Chlorantraniliprole didn＇t have significant correlation with emamectin benzoate, chlorpyrifos and phoxim. Therefore, emamectin benzoate, chlorantraniliprole and beta-cypermethrin are recommended to control oriental armyworm. Meanwhile, to postpone the occurrence and development of insecticide resistance in Shaanxi and Shanxi provinces, alternative and rotational application of insecticides between chlorantraniliprole and emamectin benzoate or chlorpyrifos is necessary.

Identification of summer nectar plants contributing to outbreaks of Mythimna separata(Walker)(Lepidoptera: Noctuidae) in North China

The presence of adequate nectar sources along the migratory pathway is a prerequisite for moths of oriental armyworm, Mythimna separata, to reach target habitats and lay eggs. Identification of plant species and assessment of their suitability as wild nectar sources along the summer migration pathway of adult M. separata are critical steps in making effective forecasts of second-and third-generation armyworm outbreaks in North China. We explored five aspects of moth-nectar source relationships：（1） Field investigations in 2014 and 2015 showed that the florescence of chaste tree, Vitex negundo var. heterophylla, a primary nectar plant in apiculture, coincided with the summer migration of oriental armyworm moths. The flowers of chaste tree opened day and night and M. separata moths were found actively flying around and resting on tree.（2） Identification of moth-borne pollen by scanning electronic microscopy（SEM） micrographs in 2015 and 2016 revealed that 15.4% of 369 moths trapped by blacklight traps, 17.1% of 557 moths captured by vertical-pointing searchlight traps and 24.4% of 311 moths caught by pheromone traps were found with pollen attached to their proboscis. A total of 228 pollen-carrying individuals out of 1237 armyworm moths were captured by the three types of traps. The three highest percentages of pollen-bearing armyworm moths were carrying pollen of Toona sp.（59.65%）, Ligustrum sp.（27.19%） and Syringa sp.（4.39%）. In total, 14 types of pollen were detected where three were identified to species, 10 to genus and one as an unknown species.（3） Analysis of geographical distribution, flowering periods and relative abundances of candidate nectar plants suggested that Chinese toon, Toona sinensis, and privet, Ligustrum lucidum, were two important nectar plant species.（4） Analysis on relationship between pollen and ovarian development showed that 14, 15 and 10 females carrying pollen of T. sinensis had ovaries at developmental stages Ⅰ, Ⅱ and Ⅲ, respectively. Many T. sinensis pollen grains were present on the proboscises of female moths.（5） Comparison of trap effectiveness between blacklight and vertical-pointing searchlight traps indicated blacklight traps caught the highest proportion（35.2%） of female in ovarian stage Ⅴ, whereas vertical-pointing searchlight traps caught the highest proportion（43.5%） of female in ovarian stage I with additional proportions of decreasing percentages for stages Ⅱ, Ⅲ and Ⅳ. Integrated analysis on these five aspects suggests that T. sinensis is an important, suitable summer nectar source that potentially influences migration and reproduction of M. separata moths andthus contributes to outbreaks of this pest. The vertical-pointing searchlight trap was the most effective tool for monitoring the migratory flight of adult M. separata. Privet, Ligustrum lucidum, and clove, Syringa sp., are possible suitable summer nectar plants that need further study to elucidate their importance. The reason why field investigation results on chaste tree differed from the results of moth-borne pollen analyses was discussed.

Analysis on the migration of first-generation Mythimna separata(Walker) in China in 2013

Mythimna separata（Walker） is an important pest which can cause serious damages to cereal crops. In the past two decades, several heavy outbreaks have taken place in northern China. In order to develop a fine-scale method of forecasting outbreaks, population data were collected in northern China using searchlight traps and ground light traps. A background weather pattern analysis and trajectory analysis were performed via the Weather Research and Forecast（WRF） and FLEXPART models. Our results showed that heavy migration of first-generation M. separata appeared in northern China in 2013. In Yanqing District, Beijing, the cumulative number of captured adults in searchlight traps was around 250000 and the daily maximum for trapped moths was 86000. During the peak period, the majority of M. separata moths arrived after 00：00 every night. The sex ratio（female：male） at each monitoring site was greater than 1 and greatly fluctuated with population dynamics. During the migration peak, prevailing downdraft winds benefited M. separata moths to land passively. Trajectory simulation showed that immigrants were from Anhui, Jiangsu and Hubei provinces and most of them could continue to fly into the northeastern regions of China. These results provide technical support for fine-scale forecasting of the outbreak of M. separata at meso-and micro-scale.

苦皮藤素Ⅴ对粘虫Mythimna separata细胞色素P450及谷胱甘肽S-转移酶的诱导作用

【目的】研究解毒酶系在苦皮藤素V代谢中的作用。【方法】利用UV-Vis分光光度计,研究了苦皮藤素V对6龄5日龄粘虫Mythimna separata的中肠及脂肪体细胞色素P450(包括P450、NADPH-P450还原酶)和谷胱甘肽S-转移酶(GST)的诱导作用。【结果】粘虫幼虫的细胞色素P450与CO完全结合后,逐渐失活变为细胞色素P420;经苦皮藤素V处理后,粘虫幼虫的中肠P450含量较未经苦皮藤素V处理的对照增加了2.4倍,NADPH-P450还原酶活性、GST活性均低于对照,而脂肪体P450含量、NADPH-P450还原酶活性、GST活性分别为对照的1.32,1.30和1.13倍。【结论】苦皮藤素V对粘虫中肠P450具有较强的诱导作用。

Effects of alternating temperatures on development and reproduction of the armyworm,Mythimna separata (Walker)

Effects of cycling and constant temperature on development and reproduction of the armyworm, Mythimna separata (Walker) were investigated by rearing the insects on the artificial diet under temperatures covering low(16-24℃ vs. 20 ℃ ), medial(20-28℃ vs. 24℃ ) and high(24-32 ℃ vs. 28 ℃) regions. In the low region, all the immature stages of the insect developed significantly faster under alternating temperatures than at corresponding constant temperatures. For the medial region, their development was little affected by variable temperatures. For the high region, developmental durations tended to be longer under cycling temperatures except egg stage.Mean fecundities of the adult moths markedly increased under cycling temperatures as compared with those at respective constant temperature in all regions.Possible reasons for effect of alternating temperatures on insect development are discussed in some details.

Association of host plant growth and weed occurrence with armyworm(Mythimna separata) damage in corn fields

To clarify association between armyworm（Mythimna separata） damage level and the corn growth and weed occurrence, we investigated corn plant height, stem diameter and vigor as well as weed coverage and biomass. The investigations were conducted at three locations of Shaanxi Province, China which were suffered seriously from armyworm. Significant correlations were found between the parameters analyzed. At stunted corn growth and presence of plenty of weeds, the armyworm damage tended to be heavy; oppositely, when corn grew well and weed density were low, armyworm harm was the minimal. Therefore, corn growing status and weed density can significantly affect armyworm damage level. Our results imply that promoting corn growth and timely removal of weeds are conducive to reducing armyworm occurrence.

Effect of a near-zero magnetic field on development and flight of oriental armyworm(Mythimna separata)

The geomagnetic field affects all living organisms on the Earth.In this study we investigated the developmental and behavioral effects of rearing Mythimna separata in a near-zero magnetic field(<500 n T)compared to the local geomagnetic field(approximately 50μT).The near-zero magnetic field produced by a Helmholtz coil system significantly lengthened larval and pupal development durations,increased male longevity,and reduced pupal weight,female reproduction,and the relative expression level of the vitellogenin(Vg)gene in newly emerged females.Moreover,the near-zero magnetic field had a considerable negative effect on the mating ratio of M.separata adults.In addition,the moths in the near-zero magnetic field displayed less flight activity late in the night than those in the Earth's normal geomagnetic field,indicating that the flight rhythm of M.separata may be affected by the near-zero magnetic field.Reduction in magnetic field intensity may have negative effects on the development and flight of oriental armyworm,with consequent additional effects on its migration.

Effects of fluoride on growth and reproduction of the army worm, Mythimna separata (Walker)

This paper deals with effects of airborne fluorine on the army worm, Mythimna separata (Walker), by rearing the larvae on the wheat foliage exposed to HF or fumigating the larvae on the artificial diet with the pollutant. Larval relative growth rate (GR) and index of population trend of the insect reduced by 5% and 11%, respectively, when the larvae were reared on the foliage taken from the wheat plants exposed to 0.87 g·dm-2·day-1 of fluorine compared with those of the control. An extra instar appeared in a majority of the larvae treated. Survival rate and GR of the larvae on the wheat plant being exposure to the same concentration of fluorine in field open-top fumigation device were 40% and 15% lower than that of the control, respectively. Similar experiment with the insect on the artificial diet also showed that direct impact of the pollutant on the army worm was greater than its indirect effect via their host plant.

粘虫(Mythimna seperata)为害小麦的产量损失和幼虫密度及生育期的关系

研究进一步确定了粘虫(Mythimna seperata)为害损失同幼虫密度和小麦生育期的关系。结果表明,在同样的幼虫密度下,生育期越晚,受害越轻。数据表明,临界受害期在5月10日前后,过了5月15日,即使全部叶片都受害,产量也不会受影响。本研究同时测定了各龄幼虫的食叶量和扬麦5号小麦的全株叶片面积,并根据上述结果分析推导出了包括幼虫数量和生育期在内的估计粘虫为害损失百分率的公式:DP=0.031L(71.23-1.480x),式中 DP=损失百分率,L=能发育到6龄并化蛹的幼虫数,X=从4月1日开始的日数。应用这个方程可很方便地估计不同时期不同幼虫数所造成的损失百分率。

Secondary effect of SO_2 pollution: enhanced growth of the army worm, Mythimna separata (Walker)

Effects of SO2 pollution on growth of the army worm, Mythimna separata, were investigated by rearing the larvae for 12 days on the wheat plants being exposed to SO2 levels ranged from 50 to 200 ppb in the field open-top fumigation devices. Larval period shortened while their mean relative growth rate (MRGR) increased as SO2 dose elevated. Similar experiment with the insect on the artificial diet excluded the possibility that SO2 pollution directly stimulated growth of the army worm.The patterns of change in amino acid hydrolysates in the host foliage suggested that contents of methionine, and perhaps, arginine in food material exposed to unpolluted air were at such levels that they could only support sub-optimum growth of the insect, and SO2 pollution increased concentrations of these limiting amino acids, resulting in better performance of the larvae.

Effects of temperature on population growth and intake of food by the army worm, Mythimna separata (Walker)

Population life tables of the army worm, Mythimna separata, were constructed and nutritional parameters of food for the larvae were examined at 5 temperatures from 16℃ to 32℃. The temperature suitable for growth and reproduction of the insect ranged from 20℃ to 28℃ with the optimum of 24℃. Their survivalship and fecundity were much poorer at 32℃ than at 16℃. Indices of population trend of the insect at these temperatures could well fitted with the parabolic curve which theoretically indicated that the population density would multiply by some 660 after one generation circle at the optimal temperature, 22.6℃, and it would decline at temperatures higher than 32.9℃ or lower than 12.3℃. Larval food intake and their AD tended to go up while their ECI and ECD to go down with a rise in temperature. These alterations in population size and feeding behavior caused by temperature would exert an important effect on their damage to crops.

Effect of Mythimna separata Digestive Fluid on Infectivity of Nuclear Polyhedrosis Virus

The dissolution of polyhedra of Mythimna separata nuclear polyhedrosis. virus by digestive fluid (pH11. 03) collected from the 5th instar M. separata larvae was studied in vitro. Observations were made at timed intervals using phase contrast microscopy and scanning electron microscopy. Under phase contrast microscopy, the polyhcdra lost their refrigence by 5 minute exposure to the digestive fluid. After exposure to the fluid for 30 minutes, all of the PIBs were dissolved. Chages of the PIBs were also observed under scanning electron microscopy, after 5 minute exposure to the fluid, damaged PIBs and PIB-derived debris were seen. After 30 minute exposure, only remains of PIBs were found. The effect of M. separata digestive fluid on the infectivity of Ms NPV was examined by nconatcs bioassay. The results indicated that virions from Ms NPV-PIBs were rapidly inactivated after 15 minute exposure to digestive fluid and all of virions were non-infectious.

Regularity of population occurrence and migration in the oriental armyworm, Mythimna separata(Walker)

The oriental armyworm,Mythimna separata（Walker）（Lepidoptera：Noctuidae）,is a major migrant pest of grain in China and other Asian countries,causing huge crop production and economic losses nationwide annually.Much of the uniquely difficult aspects of managing M.separata arises from its long-distance migratory behavior.For example,direction and timing of winds from its overwintering regions in the south largely determine where and when large influxes of migrants arrive in the north to oviposit,making prediction of infestations difficult.