The clock gene,period, influences migratory flight and reproduction of the oriental armyworm,Mythimna separata (Walker)

The oriental armyworm, Mythimna separata, is a major long-distance migratory insect pest of grain crops in China and other Asian countries. Migratory flights and reproductive behavior usually occur at night, regulated by a circadian rhythm. However, knowledge about the linkages between adult flight, reproduction, and clock genes is still incomplete. To fill this important gap in our knowledge, a clock gene (designated Msper) was identified and phylogenetic analysis indicated that the encoded protein (MsPER) was highly similar to PER proteins from other insect species. Quantitative RT-PCR assays demonstrated that significantly different spatiotemporal and circadian rhythmic accumulations of mRNA encoding MsPER occurred during development under steady 14 h : 10 h light : dark conditions. The highest mRNA accumulation occurred in adult antennae and the lowest in larvae. Msper was expressed rhythmically in adult antennae, relatively less in photophase and more entering scotophase. Injecting small interference RNA (siRNA) into adult heads effectively knocked down Msper mRNA levels within 72 h. Most siRNA-injected adults reduced their evening flight activity significantly and did not exhibit a normal evening peak of flight activity. They also failed to mate and lay eggs within 72 h. Adult mating behavior was restored to control levels by 72 h post injection. We infer that Msper is a prominent clock gene that acts in regulating adult migratory flight and mating behaviors of M. separata. Because of its influence on migration and mating, Msper may be a valuable gene to target for effective management of this migratory insect.     

Flight capacity: genetic determination and physiological constraints in a migratory moth Mythimna separata

Abstract. Flight capacity of the oriental armyworn Mythimna separata (Walker) (Lepidoptera: Noctuidae) was estimated by a new tethered flight technique. Samples of a migrant population showed a substantial proportion of 'long fliers'. Heritability of flight capacity was found to be significant ( h ²= 0.27). Female moths from a line selected for long pre-reproductive period (PRP) showed greater flight capacity than those from a short PRP line. This difference was not apparent among males. Onset of calling behaviour inhibited the expression of flight capacity in females showing a clear 'oogenesis-flight syndrome'. These results are discussed in relation to seasonal migration in eastern China and the mechanisms involved in the migratory strategy of this species.     

Molecular characterization of DNA methyltransferase 1 and its role in temperature change of armyworm Mythimna separata Walker

DNA methylation refers to the addition of cytosine residues in a CpG context (5′-cytosine-phosphate-guanine-3′). As one of the most common mechanisms of epigenetic modification, it plays a crucial role in regulating gene expression and in a diverse range of biological processes across all multicellular organisms. The relationship between temperature and DNA methylation and how it acts on the adaptability of migratory insects remain unknown. In the present work, a 5,496 bp full-length complementary DNA encoding 1,436 amino acids (named MsDnmt1) was cloned from the devastating migratory pest oriental armyworm, Mythimna separata Walker. The protein shares 36.8–84.4% identity with other insect Dnmt1 isoforms. Spatial and temporal expression analysis revealed that MsDnmt1 was highly expressed in adult stages and head tissue. The changing temperature decreased the expression of MsDnmt1 in both high and low temperature condition. Besides, we found that M. separata exhibited the shortest duration time from the last instar to pupae under 36°C environment when injected with DNA methylation inhibitor. Therefore, our data highlight a potential role for DNA methylation in thermal resistance, which help us to understand the biological role adaptability and colonization of migratory pest in various environments.     

Chromosome-level genomes of two armyworms,Mythimna separata and Mythimna loreyi,provide insights into the biosynthesis and reception of sex pheromones

Mythimna separata and Mythimna loreyi are global pests of gramineous cereals, heavily controlled with synthetic insecticides. Here, we generated two high-quality chromosome-level genome assemblies for M. separata (688 Mb) and M. loreyi (683 Mb). Our analysis identified Z and W chromosomes, with few genes and abundant transposable elements (TEs) found on the W chromosome. We also observed a recent explosion of long interspersed nuclear elements (LINEs), which contributed to the larger genomes of Mythimna. The two armyworms diverged ~10.5 MYA, with only three chromosomes have intrachromosomal rearrangements. Additionally, we observed a tandem repeat expansion of α-amylase genes in Mythimna, which may promote the digestion of carbohydrates and exacerbate their damage to crops. Furthermore, we inferred the sex pheromone biosynthesis pathway for M. separata, M. loreyi and Spodoptera frugiperda. We discovered that M. loreyi and S. frugiperda synthesized the same major constituents of sex pheromones through different pathways. Specifically, the double bonds in the dominant sex pheromone components of S. frugiperda were generated by Δ9- and Δ11-desaturase, while they were generated by Δ11-desaturase and chain-shortening reactions in M. loreyi. We also identified pheromone receptor (PR) genes and inferred their corresponding components. These findings provide a better understanding of sex pheromone communication and promote the development of a new pest control strategy involving pheromone traps, which are more effective and environmentally friendly than current strategies.     

大气CO2浓度升高对粘虫生长发育和繁殖的影响

为明晰大气CO_2浓度升高对粘虫Mythimna separata(Walker)生长发育和繁殖的影响,在人工气候箱800μL/L与400μL/L两种不同CO_2浓度下,用人工饲料连续饲养粘虫3代后,分析其发育历期、体重、存活率等种群参数的变化。结果表明:不同CO_2浓度处理下各世代的粘虫均能正常发育,各虫态的发育历期为:幼虫成虫蛹卵,幼虫1-3代的发育历期在两种CO_2浓度下差异达极显著水平(P0.001);CO_2浓度倍增条件下的蛹重和成虫体重较对照条件下明显下降;CO_2浓度倍增对粘虫的性比影响不大,但可导致粘虫雌成虫寿命缩短,单雌平均产卵量和单雌日均产卵量下降。高CO_2浓度下第1代粘虫种群的平均世代周期(T)和种群加倍时间(DT)分别延长了1.19和1.24 d,1-2代粘虫的净生殖率(R0)、内禀增长率(rm)和周限增长率(λ)呈下降趋势,至第3代略有回升,但未达到显著性差异水平(P0.05);高CO_2浓度条件下粘虫1-3代的存活率分别为44%、31%和33%,分别较对照条件下对应世代粘虫的存活率低10%、27%和22%,其中第2、3代粘虫的存活率在两种CO_2浓度条件下差异显著(P0.05),第1代粘虫在高CO_2浓度下存活率明显下降,至第3代时逐渐趋于稳定,说明随着世代的延长,粘虫对高CO_2浓度的适应能力增强。     

Reduced Mythimna separata infestation on Bt corn could benefit aphids

Use of genetically engineered plants that express insecticidal Cry proteins derived from Bacillus thuringiensis (Bt) have been proven efficacious for managing lepidopteran pests. However, in some cases herbivores that are not targeted by the Bt trait have increased in importance. It has been suggested that reduced caterpillar damage to Bt crops could lead to decreased levels of induced plant defensive compounds which might benefit other non-target herbivores. Here we investigated the potential effect of reduced damage by larvae of Mythimna separata on aphid populations in Bt corn. We compared the performance of Rhopalosiphum maidis feeding on non-Bt corn plants that had been infested by M. separata larvae or were uninfested. The results showed that caterpillar-infested corn plants significantly reduced the fitness of R. maidis leading to a prolonged nymphal development time, reduced adult longevity and fecundity compared to uninfested plants. Consequently, the population growth rate of corn aphids feeding on caterpillar-infested corn plants was significantly lower than on uninfested plants. As expected, the aphids performed significantly better on Lepidoptera-resistant Bt corn than on non-Bt corn when plants were infested with M. separata, since the caterpillars caused very little damage to the Bt plants. The current findings indicate that reduced M. separata infestation could benefit aphid development in Bt corn. Bt corn has the potential to be commercialized in China in the near future and aphids and other non-target pests should be monitored in the farming fields.     

Chitin deacetylase: A potential target for Mythimna separata (Walker) control

Chitin deacetylase (CDA) is a hydrolytic enzyme that modifies chitin into chitosan in the body of insects. In this study, we obtained a full-length complementary DNA sequence (MsCDA1) from the oriental armyworm Mythimna separata by high-throughput sequencing. MsCDA1 is 1,952 bp long and includes 1,620 bp open reading frame encoding 539 amino acids. Analysis by quantitative real time polymerase chain reaction showed that MsCDA1 expression was higher at the adult stage than at earlier developmental stages. MsCDA1 was expressed in all larval tissues examined, in which the highest expression level was found in the midgut. The RNA interference (RNAi) suppressed MsCDA1 expression levels at 12, 24, and 48 hr after injection of double-stranded RNA (1–4 μg per larva) specific to MsCDA1. Under RNAi condition, CDA enzyme activity was significantly reduced and changes an ultramicroscopic structure of M. separata peritrophic matrix especially in its microfibrillar organization exhibiting loose network. In contrast, the surface of the peritrophic matrix was relatively smooth and well organized at control or low RNAi conditions. Moreover, RNAi of MsCDA1 expression impaired larval growth and development, occasionally leading to larval death. These results demonstrate that MsCDA1 plays a crucial role in maintaining peritrophic matrix integrity in M. separata.     

Influence of green light at night on Juvenile hormone in the oriental armyworm Mythimna separata (Lepidoptera: Noctuidae)

The oriental armyworm Mythimna separata is an agricultural insect pest in Eastern Asia. Mythimna separata moths have a high phototactic response to green (520 nm) light. The biological characteristics of insects living under light of a specific wavelength at night can change and, accordingly, Juvenile hormone (JH) levels may be influenced by this light. The present study evaluates changes in the total JH levels at different developmental stages (larvae, pupae and adults) of M. separata reared under green light with different exposure periods at night (or dark period). The results show that, when the exposure time per day of the green light at night is extended, the JH levels in the final‐instar larvae (22 days) and older age pupae (8 days) are significantly reduced, and the JH levels in earlier age pupae (4 days) and adults (3, 6 and 9 days) are significantly increased, compared with groups not exposed to green light. Additionally, the JH level of male moths significantly differs from that of the female moths. We suggest that the JH level of M. separata insects could be regulated by the green light at night (or dark period). The findings of the present study will help to explain the relationship between the light environment and biological characteristics in nocturnal moths.     

Eimeria and Klossia spp. (Protozoa: Sporozoa) from Wild Mammals in Borneo*

SYNOPSIS. During a survey of parasites of Sabah mammals, coccidia oocysts were found in 9 of 22 host species examined. Sporulated oocysts of Klossia sp. from Rattus whiteheadi are described. New geographic records are reported for Eimeria callosciuri, Eimeria sabani, and Eimeria tupaiae. New host and geographic records are reported for Eimeria separata from Rattus cremoriventer, for E. tupaiae and Eimeria ferruginea from Tupaia tana, for Eimeria nieschulzi from Rattus muelleri, and for E. sabani and Klossia sp. from R. whiteheadi.     

Calling behaviour in M.convecta females under different temperature and photoperiodic conditions

Abstract. . The effects of temperature and photoperiod on calling behaviour in females of the Australian common armyworm, Mythimna corwecta (Walker) (Lepidoptera: Noctuidae), were studied in the laboratory. Age at first calling was greater at 15 and 20^oC compared with 25^oC, but there were no significant differences between 25 and 30^oC. Females kept at 10^oC did not call, but if transferred at 10 days to 20^oC they called after a period similar to the pre-calling period at constant 20^oC. Onset time of calling was earlier at 15 and 20^oC compared with 25 and 30^oC, but there were no significant differences in calling duration. At 20^oC, age at first calling was increased by shorter day length (12h and 14h, compared with 16h), and there was an interaction between daylength and moth age affecting the duration of calling. Onset times advanced with longer daylength, but peak calling was later in relation to the lengtii of the scotophase. These results are discussed in connection with migration in M.convecta. Evidence for a circadian rhythm of calling was found in females entrained for 3 and 8 days after emergence under reverse-cycle conditions and then transferred to constant darkness. However, after 56 h and 80 h respectively of darkness, calling became de-synchronized. Subsequently, it appeared to re-synchronize to a different clock, which approximately correlated with the normal photoperiods the moths had experienced during larval development.