Oviposition and larval dispersal of the common armyworm,Mythimna convecta (Walker) (Lepidoptera: Noctuidae)

The common armyworm, Mythimna convecta (Walker), is a pest of cereal crops and pasture grasses in Australia. During autumn, egglaying in grasslands commonly occurs before plant growth commences. The possible association between oviposition and dried grasses was investigated in field studies of larval distribution in a pasture and a crop habitat, and in laboratory studies of oviposition site preferences and the mode of dispersal of newly hatched first instar larvae. A comparison of a green grass pasture with and without a component of dried grass showed that highest densities of M. convecta larvea. were found in the former. In the laboratory, egg batches were laid between plant parts in close contact, such as between the stem and leaf axil and between seeds in the seed head. The crevices in dried plant material and seed-heads were significantly more acceptable for oviposition than in green foliage. Oviposition in dried plant material enables M. convecta to rapidly colonize ephemeral grasslands immediately after rains when the neonate larvae can disperse and feed on new vegetative growth. Shortly after hatching, 93% of first instar larvae used fine silken threads for aerial dispersal in 0.5 m s^-1 air currents and 43% were blown a distance of more than 1.0 m. In the field, sticky traps caught larvae dispersing in all directions from a central source, and indicated that approximately 30% were transported 2 m after 7 days of light to medium breezes. Terminal velocities of newly hatched larvae were 20–130 cm s^-1, depending on the length of the trailing thread.     

A flagellate protozoan from Mythimna separata (Lepidoptera: Noctuidae)

Abstract

A trypanosomatid flagellate, Herpetomonas sp., is recorded from an adult Mythimna separata (Walker) (Lepidoptera: Noctuidae). Promastigote, paramastigote, and opisthomastigote stages measured 3.4–12 × 1.3–4.3 μm, Giemsastained, with flagella measuring 13.8–18.7 μm. A few amastigotes (3.9–8.3 × 3.7–6.7 μm) were also observed. Fourteen larvae fed with a suspension of flagellates were found later to be infected with the protozoan. No flagellate infections were found in the hymenopteran parasitoid, Apanteles ruficus (Haliday), or the hyperparasitoid, Trichomalopsis sp.     

Parasitoids and hyperparasitoids of Mythimna convecta (Walker) (Lepidoptera: Noctuidae) larvae infesting late-maturing maize in southern New South Wales

Abstract  After several reports of late-maturing maize in Murrumbidgee valley, southern New South Wales, Australia, being severely damaged by armyworm, five crops were surveyed for the presence of larvae in April 2003. Mythimna convecta Walker (Lepidoptera: Noctuidae) was the only species successfully reared from armyworm larvae collected in the field. Ninety-six per cent of armyworm larvae collected were parasitised. Five parasitoid species, Cuphocera sp. nr pilosa (Malloch), Ceromya horma (Malloch), Tritaxys scutellate (Macquart), Chaetophthalmus sp. (Diptera: Tachinidae) and Netelia sp. (Hymenoptera: Ichneumonidae) were reared from M. convecta larvae. Cuphocera sp. nr pilosa was the most frequently encountered parasitoid being reared from 83% of M. convecta larvae collected. Examination of maize plants at each collection site showed high numbers of tachinid puparia adhering to plants. Cuphocera sp. nr pilosa was the only species reared from these puparia. Of the Cu . sp. nr pilosa puparia collected, 23–83% were parasitised by five parasitoid wasps: Trichomalopsis sp. Crawford (Hymenoptera: Pteromalidae), Brachymeria sp. Westwood, Eupelmus sp. Dalman (Hymenoptera: Chalcididae), Perilampus sp. Latreille (Hymenoptera: Perilampidae) and a species belonging to the family Diapriidae.     

A new cell line from larval fat bodies of the bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

Summary A new cell line, designated IOZCAS-Ha-I, was initiated from the fat body of larvae of Helicoverpa armigera (Lepidoptera: Noctuidae) in TNM-FH medium containing 10% fetal bovine serum. Spherical cells were predominant among the various cell types. The cell line showed a typical lepidopteran chromosome pattern ranging from 58 to 239 chromosomes in the majority of the cells, it was confirmed to have originated from the H. armigera by the DNA amplification-fingerprinting polymerase chain reaction (DAF-PCR) technique. The new cell line was only slightly susceptible to the multiple nucleocapsid nuclear polyhedrosis viruses (NPV) from H. armigera.     

In vitro calcification in human osteoblastic cell line derived from periosteum

Human osteoblastic cell cultures were established from human periosteum, identified on the basis of high alkaline phosphatase activity in the confluent state. The cells in the growing state showed a fibroblastic morphology and grew to form multiple layers. The cells could spontaneously calcify, and the process was accelerated by alpha-glycerophosphate. Minerals deposited on the cells consisted exclusively of calcium and phosphorus, and matured into hydroxyapatite crystals. The calcification was stimulated by the treatment with 1 alpha,25-dihydroxy-vitamin D3. These results indicate that the osteoblastic cells have the capacity to differentiate into osteocytes and form calcified human bone tissue in vitro.     

A new cell line from Spodoptera exigua (Lepidoptera: Noctuidae) and its differentially expressed genes

A new cell line, designated IOZCAS‐Spex XI, was established from the pupal ovaries of Spodoptera exigua (Lepidoptera: Noctuidae) in TNM‐FH medium containing 10% foetal bovine serum. The spherical cells were predominant among the various cell types. The population‐doubling time during the logarithmic phase of growth was 81.7 h. It was confirmed that the cell line originated from S. exigua by DAF‐PCR technique. Analysis of susceptibility to baculovirus showed that the new cell line was susceptible to S. exigua nucleopolyhedrovirus (SeNPV), Autographa californica multiple NPV (AcMNPV) and slightly susceptible to S. litura NPV (SpltNPV), while not permissive to Helicoverpa armigera NPV and Hyphantria cunea NPV (HcNPV). Real‐Time PCR analysis was carried out to compare some differentially expressed genes between the cell line and the primary culture. The result showed that marked significant differences were observed in the expression of the genes of SUMO‐1 activating enzyme, BCCIP‐like protein, 10 kDa HSP, CypA, receptor for activated PKC, PDI‐like protein ERp57, ALDH, DEAD box ATP‐dependent RNA helicase‐like protein (P < 0.01), while a significant difference was obtained in the expression of GST gene between the cell line and the primary culture (P < 0.05).     

In vitro Metabolism of Flupyrazofos to Plutella xylostella (Lepidoptera: Yponomeutidae) and Spodoptera exigua (Lepidoptera: Noctuidae)

In vitro metabolism study was conducted to observe the role of enzymes and its kind responsible for tolerance in metabolism of flupyrazofos to Diamond backmoth (DBM), Plutella xylostella as a susceptible insect and Beet armyworm (BAW), Spodoptera exigua as a tolerance insect. The results indicated that there were some differences between the DBM (LD₅₀, 0.54 ppm) and BAW (LD₅₀, >2,000 ppm). In the microsomal fractions, the 4th instar BAW larvae metabolized almost the same amount (about 38%) of flupyrazofos with or without NADPH, but it increased the amount of flupyrazol in the 3rd instar larvae of DBM. Flupyrazofos-oxon was slightly increased with the addition of NADPH in both species. With the addition of piperonyl butoxide (PB) or Iprobenfos (IBP), the amount of flupyrazol was decreased to one third of flupyrazol produced by NADPH in BAW and one fourth of flupyrazol produced by NADPH in DBM. Some effects of inhibitors, PB and IBP, and co-factor, NADPH, were showed when flupyrazofos was metabolized with or without them. This means that esterase and oxidase are responsible for the tolerance mechanism of flupyrazofos in DBM and BAW.     

A new cell line from the embryonic tissue of Helicoverpa armigera HBN. (Lepidoptera: Noctuidae)

A new cell line from the embryonic tissue of Helicoverpa armigera was established and designated as NIV-HA-197. It was maintained in TNM-FH medium supplemented with 10% fetal bovine serum. The cell line at passage 20 had a heterogeneous population of cells consisting of mainly epithelial-like cells (70%), followed by fibroblast-like (27%), and multinucleated giant (3%) cells. The chromosome number ranged from 45 to 185. The growth curve at passage 40 showed a fivefold increase in cell number with a population-doubling time of approximately 60 h. The cell line was found infected with the microsporidium Nosema heliothids at passage 9. Using the antiprotozoan drug Metrogyl 400 and simultaneous heat treatment, the parasite was removed from the culture. The cell line can be cryopreserved for 30 mo. The species specificity of the new cell line was determined by studying the isoenzyme profile of four enzymes, viz., lactate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, and glucose 6-phosphate dehydrogenase, and by heteroduplex analysis. Heteroduplex analysis was used to analyze the mitochondrial 16S ribosomal ribonucleic acid gene sequences along with the host insect gene sequences, and 100% homology was obtained, confirming the conspecificity of the cell line. The cell line was found to be susceptible to the baculoviruses Autographa californica multiple nucleopolyhedrovirus, Spodoptera litura multiple nucleopolyhedrovirus, and H. armigera single nucleopolyhedrovirus (HaSNPV). More than 90% of the cells were infected by HaSNPV on the seventh post infection day (PID), and 28.8 x 10(6) NPV/ml was yielded on the 10th PID. The in vitro-grown HaSNPV caused 100% mortality, when fed to the second instar H. armigera larvae, in 6 d. Cessation of feeding was observed on the second PID.     

Autumn migration of Mythimna separata (Lepidoptera: Noctuidae) over the Bohai Sea in northern China

The autumn migration of Mythimna separata (Walker) (Lepidoptera: Noctuidae) across the Bohai Sea was observed with a scanning entomological radar and a searchlight trap at Beihuang, an island located in the center of the Bohai Gulf of northern China, in 2003-2006. During the autumn migration, M. separata flew at the altitudes of 50-500 m, with a displacement speed of 4-12 m/s, toward the southwest. Variations of area density of the radar targets and of catches in the searchlight trap through the night indicated that the flight duration of M. separata was approximately 10 h. Based on these observations, M. separata that originated in northeastern China (i.e., Liaoning, Jilin, and Heilongjiang provinces and part of the Inner Mongolia autonomous region) could immigrate into eastcentral China and subsequently to southern China (i.e., Fujian, Guangdong, and Guangxi provinces) within a week for overwintering.     

New cell lines from larval fat bodies of Spodoptera exigua: characterization and susceptibility to baculoviruses (Lepidoptera: Noctuidae)

Two new cell lines, designated IOZCAS-Spex-II and IOZCAS-Spex-III, were initiated from the fat bodies of larvae of the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae) in TNM-FH medium containing 10% fetal bovine serum. The spherical cells were predominant among the various cell types and measures approximately 15 microm in diameter. The cell lines were mainly composed of tetraploid cells with chromosome numbers ranging from 116 to 131 (n=31). The cell lines were confirmed to have originated from the S. exigua by DAF-PCR technique. They were susceptible to the multiple nucleocapsid nuclear polyhedrosis viruses from S. exigua.     

The role of nectar plants in severe outbreaks of armyworm Mythimna separata (Lepidoptera: Noctuidae) in China

The period from March to mid April, when oriental armyworm Mythimna separata (Walker) moths migrate from South China to Central China over several nights, is an important window of time in annual armyworm population increase. The presence of nectar sources along the pathway of the migratory population is a prerequisite for moths to reach target habitats and lay eggs. Using flowering periods and geographic distributions, the major spring nectar plants suitable for M. separata moths were identified from among 102 species/varieties of apicultural nectar plants. The nectar plants proposed as important to M. separata include milk vetch Astragalus sinicus L., rape Brassica napa L. and six other species. Spearman's rank correlation analyses were conducted between the annual population size of M. separata and the acreages of milk vetch and rape in the daily stopover areas for migrating populations of M. separata during 1950-1979. The Spearman's coefficient between milk vetch and M. separata was 0.6259 and the correlation was highly significant (P<0.001). Further regression analysis with data from 1950-1979 and from 1980-1992 also revealed a close relationship between annual acreage of damaged crops/wheat and acreage of milk vetch. These results strongly suggest that the unprecedented enlargement in the geographic distribution of milk vetch from Central China into South China was the key factor in the frequent severe oriental armyworm outbreaks that occurred during 1966-1977. This is thought to be the first report in the world that reveals the key role of nectar sources in long distance, regional scale, migration of moths. The argument for the key role of milk vetch is supported by the simultaneous decline in the level of damage inflicted by M. separata and the acreage of milk vetch after 1980.     

Superparasitism behavior and host discrimination of Campoletis chlorideae (Ichneumonidae: Hymenoptera) toward Mythimna separata (Noctuidae: Lepidoptera)

The oriental army worm, Mythimna separata Walker (Lepidoptera: Noctuidae), was adopted as a host to keep Campoletis chlorideae Uchida (Hymenoptera: Ichneumonidae) populations in the laboratory, because it can be mass reared. However, wasp cocoon production in this system via single parasitism was not satisfactory. To improve the rearing efficiency of C. chlorideae on M. separata, we studied the effects of superparasitism on the offspring production of C. chlorideae, as well as the host discrimination ability of the adult wasps in the laboratory. The results showed that, compared with single parasitism, both double and quintuple parasitism significantly increased C. chlorideae cocoon production in M. separata without a significant effect on emergence, weight, or sex ratio of adult wasps at the P < 0.05 level. Host selection experiments suggested that C. chlorideae preferred to lay eggs on unparasitized hosts rather than freshly parasitized hosts but could distinguish neither the 24-h postparasitization hosts from the unparasitized hosts nor the freshly self-parasitized hosts from the conspecifically parasitized hosts. No-choice behavior tests indicated that C. chlorideae took significantly longer time to accept the freshly parasitized hosts containing more than two eggs compared with the unparasitized hosts. In the end, superparasitism behavior and its application in the mass rearing of this endoparasitoid are discussed.     

Primary and continuous midgut cell cultures from Pseudaletia unipuncta (Lepidoptera: Noctuidae)

Midgut epithelial cells were isolated from fifth-instar Pseudaletia unipuncta larvae by collagenase treatment of midgut tissue, and cultured in TNM-FH medium. Long-term continuous culture and maintenance of midgut cells were achieved with P. unipuncta armyworm intestinal cells. Several cells lines were obtained from these P. unipuncta primary cultures, and they have been subcultured and maintained for over 24 mo. The three major midgut cell types were present in the cultures, including stem (regenerative), columnar, and goblet cells. In vitro morphogenesis and differentiation of columnar and goblet cells from stem cells were observed. There appeared to be a cycle of cell death of goblet and columnar cells followed by their replacement from stem cells every 7-8 wk. After approximately six passages, the cell density in T-flasks appeared to be somewhat constant, reaching 10(3)-10(4) cells per milliliter of medium. The columnar cells are round to rectangular in shape and possess a brush border, while the goblet cells have a classic flask-like shape with a central cavity. Peritrophic membrane-like secretions were observed in all the culture flasks. Infection of these cells with multiply embedded nucleopolyhedrovirus was confirmed, and we conclude that these midgut cells can be used as an in vitro model system to study early events in baculovirus infection.     

一日龄粘虫不同时长吊飞对生殖及寿命的影响

【目的】本研究旨在阐明羽化后1日龄粘虫Mythimna separata(Walker)的飞行潜力以及飞行对成虫生殖和寿命的影响,从而进一步解析粘虫的迁飞行为特征。【方法】利用室内昆虫飞行磨计算机数据采集系统,研究了羽化后1日龄粘虫吊飞6、9和12 h的飞行能力差异,以及不同时长吊飞后粘虫的生殖参数和成虫寿命的变化情况。【结果】结果表明,1日龄雌雄蛾均具有一定的飞行能力,但当吊飞时间超过9 h之后,成虫的飞行时间、距离和速度均不再显著上升。除吊飞12 h成虫的飞行速度外,其它处理雌雄蛾的飞行能力之间无显著差异。羽化后1日龄不同时长吊飞对粘虫的生殖均有显著的促进作用,但随着吊飞时间的延长,这种促进作用显著降低。6、9和12 h吊飞处理均导致成虫产卵前期显著缩短,但仅吊飞6和9 h成虫的产卵量和产卵历期显著高于对照。吊飞12 h成虫的产卵量和产卵历期则显著低于吊飞6和9 h的成虫,并和对照差异不显著。吊飞12 h后,雌雄成虫寿命均显著缩短。【结论】羽化后1日龄飞行显著促进粘虫的生殖,但随着飞行时间的延长,飞行对生殖的促进效应显著减弱。     

Amplified fragment length polymorphism analysis of Mythimna separata (Lepidoptera: Noctuidae) geographic and melanic laboratory populations in China

Genetic diversity within and among three wild-type natural populations and one melanic laboratory population of Mythimna separata (Walker) (Lepidoptera: Noctuidae) were evaluated using amplified fragment length polymorphism (AFLP) analysis. Although extensive genetic diversity occurs among individuals from different geographic populations (P = 54.5%, h = 0.209, I = 0.305), the majority of the genetic diversity is within populations and not between populations (G(ST) = 0.172), indicating high gene flow (N(M) = 2.403) and suggesting that M. separata in northern China are a part of a single large metapopulation. Genetic diversity in the natural populations was significantly higher than that in the melanic laboratory population (with P = 43.4% versus P = 25.9%, h = 0.173 versus h = 0.086, and I = 0.251 versus I = 0.127), suggesting that the melanic laboratory population is narrowly genetic-based and genetically uniform. Genetic similarities based on AFLP data were calculated, and cluster analysis was preformed to graphically display groupings between individuals and populations. Individuals from the same region were not grouped together in cluster analysis of three natural populations, whereas melanic individuals from laboratory population were grouped together very well. Four subpopulations were clustered into two broad groups. Melanic laboratory population became a single group, which had apparent differentiation from the other group in which three natural subpopulations were included. These results indicated that although high genetic variability existed among the individuals of natural populations, there was little genetic differentiation among three geographic populations that could be explained by the effects of the long distance migration of the oriental armyworm in China enhanced the level of gene flow. Influences of migration on the genetic polymorphism and differentiations that make a significant contribution to evolution in this insect are reviewed.