Xin-Cheng Zhao

Autumn Migration of Mythimna separata (Lepidoptera: Noctuidae) over the Bohai Sea in Northern China

Abstract 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 ≈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.

Arrangement of Output Information from the 3 Macroglomerular Units in the Heliothine Moth Helicoverpa assulta: Morphological and Physiological Features of Male-Specific Projection Neurons

Helicoverpa assulta is exceptional among heliothine species studied so far as concerns composition of the pheromone blend. Previous reports have accordingly pointed out distinct characteristics in the male-specific olfactory pathway of this species, peripherally by an unusual distribution of 2 sensillum categories and centrally by a particular anatomical arrangement of the male-specific glomeruli constituting the macroglomerular complex (MGC). In order to determine the physiological tuning of the 3 MGC units in this species, we have characterized male-specific antennal-lobe projection neurons morphologically and physiologically by use of the intracellular recording and staining technique combined with confocal microscopy. The results show 2 projection neuron types of equal numbers, one that responds to the primary pheromone component, cis-9-hexadecenal, and arborizes in the cumulus and one that responds to the interspecific signal, cis-9-tetradecenal, and arborizes in the dorsomedial unit. A third type responded to the secondary pheromone component, cis-11-hexadecenal, and innervated the smaller ventral unit. The results complement previous findings from tracing of physiologically identified receptor neurons and determine for the first time the functional specificity of each glomerulus in the MGC of H. assulta. The results are particularly interesting because heliothine moths are attractive objects for comparative studies addressing questions concerning divergence of male-specific olfactory characteristics related to speciation.

Processing of Pheromone Information in Related Species of Heliothine Moths

In heliothine moths, the male-specific olfactory system is activated by a few odor molecules, each of which is associated with an easily identifiable glomerulus in the primary olfactory center of the brain. This arrangement is linked to two well-defined behavioral responses, one ensuring attraction and mating behavior by carrying information about pheromones released by conspecific females and the other inhibition of attraction via signal information emitted from heterospecifics. The chance of comparing the characteristic properties of pheromone receptor proteins, male-specific sensory neurons and macroglomerular complex (MGC)-units in closely-related species is especially intriguing. Here, we review studies on the male-specific olfactory system of heliothine moths with particular emphasis on five closely related species, i.e., Heliothis virescens, Heliothis subflexa, Helicoverpa zea, Helicoverpa assulta and Helicoverpa armigera.

A multisensory centrifugal neuron in the olfactory pathway of heliothine moths.

We have characterized, by intracellular recording and staining, a unique type of centrifugal neuron in the brain olfactory center of two heliothine moth species; one in Heliothis virescens and one in Helicoverpa armigera. This unilateral neuron, which is not previously described in any moth, has fine processes in the dorsomedial region of the protocerebrum and extensive neuronal branches with blebby terminals in all glomeruli of the antennal lobe. Its soma is located dorsally of the central body close to the brain midline. Mass‐fills of antennal‐lobe connections with protocerebral regions showed that the centrifugal neuron is, in each brain hemisphere, one within a small group of neurons having their somata clustered. In both species the neuron was excited during application of non‐odorant airborne signals, including transient sound pulses of broad bandwidth and air velocity changes. Additional responses to odors were recorded from the neuron in Heliothis virescens. The putative biological significance of the centrifugal antennal‐lobe neuron is discussed with regard to its morphological and physiological properties. In particular, a possible role in multisensory processes underlying the moths ability to adapt its odor‐guided behaviors according to the sound of an echo‐locating bat is considered. J. Comp. Neurol. 521:152–168, 2013.

Annual Migration of Agrotis segetum (Lepidoptera: Noctuidae): Observed on a Small Isolated Island in Northern China.

Migration behavior of the turnip moth, Agrotis segetum (Lepidoptera: Noctuidae), is not well known by far. Here, we present the data from an 11-year study on A. segetum by means of searchlight trapping and ovarian dissection on Beihuang (BH) Island, which located in the center of the Bohai Strait in northern China. The data showed a large number of A. segetum flight across the strait each year, which provides direct evidence that A. segetum is a long-distance migrant, migrating at least 40 - 60 km to reach the trapping site. The migration period during 2003-2013 ranged from 115 to 172 d. Among the catches, the proportion of females was significantly higher than that of males in each month from May to September. Ovarian dissection showed that the proportion of mated females and the proportion of sexually mature females was significantly higher than that of unmated females and sexually immature females in early summer, respectively, but conversely in autumn. The early summer populations migrate in a south-north direction, which might undertake a long-distance flight on several successive nights. The autumn populations migrate in a north-south direction, which might originate not far from the trapping site. Based on these findings, the migratory physiology of A. segetum was discussed.

Central Projections of Gustatory Receptor Neurons in the Medial and the Lateral Sensilla Styloconica of Helicoverpa armigera Larvae

Food selection behavior of lepidopteran larvae is predominantly governed by the activation of taste neurons present in two sensilla styloconica located on the galea of the maxilla. In this study, we present the ultrastructure of the sensilla styloconica and the central projection pattern of their associated receptor neurons in larvae of the heliothine moth, Helicoverpa armigera. By means of light microscopy and scanning electron microscopy, the previous findings of two morphologically fairly similar sensilla comprising a socketed conic tip inserted into a large peg were confirmed. However, the peg size of the medial sensillum was found to be significantly bigger than that of the lateral sensillum. The sensory neurons derived from each sensillum styloconicum were mapped separately using anterograde staining experiments combined with confocal laser-scanning microscopy. For determining the afferents’ target regions relative to each other, we reconstructed the labeled axons and placed them into a common reference framework. The sensory axons from both sensilla projected via the ipsilateral maxillary nerve to the suboesophageal ganglion and further through the ipsilateral circumoesophageal connective to the brain. In the suboesophageal ganglion, the sensory projections targeted two areas of the ipsilateral maxillary neuropil, one located in the ventrolateral neuromere and the other adjacent to the neuromere midline. In the brain, the axon terminals targeted the dorso-anterior area of the ipsilateral tritocerebrum. As confirmed by the three-dimensional reconstructions, the target regions of the neural projections originating from each of the two sensilla styloconica were identical.

Representation of pheromones, interspecific signals, and plant odors in higher olfactory centers; mapping physiologically identified antennal-lobe projection neurons in the male heliothine moth

The arrangement of anatomically separated systems for information about general and pheromone odorants is well documented at the initial levels of the olfactory pathway both in vertebrates and insects. In the primary olfactory center of the moth brain, for example, a few enlarged glomeruli situated dorsally, at the entrance of the antennal nerve, are devoted to information about female-produced substances whereas a set of more numerous ordinary glomeruli (OG) receives input about general odorants. Heliothine moths are particularly suitable for studying central chemosensory mechanisms not only because of their anatomically separated systems for plant odors and pheromones but also due to their use of female-produced substances in communication across the species. Thus, the male-specific system of heliothine moths includes two sub-arrangements, one ensuring attraction and mating behavior by carrying information about pheromones released by conspecifics, and the other inhibition of attraction via signal information emitted from heterospecifics. Based on previous tracing experiments, a general chemotopic organization of the male-specific glomeruli has been demonstrated in a number of heliothine species. As compared to the well explored organization of the moth antennal lobe (AL), demonstrating a non-overlapping representation of the biologically relevant stimuli, less is known about the neural arrangement residing at the following synaptic level, i.e., the mushroom body calyces and the lateral horn. In the study presented here, we have labeled physiologically characterized antennal-lobe projection neurons in males of the two heliothine species, Heliothis virescens and Helicoverpa assulta, for the purpose of mapping their target regions in the protocerebrum. In order to compare the representation of plant odors, pheromones, and interspecific signals in the higher brain regions of each species, we have created standard brain atlases and registered three-dimensional models of distinct uniglomerular projection neuron types into the relevant atlas.

Glomerular identification in the antennal lobe of the male moth Helicoverpa armigera.

This study investigates anatomical organization of the antennal lobe (AL) glomeruli of the male cotton bollworm Helicoverpa armigera by synaptic antibody staining combined with three‐dimensional reconstruction. To identify all glomeruli, their boundaries were accurately determined by means of several additional staining techniques visualizing the neuron categories forming the characteristic spherical neuropils. In total, 78–80 glomeruli were identified in the male H. armigera. The number of glomeruli was considerably larger than that previously reported in this species. Thus, compared with previous studies, we identified 15 new glomeruli, G63–G77. Most of them are located in the posterior part of the AL, which was previously considered to be a part of the protocerebrum. From the general anatomical organization of the AL glomeruli of H. armigera, we classified these neuropil structures into four groups, the macroglomerular complex, posterior complex, labial‐palp pit organ glomerulus, and ordinary glomeruli. The complete identification of glomeruli is important for future studies seeking to explore further the coding mechanisms residing within the primary olfactory center of the moth brain. J. Comp. Neurol. 524:2993–3013, 2016.

Annual Migration of Cabbage Moth, Mamestra brassicae L. (Lepidoptera: Noctuidae), over the Sea in Northern China.

The cabbage moth, Mamestra brassicae L. (Lepidoptera: Noctuidae), is a serious pest of vegetable crops throughout the world. In order to determine whether or not M. brassicae is a migrant, and if yes, what is the pattern of M. brassicae seasonal migration, a long-term study on M. brassicae from April to October in 2003–2014 was carried out by means of a searchlight trap on a small island located in the center of the Bohai Strait. The results show that a large number of M. brassicae were trapped every year on the island, which indicates that M. brassicae is a migrant and migrated at least 40–60 km across the Bohai Strait. The mean migration period of M. brassicae over the sea within one year is 151 ± 8 d in 2003–2014, with the shortest time span 78 d in 2003 and the longest 189 d in 2014, respectively. The number of M. brassicae captured, however, varies considerably between months or years. The majority of captures were female, with different levels of ovarian development and mating status. Most of the females trapped in May-July during 2010–2014 had a high mating rate and advanced level of ovarian development, suggesting that the migration of this species does not conform to the hypothesis of ‘oogenesis-flight syndrome’. The findings of the present study are beneficial to the development of forecasting systems and management strategies of M. brassicae.

Characteristics of morphology, electrophysiology, and central projections of two sensilla styloconica in Helicoverpa assulta larvae.

The medial and lateral styloconic sensilla, constituting the main taste organs of lepidopterous caterpillars, were investigated in the oligophagous species, Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae). In this paper, the two sensilla were morphologically and physiologically characterized by scanning electron microscopy and tip recordings, respectively. The central projections of their respective sensory neurons were mapped by anterograde staining experiments combined with confocal laser scanning microscopy. The results showed that the two sensilla are in general morphologically similar. However, the size of the peg on the medial sensillum is significantly greater than that of the lateral. Tobacco leaf saps, sinigrin, and nicotine elicited strong responses from neurons housed by the medial sensillum, whereas sucrose activated primarily the lateral sensillum. All stained neurons in either sensillum showed a projection pattern involving axons entering the subesophageal ganglion through the ipsilateral maxillary and passing further on through the ipsilateral circumesophageal connective to the tritocerebrum of the brain. In the subesophageal ganglion, the axons targeted two areas: the ventrolateral section and the region near the neuromere midline. One distinction between the staining patterns originating from the two sensilla, however, is that axons arising from the medial sensillum, and not the lateral, give off some additional neural branches in the subesophageal ganglion including a few arborizations surrounding a tract, plus a long process extending posteriorly along the midline. Differences in the central projections derived from the two sensilla styloconica have not been reported previously.