Changes of Estrogen in Serum and Estrogen Receptor β in the Relevant Brain Regions Following Mating Behavior of the Male Mandarin Vole Microtus mandarinus

In order to investigate the estrogen and estrogen receptor β changes after mating behavior of male mandarin vole （Microtus mandarinus）, the radioimmunoassay （RIA） and immunohistochemistry methods were used to investigate changes of the serum estrogen （E） concentrations, estrogen immunoreactive neurons （E-IRs） and estrogen receptor β immunoreactive neurons （ERβ-IRs） in the relevant brain regions following mating behavior. Fifteen sexually matured male voles were randomly divided into three groups and treated differently： （1） control group： voles were exposed to clean hard-wood shavings （n=5）, （2） exposure group： voles were exposed to the soiled bedding for more than 24h on which estrous females had been placed （n=5）, and （3） mating group： voles were placed with an estrous female for more than 24h （n=5）. The results showed circulating serum E concentrations were significantly higher in the mating group than in the exposure group and the control group, and there were no significant difference between the exposure group and the control group. E-IRs and ERβ-IRs were detected in the following brain regions related to mating behavior： the arcuate nucleus （ARC）, bed nucleus of the stria terminalis （BST）, lateral septal nucleus （LS）, medial amygdaloid nucleus （ME）, medial preoptic area （MPO） and ventromedial hypothalamic nucleus （VMH）. The results showed that there were significantly more E-IRs in the six brain regions in the mating group than in the control group and the exposure group, and there were no significant difference between the exposure group and the control group except for LS. There was no significant difference in ERβ-IRs in the six brain regions among the three groups, and there were some lighter -stained ERβ-IRs in these brain regions. The results suggested that estrogen affect mating activity of male mandarin voles, but ERβ might not play an important role in mating behavior of male mandarin voles. Instead, it might be through other receptors.

Stk2,a Mitogen-Activated Protein Kinase from Setosphaeria turcica,Specifically Complements the Functions of the Fus3 and Kss1 of Saccharomyces cerevisiae in Filamentation,Invasive Growth,and Mating Behavior

Setosphaeria turcica,an essential phytopathogenic fungus,is the primary cause of serious yield losses in corn; however,its pathogenic mechanism is poorly understood.We cloned STK2,a newly discovered mitogen-activated protein kinase gene with a deduced amino acid sequence that is 96% identical to MAK2 from Phaeosphaeria nodorum,56% identical to KSS1 and 57% identical to FUS3 from Saccharomyces cerevisiae.To deduce Stk2 function in S.turcica and to identify the genetic relationship between STK2 and KSS1/FUS3 from S.cerevisiae,a restructured vector containing the open reading frame of STK2 was transformed into a fus3/kss1 double deletion mutant of S.cerevisiae.The results show that the STK2 complementary strain clearly formed pseudohyphae and ascospores,and the strain grew on the surface of the medium after rinsing with sterile water and the characteristics of the complementary strain was the same as the wild-type strain.Moreover,STK2 complemented the function of KSS1 in filamentation and invasive growth,as well as the mating behavior of FUS3 in S.cerevisiae,however,its exact functions in S.turcica will be studied in the future research.

Mating behavior induces changes of expression of Fos protein,plasma testosterone and androgen receptors in the accessory olfactory bulb (AOB) of the male mandarin vole Microtus mandarinus

In order to investigate the neuroendocrine mechanism of the mating behavior in the adult male mandarin voles Microtus mandarinus,the radioimmunoassay(RIA)and immunohistochemistry methods were used to investigate the differences in plasma testosterone(T)concentrations and distribution of T immunoreactive neurons(T-IRs),androgen receptor immunoreactive neurons(AR-IRs)and Fos protein immunoreactive neurons(Fos-IRs)in the accessory olfactory bulb(AOB)and the main olfactory bulb(MOB)following exposure to clean hard-wood shavings(control group),soiled bedding(exposure group)or contact with an estrous female(mating group).Results showed that plasma T concentration was significantly higher in the mating group than that in the exposure group,and both the mating group and the exposure group displayed significantly higher plasma T concentration than the control group.T-IRs,AR-IRs and Fos-IRs were investigated with the immunohistochemistry method in granule cell(GC)and mitral cell(MC)of the MOB and the AOB in the three groups.There were significantly more T-IRs,AR-IRs and Fos-IRs in MC and GC of the AOB in the mating group than that in the exposure group or the control group.T-IRs,AR-IRs and Fos-IRs did not show significant differences between the exposure group and the control group.Furthermore,obvious differences in MC and GC of the MOB were not found among the three groups.The results confirm that both changes of T and AR in the AOB might be underlying mating behavior in the adult male mandarin voles.

AN OBSERVATION STUDY ON BIOLOGICAL CHARACTERS AND MATING BEHAVIOR OF DAHURIAN CASEBEARER(COLEOPHORA DAHURICA FLKV.)ADULTS

Through rearing tests and observations under laboratory conditions, biological characters and mating behavior of Dahurian casebearer (Coleophora dahurica Flkv.) adults were studied in this paper. Results discovered that the time of emergense of male adults was 24h to 36h earlier than that of female adults. The peak of sex pheromone secretion of the female adults appeared of 24h to 42h after the emergence. The reaction of male adults on the sex pheromone was most sensitive during 30h to 48h after emerged, so the earlier emergence of the male adults was favorable to mating. The phenomenon of repeated mating did not exist in female adults, but the male adults could mate for several times, with maximum of 4 times. There was a definite behavioral reaction during the mating period of male and female adults, and the certain period of feeding and mating times could affect the adults longervity to a certain degree.

Relationship between biological features and the SMVS's inhibitory effect on mating behavior of Nilaparvata lugens (Stal)

Our previous studies revealed that second malevibration signal (SMVS) restrained the matingbehavior of N. lugens, the influences of threebiological features (density, age, and wingform) on SMVS’s inhibitory effect were hereinstudied by playing back its record. The dura-tion of playback was 4 h. Except otherwisestatement, N. lugens tested were virginmacropterous males and females aged 4-6 d af-ter emergence, and the density was 5 pairs (5females and 5 males) of N. lugens per cage (4cm in diameter and 8 cm in height). The in-hibitory effect of SMVS was evaluated usingmating rate (i. e. the rate of females withspermatophore). The results were as follows:

Mating behaviors in ovoviviparous black rockfish(Sebastes schlegelii):molecular function of prostaglandin E2 as both a hormone and pheromone

Prostaglandins(PGs)are profound hormones in teleost sexual behavior,especially in mating.PGs act as pheromones that affect the olfactory sensory neurons of males,inducing the initiation of a series of mating behaviors.However,the molecular mechanism by which PGs trigger mating behavior in ovoviviparous teleosts is still unclear.In the present study,we employed the ovoviviparous black rockfish(Sebastes schlegelii),an economically important marine species whose reproductive production is limited by incomplete fertilization,as a model species.The results showed that when the dose of PGE2 was higher than 10 nmol/L,a significant(P<0.05)increase in mating behaviors was observed.Dual-fluorescence in situ hybridization indicated that PGE2 could fire specific neurons in different brain regions and receptor cells in the olfactory sac.After combining with specific neurons in the central nervous system(CNS),a series of genes related to reproduction are activated.The intracerebroventricular administration of PGE_(2) significantly increased lhb levels(P<0.05)in both sexes.Moreover,steroidogenesis in gonads was also affected,inducing an increase(P<0.05)in E_(2) levels in males and T levels in females.PGE_(2) levels were also increased significantly(P<0.05)in both sexes.The present study revealed that PGE2 can activate mating behavior in black rockfish in both hormone and pheromone pathways,leading to variations in sex steroid levels and activation of reproductive behaviors.Our results provide not only novel insight into the onset of mating behaviors in ovoviviparous teleosts but also solutions for the incomplete fertilization caused by natural mating in cage aquaculture.

MATING BEHAVIOR OF THE CAT FLEA,CTENOCEPHALIDES FELIS BOUCHE(SIPHONAPTERA:PULICIDAE) AND MALE RESPONSE TO FEMALE EXTRACT ON AN ARTIFICIAL FEEDING SYSTEM

The mating behavior of cat flea, Ctenocephalides felis (Bouche) was studied on an artificial feeding device. Male and female can mate repeatedly with same partner or different ones. In the situation of male: female ratio of 1:5, each mating lasted an average of 6.6 min, with a mean interval between matings at 2.5 min., compared to 11.1 min and 12.1 min respectively in a cell with 5 males and 1 female. As many as 48 mating events were observed for one male during an 8 h period. One female mated 27 times in 7 h with 5 males in the same cell. Newly emerged males and females can not mate before blood meal and about 24 h blood feeding is required for successful mating. Newly emerged males can not mate with fed females (fed for 48 h), but fed males can mate with newly emerged females who are feeding the blood. Significantly more male contacts and male male mating attempts were observed after the paper treated with female extract was introduced into the cell. The paper contacts and mating attempts were 16.75-32.25 times and 15.75-31.38 times, respectively, on average during a period of 20 min when different doses (FE) of extract were provided.

Adaptive strategies of overwintering adults： Reproductive diapause and mating behavior in a grasshopper, Stenocatantops splendens （Orthoptera： Catantopidae）

To understand the adaptive strategies of the overwintering adults of Steno- catantops splendens, the mechanism of maintenance and termination of the reproductive diapause, the variation in mortality between overwintering females and males, and the mating strategy of the males were investigated. The results indicated that the adult repro- ductive diapause in natural conditions was mainly regulated by photoperiod in the fall - long photoperiods promoted reproductive development and short photoperiods maintained reproductive diapause, and the sensitivity of the overwintering adults to photoperiod was over before the end of the winter. When transferred from natural conditions to controlled laboratory conditions on dates from September through February, pre-oviposition became increasingly shorter with increasingly deferred transfer dates regardless of photoperiod conditions. The adults treated with low temperature for 30 days in September through November had significantly shorter pre-oviposition, suggesting that low temperatures in winter had an important role in the termination of reproductive diapause. The female had a significantly lower supercooling point than the male, which was related to their lower mortality after winter. In addition, observations of wild populations of the species indicated that mating behavior prior to winter and the duration of pre-mating period were not affected by photoperiod; mating and sperm transfer were mostly completed by November. Compared with females only mating before winter, females mating in the spring had shorter life span, longer pre-oviposition, lower hatching rate and laid fewer egg pods while showing no significant difference with regard to ovipositional interval, per pod number of eggs and nymph dry weight.

Mating behavior and sexual selection in a polygamous beetle

Mating behavior and sexual selection in relation to morphometric traits in a polygamous beetle, Glenea cantor （F.） （Coleoptera： Cerambycidae）, were investigated. Upon encounter, a male approached a female, mounted her, grasped her terminal abdomen with his hind tarsi, and attempted to mate. Successful mating lasted about 3.5 h. Although all traits measured in females and half of traits in males were significantly correlated with mating success, the primary selection on virgin females was the genital trait, thebursa copulatrix length, and that on males was the body length and hind tarsal length. Longer bursa copulatrix accommodated a larger ejaculate, suggesting that this female trait benefits the male that first mates with the female in terms of in- creasing ejaculate size to beat subsequent males in sperm competition. Under a female-biased sex ratio, more than 20% of mat- ings failed within 20s after the male genitalia had been inserted into hers, suggesting that males assess genital features of the re- ：male before insemination and undertake cryptic male mate choice. Larger males were more capable of grasping females and achieving mating. During the premating struggle the male almost always used his hind tarsi to lift the female terminal abdomen to the position for his genitalia to insert, and as a result, males with longer hind tarsi achieved higher mating success [Current Zoolo- gy 59 （2）： 257-264, 2013].

Mating behavior of Psammotettix alienus （Hemiptera： Cicadellidae）

The Wheat dwarf virus, the causal agent of the wheat dwarf disease, is trans- mitted by leafhoppers from the genus Psammotettix and currently the main protection strategy is based on the use of insecticide treatments. Sustainable management strategies for insect vectors should include methods that are targeted to disrupt reproductive behavior and here we investigated the mating behavior of Psammotettix alineus （Dahlbom 1850） in order to determine the role of vibrational signals in intra-specific communication and pair formation. Both genders spontaneously emit species- and sex-specific calling songs that consisted of regularly repeated pulse trains and differ primarily in pulse train duration and pulse repetition time. Females preferred the conspecific male calling song. After a coordinated exchange of pulse trains, the male approached the stationary female. During the close range courtship and also immediately prior to copulatory attempts distinct male vibrational signals associated with wing flapping and wing vibrations were recorded from the substrate. In the presence of a receptive female, competing males emitted vibrational signals most likely aimed to interfere with male-female interaction. Mated females re- gained sexual receptivity after they laid eggs. Although results suggest that the viruliferous status of insects may have an effect on vibrational songs, our current results did not reveal a significant effect of virus on leafhopper performance in mating behavior. However, this study also suggests, that detailed understanding ofplant-vecto^virus interactions relevant for vector mating behavior is essential for trying new approaches in developing future control practices against plant viruses transmitted by insect vectors.

Reproductive behavior in horseshoe crabs:Does density matter?

While the four species of horseshoe crabs share many common reproductive traits with respect to their reproductive systems, they do differ with respect to their mating behavior （monogamy vs. polygynandry）. Past research has attributed these differences to a number of factors including： spawning densities, operational sex ratios （OSR＇s）, male condition （or age）, environmental and/or genetic factors, or a combination thereof. Mating behaviors in the three Asian horseshoe crab species （Tachy-pleus gigas, T. tridentatus, and Carcinoscorpius rotundicauda） with low spawning densities and 1：1 operational sex ratios are typically monogamous. In Limuluspolyphemus, mating behavior is more variable ranging fi＇om monogamy to polygynandry. Here we provide evidence, through a long term behavioral study, that variation in mating behavior is influenced by population density in L. polyphemus. Our study population on two beaches in Connecticut （Long Island Sound） have a spawning density 400 times less than that found in Delaware Bay （0.002 females/m2 vs. 0.8 females/m2） but similar operational sex ratios. Between 90%-95% of all spawning females in CT were paired with only one male, thus exhibiting monogamous behavior. In contrast, between 30 and 60% of spawning females in Delaware Bay have more than one mate and produce clutches of eggs with multiple paternities. Male condition played no role in mating behavior in CT populations. We also observed that on average 18% of the females on the spawning beaches are single. These results suggest that population density is an important condition that determines mating behavior. Also, low population density may lead to decreased mate finding ability and lost opportunities for spawning

Species Concepts as Applied to the Whitefly Bemisia tabaci Systematics:How Many Species Are There?

The worldwide distribution and extensive genetic diversity of the whitefly,Bemisia tabaci,has long been recognized.However,the levels of separation within B.tabaci and the nomenclature of the various genetic groups have been a subject of debate.Recent phylogenetic analyses indicate that B.tabaci is a complex composed of 28 morphologically indistinguishable species.In this article,we first review the debate and difficulties associated with B.tabaci＇s taxonomy and systematics,and argue for the need to apply the biological species concept in order to elucidate B.tabaci＇s systematics.We summarize the accumulated genetic and behavioural data on reproductive incompatibilities evident amongst phylogenetic mtCOI groups of B.tabaci.Crossing studies have been conducted with 14 of the 28 putative species covering 54 reciprocal inter-species pairs,and observations on mating behaviour have been conducted for seven species pairs.Data from both crossing trials and behavioural observations indicate a consistent pattern of reproductive isolation among the putative species.We then discuss the technical and conceptual complexities associated with crossing experiments and behavioural observations designed to reveal reproductive incompatibility.Finally,we elaborate on a strategy for further clarifying the pattern of reproductive isolation between B.tabaci groups and propose future research directions on the systematics of this complex.

Wing-fanning frequency as a releaser boosting male mating success— High-speed video analysis of courtship behavior in Campoplex capitator, a parasitoid of Lobesia botrana

Campoplex capitator is an ichneumonid parasitoid with a narrow host range,comprising grapevine moth pests.Despite being considered one of the possible candidates for biocontrol of Lobesia botrana,knowledge about its biology is limited and massrearing for commercial purposes is still lacking.This research provides a quantitative analysis of the C.capitator courtship and mating behavior.C.capitator mating sequence was analyzed by high-speed video recordings.Main behavioral parameters,with special reference to male wing fanning and antennal tapping,were quantified and linked with mating success.Furthermore,we analyzed the occurrence of population-level behavioral asymmetries during C capitator sexual interactions and their impact on male success.Results showed that male wing fanning was crucial to successfully approach the female.Males achieving higher mating success performed wing-fanning at higher frequencies over unsuccessful ones.After wing fanning,most of males palpated the females body with their antennae,before attempting copulation.The overall mating success was>70%,with a rather long copula duration(254.76±14.21 s).Male wing-fanning was lateralized on the left at population level,while antennal tapping displays were right-biased.Sidebiased male displays do not differ in terms of frequency and duration of their main features.This research adds basic knowledge to the C.capitator behavioral ecology.Since rearing protocols for C.capitator are being developed male wing fanning frequency may represent a useful benchmark for monitoring mate quality over time,tackling mating success reductions due to prolonged mass-rearing.

The reproductive strategy and the vibrational duet of the leafhopper Empoasca vitis

The recent description of a new vibrational mating disruption method to control the leafhopper Scaphoideus titanus Ball opened questions about its possible application to other leafhopper pests. Since the prerequisite for the method＇s successful application is a deep knowledge of the species mating behavior and the exact role of associated signals, we conducted behavioral assays on the green leafhopper Empoasca vitis Gothe, a pest of grapevine and other crops in Europe and Asia. Laser vibrometer recordings of single and paired individuals （male and female） during a 24-h period enabled us to detect and describe 2 male and 1 female signal. The pair formation starts when the female replies to a male call and a duet is established, then it continues through 2 different behavioral stages： Location and Courtship. The proper courtship begins only when the male locates the female. The latter is characterized by a significant change in temporal parameters that regards both the signals and the duet structure. Although the male calling activity and the female replying rate were the same during the 24 h, a lower number of matings was recorded during the night. We discuss the possible role of vision and of the species ecology as factors of reproductive success and mating strategy. Our conclusion is that the mechanical mating disruption technique seems feasible for future application to this species.

Phylogeny of the industrial relevant,thermophilic genera Myceliophthora and Corynascus

Species of the genus Myceliophthora and its teleomorph Corynascus have attracted increasing interest due to their potential to produce thermostable enzymes.This study re-assessed the phylogenetic relationship of 49 isolates of nine species belonging to Myceliophthora and Corynascus.One species,M.vellerea,was shown not to belong to the genus Myceliophthora and should be placed in the genus Ctenomyces.The other species belonged to two phylogenetic clusters:mesophilic fungi with the type species M.lutea and C.sepedonium,and thermophilic fungi with M.thermophila,M.hinnulea and C.thermophilus.The phylogenetic data provides no clear separation of the two genera Corynascus and Myceliophthora.To avoid confusion in future taxonomic studies,it is proposed that all existing Corynascus species be renamed to Myceliophthora,which is the old name and the one more frequently used.Furthermore,this study identified two groups within the isolates listed as M.thermophila and assigned one group(five isolates)to M.heterothallica based on AFLP analysis and mating behavior.This study provides new insights into the genetic differences within the genus Myceliophthora and will therefore be essential for the interpretation of future genomic and physiological studies of these species.

Sexual audience affects male's reproduction investment without consequences on reproductive outputs

Males evolved plastic strategies to respond to male-male competition and exhibit adaptive traits and behaviors maximizing their access to the females and limiting sperm competition.Mating behaviors allow males to express quick responses to current sexual audience,that is,the number of nearby conspecifics prone to mate.In contrast,physiological responses are frequently delayed because they are constrained by the time and resources having to be mobilized to produce and export sperm and associated products.This is especially critical in species for which males produce spermatophores.Here we investigated in what extend moth males(the tortricid moth Lobesia botrana)producing spermatophores exhibit plastic behavioral and physiological responses to different sexual audiences before and during mating and the consequences for their reproductive output.We found that males adjusted their mating behaviors and spermatophore size to a potentially elevated risk of sperm competition perceived before mating.In addition,males responded to the closed presence of females during mating by reducing their mating duration.Surprisingly,the various behavioral and physiological responses we highlighted here were not fully reflected in their reproductive performance as we did not reveal any effect on fecundity and fertility of their mate.The selective pressure exerted on males experiencing male-male competition could thus be sufficient to trigger adjustment in male mating behaviors but constrains physiological responses according to the perception of competition.