Male Pupa Research Articles

A review of the genus Neogreenia MacGillivray (Hemiptera: Coccomorpha: Qinococcidae) with descriptions of two new species.

The species of Neogreenia MacGillivray (Hemiptera: Coccomorpha: Qinococcidae) are reviewed. The genus is distributed in the Oriental and Palaearctic Regions and now contains seven species. The present study provides updated generic descriptions of all the developmental stages and a brief account of the biology and life cycle of Neogreenia. The genotype, Neogreenia zeylanica (Green, 1896), is redescribed. The following are described and illustrated for the first time: the second-instar nymph, third-instar female, and male third-instar nymph and pupa of N. zizyphi Tang; the second-instar nymph of N. lonicera Wu & Nan; and the second-instar nymph of N. sophorica Wu. Two new species are described and illustrated: Neogreenia ficus Zheng & Wu, sp. n. based on the adult female and Neogreenia tangi Zheng & Wu, sp. n. based on all female instars. These new species bring the total number of described species of Neogreenia to seven. An identification key to all the developmental stages of Neogreenia, and identification keys to species based on adult females and available nymphal instars are provided.

The Genome of the Yellow Mealworm, Tenebrio molitor: It's Bigger Than You Think.

Insects are a sustainable source of protein for human food and animal feed. We present a genome assembly, CRISPR gene editing, and life stage-specific transcriptomes for the yellow mealworm, Tenebrio molitor, one of the most intensively farmed insects worldwide. Long and short reads and long-range data were obtained from a T. molitor male pupa. Sequencing transcripts from 12 T. molitor life stages resulted in 279 million reads for gene prediction and genetic engineering. A unique plasmid delivery system containing guide RNAs targeting the eye color gene vermilion flanking the muscle actin gene promoter and EGFP marker was used in CRISPR/Cas9 transformation. The assembly is approximately 53% of the genome size of 756.8 ± 9.6 Mb, measured using flow cytometry. Assembly was complicated by a satellitome of at least 11 highly conserved satDNAs occupying 28% of the genome. The injection of the plasmid into embryos resulted in knock-out of Tm vermilion and knock-in of EGFP. The genome of T. molitor is longer than current assemblies (including ours) due to a substantial amount (26.5%) of only one highly abundant satellite DNA sequence. Genetic sequences and transformation tools for an insect important to the food and feed industries will promote the sustainable utilization of mealworms and other farmed insects.

Morphology of the preadult stages of Coronaproctus castanopsis Li, Xu & Wu (Hemiptera: Coccomorpha: Monophlebidae).

All the preadult stages of Coronaproctus castanopsis Li, Xu & Wu, 2023 are described and illustrated (first- and second-instar nymphs, third-instar male and female nymphs, and the male pupa), based on material from Zhejiang Province, China. A key to separate the developmental stages is provided.

Myrmecophily of the enigmatic stag beetleTorynognathus chrysomelinus(Coleoptera: Lucanidae: Lucaninae) with remarks on adult morphology, immature stages, and systematic position

AbstractAdults and various immature stages of the stag beetleTorynognathus chrysomelinusBomans, 1986 were found inside the nests ofPseudolasiusants in Peninsular Malaysia. This paper provides a detailed description ofT. chrysomelinus, which is the first ever record of myrmecophily in this genus and the third record in the family Lucanidae. This is also the first record of myrmecophilous Coleoptera withPseudolasiusants as a host. The description includes the illustration of the male genitalia, mouthparts, larva, and pupa of this genus for the first time. Furthermore, the systematic position ofT. chrysomelinuswas inferred by the molecular phylogenetic analysis of the family Lucanidae. The results suggest thatTorynognathusis a sister taxon to the genusAegus.

Biology and Morphometric Study of Okra Shoot and Fruit Borer <i>Earias Vitella</i> (F.)

Investigations on biology of shoot and fruit borer (Earias vitella) on okra revealed that the freshly laid eggs were spherical, bluish green and finally become brownish. These measured 0.52± 0.02 x 0.51± 0.02 mm.The incubation period was 3.35± 0.76 days, and larval stage passed through four instars. The first,second, third and fourth instar was 2.16± 0.71, 1.43± 0.50, 2.26± 0.73 and 3.30±1.02 days, respectivelywith the total larval period of 9.13± 1.71 days. These were measured 1.77± 0.28, 2.60± 0.40, 6.45± 0.56 and14.15± 2.12 mm long respectively and 0.27± 0.03, 0.36± 0.08, 1.24± 0.07 and 3.23± 0.51 mm, respectivelyin breadth. The pre-pupa lasted for 1.43± 0.50 days. The male and female pupa measured 9.78± 0.46 mmand 3.14± 0.31 mm, 10.14± 0.69 mm and 3.37± 0.09 mm, respectively, with duration of 7.90± 0.92 and7.53± 0.50 days, respectively. The adult male measured 9.13± 0.41 mm in length and 21.02± 0.98 mm inbreadth with expanded wings. Whereas, female 10.71± 1.14 mm in length and 22.30± 1.48 mm in breadthwith expanded wings. The preoviposition, oviposition and post-oviposition periods were 1.50± 0.68, 6.00±1.33 and 4.30± 1.53 days, respectively. The fecundity was 265.00± 86.75 eggs/ female. The total lifecycleof male was 30.51± 6.00 days, while in female it was 32.60± 5.59 days.

Complete mitogenome and phylogenetic significance of Metoecus javanus (Pic, 1913) (Coleoptera: Ripiphoridae) from Southwest China, with notes on morphological traits of adult and immature stages.

Wedge-shaped beetles (Ripiphoridae) not only exhibit enigmatic morphological and biological traits but also disputable phylogenetic positions. At present, however, genetic information regarding this family remains limited. In this study, we report on the complete mitogenome of one ripiphorid beetle, Metoecus javanus (Pic, 1913), from Southwest China, as well as its different developmental stages, populations, and morphological variability. The complete mitogenome of M. javanus was 16 109 bp in length, containing 13 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and a noncoding control region. Of the 37 genes, 23 were located on the majority strand (J-strand) and 14 were located on the minority strand (N-strand). All PCGs started with "ATN" (N represents A, T, G, and C), and terminated with "TAA", except for NAD1 with "TAG" and COX2 with a single "T". The five most used codons in the PCGs were UUA(L), UCU(S2), CCU(P), UCA(S2), and GGA(G), indicating a strong bias toward A + T-rich codons. All 22 tRNAs showed typical cloverleaf structures, except trnS1, which lacked a dihydrouridine (DHU) stem. The control region contained five types of tandem repeats, with the repeat units ranging from 17 to 24 bp. Phylogenetic analysis of the concatenated set of 13 PCGs and two rRNAs (rrnL and rrnS) of M. javanus and 17 other Tenebrionoidea species indicated that M. javanus did not cluster with Pelecotoma fennica (Ripiphoridae: Pelecotominae), another wedge-shaped beetle, but was located at the base of the Mordellidae + P. fennica clade. This reconstruction supported the paraphyly of Ripiphoridae with respect to Mordellidae. Using the mitogenome COX1 data, wedge-shaped beetles from different stages (male adult, female adult, and pupa), different geographical populations (Nujiang and Lincang), and different wasp hosts (Vespidae: Vespa velutina Lepeletier, 1836 and Vespa bicolor Fabricius, 1787) were identified as a same species (i.e., M. javanus). Based on morphological observations of all specimens, we identified and described variability in the adult pronotum, male genitalia, and pupa of M. javanus. The present results provide important genetic and morphological information for further investigations on the phylogenetic position of Ripiphoridae and its evolutionary diversity.

Biology of Pink Bollworm Pectinophora gossypiella (Saunders) (Gelechiidae: Lepidoptera) under Laboratory Conditions

The present study on biology of pink bollworm on two modified artificial diets separately was carried out in the Toxicology laboratory, Department of Agriculture Entomology, Post Graduate Institute, Dr PDKV, Akola during October to February 2018–19 and 2019–20 with controlled conditions such as 25±2oC temperature 65±5% RH and photoperiod of 14:10 h light: dark. Two modified artificial diets were prepared separately with some modifications in the ingredients, pink bollworm larvae are reared on two above said modified artificial diets and observations were recorded on different biological parameters like egg incubation period (days), larval period (days), pupal period (days), adult emergence (%), adult male and female longevity(days), preoviposition period (days), oviposition (days), fecundity(no of eggs per female), total life cycle(days) and on morphometry of different stages pink bollworm like egg, first, second, third and fourth instar larval stages, male and female pupa and male and female adult moth. Our results indicated that pink bollworm has four larval instars with mean larval duration of 18.60±0.54 days in modified artificial diet 1 and 17.72±0.36 days in modified artificial diet 2 population respectively. Total life cycle from egg to adult was 46.74±0.31 and 54.71±0.62 days in modified artificial diet 1 and modified artificial diet 2 population respectively. Total life cycle of pink bollworm was completed somewhat earlier in in modified artificial diet 1 in comparison to modified artificial diet 2. Our results showed that there was only a little difference in the biology of pink bollworm on both modified artificial diets, and hence both modified artificial diets can be used for laboratory rearing purposes.

Multiple Parasitism by Strepsipteran Insects Accelerates Timing of Strepsipteran Adult Emergence

Superparasitism (multiple parasites in a single host) is one of the most significant examples of density-dependent intraspecific competition among insect parasitoids. In this study, we showed that multiple parasitism by Blissoxenos esakii (Insecta, Strepsiptera) induce earlier adult emergence than solitary parasitism (single parasite in a host). Superparasitism creates conditions wherein multiple strepsipteran larvae must compete for the limited (two) exit spaces on the host body for their emergence as adults. As a result, selection might have favored strepsipterans that shortened the period of larval growth, so that they can emerge earlier. Thus, superparasitism causes a dramatic change in the life history of strepsipteran. Photo 1. Dwarf bamboos are common in Yamanashi, Japan. Macropes obnubilus, the host bug of Blissoxenos esakii, are found under the leaf sheath of a dwarf bamboo. Photo credit: Yuta Nakase. Photo 2. Host bug Macropes obnubilus. Bar = 1 mm. Photo credit: Yuta Nakase. Photo 3. Host bug Macropes obnubilus with its wings removed. Two Blissoxenos esakii (a female adult on the left and a male pupa on the right as one faces, respectively) extrude the cephalothoraxes of their puparia from the membranous area between the thorax and abdomen of M. obnubilus. Bar = 1 mm. Photo credit: Yuta Nakase. Photo 4. Abdomen of a dissected M. obnubilus bug with two B. esakii (a female larva on the left and a male pupa on the right as one faces, respectively). Occasionally, three or more B. esakii individuals parasitize one host. Different individual from the one in Photo 3. Photo credit: Yuta Nakase. These photographs illustrate the article “Parasitism by multiple strepsipterans accelerates timing of adult parasite emergence” by Yuta Nakase, Yuichiro Fukumasu, Tsubasa Toji, Natsumi Ishimoto, and Takao Itino pulished in Ecology. https://doi.org/10.1002/ecy.3811

Sex and stress modulate pupal defense response in tobacco hornworm.

In insects, larval and adult defenses against predators have been well studied. However, pupal (also known as resting stage) defenses have been overlooked and not examined thoroughly. Although some pupa possess antipredator strategies such as hairs, spines, cryptic coloration, and exudation of chemicals, few studies have tested these responses and the factors affecting them. Here, we investigated the behavioral responses in tobacco hornworm Manduca sexta that pupates in soil by introducing an external stimulus using vibrations from an electric toothbrush to mimic predation. We observed that M. sexta made violent wriggling (twitching), followed by pulsating movements in response to the vibrational stimulus. Detailed examination showed that these twitches and pulsating events occurred more frequently and for longer periods of time in male pupa and were dependent on the magnitude of the stress (high and low frequency). However, when we estimated the angular force exerted by pupa using radian and angular momentum of twitches, it was found to be independent of pupal sex. A follow-up experiment on possible cascading effects of stress exposure on eclosion success revealed that low- and high-frequency stress exposure didn't cause any of the common defects in eclosed adults. Our study clearly demonstrates that the so-called defenseless pupal stage uses a wide range of measurable defense behaviors that can actively defend against predators and should be examined further-linking observed behavior with underlying mechanisms.

Sublethal effects of chlorfenapyr on Plutella xylostella (Lepidoptera: Plutellidae).

The diamondback moth (DBM), Plutella xylostella (L.), is the most destructive pest of cruciferous vegetables worldwide. Chlorfenapyr is an important insecticide for controlling DBM. The impacts of three sublethal doses (LC1 , LC10 and LC30 ) of chlorfenapyr on the chlorfenapyr-exposed DBM individuals and their unexposed F1 and F2 offspring were investigated in order to reveal the non-lethal deleterious effects of chlorfenapyr and its potential hormetic effects. LC1 significantly increased female pupa weight of F0 and F1 generations, and F0 fecundity as well as F1 gross reproduction rate (GRR). The LC1 -elicited rise in emergency rate and fecundity was significantly greater in F0 than in F1 . By contrast, LC30 significantly decreased age-specific survival rates, pupation rate, male pupal weight, emergence rate and fecundity of F0 and F1 generations as well as female adult proportion and GRR, net reproduction rate (R0 ), intrinsic rate of increase (rm ) and finite rate of increase (λ) of F1 generation. The LC30 -induced reductions in pupation rate, adult emergence rate, male and female pupa weight, and fecundity were greater in F1 than in F0 . While LC10 elicited only a mild inhibition (extension of pupal duration) in F0 , it yielded both deleterious (drops in female proportion and age-specific survivals) and hormetic effects (ups in male longevity and female fecundity) in F1 . The results demonstrate that the sublethal effects of chlorfenapyr on DBM vary from inhibition to stimulatory hormesis, depending on the dose and generation. © 2022 Society of Chemical Industry.

A new species of the Arthuri Complex of the Strodei Subgroup of Nyssorhynchus (Diptera: Culicidae).

Nyssorhynchus (Nyssorhynchus) rondoniensis, a new species of the Arthuri Complex of the Strodei Subgroup, is described and validated using morphological characters of the adult male and female, male genitalia, fourth-instar larva and pupa. The new species is recorded in the municipalities of Campo Novo de Rondnia and Monte Negro, Rondnia State, Brazil. Based on DNA sequence data, the new species (as Ny. arthuri species C) was found to belong to a separate lineage within the Arthuri Complex. Morphological characteristics of the male genitalia and fourth-instar larva confirmed that the new species shared morphological similarities with other species of the Arthuri Complex, but it can be distinguished by characteristics of the male genitalia, adult female and larva. Nyssorhynchus rondoniensis may be involved in malaria transmission because females can be easily misidentified as Ny. oswaldoi (Peryass, 1922) s.l. or Ny. konderi (Galvo Damasceno, 1942) s.l. Both species were previously hypothesized to be local vectors in Acre and Rondnia States.

Redescription of Wyeomyia (Decamyia) felicia (Dyar amp; Nez Tovar, 1927) and description of a new species of the subgenus from Trinidad (Diptera: Culicidae).

Mosquito species of the subgenus Decamyia Dyar, 1919 of genus Wyeomyia Theobald, 1901 are known to have immature stages mostly associated with Heliconia plants. Currently, the subgenus includes five species, some of which are poorly characterized. Here, we redescribe and illustrate the larval, pupal and adult life stages of Wy. felicia (Dyar Nez Tovar, 1927). In addition, Wyeomyia monticola, a new species of Decamyia from Trinidad, close to Wy. felicia, is described. The new species can be distinguished by morphological characters of the male genitalia, larva and pupa.

A new species of the Nuneztovari Complex of Nyssorhynchus (Diptera: Culicidae) from the western Brazilian Amazon.

Nyssorhynchus (Nyssorhynchus) jamariensis, a new species of the Nuneztovari Complex, previously known as Anopheles (Nyssorhynchus) nuneztovari A, is described and validated using morphological characters of the adult male and female, male genitalia and immature stages. The species is recorded from the western Brazilian Amazon, where it was collected in pastures in the vicinity of the Jamari River, municipality of Monte Negro, Rondnia State, Brazil. Illustrations of the male genitalia, fourth-instar larva and pupa are provided. Nyssorhynchus jamariensis may be involved in malaria transmission, but its vector status needs further investigation.

A new coccoid family (Hemiptera: Coccomorpha) for an unusual species of scale insect on Podocarpus macrophyllus (Podocarpaceae) from southern China.

A new family, Qinococcidae Wu, fam. n., is established for the species Qinococcus podocarpus gen. n. and sp. n., collected from southern China under the bark of Podocarpus macrophyllus (Podocarpaceae). The adult female and male, egg, and all immature instars including the male pupa of Q. podocarpus sp. n. are described and illustrated. The new family is diagnosed mainly on the adult male possessing an inverted -like post-tergite and unusual venation of the fore wing, with a pterostigma, posterior cubital vein and bifurcate anterior flexing patch. A key to all the developmental stages is provided.

Identification of male and female pupal characteristics of Zeugodacus cucurbitae (Coquillett) via machine vision.

Images of original pupae of Zeugodacus cucurbitae (Coquillett) were normalized, grayed, and segmented to identify male and female pupae of this species via machine vision. The image of each pupa was divided into 25 small areas. The differences in surface texture features in each small area within 11 days were compared. The texture characteristics of both male and female pupae were screened by combining the eclosion of both sexes of Z. cucurbitae (Coquillett). Results indicated that the pectinate setae on the abdominal backplane could be used as a basis for the identification of the male and female pupa of Z. cucurbitae (Coquillett). Moreover, machine vision correctly identified these characteristics with an accuracy of 96.0%. This study lays a foundation for the identification of male and female pupae using machine vision and also for the comprehensive control of Z. cucurbitae (Coquillett).

Taxonomic notes on the genus Cyphococcus Laing (Hemiptera: Coccomorpha: Coccidae), with description of a new species from India.

The adult female, first-instar nymph, male pupa and adult male of a new species of the genus Cyphococcus Laing (Hemiptera: Coccomorpha: Coccidae) from India, C. williamsi Joshi Rajgopal sp. n., are described and illustrated. This is the first record of the genus Cyphococcus and the subfamily Cyphococcinae from the Oriental region. Information on distribution, biology, host plants, associated coccinellid predators and hymenopteran parasitoids is given.

Distributions Pattern of Protective Silken Discs Weaved by Larvae of Sugarcane Top Shoot Borer Scirpophaga excerptalis (Walker)

The distribution pattern of protective silken discs weaved by larvae of five broods (generations) of top shoot borer Scirpophaga excerptalis (Walker) was studied in three early maturing varieties (Co 0238, CoPk 09151 and CoLk 94184) and two mid late varieties (CoLk 13204 and CoLk 8102). The mean number of silken discs guarding the female pupa was more in first, second, third, fourth and fifth broods than male pupa in all varieties. The ranges of the discs protecting the female pupae in all broods (first, second, third, fourth and fifth) varied 3-12, 2-21, 2-17, 3-13 and 2-9, respectively. The male pupae were surrounded by 5.23 (CoLk 8102), 6.93, 8.73, 6.67 and 3.92 (CoPk 05191) discs, respectively which were higher than the other varieties in respective broods. The results allow us to better understand the distribution pattern of silken discs weaved by larvae of different broods of S. excerptalis in early and mid-late varieties.

Morphological Description of the Female, Pupa, and Fourth-Instar Larva and Redescription of the Male of Culex (Melanoconion) aliciae (Diptera: Culicidae).

The female, pupa, and fourth-instar larva of Culex (Melanoconion) aliciae are described for the first time, and the male is redescribed. The cibarium, female and male genitalia, pupa, and fourth-instar larva are illustrated. Information on distribution, bionomics, and taxonomy is also included. Males of Cx. aliciae Duret and Cx. martinezi Casal & Garcia are indistinguishable, except for the appearance of the ninth tergal lobes. The female of Culex martinezi has not been described before. The females of Culex aliciae can be recognized by characteristics of the occipital scales, cibarium, and genitalia. The larva of Cx. aliciae and Cx. martinezi can be distinguished from characters of the thorax and abdomen.

Identification and validation of stage-specific reference genes for gene expression analysis in Callosobruchus maculatus (Coleoptera: Bruchidae)

In light of a number of recent studies highlighting the increasing research interest in bruchids, it is crucial to validate suitable reference genes that could be used in quantitative gene expression studies. Callosobruchus maculatus is a serious pest of stored grains and field legumes in which reference genes have not been assessed and validated to date. The present study aimed to identify and validate reference genes in different developmental stages of C. maculatus shortlisted from commonly used reference genes such as VATPase, TRIP12, TBP, TF11D, ACTIN, GST, ANNEXIN, PTCD3, RPL32, and β -Tub in various insects. Dedicated algorithms like GeNorm, NormFinder, and BestKeeper were used to analyze the stability of these candidate genes, which revealed GST for third instar, ANNEXIN and PTCD3 for the fourth instar, TF11D and VATPase for male pupa, RPL32 and β-tub for female pupa, β-tub and TBP for adult male and VATPase and GST for adult females as suitable reference genes for expression studies in C. maculatus. The final comprehensive ranking using RefFinder identified GST and TBP as the best reference genes for all the developmental stages of C. maculatus. To the best of our knowledge, this is the first report which evaluates and validates stable reference genes in C. maculatus. The information of stage-specific gene expression, generated in this study will be useful for future molecular, physiological, and biochemical studies on C. maculatus and other closely related bruchids.

Morphology, biology, and damage behaviors of nutsedge borer (Bactra venosana Zeller) in Tra Vinh Province, Vietnam

Abstract. Nguyen HU, Phan QN, Dang HT, Son TTN, Nguyen HN, Nguyen THT. 2021. Morphology, biology, and damage behaviors of nutsedge borer (Bactra venosana Zeller) in Tra Vinh Province, Vietnam. Biodiversitas 23: 733-741. Nutsedge borer, Bactra venosana Zeller (Lepidoptera: Tortricidae), is a common pest of sedge in Tra Vinh Province. To control this species, its morphology, biology, and behaviors were investigated in Tra Vinh Province. The study was conducted with the aim of determining the information related to the Bactra venosana, as a basis for developing a strategy to manage this species by means of safety solutions. The study was carried out by surveying the behavior of nutsedge borer in sedge fields and determining their morphological and biological characteristics under laboratory conditions. The results showed that the eggs were 1.06 ± 0.17 mm (in length), 0.84 ± 0.16 (in width), oval, and white after turning pale yellow. Larvae progressed through 5 instars with body length and width of 1.20 ± 0.18 mm to 11.3 ± 1.23 mm and from 0.25 ± 0.05 to 1.05 ± 0.25 mm, respectively. Newly hatched larvae were dark yellow, and pupae were pale green. The male pupa was 7.75 ± 0.68 mm long, while the female was 8.46 ± 0.93 mm long, light green, and dark brown about to mature. The male adult of B. venosana was usually gray-brown and 7.70 ± 0.60 mm in body length, whereas the female adult was silver-gray, 8.41 ± 0.86 mm. The life cycle passed 35.9 ± 2.55 days in laboratory conditions. Under field conditions, eggs were laid scattered on the surface of the host plants. Larvae bored inside the stems of sedge. Newly damaged sedge plants had no symptoms at first whose bodies got soft, turned from light green to pale yellow, and gradually wilted and died.