Optimal cold storage protocol for Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae), a biological control agent for coleopteran pests in stored products.

BACKGROUND: Anisopteromalus calandrae (Howard) is a solitary ectoparasitoid with wide-ranging potential applications as a natural biological control agent against various coleopterous pests in food warehouses. Implementing an effective cold storage program is crucial for extending the shelf life of biological control agents and ensuring their stable and abundant supply. Herein, we attempted to determine the optimal cold storage conditions for Anisopteromalus calandrae by investigating the effect of cold storage at three different temperatures (7, 13, and 19 °C) for 7, 21, and 35 days on four developmental stages (late-instar larvae, early-stage pupae, mid-stage pupae, and 2-day-old adults). Additionally, we explored the maximum cold storage potential by observing early-stage pupae stored at 13 °C for various durations (30, 60, 90, 120, and 150 days). RESULTS: The most suitable cold storage temperature for the early-stage pupae of Anisopteromalus calandrae was 13 °C, and the highest adult emergence rate (98.3%) was after 90 days of storage at 13 °C. Furthermore, we did not find any significant effect on longevity (female: 44.3 days; male: 38.1 days) or fecundity (121.7 wasps). The female ratio ranged from 43.5% to 50.8%. More importantly, cold storage did not adversely affect the developmental duration or fecundity of the offspring. CONCLUSION: This study offers crucial insights for managing Anisopteromalus calandrae populations under laboratory conditions and lays the foundation for potential industrial production and development. © 2023 Society of Chemical Industry.

Identification and immune analysis of antimicrobial peptides from the cigarette beetle (Lasioderma serricorne).

Antimicrobial peptides (AMPs) in insects are endogenous peptides that are effector components of the innate defense system of the insect. AMPs may serve as antimicrobial agents because of their small molecular weight and broad-spectrum antimicrobial activity. In this study, we performed transcriptome analysis of cigarette beetle (Lasioderma serricorne) larvae, parasitized by the ectoparasitic wasp, Anisopteromalus calandrae. Several AMP genes were significantly upregulated following A. calandrae parasitism, postulating the hypothesis that the parasitization enhanced the host's resistance against pathogenic microorganisms through the regulation of host AMP genes. Specifically, 3 AMP genes (LsDef1, LsDef2, and LsCole) were significantly upregulated and we studied their immune function in L. serricorne. Immune challenge and functional analysis showed that LsCole was responsible for the immune response against Gram-negative and Gram-positive bacteria, while LsDef1 and LsDef2 were involved in insect defense against Gram-positive bacteria. Purified recombinant LsCole exhibited antimicrobial activities against the Gram-negative bacterium Escherichia coli and the Gram-positive bacterium Staphylococcus aureus. LsDef2 showed an antibacterial effect against S. aureus. LsCole and LsDef2 exhibited antibiofilm activity against S. aureus. The 2 AMPs disrupted cell membranes and caused leakage of S. aureus cell contents. The results indicated that the 3 AMPs in L. serricorne are involved in the innate immunity of this pest insect. These AMPs may have potential as antimicrobial agents for bacterial infection chemotherapy. Hence, data are discussed in relation to new control strategies with greater biosafety against pest insects with use of microbial biocontrol agents in combination with RNA interference against the insect's defensive AMP genes.

Six new species of the leafhopper subgenus Pediopsoides (Pediopsoides) (Hemiptera, Cicadellidae, Eurymelinae, Macropsini) from China.

The nominate subgenus Pediopsoides (Pediopsoides) Matsumura, 1912 is widely distributed in the Oriental region but has high species diversity only in southern China. The present paper describes and illustrates six new species of Pediopsoides (Pediopsoides) namely, P. (P.) ailaoshanensis Li & Dai, sp. nov., P. (P.) quadrispinosus Li & Dai, sp. nov., P. (P.) flavus Li & Dai, sp. nov., and P. (P.) pianmaensis Li & Dai, sp. nov. all collected in Yunnan Province, southwestern China, P. (P.) maoershanensis Li & Dai, sp. nov., found in Guangxi Autonomous Region, southern China, and P. (P.) huangi Li & Dai sp. nov., from Taiwan, previously incorrectly recorded as a new name by Li & Dai, 2018 in Dai et al. 2018: 203 for P. (P.) femorata Huang & Viraktamath, 1993 (nec Pediopsisfemorata Hamilton, 1980). Two new junior synonyms of Sispocnis Anufriev, 1967 are proposed, i.e., Digitalis Liu & Zhang, 2002, syn. nov. and Neosispocnis Dmitriev, 2020, syn. nov.

Descriptions of two new Idiocerini leafhoppers of the genus Idioscopus (Hemiptera, Cicadellidae) from China.

Two new species of the leafhopper genus Idioscopus Baker are described from China: Idioscopusbihamulus sp. nov. and I.ventrispinus sp. nov., the latter recorded on a species of Myrica L. (Myricaceae) as its host plant. A key and checklist to species of the genus from China are provided and Idioscopustaiwanus Huang & Maldonado-Capriles, 1992 is placed as a junior synonym of Idioscopusclypealis (Lethierry, 1889), syn. nov.

Comparison of mitogenomes of three Petalocephala species (Hemiptera: Cicadellidae: Ledrinae) and their phylogenetic analysis.

Ledrinae is a unique group of leafhoppers with a distinct appearance. Petalocephala is the largest Ledrinae genus that is difficult to identify except by dissecting the male genitals. To date, research on Ledrinae is relatively less compared with other leafhoppers. Therefore, to better understand this group, we sequenced and analyzed three complete Petalocephala mitochondrial genomes. We comparatively analyzed these general Petalocephala genomic features (including size, AT content, AT/GC skew, 13 protein-coding gene nucleotide compositions, etc.), and predicted 22 transfer RNA secondary structures. We obtained highly consistent phylogenetic results within Cicadellidae based on mitogenomic data using the maximum likelihood and Bayesian methods. Our results showed that all subfamilies were monophyletic and had a high node support rate, and there was a sister group relationship between Ledrinae and all other leafhopper groups. Furthermore, treehoppers were found to originate from leafhoppers and showed sister group relationships with Megophthalminae. Within Ledrinae, all phylogenetic trees supporting phylogenetic relationships were as follows: ([P. dicondylica + P. gongshanensis] + [Tituria pyramidata + [Ledra auditura + P. gongshanensis]]) Based on the complete mitogenome phylogenetic analysis and the comparison of morphological characteristics, we propose that Petalocephala is not monophyletic.

Description of two new species of the genus Siniassus (Hemiptera: Cicadellidae: Iassinae: Hyalojassini) from China.

Two new species of leafhoppers: Siniassus gracilitus sp. nov. and S. nigrimaculatus sp. nov., of tribe Hyalojassini, subfamily Iassinae (Hemiptera: Cicadellidae) are described and illustrated from China. A key to Siniassus species and updated checklist with distributions are provided.

Description of three new species of the leafhopper genus Idioscopus Baker, (Hemiptera: Cicadellidae: Eurymelinae) from Yunnan, China.

Three new species of the leafhopper genus Idioscopus Baker, 1915 from China: I. furcaprocessus sp. nov., I. longiprocessus sp. nov., and I. serratastylus sp. nov. are described and photographed based on specimens collected in Yunnan Province of southern China. These three new species can be recognised mainly by the coloration and male genitalia.

The complete mitochondrial genome of Oenopia sauzeti (Coleoptera: Coccinellidae).

Oenopia sauzeti Mulsant are predators of various aphids, such as Myzus persicae, Rhopalosiphum maidis, and Sitobion avenae. We sequenced the mitochondrial genome (mitogenome) of O. sauzeti. Consistent with other mitogenomes of Coleoptera, the circular mitogenome of O. sauzeti consists of 17,630 bp, including 13 PCG genes, 2 rRNA genes, and 22 tRNA genes. The mitogenome differs in 2 long non-coding regions of 1285 bp and 1843 bp. The 22 tRNA genes products are folded into the typical cloverleaf secondary structure, except trnS1, which lacks the dihydrouridine (DHU) arm. The base composition is AT-biased (80.1%). The phylogenetic tree confirms monophyly at the genus level within the Coccinellinae and supports O. sauzeti as a sister group to Aiolocaria hexaspilota.

Mitogenomics of five Olidiana leafhoppers (Hemiptera: Cicadellidae: Coelidiinae) and their phylogenetic implications.

Similar morphological characteristics and limited molecular data of Olidiana resulted in their unknown phylogenetic statuses and equivocal relationships. To further understand the genus Olidiana, we sequenced and annotated five Olidiana complete mitochondrial genomes (mitogenomes). Our results show that Olidiana mitogenomes range from 15,205 bp to 15,993 bp in length and include 37 typical genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) and a control region. Their nucleotide composition, codon usage, features of control region, and tRNA secondary structures are similar to other members of Cicadellidae. We constructed the phylogenetic tree of Cicadellidae using the maximum likelihood (ML) and Bayesian inference (BI) methods based on all valid mitogenome sequences. The most topological structure of the obtained phylogenetic tree is consistent. Our results support the monophyletic relationships among 10 subfamilies within Cicadellidae and confirm Iassinae and Coelidiinae to be sister groups with high approval ratings. Interestingly, Olidiana was inferred as a paraphyletic group with strong support via both ML and BI analyses. These complete mitogenomes of five Olidiana species could be useful in further studies for species diagnosis, evolution, and phylogeny research within Cicadellidae.

Two new species of the leafhopper genus Calodia Nielson (Hemiptera, Cicadellidae, Coelidiinae) from China, with a checklist and key to Chinese species.

Two new species of the leafhopper genus Calodia Nielson are described and illustrated: C. quadrimacula sp. nov. from Guizhou and Yunnan Provinces and C. zuoae sp. nov. from Yunnan Province, China. A checklist along with distribution and a key to species based on male genitalia of the genus Calodia from China are provided. Olidiana nigritibiana (Li), comb. nov. (earlier in the genus Calodia) is proposed. At present, this genus comprises 45 known species worldwide, of which 19 species are recorded from China.

A new genus and new species of the leafhopper tribe Idiocerini (Hemiptera: Cicadellidae: Eurymelinae).

A new genus of the leafhopper tribe Idiocerini (Hemiptera: Cicadellidae: Eurymelinae) Longiaedeagus gen. nov. with type species Longiaedeagus flavofasciatus sp. nov. from China is described and illustrated. This new genus can be distinguished from other leafhopper genera by the broad style with truncate apex, and aedeagus with a long, unpaired retrorse preapical process.

Complete mitochondrial genome of Mukaria splendida Distant (Hemiptera: Cicadellidae: Deltocephalinae: Mukariini) and phylogenetic analysis.

The complete mitochondrial genome (mitogenome) of Mukaria splendida Distant, 1908 (Hemiptera: Cicadellidae: Deltocephalinae) was first reported in Mukariini. The length of this mitogenome is 16,711 bp, which has an A + T content of 79% (A = 44.5%, T = 34.5%, G = 8.8%, C = 12.2%). A total of 37 genes were annotated [13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNA), and 2 ribosomal RNA genes (rRNA)]. Among the 37 genes, 4 protein coding genes (ND1, ND4, ND4L, ND5), 8 tRNA genes (trnQ, trnC, trnY, trnF, trnH, trnP, trnL2, trnV), and 2 rRNA (12S rRNA, 16S rRNA) were encoded by N chain, and the remaining genes were encoded by J chain. Overall, there were 14 gene overlaps and 9 gene gaps in the mitochondrial genome of this species. All PCGs were started with ATD (ATA/ATT/ATG), and stopped with TAR, except ATP6, which ends with single T. The phylogenetic analysis confirms that M. splendida clustered with other Deltocephalinae species.

Complete mitochondrial genome of Penthimia melanocephala Motschulsky, 1863 (Hemiptera: Cicadellidae: Deltocephalinae).

Penthimia is the largest genus in the tribe Penthimiini of the subfamily Deltocephalinae. To date, there are no available mitogenome sequences from Penthimia. In this study, we have sequenced and annotated the complete mitogenome of Penthimia melanocephala (Motschulsky 1863), the mitogenome is 15,308 bp in length, which including 13 protein-coding genes (PCGs), 2 ribosomal RNA genes (rRNAs), 22 transfer RNAs (tRNAs) and a long non-coding region (control Region), base composition of whole sequences are A (50.4%), C (15.3%), G (10.0%), and T (24.3%). All PCGs started with the typical ATN codon and stopped with the typical TAN codon except for COII and COIII stopped with single T. Within phylogenetic tree, Deltocephalinae members were clustered into a clade. The complete mitogenome of P. melanocephala can provide essential DNA molecular data for further evolutionary and phylogenetic analysis.

Three new species of the leafhopper genus Thagria Melichar (Hemiptera: Cicadellidae: Coelidiinae) from China.

Three new species of the leafhopper genus Thagria from China, T. trifurcata sp. nov., T. denticosta sp. nov. and T. fimbriata sp. nov., are described, illustrated and photographed. In addition, T. lateralis Fan Li, 2017 is placed as a new synonym of T. menglana Wang Zhang, 2017.

The Phylogenetic Implications of the Mitochondrial Genomes of Macropsis notata and Oncopsis nigrofasciata.

Macropsinae are forest pests that feed on woody plants. They can damage the growth of trees and crops, and some species can also spread plant pathogens. Due to their widespread effects, these leafhoppers are of great economic significance, which is why there is a need to study their genomes. To fill the gap in the mitochondrial genomic data of the subfamily Macropsinae, we sequenced the complete mitochondrial genomes of Macropsis notata and Oncopsis nigrofasciata (which were 16,323 and 15,927 bp long, respectively). These two species are representative species of the leafhoppers group (Cicadellidae); the mitochondrial genomes of these species range from a length of 15,131 bp (Trocnadella arisana) to 16,811 bp (Parocerus laurifoliae). Both mitogenomes contained 37 typical insect mitochondrial genes and a control region; there were no long non-coding sequences. The genes within the mitogenome were very compact. The mitogenomes from both species contained two kinds of parallel repeat units in the control region. The whole mitogenomes of Macropsinae showed a heavy AT nucleotide bias (M. notata 76.8% and O. nigrofasciata 79.0%), a positive AT Skew (0.15 and 0.12), and a negative GC Skew (-0.14 and -0.08). Upon comparative ML and BI analysis, some clade relationships were consistent among the six trees. Most subfamilies were reconstructed into monophyletic groups with strong support in all analyses, with the exception of Evacanthinae and Cicadellinae. Unlike the results of previous research, it was shown that although all Deltocephalinae species are grouped into one clade, they were not the sister group to all other leafhoppers. Further, Cicadellinae and Evacanthinae were occasionally reconstructed as a polyphyletic and a paraphyletic group, respectively, possibly due to the limited numbers of samples and sequences. This mitogenome information for M. notata and O. nigrofasciata could facilitate future studies on the mitogenomic diversity and evolution of the related Membracoidea, and eventually help to control their effects on plants for the betterment of society at large.

Complete mitogenome of Olidiana ritcheriina (Hemiptera: Cicadellidae) and phylogeny of Cicadellidae.

BACKGROUND: Coelidiinae, a relatively large subfamily within the family Cicadellidae, includes 129 genera and ∼1,300 species distributed worldwide. However, the mitogenomes of only two species (Olidiana sp. and Taharana fasciana) in the subfamily Coelidiinae have been assembled. Here, we report the first complete mitogenome assembly of the genus Olidiana. METHODS: Specimens were collected from Wenxian County (Gansu Province, China) and identified on the basis of their morphology. Mitogenomes were sequenced by next-generation sequencing, following which an NGS template was generated, and this was confirmed using polymerase chain reaction and Sanger sequencing. Phylogenic trees were constructed using maximum likelihood and Bayesian analyses. RESULTS: The mitogenome of O. ritcheriina was 15,166 bp long, with an A + T content of 78.0%. Compared with the mitogenome of other Cicadellidae sp., the gene order, gene content, gene size, base composition, and codon usage of protein-coding genes (PCGs) in O. ritcheriina were highly conserved. The standard start codon of all PCGs was ATN and stop codon was TAA or TAG; COII, COIII, and ND4L ended with a single T. All tRNA genes showed the typical cloverleaf secondary structure, except for trnSer, which did not have the dihydrouridine arm. Furthermore, the secondary structures of rRNAs (rrnL and rrnS) in O. ritcheriina were predicted. Overall, five domains and 42 helices were predicted for rrnL (domain III is absent in arthropods), and three structural domains and 27 helices were predicted for rrnS. Maximum likelihood and Bayesian analyses indicated that O. ritcheriina and other Coelidiinae members were clustered into a clade, indicating the relationships among their subfamilies; the main topology was as follows: (Deltocephalinae + ((Coelidiinae + Iassinae) + ((Typhlocybinae + Cicadellinae) + (Idiocerinae + (Treehopper + Megophthalminae))))). The phylogenetic relationships indicated that the molecular taxonomy of O. ritcheriina is consistent with the current morphological classification.

Structural and phylogenetic implications of the complete mitochondrial genome of Ledra auditura.

We sequenced and annotated the first complete mitochondrial genome (mitogenome) of Ledra auditura (Hemiptera: Cicadellidae: Ledrinae) and reconstructed phylogenetic relationships among 47 species (including 2 outgroup species) on the basis of 3 datasets using maximum likelihood (ML) and Bayesian inference (BI) analyses. The complete L. auditura mitogenome (length, 16,094 bp) comprises 37 genes [13 protein-coding genes (PCGs), 22 tRNAs, and 2 rRNAs], 1 control region, and 2 long non-coding regions. The first long non-coding region (length, 211 bp) is located between tRNA-I and tRNA-Q and the second region (length, 994 bp) between tRNA-S2 and ND1. All PCGs show ATN (Met/Ile) as their start codon and TAR as their stop codon. Except tRNA-S1 (AGN), which lacks the dihydrouridine arm, all tRNAs can fold into the typical cloverleaf secondary structure. The complete L. auditura mitogenome shows a base composition bias of 76.3% A + T (A = 29.9%, T = 46.4%, G = 13.3%, and C = 10.5%), negative AT skew of -0.22, and positive GC skew of 0.12. In ML and BI analyses, L. auditura was clustered with Evacanthus heimianus (Hemiptera: Cicadellidae: Evacanthinae) with strong branch support.

Key to species of the leafhopper genus Bhatia Distant, 1908 (Hemiptera: Cicadellidae: Drabescini) with description of a new species from China.

A new species of Bhatia Distant, B. longiradiata sp. nov. is described from China (Guangdong, Guangxi). A key to the Chinese species of the genus is given.

Review of the leafhopper subgenus Pediopsoides (Sispocnis) (Hemiptera, Cicadellidae, Eurymelinae, Macropsini) with description of two new species from China.

The leafhopper subgenus Pediopsoides (Sispocnis) Anufriev, 1967 is reviewed and the type species is fixed as Bythoscopuskogotensis Matsumura, 1912. Six valid species of the subgenus are recognized including two new species described and illustrated here, Pediopsoides (Sispocnis) rectus Li, Li & Dai, sp. nov. and P. (S.) triangulus Li, Li & Dai, sp. nov. from Sichuan Province of Southwestern China. Additionally P. (S.) heterodigitatus Dai & Zhang, 2009 is proposed as a junior synonym of P. (S.) aomians (Kuoh, 1981) based on examination of many specimens. A key to species of the subgenus is also provided for identification.

Three new species in the leafhopper tribe Drabescini (Hemiptera, Cicadellidae, Deltocephalinae) from southern China.

Three new species of the leafhopper tribe Drabescini: Drabescusbilaminatus sp. n., Drabescusmultipunctatus sp. n., and Paraboloponarobustipenis sp. n. are described and illustrated from southern China. A key and checklist to the species of Parabolopona are also provided.