First report of Pseudopithomyces palmicola causing leaf spot on tobacco in China.

In July 2023, a new leaf spot disease emerged on tobacco leaves in Meitan County, Guizhou Province, China (27°20'18" - 28°12'30"N, 107°15'36" - 107°41'08"E, average altitude 972 meters). Initially, the symptoms showed raised yellow-brown spots; subsequently, the lesions expanded and became broken and perforated, leading to a significant loss of economic value, the prevalence rate exceeded 30%. For isolation, two tissue fragments (0.2 × 0.2 cm) of symptomatic leaves were sterilized in 75% ethanol for 30 s, 3% NaClO for 2 min, and were washed 3 times in sterilized distilled water, and were subsequently inoculated on potato dextrose agar (PDA), and incubated at 28°C for 9 days in the dark. The two strains CW16 and CW28 were isolated using the single hyphae method (Nouri et al. 2023). Both strains formed pale to yellow white colonies on PDA. Conidia had three constricted transverse septa and 1 to 2 longitudinal septa in the central cells, with thick and hyaline conidiophores and mostly globose, pale brown conidia with slightly constricted septa, their average size were measured as 13.4-22.4×8.358-13.347 µm (n = 50). Genomic DNA was extracted from the isolated strains CW16 and CW28. The internal transcribed spacer regions 1 and 2 as well as 5.8S nuclear ribosomal RNA (ITS), large subunit nrRNA (LSU), and partial DNA-directed RNA polymerase II second largest subunit (RPB2) genes were amplified using primers (Cui et al. 2023). The sequences had been deposited in GenBank under accession numbers ITS: PP024201, PP024205; LSU: PP024207, PP024209; RPB2: PP060480, PP060481. The sequences analysis revealed a high similarity of 99.74 to 100% between strains CW16 and CW28 with P. palmicola isolate KM42 (ITS OQ875842, LSU OQ875844, RPB2 OQ883943) in GenBank. Using BLAST for homology matching, two isolates (CW16, CW28) and with the sequences of the ten type isolates from GenBank, phylogenetic analysis was conducted using the Maximum Likelihood method in MEGA (11.0) software based on ITS, LSU and RPB2 sequences, which showed that strains CW16, CW28 clustered in the same score as the Pseudopithomyces palmicola, confirming the morphological and molecular characteristics identification. The pathogenicity tests were conducted on healthy tobacco plants with 4-5 leaves (Fig. S1B), the isolated strains, CW16 and CW28, were used to inoculate the healthy tobacco leaves, while blank PDA was used as a control. All plants were maintained in a greenhouse at 28°C with a relative humidity of 90%. After 9 days, necrotic spots were observed on all tobacco leaves inoculated with CW16 and CW28 fungal plugs, while the blank PDA-inoculated tobacco leaves showed no symptoms. Based on morphological and molecular characteristics, the same pathogen P. palmicola was identified from the inoculated leaves, fulfilling Koch's postulates. This study represents the first reported of tobacco leaf spot caused by P. palmicola in China and provides a theoretical basis for future prevention and control measures.

Intraguild species presence alters Aphidoletes aphidimyza (Diptera: Cecidomyiidae) and Aphidius gifuensis (Hymenoptera: Braconidae) foraging responses.

The predatory gall midge, Aphidoletes aphidimyza (Rondani), and tobacco aphid cocoon wasp, Aphidius gifuensis Ashmead, are important natural enemies of Myzus persicae (Sulzer) (Hemiptera: Aphididae). Predation by A. aphidimyza and A. gifuensis can regulate M. persicae; however, how interspecific interference competition affects their foraging efficiency is unknown. Here, we investigated the consumption and parasitization abilities of A. aphidimyza 3rd instar larva and A. gifuensis adults under various conditions. Consumption of parasitized aphids by A. aphidimyza 3rd instar larvae was significantly lower than that of nonparasitized controls, with a substantial increase in handling time. The presence of A. gifuensis adults did not significantly affect the predation capacity of A. aphidimyza larvae. Relative to controls, A. aphidimyza larvae predation trace (PT) and imago activity significantly decreased A. gifuensis parasitism rates at different aphid densities. Further, A. aphidimyza larvae PT increased the A. gifuensis handling time of M. persicae, whereas the presence of A. aphidimyza adults had the opposite effect. Coexistence with heterospecific natural enemies reduced the parasitic capacity of A. gifuensis, whereas A. aphidimyza larvae predation capability was influenced to a lesser extent. Our results demonstrate that intraguild interactions strongly influence the predatory and parasitic efficacy of A. aphidimyza and A. gifuensis, although the effect on A. gifuensis was more pronounced. For effective biological control of M. persicae using A. aphidimyza and A. gifuensis, we recommend releasing A. aphidimyza first to mitigate intraguild predation and enhance the overall success of the pest control program.

Safety evaluation and sublethal effect of broflanilide on Aphidius gifuensis.

Ensuring the safety of insecticides to natural enemy insects of pests is crucial for integrating chemical and biological control strategies. Broflanilide, a novel meta-diamide insecticide, exhibits high insecticidal activity against Myzus persicae (Sulzer) (Hemiptera: Aphididae). To integrate chemical and biological control against M. persicae, we assessed the toxicity of broflanilide to Aphidius gifuensis, and evaluated its safety and sublethal effects. The LC10, LC25, and LC50 values of broflanilide against A. gifuensis were 0.733 mg/L, 1.613 mg/L, and 3.852 mg/L, respectively. The selectivity toxicity ratio of broflanilide to A. gifuensis was 1.516, indicating higher toxicity to M. persicae compared to A. gifuensis. The risk quotient of broflanilide to A. gifuensis adults was 6.18. The percent reduction in the emergence of the parasitoid pupae was -1.15, with a risk grade of 1. The sublethal concentration of broflanilide had no significant influence on the intrinsic rate of increase (r), finite rate of increase (λ), net reproductive rate (R0), and mean fecundity (F) of A. gifuensis in the F1 generation. The mean generation time (T) increased by 0.51 days and 0.39 days in the LC10 and LC25 treatments, respectively; the difference between LC10 treatment and the control was significant, while the difference between LC25 treatment and the control was not significant. The results showed that the sublethal concentration of broflanilide did not have a significant inhibitory effect on the population growth of A. gifuensis.

Morphology and distribution of sensilla on antennae and mouthparts of the adult bruchid beetles, Bruchidius coreanus (Coleoptera: Bruchidae).

Bruchidius coreanus is a serious pest on Gleditsia sinensis Lam during seed storage, causing significant losses to their yield in southwest China. To gain insight into their behavioral mechanisms, the external morphology, ultrastructure, and distribution of sensilla on antennae, maxillary palps, and labial palps of both male and female B. coreanus were observed using a scanning electron microscope. The results revealed that both male and female adults had serrated antennae comprising a scape, a pedicel, and nine flagellomeres (F1-F9). There were eight types and seven subtypes of antenna sensilla observed in both sexes, including Böhm sensilla (BS), two subtypes of sensilla chaetica (SC1 and SC2), two subtypes of sensilla trichodea (ST1 and ST2), three subtypes of sensilla basiconica (SB1, SB2, and SB3), sensilla auricillica (SA), sensilla styloconicum (SS), capitate pegs (CP), and sensilla cavity (SCa). The average length of BS and ST (ST1 and ST2) showed significant differences between males and females. Furthermore, the number of SC (SC1 and SC2), ST1, and SCa differed significantly between the sexes. Four types of sensilla were found on the maxillary palps and labial palps, with the length of ST on these palps significantly differing between males and females. Additionally, SS on male labial palps was significantly longer than in females. The number of SC significantly differed between the male and female maxillary palps and labial palps, while ST and SS showed significant differences in the maxillary palps. These findings will contribute to further electrophysiological recording and behavioral research. RESEARCH HIGHLIGHTS: The external morphology and distribution of various sensilla on the antennae, maxillary palps, and labial palps of Bruchidius coreanus were described. Eight types and seven subtypes of antenna sensilla were observed on the antennae, while four types of sensilla were observed on the maxillary palps and labial palps. The capitate pegs were found exclusively on the antennae of female B. coreanus.

First Report of Leaf Spot on Zanthoxylum bungeanum Caused by Nigrospora sphaerica in China.

Zanthoxylum bungeanum Maxim., a deciduous shrub in Zanthoxylum genus of the Rutaceae family, has not only highly economical values as condiment and medicine, but also significantly ecological values in soil and water conservation. In March 2023, a typical leaf spot disease on Z. bungeanum (Variety "Xiao Qingjiao") was observed in the field with an area of 26.68 ha with 35% incidence and 25.4% disease intensity in Zhenfeng County (25°38'57.60″ N, 105°64'98.64″ E, 1,156 m), Guizhou Province, China. The symptom leaves showed as irregularly shaped necrotic lesions, brown to dark brown with black margin. 30 samples with typical symptoms were collected and cut into 0.5 cm × 0.5 cm pieces. Their surfaces were disinfected with 1.5% NaClO for 2 min followed by 75% ethanol for 35 s, rinsed three times with sterile distilled water, finally incubated on PDA plates at 27°C. A total of 36 isolates were obtained through single-spore cultivation. The colonies on PDA were fluffy with abundant aerial mycelia and covered the whole plates (diameter 90 mm) in 7 days. Conidia were brown to black, single-celled, smooth, spherical or oblate, 12.0-17.0 × 12.5-18.5 µm (av. = 14.5 × 15.5 µm, n = 50) and grew on a colorless transparent vesicle at the apical cell of conidiophores. The morphological characteristics were similar with N. sphaerica (Wang et al. 2017). The 5.8S DNA (ITS), translation elongation factor 1-alpha (TEF1-α) and ß-tubulin (TUB2) genes were amplified with primers ITS4/ITS5, EF1-728F/EF2, and BT2A/BT2B, respectively (White et al. 1990; Carbone and Kohn 1999, O'Donnell et al. 1998; Glass and Donaldson 1995). The ITS, TEF1-α and TUB2 sequences of two randomly selected isolates, GUCC 21-187 and GUCC 21-235, had > 99% nucleotide identities (ITS: 99.60% (504/506 bp, OR646539) and 99.61% (506/508 bp, OR640300); TEF: 100% (470/470 bp, OR654285) and 100.00% (471/471 bp, OR654286); TUB: 100.00% (408/408 bp, OR661269) and 99.52% (411/413 bp, OR661270), respectively) with those sequences of N. sphaerica (LC 7294) in GenBank (KX985932, KY019397 and KY019602, respectively). The phylogenetic tree based on sequences of ITS, TEF1-α and TUB2 indicated that GUCC 21-187 and GUCC 21-235 were most closely related to N. sphaerica (LC 7294), supported with 100%/100%/1 bootstraps. Based on morphological characteristics and molecular datasets analyses, the isolates were identified as N. sphaerica. 10 healthy 2-years-old Z. bungeanum plants were sprayed with conidial suspensions (1 × 106 conidia/mL) of the isolates and the other 5ere sprayed with sterile water as the controls, all the treated plants were cultivated in a glasshouse at 25°C under 85% relative humidity. Typical leaf spot symptoms appeared on inoculated Z. bungeanum plants after 8 days, while the control plants remained asymptomatic. N. sphaerica was re-isolated from the lesions of inoculated plants and identified by morphological and molecular identification. Pathogenicity test was performed three times with analogous results, fulfilling Koch's postulates. N. sphaerica had been reported as a common pathogen on a variety of plants including sugarcane, kiwifruit and blueberry (Cui et al. 2018; Chen et al. 2016; Wright et al. 2008). To our knowledge, this is the first report of leaf spot disease caused by N. sphaerica on Z. bungeanum in China. Our report would be helpful to Z. bungeanum growers to recognize this leaf spot disease, and corresponding measures could be taken to minimize or avoid the economic losses caused by it.

Selection and validation of optimal reference genes for RT-qPCR analyses in Aphidoletes aphidimyza Rondani (Diptera: Cecidomyiidae).

Aphidoletes aphidimyza is a predator that is an important biological agent used to control agricultural and forestry aphids. Although many studies have investigated its biological and ecological characteristics, few molecular studies have been reported. The current study was performed to identify suitable reference genes to facilitate future gene expression and function analyses via quantitative reverse transcription PCR. Eight reference genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), RPS13, RPL8, RPS3, α-Tub, ß-actin, RPL32, and elongation factor 1 alpha (EF1-α) were selected. Their expression levels were determined under four different experimental conditions (developmental stages, adult tissues, sugar treatment, and starvation treatment) using qRT-PCR technology. The stability was evaluated with five methods (Ct value, geNorm, NormFinder, BestKeeper, and RefFinder). The results showed that GAPDH, RPL32, and EF1-α were ranked as the best reference gene combinations for measuring gene expression levels among different developing stages and in various starvation treatments. RPL8 and RPS3 were recommended to normalize the gene expression levels among different adult tissues. RPL32, ß-actin, and EF1-α were recommended sugar-feeding conditions. To validate the utility of the selected reference pair, RPL8, and RPS3, we estimated the tissue-biased expression level of a chemosensory protein gene (AaphCSP1). As expected, AaphCSP1 is highly expressed in the antennae and lowly expressed in the abdomen. These findings will lay the foundation for future research on the molecular physiology and biochemistry of A. aphidimyza.

Pest management of postharvest potatoes: lethal, sublethal and transgenerational effects of the ectoparasitic mite Pyemotes zhonghuajia on the potato worm Phthorimaea operculella.

BACKGROUND: Potato, Solanum tuberosum, is one of the most important food crops in the world, playing a significant role in global food security. However, many potato industries and farms may suffer losses of tuber yield and quality in storage due to lepidopteran pests. Here, we evaluated the effectiveness of an ectoparasitic idiobiont mite Pyemotes zhonghuajia in the biological control of the potato tuber moth (PTM) Phthorimaea operculella by determining the lethal, sublethal (nonconsumptive) and transgenerational effects of P. zhonghuajia of various population densities and exposure durations on PTM survival, development and reproduction. RESULTS: Pyemotes zhonghuajia females were capable of killing all instar stages of PTM, while resistance to mite parasitism increased with the development of PTM life stage. The mortality of mature larvae (i.e., fourth instar) and pupae increased with increasing mite density and exposure duration. P. zhonghuajia imposed significant negative sublethal impacts on PTM pupation rate, female fecundity and adult longevity but not on immature development. The sublethal stress was transgenerational, resulting in lower reproduction in the offspring generation. CONCLUSION: P. zhonghuajia induces lethal, sublethal and transgenerational effects and significantly decreases PTM survival and reproductive out, demonstrating its high efficiency in the biological control of PTM. Our study provides insight into the mechanisms underlying the nonconsumptive effects of parasitism in an ectoparasite-host system and delivers critical information for the design and implementation of augmentative releases of P. zhonghuajia in the biological control of PTM in potato storage. © 2023 Society of Chemical Industry.

Effects of cold storage after cold acclimation on the fitness of Habrobracon hebetor (Hymenoptera: Braconidae).

Habrobracon hebetor (Say) (Hymenoptera: Braconidae) is a cosmopolitan, idiobiont, and gregarious ectoparasitoid, which can parasitize the larvae of several pyralid and noctuid moths. However, adult parasitoids require cold storage to ensure that adequate individuals are available when a pest outbreak occurs. To understand the effects of cold storage after acclimation on offspring fitness of H. hebetor, the development, fecundity, population parameters, and paralysis rate of the F1 generation were evaluated using an age-stage, two-sex life table. Four pairing treatments were used in this study, with refrigerated males and females (ReF×ReM), unrefrigerated females and refrigerated males (UnF×ReM), refrigerated females and unrefrigerated males (ReF×UnM), and unrefrigerated females and males (UnF×UnM, control). Cold storage after acclimation had no significant effect on the fecundity or oviposition period of F0-generation H. hebetor. Moreover, the survival rate (Sa = 61.43%), proportion of females (Nf/N = 0.41), intrinsic rate of increase (r = 0.3450), finite rate of increase (λ = 1.4121), net reproduction rate (R0 = 149.47), and net paralysis rate (C0 = 74.52) of ReF×UnM and UnF×UnM (Sa = 50.00%, Nf/N = 0.34, r = 0.3297, λ = 1.3881, R0 = 155.69, C0 = 62.90, respectively) treatments were significantly higher than those of the ReF×ReM treatment (Sa = 45%, Nf/N = 0.16, r = 0.2277, λ = 1.2558, R0 = 68.81, C0 = 31.61, respectively) (except for the Sa of UnF×UnM treatment), and there was no significant difference between the 2 treatments. Overall, it is advisable to avoid simultaneous cold storage of female and male parasitoids or to add unrefrigerated males appropriately when using cold-stored parasitoids to control pests.

A new species of Neohydatothrips (Thysanoptera: Sericothripinae) from China.

Neohydatothrips stachyurus sp. n. is described from Guizhou, China. Morphologically, this new Sericothripinae species is characterized by the shape of blotch on pronotum and the distribution of microtrichia on abdominal segments. The distribution of Neohydatothrips species from China also is discussed.

Leptosphaerulina americana causing holing disease on tobacco in China.

Tobacco (Nicotiana tabacum) is one of the most important industrial crops in the world. Its leaves are the main raw material for cigarettes, but they are often threatened by fungal pathogens in the production process (Wang et al. 2022). From May to June 2022, a disease of tobacco (cv K326) (15% of plants) in a 0.3-ha field in Jingxi of Guangxi Province showed symptoms of local necrosis and perforation of middle and basal leaves (Fig S1). Pieces of leaf tissue (3 × 3 mm) were excised from the edge of the necrotic lesion of each plant, treated with 75% ethanol for 10 s, soaked in 2% NaClO solution for 1-2 min, rinsed with sterile water for three times, and then plated on potato dextrose agar（PDA）medium and incubated at 28°C. Isolate TJYA13 was used for subsequent studies. After 8 days, the colony margin was yellowish brown and irregular, the center was black and plicated. The isolate TJYA13 was incubated on oatmeal agar medium at 28°C for 4 days, and many pseudothecia were observed embedded on the surface of the medium. Pseudothecium was globose or subglobose, dark brown, and size was 184.7-304.7 µm × 187.5-340.5 µm (n=20). Ascospores were usually wrapped by the saccate ascus in pseudothecium, cylindrical or ellipsoidal, with 5-6 transverse septa, and size was 12.2-18.5 µm × 35.6-51.8 µm (n=80). The morphological characteristics of ascospores were consistent with a Leptosphaerulina species (Hou et al. 2020). For accurate identification, the genomic DNA of isolate TJYA13 was extracted with Ezup Column Fungi Genomic DNA Purification Kit (Sangon, Shanghai, China). The ITS region, 28s ribosomal RNA (LSU), ß-tubulin (TUB), and RNA polymerase II second largest subunit (RPB2) were amplified with primers ITS1/ITS4 (Gardes and Bruns 1993; White et al. 1990), LROR/LR7 (Rehner and Samuels 1994), Btub2Fd/Btub4Rd (Woudenberg et al. 2009), and RPB2-5F2/fRPB2-7cR (Liu et al. 1999), respectively and sequenced at Sangon Biotech (Sichuan, China). The sequences were deposited in GenBank (accession nos. OP926927, OP926933, OP939419, OP939422). The phylogenetic analysis grouped the isolate TJYA13 within the L. americana clade (Fig S2) (Hou et al. 2020). Pathogenicity of the isolate TJYA13 was verified on four healthy tobacco plants (cv K326). The mycelial plugs were inoculated on leaves sterilized with 75% ethanol, and control plants were inoculated with sterile PDA plugs. Plants were incubated at 28 ℃ and 78% humidity. After 10 days, the leaves inoculated with mycelial plugs had symptoms similar to those in the field, but there were no symptoms on the control leaves. L. americana were reisolated from the leaves inoculated with the mycelial plugs. To the best of our knowledge, this is the first report of L. americana causing holing disease on tobacco in China. This disease may reduce yields and lower quality of flue-cured tobacco leaf. Therefore, the emergence of tobacco holing disease should be noted to prevent potential damage to tobacco production in Guangxi. Reference 1. Hou L. W., et al. 2020. Stud. Mycol. 96: 309-396 2. Liu, Y. J., et al. 1999. Mol. Biol. Evol. 16:1799. 3. Rehner, S. A., and Samuels, G. J. 1994. Mycol. Res. 98:625. 4. Wang H. et al. 2022. Microorganisms. 10: 1890. 5. White, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. 6. Woudenberg, J. H. C., et al. 2009. Persoonia 22:56. The author(s) declare no conflict of interest. Funding: Funding was provided by Guangxi Zhuang Autonomous Region Tobacco Monopoly Bureau (grant no. 202,145,000,024,006). Tobacco (Nicotiana tabacum) is one of the most important industrial crops in the world. Its leaves are the main raw material for cigarettes, but they are often threatened by fungal pathogens in the production process (Wang et al. 2022). From May to June 2022, a disease of tobacco (cv K326) (15% of plants) in a 0.3-ha field in Jingxi of Guangxi Province showed symptoms of local necrosis and perforation of middle and basal leaves (Fig S1). Pieces of leaf tissue (3 × 3 mm) were excised from the edge of the necrotic lesion of each plant, treated with 75% ethanol for 10 s, soaked in 2% NaClO solution for 1-2 min, rinsed with sterile water for three times, and then plated on potato dextrose agar（PDA）medium and incubated at 28°C. Isolate TJYA13 was used for subsequent studies. After 8 days, the colony margin was yellowish brown and irregular, the center was black and plicated. The isolate TJYA13 was incubated on oatmeal agar medium at 28°C for 4 days, and many pseudothecia were observed embedded on the surface of the medium. Pseudothecium was globose or subglobose, dark brown, and size was 184.7-304.7 µm × 187.5-340.5 µm (n=20). Ascospores were usually wrapped by the saccate ascus in pseudothecium, cylindrical or ellipsoidal, with 5-6 transverse septa, and size was 12.2-18.5 µm × 35.6-51.8 µm (n=80). The morphological characteristics of ascospores were consistent with a Leptosphaerulina species (Hou et al. 2020). For accurate identification, the genomic DNA of isolate TJYA13 was extracted with Ezup Column Fungi Genomic DNA Purification Kit (Sangon, Shanghai, China). The ITS region, 28s ribosomal RNA (LSU), ß-tubulin (TUB), and RNA polymerase II second largest subunit (RPB2) were amplified with primers ITS1/ITS4 (Gardes and Bruns 1993; White et al. 1990), LROR/LR7 (Rehner and Samuels 1994), Btub2Fd/Btub4Rd (Woudenberg et al. 2009), and RPB2-5F2/fRPB2-7cR (Liu et al. 1999), respectively and sequenced at Sangon Biotech (Sichuan, China). The sequences were deposited in GenBank (accession nos. OP926927, OP926933, OP939419, OP939422). The phylogenetic analysis grouped the isolate TJYA13 within the L. americana clade (Fig S2) (Hou et al. 2020). Pathogenicity of the isolate TJYA13 was verified on four healthy tobacco plants (cv K326). The mycelial plugs were inoculated on leaves sterilized with 75% ethanol, and control plants were inoculated with sterile PDA plugs. Plants were incubated at 28 ℃ and 78% humidity. After 10 days, the leaves inoculated with mycelial plugs had symptoms similar to those in the field, but there were no symptoms on the control leaves. L. americana were reisolated from the leaves inoculated with the mycelial plugs. To the best of our knowledge, this is the first report of L. americana causing holing disease on tobacco in China. This disease may reduce yields and lower quality of flue-cured tobacco leaf. Therefore, the emergence of tobacco holing disease should be noted to prevent potential damage to tobacco production in Guangxi.

Transgenerational effects of multiple mating in Spodoptera litura Fabricius (Lepidoptera: Noctuidae).

Polyandrous mating can result in sexual conflict and/or promote the evolution of mating patterns. Does multiple mating by females support the genetic benefits hypothesis and can it be validated as an evolutionary strategy? If we are to decipher the consequences of sexual interactions and understand the interplay of sexual conflict and multiple generational benefits, the transgenerational effects need to be followed over multiple generations. We investigated the effects of three mating patterns, single mating, repeated mating, and multiple mating, on parental Spodoptera litura copulation behavior, and then identified the impact on the development, survival, and fecundity of the F1 and F2 generations. Fecundity was not significantly affected in the F1 generation but was substantially enhanced in the F2 generation. There was a reversal of offspring fitness across the F2 generations from the F1 generations in progeny produced by multiple mating. In addition, the intrinsic rate of increase, finite rate of increase and net reproductive rate in the F1 generation the multiple mating treatment was significantly lower than in the single mating treatment, but there was no apparent effect on the F2 generation. Repeated mating had no significant effects on progeny fitness. We postulate that multiple mating imposes cross-transgenerational effects and may ultimately influence multigenerational fitness in S. litura.

Three new species of Atkinsoniella (Arthropoda, Insecta, Hemiptera, Cicadellidae, Cicadellinae) from China, with an updated checklist to the known species worldwide.

The sharpshooter genus Atkinsoniella Distant, 1908 includes 99 valid species worldwide. Here, three new species from China are described and illustrated: Atkinsoniellastenopyga, A.wangi, and A.yingjiangensisspp. nov. An updated checklist of the known Atkinsoniella species worldwide based on the data of previous literature and studied materials is also provided. All the type specimens of three new species are deposited at the Institute of Entomology, Guizhou University, Guiyang, China.

The complete mitochondrial genome of Picromerus lewisi Scott 1874 (Hemiptera: Pentatomidae: Asopinae).

Picromerus lewisi Scott (Hemiptera: Pentatomidae) is a widely used natural enemy, through this study, we proved that its complete mitochondrial genome of it had similar characteristics to those of other Hemiptera. The mitogenome of P. lewisi is a circular molecule of 18,123 bp with 74.0% A + T content, containing 13 protein-coding genes (PCGs), 22 tRNAs, 2 rRNAs, and one control region. Phylogenetic tree based on 13 PCGs from 17 Panheteroptera species (two species of the Cimicomorpha are used as outgroup, 15 species belong to the Pentatomomorpha) suggested that P. lewisi has a closer relationship with E. thomsoni within Pentatomidae family.

Complete mitochondrial genome of Gortyna guizhouensis Wu, Yang, and Han, 2022 (Lepidoptera, Noctuidae) from China.

Gortyna guizhouensis Wu, Yang, and Han, 2022 (Lepidoptera, Noctuidae) is a newly discovered moth species that feeds on tobacco piths during the larval stage. In this study, we sequenced and analyzed the complete mitochondrial genome (mitogenome) of G. guizhouensis larvae via next-generation sequencing. The mitogenome was 15,441 bp long with an overall A + T content of 79.1%, 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA subunit genes and one control region. The phylogenetic tree, based on the nucleic acid sequences of 13 shared PCGs of 20 Noctuidae species, revealed that G. guizhouensis is in a well-supported clade with Striacosta albicosta.

Screening for broad-spectrum antimicrobial endophytes from Rosa roxburghii and multi-omic analyses of biosynthetic capacity.

Plants with certain medicinal values are a good source for isolating function-specific endophytes. Rosa roxburghii Tratt. has been reported to be a botanical source of antimicrobial compounds, which may represent a promising candidate for screening endophytic fungi with antimicrobial potential. In this study, 54 endophytes were isolated and molecularly identified from R. roxburghii. The preliminary screening using the plate confrontation method resulted in 15 different endophytic strains showing at least one strong inhibition or three or more moderate inhibition against the 12 tested strains. Further re-screening experiments based on the disc diffusion method demonstrated that Epicoccum latusicollum HGUP191049 and Setophoma terrestris HGUP190028 had excellent antagonistic activity. The minimum inhibitory concentration (MIC) test for extracellular metabolites finally indicated that HGUP191049 had lower MIC values and a broader antimicrobial spectrum, compared to HGUP190028. Genomic, non-target metabolomic, and comparative genomic studies were performed to understand the biosynthetic capacity of the screened-out endophytic fungus. Genome sequencing and annotation of HGUP191049 revealed a size of 33.24 megabase pairs (Mbp), with 24 biosynthetic gene clusters (BGCs), where the putative antimicrobial compounds, oxyjavanicin, patulin and squalestatin S1 were encoded by three different BGCs, respectively. In addition, the non-targeted metabolic results demonstrated that the strain contained approximately 120 antimicrobial secondary metabolites and was structurally diverse. Finally, comparative genomics revealed differences in pathogenicity, virulence, and carbohydrate-active enzymes in the genome of Epicoccum spp. Moreover, the results of the comparative analyses presumed that Epicoccum is a promising source of antimicrobial terpenes, while oxyjavanicin and squalestatin S1 are antimicrobial compounds shared by the genus. In conclusion, R. roxburghii and the endophytic HGUP191049 isolated from it are promising sources of broad-spectrum antimicrobial agents.

Population Parameters and Feeding Preference of Spodoptera litura (Lepidoptera: Noctuidae) on Different Asparagus officinalis Tissues.

Spodoptera litura is an important pest that seriously affects Asparagus officinalis production. To clarify the population characteristics and feeding preference of S. litura on different asparagus tissues, asparagus stems and leaves were selected as the research objects, related studies were conducted by constructing the life table and the feeding preference experiment. The results showed that S. litura could complete its development and reproduction normally on asparagus stems or leaves. Although the adult longevity and fecundity of S. litura on the two types of tissues were not significantly different, the development duration of larvae and pupae, and total preoviposition period on leaves were significantly longer than those raised on stems. The intrinsic rate of increase and finite rate of increase were 0.186 d-1 and 1.204 d-1 on stems, which were significantly higher than those fed on leaves (0.161 d-1 and 1.175 d-1). The mean generation time on stems (32.88 d) was significantly lower than on leaves (36.88 d). It indicated that stems were more suitable for its population growth. In the feeding preference, the third and fifth instar larvae preferred to feed on leaves, and other instar larvae (except for the sixth instar of 2.5 h) had no significant difference. These results will provide a theoretical reference for further research and forecasting and integrated control.

Description and complete mitochondrial genome of Atkinsoniella zizhongi sp. nov. (Hemiptera: Cicadellidae: Cicadellinae) from China and its phylogenetic implications.

A new species, Atkinsoniella zizhongi sp. nov. of the subfamily Cicadellinae, was described and illustrated from China. The new species is similar to A. nigrominiatula (Jacobi, 1944), A. limba Kuoh, 1991, A. dormana Li, 1992, A. peaka Yang, Meng et Li, 2017, and A. divaricata Yang, Meng et Li, 2017. But the characteristics of aedeagus and pygofer process can be used to distinguish them easily. The complete mitochondrial genome of the paratype was sequenced and assembled. The mitogenome of A. zizhongi sp. nov. was 16,483 bp in length, with an A+T content of 75.9%, containing 37 typical genes and a control region (CR). The gene order was consistent with the inferred insect ancestral mitochondrial genome. All of the PCGs were determined to have the typical stop codon TAA or TAG, while COX2 and ND5 ended with incomplete termination codons T and TA, respectively. In addition, phylogenetic trees were reconstructed based on PCGs and rRNAs using both the maximum likelihood (ML) and Bayesian inference (BI) methods. The results showed that the intergeneric and interspecific relationships within the subfamily Cicadellinae were completely consistent in all of the phylogenetic trees, except that the different interspecific relationships within the genus Bothrogonia were detected in the ML analysis based on the amino acid sequences. This study enriches the species diversity of Cicadellinae and further promotes research on its phylogeny.

A Long-Read Genome Assembly of a Native Mite in China Pyemotes zhonghuajia Yu, Zhang & He (Prostigmata: Pyemotidae) Reveals Gene Expansion in Toxin-Related Gene Families.

Pyemotes zhonghuajia Yu, Zhang & He (Prostigmata: Pyemotidae), discovered in China, has been demonstrated as a high-efficient natural enemy in controlling many agricultural and forestry pests. This mite injects toxins into the host (eggs, larvae, pupae, and adults), resulting in its paralyzation and then gets nourishment for reproductive development. These toxins have been approved to be mammal-safe, which have the potential to be used as biocontrol pesticides. Toxin proteins have been identified from many insects, especially those from the orders Scorpions and Araneae, some of which are now widely used as efficient biocontrol pesticides. However, toxin proteins in mites are not yet understood. In this study, we assembled the genome of P. zhonghuajia using PacBio technology and then identified toxin-related genes that are likely to be responsible for the paralytic process of P. zhonghuajia. The genome assembly has a size of 71.943 Mb, including 20 contigs with a N50 length of 21.248 Mb and a BUSCO completeness ratio of 90.6% (n = 1367). These contigs were subsequently assigned to three chromosomes. There were 11,183 protein coding genes annotated, which were assessed with 91.2% BUSCO completeness (n = 1066). Neurotoxin and dermonecrotic toxin gene families were significantly expanded within the genus of Pyemotes and they also formed several gene clusters on the chromosomes. Most of the genes from these two families and all of the three agatoxin genes were shown with higher expression in the one-day-old mites compared to the seven-day-pregnant mites, supporting that the one-day-old mites cause paralyzation and even death of the host. The identification of these toxin proteins may provide insights into how to improve the parasitism efficiency of this mite, and the purification of these proteins may be used to develop new biological pesticides.

A chromosome-level genome assembly of Neotoxoptera formosana (Takahashi, 1921) (Hemiptera: Aphididae).

Neotoxoptera formosana (Takahashi), the onion aphid, is an oligophagous pest that mainly feeds on plants from the Allium genus. It sucks nutrients from the plants and indirectly acts as a vector for plant viruses. This aphid causes severe economic losses to Allium tuberosum agriculture in China. To better understand the host plant specificity of N. formosana on Allium plants and provide essential information for the control of this pest, we generated the entire genome using Pacific Biosciences long-read sequencing and Hi-C data. Six chromosomes were assembled to give a final size of 372.470 Mb, with an N50 scaffold of 66.911 Mb. The final draft genome assembly, from 192 Gb of raw data, was approximately 371.791 Mb in size, with an N50 contig of 24.99 Kb and an N50 scaffold of 2.637 Mb. The average GC content was 30.96%. We identified 73 Mb (31.22%) of repetitive sequences, 14,175 protein-coding genes, and 719 noncoding RNAs. The phylogenetic analysis showed that N. formosana and Pentalonia nigronervosa are sister groups. We found significantly expanded gene families that were involved in the THAP domain, the DDE superfamily endonuclease, zinc finger, immunity (ankyrin repeats), digestive enzyme (serine carboxypeptidase) and chemosensory receptor. This genome assembly could provide a solid foundation for future studies on the host specificity of N. formosana and pesticide-resistant aphid management.

A new species of the genus Gortyna Ochsenheimer, 1816 from China, with notes on the larval feeding habits (Lepidoptera, Noctuidae, Noctuinae: Apameini).

A new species of the genus Gortyna Ochsenheimer, 1816, G. guizhouensis sp. n., is described from Guizhou Province, China. Adults and genitalic structures of the new species and related species are illustrated. Observations of feeding habits and host plant of the larva are also provided.