Performance and preference of larval parasitoid, Microplitis pallidipes (Hymenoptera: Braconidae), on 2 Spodoptera pest species.

Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) and Spodoptera litura (Fabricius) are the main pests on corn (Poaceae: Gramineae). The performance of the larval wasp, Microplitis pallidipes Szépligeti (Hymenoptera: Braconidae), was reported on S. frugiperda and S. litura. In this study, we evaluated host selectivity, constructed an age-stage, 2-sex life table, and assessed the pest control potential of M. pallidipes against these 2 pests under laboratory conditions. In a 2-choice host preference experiment, M. pallidipes exhibited a stronger preference for S. frugiperda over S. litura and a distinct preference for second instars. We also investigated the parasitism of females that were either unfed or fed with 10% honey-water solution under different host densities and found that the highest parasitism rate was observed when M. pallidipes were fed with honey-water solution on the first day after mating and a presented female wasp:host ratio of 1:90. In a nonselective assay, M. pallidipes successfully completed a full generation on both hosts. However, the parasitoids exhibited higher fitness and population growth potential when reared on S. frugiperda, with a net reproductive rate (R0) of 24.24, an intrinsic rate of increase (r) of 0.20 per day, a finite rate of increase (λ) of 1.23 per day, and a mean generation time (T) of 15.69 days. This study elucidates the performance of M. pallidipes on 2 Spodoptera host species and offers insights into its biological control potential on lepidopteran pests.

Seasonal migration pattern of an important rice pest, Nilaparvata lugens (Hemiptera: Delphacidae), on Hainan Island, China.

The brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae) (BPH), is a major, regionally migratory pest of rice in Asia. Despite intensive migratory studies, the seasonal migratory pattern of this species in the year-round breeding region (i.e., Hainan Island) remains largely unknown. To understand the migration dynamics, we conducted relevant research based on BPH light trap catches on Hainan in 2017-2022. Results showed that the occurrence dynamics of BPH in Hainan oscillated in different years and seasons. Overall, there are 4 migration peak periods on Hainan, with outbound peak periods in April-May and August-September and inbound peak periods in June and October. Trajectory and wind fields showed that in August-September, Hainan had complex meteorological conditions, and the outbound paths of BPH were variable, mainly landing in southeastern Guangxi and southwestern Guangdong. In contrast, April-May was influenced by southeasterly winds, with a wider outbound range and a further distance, landing along northeastern Vietnam from west to east toward Guangxi and Guangdong areas with time. Wind-rain fields, trajectory, and hydrogen isotope showed that in June, southwesterly winds transported BPH northward from the Indochina peninsula, where it was hindered by rainfall in the Leizhou Peninsula area and the migrants will be "rained out" into Hainan. Identically, in October, the interactions between northeasterly winds and rainfall belts on the southwest shore of Hainan increased the opportunities for BPH to migrate southward to reach Hainan. Our results suggest that Hainan is not only an important source of BPH migrating from East Asia but also plays the role of a bridge.

Knockdown of the ecdysone receptor disrupts development and causes mortality in the melon fly, Zeugodacus cucurbitae.

Cucurbits are important economic plants that are attacked by numerous pests, among which the melon fly Zeugodacus cucurbitae is extremely problematic. New sustainable pest control strategies are necessary to replace chemical insecticides that are harmful to the environment, human health and nontarget species. The RNA interference (RNAi) technology is one of the most promising tools due to high efficiency and species specificity. We developed an RNAi strategy targeting the ecdysone receptor (ECR) of Z. cucurbitae, which plays an important role in moulting and reproduction. We identified, described and isolated the ECR gene of Z. cucurbitae and measured its expression pattern across developmental stages and tissues. ZcECR knockdown via dsZcECR ingestion caused a significant larval mortality and abnormal phenotypes in pupae and adults. About 68% of larvae fed with a dsZcECR-treated diet failed to enter the pupal stage and died. In addition, ZcECR knockdown dramatically reduced pupal weight (by 3.24 mg on average) and fecundity (by about 23%). RNAi targeting the ECR gene is therefore a promising method to control Z. cucurbitae, paving the way for the development of novel sustainable and highly specific control strategies.

Ecology, invasion history and biodiversity-driven management of the coconut black-headed caterpillar Opisina arenosella in Asia.

The coconut black-headed caterpillar (BHC), Opisina arenosella Walker (Lepidoptera: Xyloryctidae) is an important herbivore of palm trees that originates in South Asia. Over the past decades, O. arenosella has spread to several countries in Eastern and Southeast Asia. BHC larval feeding can cause severe defoliation and occasional plant death, resulting in direct production losses (e.g., for coconut) while degrading the aesthetic value of urban and rural landscapes. In this review paper, we systematically cover taxonomy, bio-ecology, invasion history and current management of O. arenosella throughout Asia. Given that O. arenosella is routinely controlled with insecticides, we equally explore options for more sustainable management through agroecological and biodiversity-based tactics e.g., cultural control or biological control. Also, recent advances in chemical ecology have unlocked lucrative opportunities for volatile-mediated monitoring, mating disruption and mass-trapping. Substantial progress has been made in augmentation biological control, with scheduled releases of laboratory-reared parasitoids lowering BHC infestation pressure up to 95%. Equally, resident ants provide 75-98% mortality of BHC egg masses within the palm canopy. Biological control has been effectively paired with sanitary measures and good agronomy (i.e., proper fertilization, irrigation), and promoted through participatory farmer training programs. Our comprehensive listing of non-chemical preventative and curative tactics offer bright prospects for a more environmentally-sound, biodiversity-driven mitigation of a palm pest of regional allure.

First Report of Alternaria alternata Causing White Leaf Spot on Allium tuberosum in China.

Puding County is the major Allium tuberosum growing area in Guizhou Province of China. In 2019, white leaf spots were observed on Allium tuberosum in Puding County (26.31°N, 105.64°E). The white spots, ranging from elliptic to irregular in shape, first appeared on leaf tips. With disease aggravation, spots gradually coalesced, forming necrotic patches with yellow margins causing leaf necrosis; sometimes there was gray mold on dead leaves. The incidence of the diseased leaf rate was estimated to be 27-48%. To identify the pathogenic agent, 150 leaf tissues (5 mm × 5 mm) were obtained from disease-healthy junctions of 50 diseased leaves. Leaf tissues were disinfected in 75% ethanol for 30 s, soaked in 0.5% sodium hypochlorite for 5 min, and flushed three times with sterile water, before being placed on potato dextrose agar (PDA) in the dark at 25 °C. When colonies appeared, the mycelial tips were picked and placed on new PDA. Purified fungus was obtained after repeating this last step several times. The colonies were grayish-green with white round margins. Conidiophores (2.7-4.5 µm × 27-81 µm) were brown, straight, or flexuous with branches and septa. Conidia (8-34 µm × 5-16 µm) were brown, with 0-5 transverse septa and 0-4 longitudinal septa. The 18S nuclear ribosomal DNA (nrDNA; SSU), 28S nrDNA (LSU), RNA polymerase II second largest subunit (RPB2), internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation elongation factor 1-alpha (TEF-α) (Woudenberg et al. 2013) were amplified and sequenced. The sequences were deposited in GenBank (ITS: OP703616, LSU: OP860684, SSU: OP860685, GAPDH: OP902372, RPB2: OP902373, TEF1-α: OP902374). According to BLAST analysis, the ITS, LSU, GAPDH, RPB2, SSU, and TEF1-α of the straishowed 100% (689 of 731 base pairs; bp), 100% (916 of 938 bp), 100% (579 of 600 bp), 100% (946 of 985 bp), 100% (1093 of 1134 bp), and 100% (240 of 240 bp) sequence identity to those of Alternaria alternata (ITS: LC440581.1, LSU: KX609781.1, GAPDH: MT109295.1, RPB2: MK605900.1, SSU: ON055699.1 and TEF1-α: OM220081.1). A phylogenetic tree was constructed using PAUP4 and the maximum parsimony method with 1000 replicas of bootstrapping for all datasets. According to morphological characteristics and phylogenetic analysis, FJ-1 was identified as Alternaria alternata (Simmons 2007, Woudenberg et al. 2015). The strain was preserved in the Agricultural Culture Collection of China (preservation number: ACC39969). To determine the pathogenicity of Alternaria alternata against Allium tuberosum, wounded healthy leaves were inoculated with a conidial suspension (106 conidial/mL) and round mycelial plugs (4mm). Sterile agar PDA plugs with no mycelium or sterile water were inoculated as negative controls. Three days later, white spots appeared on the wounded leaves inoculated with mycelial plugs or conidial suspension. However, the symptoms caused by conidial suspensions were weaker than those caused by mycelial plugs. No symptoms were observed in the control group. The experimental symptoms were consistent with the phenomena observed in the field. The same fungus was reisolated from necrotic lesions and identified as Alternaria alternata using the method described above. To our knowledge, this is the first report of Alternaria alternata causing white leaf spots on Allium tuberosum in China, a disease seriously affected the yield and quality of Allium tuberosum and caused economic losses to farmers. Reference: Simmons EG (2007) Alternaria: an identification manual. CBS Fungal Biodiversity Centre, Utrecht, the Netherlands. Woudenberg JHC, Groenewald JZ, Binder M, Crous PW ( 2013) Alternaria redefined. Stud Mycol, 75: 171-212. https://doi.org/10.3114/sim0015. Woudenberg JHC, Seidl MF, Groenewald JZ, Vries M de, Stielow JB, Thomma BPHJ, Crous PW (2015) Alternaria section Alternaria: Species, formae speciales or pathotypes? Stud Mycol, 82:1-21. https://doi.org/10.1016/j.simyco.2015.07.001.

Characterization of the complete mitochondrial genome and phylogenetic analysis of Cnaphalocrocis patnalis (Bradley 1981) (Lepidoptera: Crambidae).

Cnaphalocrocis patnalis (Bradley 1981) is a major pest that threatens the safety of rice production in the world. In the present study, we determined the complete mitogenome of C. patnalis. This mitogenome was 15,305 bp in length (GenBank accession No. OL449028), which contained two ribosomal RNA genes, 22 transfer RNAs, 13 protein-coding genes (PCGs) and one non-coding AT-rich region with a length of 344 bp. All the 22 tRNA genes displayed a typical clover-leaf structure, except for trnS1. Twelve PCGs were initiated by ATN codons, and COX1 started with TTG. All the PCGs used the typical stop codon 'TAA' and 'TAG.' Phylogenetic tree demonstrated that C. patnalis belongs to the family Crambidae.

Safety Evaluation of Chemical Insecticides to Tetrastichus howardi (Hymenoptera: Eulophidae), a Pupal Parasitoid of Spodoptera frugiperda (Lepidoptera: Noctuidae) Using Three Exposure Routes.

Spodoptera frugiperda has become a major pest in many crops worldwide. The main control strategies are biological and chemical controls. However, pesticides have varying degrees of toxicity to parasitic wasps in the field. To integrate chemical and biological controls, we evaluated the safety of insecticides to Tetrastichus howardi, an important pupal parasitoid of S. frugiperda. This study assessed the toxicity of six major control insecticides (emamectin benzoate, chlorfenapyr, indoxacarb, chlorantraniliprole, bisultap, and lufenuron) to T. howardi based on risk quotient. The results showed that indoxacarb had the lowest risk quotient (RQ = 7.43). Then the side effects of three sublethal concentrations (LC20, LC30, LC40) of indoxacarb were tested using three methods (1. Adult exposure to pesticide residues on the glass tube; 2. Adult exposure to pesticide residues on the host; 3. Larval exposure to pesticides through host exposure). Overall, T. howardi had a lower parasitism rate and emergence rate with the higher pesticide concentrations. Furthermore, among three methods, the adult exposure to pesticide residues on the glass tube was the most efficient in inhibiting the parasitism rate, and impairing the emergence rate and the offspring female/male ratio. This study guides a more scientific and comprehensive application of pesticides and releases natural enemies in the field.

Complete mitochondrial genome of nipa palm hispid beetle Octodonta nipae Maulik (Coleoptera: Chrysomelidae: Cassidinae).

Octodonta nipae (Maulik 1921) is a dangerous forestry quarantine pest, which mainly harms palms. In the present study, we determined complete mitogenome of O. nipae. This mitogenome was 15,397 bp in length (GenBank Accession no. MW802252), which contained 2 ribosomal RNA genes, 22 transfer RNAs, 13 protein-coding genes (PCGs) and one non-coding AT-rich region with the length of 883 bp. All of the 22 tRNA genes displayed a typical clover-leaf structure, with the exception of tRNAPhe, tRNALeu, tRNAAsn, tRNAPro and tRNAThr. Twelve PCGs were initiated by ATN codons, and NAD1 started with TTG. Ten PCGs used the typical stop codon 'TAA' and 'TGA', while three PCGs (COX2, COX3, NAD4) used the incomplete stop codons 'TA' or 'T'. Phylogenetic tree demonstrated that O. nipae belongs to the family Chrysomelidae and closer to the superfamily Cassidinae.

Characterization of the mitochondrial genome of Tetrastichus howardi (Olliff 1893) (Hymenoptera: Eulophidae).

The mitochondrial genome has been widely used in the study of phylogeny and species-level evolution. Here, we sequenced and analyzed the full mitogenome of Tetrastichus howardi, an important natural enemy of many lepidopteran pests. The complete mitochondrial genome has 14,791 nucleotides, containing 13 protein-coding genes (PCGs), 2 ribosomal RNA genes (rRNAs), 22 transfer RNA genes (tRNAs), and a partial control region. All the 13 PCGs started with typical ATN (ATA, ATG, and ATT) codon. Among 13 PCGs, nine genes terminated with the stop codon TAA and four genes terminated with T. Our study provides information on comparative mitogenomics of Eulophidae.

RNAi-Mediated Knockdown of Imaginal Disc Growth Factors (IDGFs) Genes Causes Developmental Malformation and Mortality in Melon Fly, Zeugodacus cucurbitae.

This study reports the first successful use of oral feeding dsRNA technique for functional characterization of imaginal disc growth factors (IDGFs) genes (IDGF1, IDGF3_1, IDGF4_0, IDGF4_1, and IDGF6) in melon fly Zeugodacus cucurbitae. Phylogenetic and domain analysis indicates that these genes had high similarity with other Tephritidae fruit flies homolog and contain only one conserved domain among these five genes, which is glyco-18 domain (glyco-hydro-18 domain). Gene expression analysis at different developmental stages revealed that these genes were expressed at larval, pupal, and adult stages. To understand their role in different developmental stages, larvae were fed dsRNA-corresponding to each of the five IDGFs, in an artificial diet. RNAi-mediated knockdown of IDGF1 shows no phenotypic effects but caused mortality (10.4%), while IDGF4_0 caused malformed pharate at the adult stage where insects failed to shed their old cuticle and remained attached with their body, highest mortality (49.2%) was recorded compared to dsRNA-green fluorescent protein (GFP) or DEPC. Silencing of IDGF3_1 and IDGF4_1 cause lethal phenotype in larvae, (17.2%) and (40%) mortality was indexed in Z. cucurbitae. IDGF6 was mainly expressed in pupae and adult stages, and its silencing caused a malformation in adult wings. The developmental defects such as malformation in wings, larval-larval lethality, pupal-adult malformation, and small body size show that IDGFs are key developmental genes in the melon fly. Our results provide a baseline for the melon fly management and understanding of IDGFs specific functions in Z. cucurbitae.

The complete mitochondrial genome of an egg parasitoid (Trichogramma chilonis).

Trichogramma chilonis Ishii is an important natural enemy of several lepidopterous pests on crops. In this study, we sequenced the complete mitochondrial genome of T. chilonis (GenBank accession number MW789210). The length of the complete mitochondrial genome was 16,147 bp, containing 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and a non-coding control region. The overall base composition of the genome in descending order was 44.8% T, 41.8% A, 9.0% G and 4.5% C, with a significant AT bias of 86.6%. Phylogenetic analysis indicated that T. chilonis had a close relationship with Trichogramma ostriniae.

The mitochondrial genome of a parasitic wasp, Chouioia cunea Yang (Hymenoptera: Chalcidoidea: Eulophidae) and phylogenetic analysis.

Chouioia cunea Yang 1989 is a parasitic wasp and natural enemy of several lepidopteran pests during their pupal stage. In this study, we sequenced and analyzed the mitochondrial genome of C. cunea, and obtained a complete DNA molecule that is 14,930 bp in size with 13 protein-coding genes (PCGs), two ribosomal RNA genes (rRNAs), and 22 transfer RNA genes (tRNAs) (GenBank accession number MW192646). All the 13 PCGs started with typical ATN (ATA, ATG, and ATT) and terminated with the stop codon TAA or TAG. Phylogenetic analysis showed that C. cunea formed the sister group with Tamarixia radiata, which belongs to the same family.

Complete mitochondrial genome of a parasitic wasp Microplitis pallidipes (Hymenoptera: Braconidae: Microgastrinae).

The complete mitochondrial genome of Microplitis pallidipes contains 15931 bp, with an A + T content of 86.5% and consists of 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a control region (GenBank accession no. MN892396). All of the 22 tRNA genes displayed an usual clover-leaf structure. Gene rearrangement events occurred in this species, there are eight tRNA genes changed their positions or/and directions. 13 PCGs started with ATN. Ten PCGs used the typical stop codon 'TAA' and 'TAG', three PCGs terminated with incomplete stop codons (TA). Phylogenetic analyses within the microgastroid complex were performed based on mitochondrial protein-coding genes.

The mitochondrial genome of an ectoparasitoid wasp, Habrobracon hebetor (Hymenoptera: Braconidae: Braconinae).

In this study, we sequenced the mitogenome of Habrobracon hebetor, and obtained almost complete mitogenome of it. The mitogenome contains 14,629 bp which consists of 13 protein-coding genes (PCGs), 20 transfer RNA genes (trnI and trnM are absent), and 2 ribosomal RNA genes (GenBank accession no. MT558946). Gene rearrangement events occurred in this species, five tRNA genes with changes in positions or/and directions are found. All of 13 PCGs started with ATN. Eight PCGs used the typical stop codon 'TAA', five PCGs terminated with incomplete stop codons (T). Phylogenetic analyses within the Cyclostomes were performed based on mitochondrial PCGs.