Xuncong Ji

The tea mosquito bug, Helopeltis theivora Waterhouse, (Hemiptera: Miridae) is a significant sap-sucking pest in tropical tea plantations that causes substantial losses in tea production on Hainan Island, China. The morphological and ultrastructural characteristics of H. theivora antennae have not been elucidated. Here, we used several microscopy techniques (SDM, SEM, and TEM) to investigate the morphology as well as the setae and sensilla on the antennae of nymphs and adults of H. theivora. SDM observations indicated that the antennae of H. theivora were filamentous in shape and included four segments: scape, pedicel, flagellum I, and flagellum II. The length of the antenna was approximately twice that of the body and the setae were enriched in flagellum II. The SEM results showed that there were a total of six types of sensilla on the antenna of H. theivora, including the sensilla chaetica (SCh), sensilla trichoidea (ST), sensilla basiconica (SB), sensilla coeloconica (SCo), sensilla mammilliformia (SM), and Böhm’s bristles (BB). In particular, there were three subtypes (I, II, and III) of different lengths in SCh and SB, and two subtypes of straight (I) and curved (II) sensilla in ST. The TEM results indicated that diverse internal structures were present in SCh, ST, SB, and SCo, suggesting different functions and different sensory mechanisms of these four main sensilla in the orientation behavior of H. theivora on tea plants. These findings provide a theoretical basis for further exploration of the olfactory orientation of H. theivora in tropical tea plantations and pave the way for the development of semiochemical-based control options in the future.

In this study, the migratory agricultural pest Spodoptera frugiperda was exposed to three periodic short-term heat stress regimes at 37 °C, 40 °C, and 43 °C (2 h daily), with a constant 26 °C control. We systematically evaluated the effects of periodic thermal stress on developmental traits across all life stages. Combined with 16S rRNA high-throughput sequencing, we analyzed the structural and functional characteristics of the gut bacterial community in adults under heat stress. The results demonstrated that 37 °C exposure accelerated egg-to-adult development, whereas 43 °C markedly extended it. Additionally, 43 °C heat stress suppressed pupation and eclosion rates. Increasing stress temperatures were negatively correlated with pupal weight and body size in both sexes. Notably, 43 °C heat stress caused complete loss of hatching ability in offspring eggs, thereby rendering population reproduction unattainable. 16S rRNA sequencing revealed that Proteobacteria (>90%) dominated the gut bacterial community at the phylum level across all treatments. Under 43 °C heat stress, although female and male adults exhibited an increase in specific bacterial species within their gut bacteria, Alpha diversity analysis revealed no significant differences in the diversity (Shannon index) and richness (Chao index) of gut bacterial communities between sexes under temperature treatments. PICRUSt2 functional prediction indicated that metabolic pathways, biosynthesis of secondary metabolites, and microbial metabolism in diverse environments constituted the dominant functions of gut bacteria in both sexes, while heat stress exerted minimal effects on the functional profiles of gut bacteria in S. frugiperda. These findings not only provide a theoretical basis for predicting summer population dynamics and formulating ecological control strategies for S. frugiperda but also offer critical insights into the adaptive interactions between this pest and its gut bacterial community under heat stress. The results lay a foundation for further exploring the interactions between insect environmental adaptability and bacterial symbiosis.

Chelonus formosanus is a parasitic wasp capable of parasitizing various Noctuidae pests, including the highly invasive Spodoptera frugiperda, and it demonstrates strong pest control potential. Both egg and larval stages primarily occur within the host organism, and the total developmental time from egg to adult is approximately 19.62 days. To investigate the microbial communities at different stages, we performed 16S rDNA sequencing (V1–V9 region) using PacBio sequencing and identified 404 bacterial species belonging to 61 classes, 116 orders, 182 families, and 308 genera across larval, pupal, female, and male adult stages. Bacterial diversity and richness varied across the stages, with Enterobacter and Enterococcus dominating in larvae and pupae and Pseudomonas emerging as dominant in female adults. In contrast, male adults predominantly resided with Ralstonia and Achromobacter. The predicted functions of bacteria within C. formosanus at different developmental stages are predominantly marked by the high abundance of metabolic pathways. This study provides a comprehensive understanding of the morphology of C. formosanus and contributes to the practical control of host pests. Additionally, our findings preliminarily characterized the microbial community of various developmental stages, laying the groundwork for its functional study.

The tea mosquito bug, Helopeltis theivora Waterhouse (Hemiptera: Miridae), is a devastating piercing-sucking pest in tropical tea plantations. The Hainan Dayezhong (HNDYZ) is a large-leaf tea cultivar widely cultivated around the Hainan tea region in South China. However, information regarding the feeding damage of H. theivora on the HNDYZ tea plant remains scarce. Here, we first describe the morphology of H. theivora in Hainan tea region. Subsequently, we investigate the feeding biology of H. theivora on HNDYZ tea shoots under laboratory conditions. Additionally, we survey the infestations of H. theivora in a small-leaf Jinxuan tea plantation and three large-leaf HNDYZ tea plantations under varying shaded conditions. The results indicated that the morphological features of eggs, nymphs, and adults of H. theivora in the Hainan tea region were similar to those of the same species reported in other tropical tea regions. Nymphs and adults of H. theivora primarily fed on tender leaves and produced a subcircular spot within 2 to 4 minutes. This feeding spot would gradually turn dark brown within 24 hours. Furthermore, the adjacent scattered spots would connect after 48 hours, resulting in a necrotic patch on the leaves by 72 hours. The peak feeding time for H. theivora occurred at night, specifically from 7:00 PM to 1:00 AM. The most preferred feeding site was at the second leaf position, accounting for 70.94 ± 3.68% of daily feeding spots. During the feeding peak, adults H. theivora produced more feeding spots than nymphs, with females and 5th-instar nymphs creating the largest feeding areas among all life stages. Field investigations showed that damage caused by H. theivora on the large-leaf HNDYZ tea cultivar was significantly greater than that on the small-leaf Jinxuan tea cultivar. More serious infestations of H. theivora were observed in the high-shade HNDYZ tea plantation compared to the medium-shade and no-shade HNDYZ tea plantations. This suggests that the different tea cultivars and shade conditions in tea plantations may influence the population of H. theivora in the field. These findings provide new insights for further research related to the feeding strategy of H. theivora on the HNDYZ tea cultivar.

Chelonus formosanus is an important parasitic natural enemy of Lepidopteran insects and has great value in the biological control of insect pests. To screen the reference genes in the real-time fluorescence quantitative PCR (qRT-PCR) assay of Chelonus formosanus, six candidate reference genes were selected based on transcriptome data, including GAPDH, ACT, TU, TF, HSP, and RP. The expression stability of these six internal reference genes was determined by qRT-PCR in different tissues (head, chest, abdomen) and at different temperatures (11℃and 41℃), as well as with two insecticides (dinotefuran and chlorantraniliprole). The BestKeeper, GeNorm, NormFinder, Delta, and RefFinder were used to evaluate the expression stability of each gene after treatments. The results show that ACT is the most stable gene in different tissue assays. TU was the most stable reference gene in different insecticide experiments. The GAPDH was the most unstable gene in tissue assays and insecticide treatments. Furthermore, the expression of RP was the most stable under high and low temperature stresses, while the HSP was significantly affected by temperature treatments. Taken together, this study provides a basis for selecting reference genes under different conditions. It is also conducive to obtaining accurate and reliable data in the gene expression experiments of Chelonus formosanus.

Cowpea (Vigna unguiculata L.), a significant vegetable crop in China, holds particular prominence in the tropical island of Hainan. This region serves as the primary production area for the winter cultivation of cowpea. Phytoplasmas are an idiopathic parasitic pathogen and cannot be cultured in vitro. It is mainly transmitted by the insect vectors with the piercing and sucking mouthparts, such as leafhoppers, plant hoppers, and psyllids. (Kumari et al. 2019). On September 11, 2023, typical characteristics of phytoplasma diseases on cowpeas were observed in the experimental base of Hainan Academy of Agricultural Sciences (20°0'38.6964″N, 110°21'35.4024″E, Haikou City, Hainan Province, China), including reduced leaf size, chlorosis, and the development of broom-like branch deformities reminiscent, as depicted in Figure 1. At the same time, we found a large number of leafhoppers near the diseased plants, and we speculated that leafhoppers are the insect carriers that spread the disease. Following an on-site investigation, it was determined that the disease incidence ranges from 10% to 15%, leading to a consequential decrease of about 10% in yield, which is a potential disease that seriously threatens the cowpea industry in Hainan. Ten disease and healthy samples were meticulously collected and subsequently preserved at -80°C within the laboratory refrigerator. Three disease samples denoted as HNNKY-1, HNNKY-2, and HNNKY-3, were randomly chosen, and total DNA extraction was carried out employing the NuClean Plant Genomic DNA Kit (CWBIO, Taizhou, China), while three healthy samples were randomly selected as control. The 16S rRNA gene was amplified by PCR using the primer pairs P1/P7 (Schneider et al. 1995) and R16F2n / R16R2 (Lee et al. 1993) and the secA gene was amplified by PCR using the primer pairs secAfor1/secArev3 (Hodgetts et al. 2008). After agarose gel electrophoresis analysis, no DNA fragments were observed in the healthy leaf samples, whereas all three disease samples yielded amplification products. The PCR products were subsequently sequenced by Hainan Nanshan Biotech Co., Ltd., Haikou, China. After sequence analysis, it was found that the 16S rRNA gene and secA gene sequences HNNKY-1, HNNKY-2, and HNNKY-3 were identical to each other. We selected two gene sequences of strain HNNKY-3 to submission to the GenBank database, The length of the 16S rRNA gene sequence is 1193 base pairs, identified by the accession number OR666421, while the secA gene sequence is 825 base pairs in length, associated with the accession number OR661282. The phytoplasma strain HNNKY-3 was named 'Vigna unguiculata' witches'-broom phytoplasma. A BLAST analysis of the 16S rRNA gene revealed that strain HNNKY-3 displayed a 100% sequence match with 'Emilia sonchifolia' witches'-broom phytoplasma (MT420682), Peanut witches'-broom phytoplasma (OR239773), and 'Raphanus sativus' witches'-broom phytoplasma (OK491387). All of these phytoplasmas were classified within the 16SrII group. Based on the BLAST analysis of partial secA gene sequences, it was discerned that sequence homogeneity ranged from 99.27% to 99.74% among the studied sequences. These sequences were collectively classified as members of the 16SrII group. In addition, a phylogenetic tree was constructed by MEGA 11 (version 11.0.13) based on the 16Sr RNA gene and secA gene by the neighbor-joining method (Tamura et al. 2004). The results demonstrated the clustering of HNNKY-3 phytoplasma strains within the 16SrII group, as illustrated in Figures 2 and 3. A virtual RFLP analysis based on the 16S rRNA gene fragment of HNNKY-3 was conducted using the interactive online phytoplasma classification tool, iPhyClassifier (Zhao et al. 2009). The results indicated that the phytoplasma strain was the same as the reference pattern of the onion yellows phytoplasma of 16SrII-A (GenBank accession: L33765), and the similarity coefficient was 1.00. To best of our knowledge, this is the inaugural documentation of 16SrII Group-related phytoplasma infecting cowpea in Hainan, China, and lays the groundwork for further research on the dissemination of cowpea phytoplasma disease within China.

Spodoptera frugiperda is a migratory agricultural pest with fast‐spreading speed, long migration distance, and wide host range, which seriously threatens the safety of economic crops. To predict the trends of S. frugiperda and its parasitoid wasp Trichogramma pretiosum in their habitats under current and future climatic conditions, based on MaxEnt model and geographic distribution data of their historical occurrence, we project the feasibility of introducing T. pretiosum to control S. frugiperda by evaluating on their potential global distribution. The results show that, under the current greenhouse gas concentration, the potential distribution area of S. frugiperda is concentrated in 50° N‐30° S, with a total area of 1.74 × 10⁶km², and the potential distribution area of T. pretiosum in the whole world is 2.91 × 10⁶km². The suitable areas of T. pretiosum cover almost all the suitable areas of S. frugiperda, which indicates that T. pretiosum can be introduced to control S. frugiperda. The results of this study can provide a theoretical basis for the monitoring and early warning of S. frugiperda and the use of T. pretiosum to control S. frugiperda.

Aphis craccivora (Hemiptera: Aphididae) is an important pest affecting various crops worldwide. However, only few studies have been conducted on the infection of A. craccivora by Lecanicillium and related insecticidal mechanisms. We investigated the infection process of A. craccivora by Lecanicillium araneicola HK-1 using fluorescence microscopy and scanning electron microscopy (SEM), and our results indicated that the conidia of strain HK-1 easily attached to the feet and dorsum of A. craccivora. The activities of chitinase and extracellular protease were induced in the aphid after treatment with HK-1. A bioassay on A. craccivora showed that the median lethal concentration (LC₅₀) of the fungus crude extract was 24.00 mg mL⁻¹ for 24 h of treatment. Additionally, the results showed that the crude extract disrupted the enzyme system of A. craccivora, inducing the inhibition of carboxylesterase (CarE) and the induction of glutathione S-transferase (GST) and acetylcholinesterase (AChE). Combining these results with those of a gas chromatography–mass spectrometry (GC-MS) analysis, it is suggested that p-cymene, hymecromone, 9,12-octadecadienoic acid (Z, Z) methyl ester, and 9,12-octadecadienoic acid (Z, Z) may be connected to the insecticidal effects we observed. This study provides a theoretical basis for the use of L. araneicola HK-1 as a potential biological control agent.

Colletotrichum capsici is one of the most important pathogens on chili peppers. Unreasonable application of chemical fungicides will lead to threats to human and animal health, environmental damage, and increased fungicide resistance to pathogens. As an alternative strategy, biological control has been paid more and more attention by academics. In this study, the Yb-1 strain was isolated from healthy cucumber leaves, which could significantly restrict the mycelium growth of C. capsici and Botrytis cinerea, with inhibition rates of 93.49% and 74.03%, respectively. Strain Yb-1 was identified as Bacillus velezensis by morphological features and 16S rRNA gene, gyrA gene, and gyrB gene sequence analysis. When pepper seeds were treated with different concentrations of bacterial suspension, we found that a medium-concentration treatment (10⁴ CFU/mL and 10⁵ CFU/mL) could promote seed germination and growth, while high-concentration treatments (10⁸ CFU/mL) limited seed germination and growth. In addition, the root-irrigation method, acupuncture-leaf method, and leaf-cutting method were used to evaluate the safety of strain Yb-1 on peppers. The results indicated that Yb-1 did not affect the normal growth of peppers. The results of further field trials showed that the control effect of strain Yb-1 on pepper anthracnose was 59.45%. Thus, the strain B. velezensis Yb-1 has great biocontrol potential for pepper anthracnose and promotes plant growth.