Chitin deacetylase (CDA) is a chitin degradation enzyme that strictly catalyzes the deacetylation of chitin to form chitosan, which plays an important role in regulating growth and development, as well as the immune response. In this study, a chitin deacetylase 3 gene (CDA3) was identified with a complete open reading frame (ORF) of 1362 bp from the genome database of Diaphorina citri, encoding a protein of 453 amino acids. Spatiotemporal expression analysis suggested that D. citri CDA3 (DcCDA3) had the highest expression level in the integument and third-instar nymph stage. Furthermore, DcCDA3 expression level can be induced by 20-hydroxyecdysone (20E). Injection of Escherichia coli and Staphylococcus aureus induced the upregulation of DcCDA3 in the midgut, while DcCDA3 was downregulated in the fat body. After silencing DcCDA3 by RNA interference, there was no influence on the D. citri phenotype. In addition, bactericidal tests showed that recombinant DcCDA3 inhibited gram-positive bacteria, including S. aureus and Bacillus subtilis (B. subtilis). In conclusion, our results suggest that DcCDA3 might play an important role in the immune response of D. citri.

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Liviidae), transmits the citrus greening pathogen 'Candidatus Liberibacter asiaticus' (CLas) by feeding on citrus phloem sap. Because phloem sap is rich in sugars but low in amino acids, ACP sucks large quantities and excretes most of it as honeydew. We studied the chemical composition of ACP honeydew on various host plants. Honeydew samples were analyzed with gas chromatography-mass spectrometry. Fourteen sugars, 13 amino acids, and six organic acids were detected in the honeydew of ACP. Sugars composed about 95% of the total compounds. Sucrose and trehalose were the predominant sugars, composing about 58 and 23% of the total sugars, respectively. Proline, asparagine, aspartic acid, and glutamic acid were the most abundant amino acids in ACP honeydew. The host plant and its infection with CLas had some effect on the honeydew composition. Glucose, chiro-inositol, myo-inositol, inositol, maltose, and turanose were lower in honeydew collected from CLas-infected citrus compared to that collected from non-infected trees. In CLas-infected citrus (pineapple sweet orange, Citrus sinensis L. Osbeck) and Bergera koenigii (L.) Spreng. [curry leaf tree (both Rutaceae)] honeydews, valine, alanine, serine, glutamine, glycine, and the organic acids were lower than in honeydew from healthy citrus. Mannose, galactose, inositol, mannitol, an unknown disaccharide, and proline were higher in the honeydew collected from B. koenigii than in honeydew collected from healthy citrus (pineapple sweet orange), whereas fructose, chiro-inositol, myo-inositol, trehalose, and lactic acid were lower. The findings of this study help us understand the metabolism and the nutrient needs of ACP that transmits CLas, the pathogen of huanglongbing in citrus.

Huanglongbing (HLB) is the most devastating citrus disease worldwide, and suppression of the Asian citrus psyllid (Diaphorina citri) is regarded as an effective method to inhibit the spread of HLB. In this study, we isolated a strain named as Serratia marcescens KH-001 from D. citri nymphs suffering from disease, and evaluated its killing effect on D. citri via toxicity test and effect on microbial community in D. citri using high-throughput sequencing. Our results indicated that S. marcescens KH-001 could effectively kill 83% of D. citri nymphs, while the fermentation products of S. marcescens KH-001 only killed 40% of the D. citrinymphs. High-throughput sequencing results indicated that the S. marcescens KH-001 increased the OTU numbers from 62.5 (PBS buffer) to 81.5, while significantly lowered the Shannon index compared with Escherichia coli DH5α (group E) (p < 0.05). OTU analysis showed that the S. marcescens KH-001 had significantly reduced the relative abundance of endosymbionts Wolbachia, Profftella, and Carsonella in group S compared with that in other groups (p < 0.05). Therefore, the direct killing effect of the fermentation products of S. marcescens KH-001 and the indirect effect via reducing the numbers of endosymbionts (Wolbachia, Profftella, and Carsonella) of D. citri endow S. marcescens KH-001 a sound killing effect on D. citri. Further work need to do before this strain is used as a sound biological control agents.

No abstract

Tapinoma melanocephalum is a ubiquitous invasive species and widely distributed in subtropical and tropical regions around the world. Here, we sequenced and annotated the complete mitochondrial genome (mitogenome) of T. melanocephalum. This mitogenome was 15,499 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and 2 ribosomal RNA unit genes (rRNAs). Compared to other Formicidae species, gene order of T. melanocephalum was not conserved and one tRNA cluster trnW-trnC-trnY converted to trnW-trnY-trnC. The whole mitogenome exhibited heavy AT nucleotide bias (79.5%). All PCGs started with the standard ATN codons. Except for cox1 and nad5 end with the incomplete codon TÀ, all PCGs terminated with the stop codon TAA. Phylogenetic analysis showed that T. melanocephalum got together with three same subfamily Dolichoderinae species and one Dorylinae species, indicating the close relationship of Dolichoderinae and Dorylinae.

Tamarixia radiata plays an important role in biological control of the psyllid Diaphorina citri Kuwayama, vector of the huanglongbing (HLB). In this study, we sequenced and analyzed the mitochondrial genome (mitogenome) of T. radiata , the first mitogenome of species in the family Eulophidae. This mitogenome was 14,752 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs) and two ribosomal RNA unit genes (rRNAs). All 13 PCGs were initiated by the ATN (ATG, ATT, and ATA) codon. Twelve PCGs terminate with the stop codon TAA or TAG except for nad1 which end with the incomplete codon T−. Phylogenetic analysis showed that T. radiata got together with three Pteromalidae species, indicating the close relationship of Eulophidae and Pteromalidae.

BACKGROUND Asian citrus psyllid (ACP), also known as Diaphorina citri, is the natural vector of Candidatus Liberibacter asiaticus (CLas), which is responsible for Huanglongbing (HLB), a devastating citrus disease. Previously, the pathogen was successfully excluded from diseased citrus plants by using the indigenous endophyte Bacillus subtilis L1‐21. However, the pathogen elimination and colonization potential of B. subtilis L1‐21 in the carrier vector ACP, as well as the recruitment of native microbial communities of psyllid in the presence of endophytes, are still unknown. RESULTS Initially, we suggested that endophyte L1‐21 reduced the CLas copies in ACP from 6.58 × 106 to 5.04 × 104 per insect after 48 h, however, the pathogen copies remained stable in the negative control. The endophyte was stable for 48 h after application. Among the bacterial genera those highlighted in ACP were Candidatus Liberibacter, Pseudomonas, Candidatus Profftella, Methylobacterium‐Methylorubrum, Pantoea, Curtobacterium, Wolbachia, Actinomycetospora, and Bacillus. Interestingly, B. subtilis L1‐21 easily colonizes the midgut of ACP but cannot be detected in eggs. When ACP with endophyte L1‐21 was allowed to feed on new citrus leaves, the highest colonization was observed. We also found that psyllids carrying endophyte L1‐21 after feeding on citrus leaves reduced the CLas copies in leaves on the 0, 3rd and 5th day from 8.18 × 10,4 2.6 × 10,3 and 0 pathogen copies/g fresh midvein, respectively. CONCLUSIONS We propose that B. subtilis L1‐21 is a native endophyte in citrus and psyllid, which efficiently reduces the CLas pathogen in both citrus and psyllids, provides a more protective effect by increasing the number of cultivable endophytes, and successfully colonizes the midgut of ACP. © 2022 Society of Chemical Industry.

Larvae of the geometrid moth Milionia basalis feed exclusively on the podocarp tree, Podocarpus macrophyllus. In this study, we sequenced and analyzed the complete mitochondrial genome (mitogenome) of M. basalis. This mitogenome was 15,901 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and two ribosomal RNA unit genes (rRNAs). The whole mitogenome exhibited heavy AT nucleotide bias (82.2%). Gene order was conserved and identical to most other previously sequenced Geometridae. Most PCGs of M. basalis had the conventional start codons ATN, with the exception of nad1 (TTG) and cox1 (CGA). Except for four genes (cox1, cox2, nad5, and nad4) end with the incomplete stop codon Tall ll other PCGs terminated with the stop codon TAA. Phylogenetic analysis showed that M. basalis got together with Apocheima cinerarius, Jankowskia athlete, and four Biston species (B. panterinaria, B. perclara, B. suppressaria, and B. thibetaria).