Loquat (Rhaphiolepis biabas, heterotypic synonym: Eriobotrya japonica) is an important edible and medicinal plant that is widely cultivated on 133 thousand hectares (recorded in 2022) in China. A s...

Autophagy controls insect development and can be targeted for pest control in agriculture. In the present study, starvation‐induced autophagy is investigated in the insect species Spodoptera frugiperda . Bioinformatics analysis and a search of the EST database (http://bioweb.ensam.inra.fr/spodobase) identifies a putative ATG8 gene of S. frugiperda . To generate a biomarker of autophagosome, the DNA sequence encoding the open reading frame of this gene is amplified and cloned into a pIEX‐4‐ mCherry‐EGFP‐SfATG8 recombinant vector. Sf9 cells are then transfected with this expression vector and starved in phosphate‐buffered saline solution for 4 h to induce autophagy, which is examined by LysoTracker staining (Life Technologies, Grand Island, New York), western blotting and fluorescence microscopy. The results obtained show that starvation stimulates lipidation of SfATG8‐PE and the formation of autophagosomes, providing a foundation for further research with respect to autophagy in insects.

The Ankyrin-repeat proteins (ANKs) play a key role in plant development and in response to abiotic stress. This research identified family members of the ANK genes in Sorghum bicolor at the whole-genome level, analyzed their sequence characteristics, evolutionary relationships, and expression patterns, and provided a scientific basis for elucidating the functionality of SbANK genes and for salt-tolerant breeding. Using bioinformatics methods, this study conducted a comprehensive identification of the SbANK gene family, analyzing its physicochemical properties, domain composition, chromosomal distribution, colinearity relationships, promoter cis-acting elements, and conserved protein motifs. Transcriptomic data and qRT-PCR were used to detect changes in their expression under salt stress. A total of 186 ANK family members were identified in the Sorghum bicolor genome, classified into 13 subfamilies and unevenly distributed across 10 chromosomes. Intra-species colinearity analysis revealed 7 pairs of duplicated genes, while inter-species colinearity analysis showed that S. bicolor and Oryza sativa share 88 pairs of orthologs, far exceeding the number found in Arabidopsis thaliana (11 pairs). Promoter analysis indicated that SbANK genes are enriched with cis-acting elements associated with hormone responses (particularly MeJA elements, accounting for 51.7%) and stress responses (particularly anaerobic-inducible elements, accounting for 60.9%). Transcriptomic expression analysis revealed that SbANK genes exhibit distinct tissue specificity, with the ANK-IQ subfamily highly expressed in leaves and the ANK-M subfamily showing the most widespread response under salt stress. Expression levels of the 10 candidate genes showing the most significant responses to salt stress were analyzed using qRT-PCR. The results indicated that SbANK91, SbANK135, and SbANK136 were significantly upregulated under 200 mmol/L NaCl treatment. The SbANK family is distinguished by a large number of member genes and structural diversity, with the ANK-M subfamily being the primary group responding to salt stress. SbANK91, SbANK135, and SbANK136 are identified as putative candidate genes for salt stress responses.

Antimicrobial peptides (AMPs) are regarded as promising candidates for the post-antibiotic era because of their broad-spectrum bactericidal activity and low propensity to induce bacterial resistance. However, their small molecular weight and intrinsic cytotoxicity toward the expression host result in poor soluble expression and low yields when unmodified AMPs are produced directly in prokaryotic systems. In this study, the coding sequences of three AMPs (Swine_2, P3-3R-8I, and QLX-3DV-1) were assembled by primer-complementary PCR, and nine recombinant expression vectors covering three fusion-tag systems were constructed by restriction digestion and ligation, namely pET32a-Swine_2, pET32a-P3-3R-8I, pET32a-QLX-3DV-1, pET32a-GSHHW-Swine_2, pET32a-GSHHW-P3-3R-8I, pET32a-GSHHW-QLX-3DV-1, pET3C-SUMO-Swine_2, pET3C-SUMO-P3-3R-8I, and pET3C-SUMO-QLX-3DV-1. The resulting plasmids were transformed into Escherichia coli BL21(DE3), and the nine fusion proteins were expressed under IPTG induction and auto-induction. The fusion proteins were purified by Ni-NTA affinity chromatography; the target AMPs were released by enterokinase digestion, SNAC-mediated chemical cleavage, or SUMO protease digestion, and the peptides were recovered by ultrafiltration. The antibacterial activity of the recovered peptides against E. coli and Staphylococcus aureus was systematically compared using the agar diffusion assay and the broth microdilution method. All three strategies yielded bioactive AMPs. For a given peptide backbone, the three preparations showed identical MIC values, whereas the peptides released by SUMO protease produced the largest inhibition zones. Because the three preparations of any one backbone share an identical primary sequence and an identical MIC, this difference cannot be attributed to a higher intrinsic potency of the peptide; it is treated here as a semi-quantitative observation. This study provides a complete technical workflow and a methodological reference for the engineered production of AMPs.