Perilla frutescen is used as traditional food and medicine in East Asia. Its seeds contain high levels of α-linolenic acid (ALA), which is important for health, but is scarce in our daily meals. Previous reports on RNA-seq of perilla seed had identified fatty acid (FA) and triacylglycerol (TAG) synthesis genes, but the underlying mechanism of ALA biosynthesis and its regulation still need to be further explored. So we conducted Illumina RNA-sequencing in seven temporal developmental stages of perilla seeds. Sequencing generated a total of 127 million clean reads, containing 15.88 Gb of valid data. The de novo assembly of sequence reads yielded 64,156 unigenes with an average length of 777 bp. A total of 39,760 unigenes were annotated and 11,693 unigenes were found to be differentially expressed in all samples. According to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, 486 unigenes were annotated in the “lipid metabolism” pathway. Of these, 150 unigenes were found to be involved in fatty acid (FA) biosynthesis and triacylglycerol (TAG) assembly in perilla seeds. A coexpression analysis showed that a total of 104 genes were highly coexpressed (r > 0.95). The coexpression network could be divided into two main subnetworks showing over expression in the medium or earlier and late phases, respectively. In order to identify the putative regulatory genes, a transcription factor (TF) analysis was performed. This led to the identification of 45 gene families, mainly including the AP2-EREBP, bHLH, MYB, and NAC families, etc. After coexpression analysis of TFs with highly expression of FAD2 and FAD3 genes, 162 TFs were found to be significantly associated with two FAD genes (r > 0.95). Those TFs were predicted to be the key regulatory factors in ALA biosynthesis in perilla seed. The qRT-PCR analysis also verified the relevance of expression pattern between two FAD genes and partial candidate TFs. Although it has been reported that some TFs are involved in seed development, more direct evidence is still needed to verify their function. However, these findings can provide clues to reveal the possible molecular mechanisms of ALA biosynthesis and its regulation in perilla seed.

This study aimed to investigate the protective value of Eucommia ulmoides extract (EUE) on chicks under cold stress. A total of 21 compounds were identified in EUE using mass spectrometry (LC-MS). Ninety chicks were divided into a control group (CS) fed a basal diet and an experimental group supplemented with EUE, exposed to 10 ± 1 °C for 8 h per day. Results showed, compared with the CS group, the body weights (BW) (p < 0.01) and average daily gains ADG (p < 0.05) of the EUE group were increased throughout the study period. Chicks fed EUE had higher AFI (0–7 d, p < 0.001) and lower feed-to-gain ratios (F/G) (0–15 d, p < 0.001). EUE increased the activities of superoxide dismutase (SOD) (15 d, p < 0.05) and glutathione peroxidase (GSH-Px) (7 d, p < 0.05), whereas it decreased malondialdehyde (MDA) (15 d, p < 0.01). The contents of IgA (7 d, p < 0.05), IgG (7 d; 15 d, p < 0.01), and IgM (15 d, p < 0. 001) were higher in the EUE group. Dietary EUE could also reduce chick organ damage. Overall, EUE as a natural feed additive can improve the growth performance, antioxidant capacity, and immune level, and reduce the organ damage of cold-stressed chicks.

Chick's susceptibility to heat stress often leads to growth retardation, immune function impairment, disease, and mortality. This thesis explores the potential ameliorative effect of 0.8% Eucommia ulmoides extract (EUE) into the diet of heat-stressed chicks in a 15-d feeding trial. The investigation reveals that feeding EUE significantly enhances the BW, ADG, AFI, and F/G of chicks experiencing heat stress. Additionally, the EUE groups exhibited higher levels of T-AOC (at 7 and 15d), SOD (at 15 d), GSH-Px (at 15 d), as well as lower MDA concentrations (at 7 and 15d) in chick serum. Pathological changes and H&E staining revealed that EUE effectively improved tissue damage in the duodenum, heart, and stomach induced by heat stress in the chicks. The EUE groups also showed higher levels of IgA (at 7 d), IgG and IgM (at 7 and 15 d). RNA-seq and WGCNA analysis revealed that EUE mitigates cellular damage and losses in heat-stressed chicks primarily through pathways involving signal transduction, protein synthesis and degradation, as well as cell cycle regulation, particularly the latter. This investigation serves as a fundamental and cognitive framework for the development and application of Eucommia ulmoides feed additives aimed at safeguarding the well-being of chicks in adverse environmental conditions.

Adventitious roots (ARs), developing from non-root tissue, play an important role in some plants. Here, the molecular mechanism of AR differentiation in Lotus japonicus L. (L. japonicus) with the transformed chicken interferon alpha gene (ChIFNα) encoding cytokine was studied. ChIFNα transgenic plants (TP) were identified by GUS staining, PCR, RT-PCR, and ELISA. Up to 0.175 μg/kg rChIFNα was detected in TP2 lines. Expressing rChIFNα promotes AR development by producing longer roots than controls. We found that the effect was enhanced with the auxin precursor IBA treatment in TP. IAA contents, POD, and PPO activities associated with auxin regulation were higher than wild type (WT) in TP and exogenous ChIFNα treatment plants. Transcriptome analysis revealed 48 auxin-related differentially expressed genes (DEGs) (FDR < 0.05), which expression levels were verified by RT-qPCR analysis. GO enrichment analysis of DEGs also highlighted the auxin pathway. Further analysis found that ChIFNα significantly enhanced auxin synthesis and signaling mainly with up-regulated genes of ALDH, and GH3. Our study reveals that ChIFNα can promote plant AR development by mediating auxin regulation. The findings help explore the role of ChIFNα cytokines and expand animal gene sources for the molecular breeding of growth regulation of forage plants.

The formation of adventitious roots (ARs) plays an important role in grass growth and production. Chicken alpha interferon (ChIFNα) is an animal glycoprotein with broad activities of immune control, gene induction, growth and development regulation. ChIFNα gene was transformed to Lotus japonicus and its expression was found to promote AR development. The primordia formation occurred 3–5 days earlier, and more root hairs appeared in the transgenic plants (TP) than in the wild-type parents (WT). Data of RNA sequences from TP and WT in vitro hydroponic culture were analyzed. Overall, the intersection of the differentially upregulated genes at 0–12 days included 278 genes in TP and 244 genes in WT plants. Gene Ontology analysis indicated that the differentially expressed genes (DEGs) in TPs were enriched in response to biotic stimulus, metabolic process, sucrose transport, carbon fixation and signal transduction. Further, Kyoto Encyclopedia of Genes and Genomes enrichment analysis found carbon metabolism, carbon fixation in photosynthetic organisms and photosynthesis enriched in TPs. Co-expression analysis of the DEGs showed that 35 and 18 transcription factors (TFs) were involved in ARs formation in TP and WT plants, respectively. However, the TFs C3H and C2C2-GATA showed the opposite regulatory pattern with ChIFNα expression. Together, these data suggest that ChIFNα promotes AR development by modulating specific pathways and TFs. These results provide new insight into the mechanism by which animal interferon modulates plant growth and lays the foundation for cultivating new forage germplasm using animal cytokines.