Pan-genome of Citrullus genus highlights the extent of presence/absence variation during domestication and selection.

The rich genetic diversity in Citrullus lanatus and the other six species in the Citrullus genus provides important sources in watermelon breeding. Here, we present the Citrullus genus pan-genome based on the 400 Citrullus genus resequencing data, showing that 477 Mb contigs and 6249 protein-coding genes were absent in the Citrullus lanatus reference genome. In the Citrullus genus pan-genome, there are a total of 8795 (30.5%) genes that exhibit presence/absence variations (PAVs). Presence/absence variation (PAV) analysis showed that a lot of gene PAV were selected during the domestication and improvement, such as 53 favorable genes and 40 unfavorable genes were identified during the C. mucosospermus to C. lanatus landrace domestication. We also identified 661 resistance gene analogs (RGAs) in the Citrullus genus pan-genome, which contains 90 RGAs (89 variable and 1 core gene) located on the pangenome additional contigs. By gene PAV-based GWAS, 8 gene presence/absence variations were found associated with flesh color. Finally, based on the results of gene PAV selection analysis between watermelon populations with different fruit colors, we identified four non-reference candidate genes associated with carotenoid accumulation, which had a significantly higher frequency in the white flesh. These results will provide an important source for watermelon breeding.

A novel sesquiterpene lactone fraction from Eupatorium chinense L. suppresses hepatocellular carcinoma growth by triggering ferritinophagy and mitochondrial damage.

BACKGROUND: Hepatocellular carcinoma (HCC) is an aggressive tumor with limited treatment options, and it is the third leading cause of cancer-related deaths. Hence, novel therapeutic strategies are required to treat HCC. Eupatorium chinense L. is a traditional Chinese medicine (TCM) that can effectively neutralize heat and smoothen the flow of "Qi" through the liver. However, the anti-HCC effects of Eupatorium chinense L. remain unknown. PURPOSE: The present study investigated the anti-HCC effects and the underlying mechanisms of the electrophilic sesquiterpenes isolated from E. chinense L. (EChLESs) in the regulation of ferroptosis and apoptosis in HCC cells. STUDY DESIGN/METHODS: Cell viability was assessed by the MTT assay. Cell apoptosis was confirmed by flow cytometry and western blotting assay. Ferroptosis was assessed by flow cytometry, transmission electron microscopy, and western blotting assay. Ferritinophagy was detected by acridine orange staining and western blotting assay. Small interfering RNA of nuclear receptor coactivator 4 (NCOA4) was used to confirm the role of ferritinophagy in the therapeutic effect of EChLESs on HCC cells. A mouse xenograft model was constructed to determine the inhibitory effect of EChLESs on HCC in vivo. RESULTS: EChLESs induced apoptosis by disrupting mitochondrial membrane potential depolarization and mitochondrial reactive oxygen species. EChLESs induced ferroptosis as noted by a significant increase in mitochondrial disruption, lipid peroxidation, and intracellular iron level and decreased glutathione level. The apoptosis inhibitor Z-VAD-FMK and lipid reactive oxygen species scavenger ferrostatin 1 attenuated EChLESs-induced cell death. NCOA4-mediated ferritinophagy through autophagic flux was the crucial pathway for ferroptosis induced by EChLESs. NCOA4 knockdown alleviated EChLESs-induced cell death. EChLESs controlled the expression of NCOA4 at the transcriptional and post-transcriptional levels. In the in vivo experiment, EChLESs suppressed HCC growth in the xenograft tumor mouse model. CONCLUSION: EChLESs enhances cell apoptosis through mitochondrial dysfunction and ferroptosis through NCOA4-mediated ferritinophagy. Thus, Eupatorium chinense L. could be a potential TCM for treating HCC.

The main component of the aphid alarm pheromone (E)-ß-farnesene affects the growth and development of Spodoptera exigua by mediating juvenile hormone-related genes.

The (E)-ß-farnesene (EßF) is one of the most important secondary metabolites in some plants and provides indirect defense against aphids. However, the direct effect of EßF against pests is still unclear. In this study, various concentrations of EßF (0.16, 0.8, and 4 g/kg) were provided in an artificial diet to determine the direct effects of EßF on Spodoptera exigua. The results showed that an artificial diet containing 4 g/kg of EßF reduced the final survival of the S. exigua larvae and per female fecundity of adults significantly when compared with CK and SC controls (p < 0.05), then ultimately it also significantly affected the intrinsic rate of increase (p < 0.05). Furthermore, the results of the EßF bioassay in an artificial diet also indicated that the proliferation of the S. exigua population was inhibited by the ingestion of EßF in a dose-dependent manner. Combined differential RNA-seq data and RT-qPCR analysis, it was found that four key genes involved in juvenile hormone degradation significantly upregulated in S. exigua larvae treated by EßF at a dose of 0.8 and 4 g/kg when compared with two controls (p < 0.05). This indicated that EßF could disturb the normal function of juvenile hormones and reduce the survival rate of S. exigua larvae. Additionally, two key genes that regulate per fecundity of S. exigua females, including SeVg and SeVgR, were significantly downregulated in adult females (p < 0.05) when they were treated with 0.8 and 4 g/kg of EßF at the larval stage, relative to the expression of these genes after treatment with controls. These findings suggested that EßF first disturbed the normal function of juvenile hormone by upregulating key degradation genes, and then inhibited the expression of SeVg/SeVgR genes and proteins, thus reducing the population size of S. exigua by increasing larval mortality and inhibiting per female fecundity.

Effects of biochar and dicyandiamide combination on nitrous oxide emissions from Camellia oleifera field soil.

Greenhouse gas emissions from agricultural soils contribute substantially to global atmospheric composition. Nitrous oxide (N2O) is one important greenhouse gas induces global warming. Nitrification inhibitors (NI) or biochar can be effective soil N2O emission mitigation strategies for agricultural soils. However, due to differences in crop physiological traits or agricultural management, the effectiveness of mitigation strategies varies among agricultural systems. Camellia oleifera is a woody oil plant widely grown and requires intensive N input, which will potentially increase N2O emissions. Thereby, mitigation of N2O emissions from C. oleifera field soil is vital for sustainable C. oleifera development. Besides NI, incorporation of C. oleifera fruit shell-derived biochar into its soil will benefit waste management and simultaneous mitigation of N2O emissions but this has not been investigated. Here, we conducted two studies to examine effects of biochar addition and NI (dicyandiamide, DCD) application on N2O emissions from C. oleifera field soil with different N (urea or NH4NO3) and incubation temperatures. Biochar effects on nitrification rates varied among N treatments. Biochar applied in combination with DCD further reduced nitrification rates (for urea treatment, decreased from 1.1 to 0.3 mg kg-1 day-1). Biochar addition consistently increased soil N2O emissions (for urea treatment, increased from 0.03 to 0.08 ng g-1 h-1) and their temperature sensitivity. DCD application reduced soil N2O emissions with greater reductions with urea application. In future cultivation of intensively managed C. oleifera gardens, NI should be applied to mitigate N2O emissions if biochar is added, especially when urea is used.