Pithiness is one of the physiological diseases of radishes, which is accompanied by the accumulation of reactive oxygen species (ROS) during the sponging of parenchyma tissue in the fleshy roots. A respiratory burst oxidase homolog (Rboh, also known as NADPH oxidase) is a key enzyme that catalyzes the production of ROS in plants. To understand the role of Rboh genes in radish pithiness, herein, 10 RsRboh gene families were identified in the genome of Raphanus sativus using Blastp and Hmmer searching methods and were subjected to basic functional analyses such as phylogenetic tree construction, chromosomal localization, conserved structural domain analysis, and promoter element prediction. The expression profiles of RsRbohs in five stages (Pithiness grade = 0, 1, 2, 3, 4, respectively) of radish pithiness were analyzed. The results showed that 10 RsRbohs expressed different levels during the development of radish pithiness. Except for RsRbohB and RsRbohE, the expression of other members increased and reached the peak at the P2 (Pithiness grade = 2) stage, among which RsRbohD1 showed the highest transcripts. Then, the expression of 40 genes related to RsRbohD1 and pithiness were analyzed. These results can provide a theoretical basis for improving pithiness tolerance in radishes.

Chinese cabbage (Brassica rapa L. ssp. pekinensis) is an important food crop. However, its growth and development are commonly impacted by black spot disease. To examine the response mechanisms of Chinese cabbage to black spot disease, transcriptome and metabolome sequencing were performed on the leaves of Chinese cabbage genotypes J405 (resistant) and B214 (susceptible), 48 h post-infection (hpi) with Alternaria brassicicola. Expression of essential genes in the jasmonic acid, cytokinin, and auxin signaling pathways of both Chinese cabbage genotypes was inhibited. The expression of the pathogenesis-related protein 1 (PR1) gene mediated by the salicylic acid pathway is inhibited in the Chinese cabbage genotype B214. The basic endochitase B (CHIB) gene in the ethylene pathway of both Chinese cabbage genotypes was upregulated. The accumulation of reactive oxygen species in the disease spots of Chinese cabbage genotype J405 was greater than in genotype B214. The respiratory burst oxidase (RBOH) gene in the reactive oxygen species metabolic pathway was significantly upregulated in genotype J405, while no change was observed in genotype B214. We found that oxidation-reduction-related genes such as type-2 peroxiredoxin genes, NADPH-dependent thioredoxin reductase genes, glutathione peroxidase genes, and glutathione S-transfer genes were differentially expressed across both Chinese cabbage genotypes at 48 hpi. Metabolomics demonstrated that delta-tocopherol and S-hexyl glutathione were all downregulated in genotype J405, while they were upregulated in genotype B214. This approach also identified differential expression of genes in the carotenoid biosynthesis pathway, the glycinebetaine biosynthesis pathway, as well as in the specific sulfur glycoside metabolism pathway. These findings indicate that ethylene signaling is important in the hormone signaling regulatory network-mediated disease resistance and defense in Chinese cabbage. When facing pathogen infection, hormone transduction pathways associated with growth and development in Chinese cabbage are inhibited. The accumulation of reactive oxygen species and the outbreak of various secondary metabolites may endow the Chinese cabbage genotype J405 with increased resistance to black spot disease.

Background: Left ventricular remodeling (LVR) in patients with ST-segment elevation myocardial infarction (STEMI) may lead to poor prognosis in which circulating exosome miRNAs play a critical role. The aim of the present study is to identify specific exosome miRNAs for LVR in patients with STEMI. Method: Plasma exosome miRNAs were assessed in 20 patients (90% male, mean age of 66.95±1.65 years) 3–6 months after STEMI and 24 healthy individuals (83% male, mean age of 33.2±0.93 years) by using qPCR. Of the 20 patients, 8 had post-STEMI LVR according to echocardiographic evaluation, and the others did not. Clinical biochemical data including total cholesterol, HDL-C, LDL-C, LDH and NT-pro-BNP were collected from the patients with STEMI at same time as exosome miRNAs assessment. Specific exosome miRNAs for LVR were identified by using qPCR. Correlations between the dysregulated exosome miRNAs and the clinical biochemical parameters in patients with STEMI were analyzed using spearman correlation test. Results: Five exosome miRNAs including hsa-miR-181a-3p (p<0.05, fold change = 0.59), let-7d-3p (p=0.01, fold change = 0.51), hsa-miR-224-5p (p<0.01, fold change = 0.11), hsa-miR-23a-3p (p<0.01, fold change = 1.42) and miR-874-3p (p<0.01, fold change = 0.48) were dysregulated in the post-STEMI patients comparing with the healthy individuals. Among them, the exosome miR-181a-3p (p=0.01, fold change = 0.09) and let-7d-3p (p=0.01, fold change = 0.16) were significantly lower expressed in patients with LVR compared to those without (Figure 1). There was no significant difference in expression of the other three miRNAs between patients with and without LVR. Exosome hsa-miR-874-3p positively associated with LDH (p<0.01, r=0.50) in all the patients with STEMI. In vitro cell culture confirmed that the miR-874-3p mimics upregulated expression of apoptosis related gene BMF (p<0.05, fold change = 1.7) in cardiomyocyte. Exosome hsa-miR-23a-3p and hsa-miR-224-5p positively correlated with both HDL-C (p<0.01, r=0.61; p=0.02, r=0.50) and LDL-C (p=0.02, r=0.50; p<0.05, r=0.52) in all patients with STEMI. No correlation between the dysregulated exosome miRNAs and cholesterol or NT-ProBNP was observed (Figure 2). Conclusions: Circulating exosome miR-181a-3p and let-7d-3p might play a potential role in LVR in patients 3–6 months after STEMI. Exosome hsa-miR-874-3p might be associated with cardiomyocyte injury. Hsa-miR-23a-3p and hsa-miR-224-5p demonstrated an activity in regulation of lipid metabolism and biosynthesis in patients with STEMI. Funding Acknowledgement: Type of funding sources: Public hospital(s). Main funding source(s): This work was supported by grants from the 3×3 Clinical Scientist Fund of Sun Yat-sen Memorial Hospital