First Report of Colletotrichum siamense Causing Leaf Spot on Jasminum sambac in China.

Jasmine (Jasminum sambac (L.) Aiton) is cultivated as a commercial floricultural crop in many countries around the world (Gao et al., 2020). From June to August 2020, leaf spots on jasmine were observed on a jasmine plantation in Hengzhou of Guangxi province. Over 40% of the plants in 6 ha fields were infected. This disease was prevalent in jasmine production area of China (Chen et al., 2012; Du et al., 2020). Symptoms began as chlorotic regions (from 5 to 10 mm in diameter) with light brown necrotic centers, which gradually expanded to the entire leaf. Eventually, the disease leaded to defoliation and dieback. The edges of the affected parts from diseased leaves were cut into pieces (3 mm2). Pieces were treated with 75% ethanol for 10 s, soaked in 2% NaClO solution for 1 min, washed three times with sterile water, and then incubated on potato dextrose agar (PDA) plates at 28℃ for 5 days in the dark. Fungal cultures that showed similar morphological characteristics were isolated, and three representative isolates (HL6-1 to HL6-3) were purified following Mo et al. (2018). The cultures on PDA changed from white to dark grey after 7 days and produced conidiomata after 14 days. Conidia were hyaline, one-celled, guttulate, cylindrical, of 12.07 to 18.09 × 4.04 to 8.05 µm, 13.17 to 16.35 × 4.22 to 6.13 µm and 10.11 to 22.17 × 3.65 to 8.1 µm for HL6-1, HL6-2 and HL6-3, respectively. Gray-brown or dark brown appressoria formed from conidia were subglobose or elliptical. Conidial appressoria and mycelial appressoria were 5.53 to 13.96 × 3.58 to 13.95 µm and 4.24 to 14.01 × 2.4 to 10.86 µm. Genomic DNA was extracted from three isolates and the partial internal transcribed spacer (ITS) regions, intergenic region of apn2 and MAT1-2-1 (ApMAT), and fragments of actin (ACT), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), chitin synthase (CHS-1), and ß-tubulin (TUB2) genes were amplified, sequenced and submitted to GenBank (ITS, ON115173 to ON115175; ApMat, ON156517 to ON156519; ACT, ON146469 to ON146471; GAPDH, ON156502 to ON156504; CHS-1, ON156507 to ON156509; TUB-2, ON156512 to ON156514). Phylogenetic tree was constructed with MrBayes v. 3.2.6 and MEGA v. 10.1.5 based on the concatenation of multiple sequences. Three isolates were grouped with strain C. siamense ICMP 18578. Results indicated three isolates were identified as Colletotrichum siamense Prihastuti, L. Cai & K.D. Hyde. To confirm the pathogenicity of the three isolates, four sets (five plants per set) of 160 healthy leaves of 2-year-old plants (J. sambac, eight leaves per plant) were slightly scratched with a sterilized toothpick at each of eight locations. Conidial suspension (1×106 conidia/mL) in 0.1% Tween 20 were inoculated onto each wounded spot of three sets as the treatment groups, while wounded leaves treated with sterile water as the control. All plants were covered with plastic bags and cultivated in phytotron (12 h/12 h light/dark, 28°C). After 7 days, irregular chlorotic regions with brown lesions were observed on inoculated leaves while no symptoms on controls. The same fungi were reisolated from inoculated leaves and confirmed by morphological and molecular identification, fulfilling Koch's postulates. Colletotrichum siamense has been associated with leaf anthracnose of J. sambac in Vietnam (Wikee et al., 2011) and J. mesnyi in China (Zhang et al., 2019). To our knowledge, this is the first report of C. siamense causing jasmine anthracnose in China, which provides a reference for the management of this disease.

GDF15 Promotes Cardiac Fibrosis and Proliferation of Cardiac Fibroblasts via the MAPK/ERK1/2 Pathway after Irradiation in Rats.

Ionizing radiation exposure is associated with a risk of cardiac fibrosis; however, the underlying molecular mechanism remains unclear. Growth/differentiation factor-15 (GDF15), a fibroblast factor, is a divergent member of the transforming growth factor ß superfamily. Next-generation sequencing analyses has revealed that Gdf15 is increased in cardiac fibroblasts during radiation-induced fibrosis. However, the role of Gdf15 in cardiac fibrosis remains unclear. In this study, we demonstrated that the upregulated expression of GDF15 in newborn rat cardiac fibroblasts and adult rats after irradiation could induce fibrosis, which was confirmed by the increased cell proliferation rate and the increased expression of fibrosis markers (Col1α and αSMA) in newborn rat cardiac fibroblasts after transfection with Gdf15 in vitro. Conversely, the downregulation of GDF15 inhibited cardiac fibrosis, as confirmed by G2/M-cell cycle arrest, suppression of cell proliferation, and low levels of Col1α and αSMA expression. We also found that suppressing the expression of Gdf15 in cardiac fibroblasts could lead to a decrease in CDK1 and inhibit phosphorylation of ERK1/2. Thus, GDF15 might promote cardiac fibroblast fibrosis through the MAPK/ERK1/2 pathway and thus contribute to the pathogenesis of radiation-induced heart disease.

miR-21 is upregulated, promoting fibrosis and blocking G2/M in irradiated rat cardiac fibroblasts.

BACKGROUND: Radiation exposure of the thorax is associated with a greatly increased risk of cardiac morbidity and mortality even after several decades of advancement in the field. Although many studies have demonstrated the damaging influence of ionizing radiation on cardiac fibroblast (CF) structure and function, myocardial fibrosis, the molecular mechanism behind this damage is not well understood. miR-21, a small microRNA, promotes the activation of CFs, leading to cardiac fibrosis. miR-21 is overexpressed after irradiation; however, the relationship between increased miR-21 and myocardial fibrosis after irradiation is unclear. This study was conducted to investigate gene expression after radiation-induced CF damage and the role of miR-21 in this process in rats. METHODS: We sequenced irradiated rat CFs and performed weighted correlation network analysis (WGCNA) combined with differentially expressed gene (DEG) analysis to observe the effect on the expression profile of CF genes after radiation. RESULTS: DEG analysis showed that the degree of gene changes increased with the radiation dose. WGCNA revealed three module eigengenes (MEs) associated with 8.5-Gy-radiation-the Yellow, Brown, Blue modules. The three module eigengenes were related to apoptosis, G2/M phase, and cell death and S phase, respectively. By blocking with the cardiac fibrosis miRNA miR-21, we found that miR-21 was associated with G2/M blockade in the cell cycle and was mainly involved in regulating extracellular matrix-related genes, including Grem1, Clu, Gdf15, Ccl7, and Cxcl1. Stem-loop quantitative real-time PCR was performed to verify the expression of these genes. Five genes showed higher expression after 8.5 Gy-radiation in CFs. The target genes of miR-21 predicted online were Gdf15 and Rsad2, which showed much higher expression after treatment with antagomir-miR-21 in 8.5-Gy-irradiated CFs. Thus, miR-21 may play the role of fibrosis and G2/M blockade in regulating Grem1, Clu, Gdf15, Ccl7, Cxcl1, and Rsad2 post-irradiation.

Plasma miR-1273g-3p acts as a potential biomarker for early Breast Ductal Cancer diagnosis.

Circulating miRNAs presenting in plasma in a stable manner have been demonstrated their potential role as a promising biomarkers in many human diseases, such as Alzheimer's disease, melanoma and ovarian carcinoma. However, few circulating miRNAs could be used for breast ductal cancer diagnosis. Here, we identified miR-1273g-3p as a biomarker for detecting breast ductal cancer. We detected miR-1273g-3p levels in the plasma of 39 sporadic breast ductal cancer patients and 40 healthy donors by Stem-loop Quantitative Real-time PCR (qRT-PCR). The results showed the plasma miR-1273g-3p level were significantly up-regulated in breast ductal cancer patients compared with healthy donors (p=0.0139). Receiver operating characteristic (ROC) curve also revealed the significantly diagnostic ability of miR-1273g-3p in patients (p=0.0414). In addition, the plasma level of miR-1273g-3p was closely related to IIIB-IIIC TNM stage. We also confirmed the higher expression level of miR-1273g-3p in breast cancer cell lines MCF-7 (4.872±0.537) than normal breast cells (Hs 578Bst). Taken together, miR-1273g-3p could represent as a potential biomarker for early breast ductal cancer diagnosis.