A novel botybirnavirus with a unique satellite dsRNA causes latent infection in Didymella theifolia isolated from tea plants

© 2023 The Author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY), https://creativecommons.org/licenses/by/4.0/The unique, recently discovered fungus Didymella theifolia specifically infects local varieties of tea plant Camellia sinensis in China, and therefore, the characterization of its mycoviruses is important. Three double-stranded (ds) RNAs (1, 2, and 3, with 6,338, 5,910, and 727 bp in size, respectively) were identified in the avirulent D. theifolia strain CJP4-1, which exhibits normal growth and morphology. Characterization of these double-stranded RNAs (dsRNAs) revealed that the two largest elements are the genomic components of a novel botybirnavirus, tentatively named Didymella theifolia botybirnavirus 1 (DtBRV1). Conversely, dsRNA3 shares no detectable similarity with sequences deposited in public databases but has high similarity with the 5′-terminal regions of dsRNAs 1 and 2 and contains a duplicated region encoding a putative small peptide. All three dsRNAs are encapsidated in isometric virions ca. 40 nm in diameter, supporting the notion that dsRNA3 is a DtBRV1 satellite. SDS-polyacrylamide gel electrophoresis in combination with peptide mass fingerprint analysis revealed that the DtBRV1 capsid protein consists of polypeptides encoded by the 5′-terminal regions of both genomic components dsRNA1 and dsRNA2. Vertical transmission of DtBRV1 through conidia is efficient, while its horizontal transmission from CJP4-1 to other strains was not detected. DtBRV1, with or without dsRNA3, has no obvious effects on fungal growth and virulence, as illustrated following transfection of the virulent D. theifolia strain JYC1-6. In summary, DtBRV1 exhibits unique molecular traits and contributes to our understanding of mycovirus diversity.Peer reviewe

Bizarre leiomyoma of the scrotum: A case report and review of the literature

Bizarre leiomyomas of the scrotum are rare benign tumors that are often misdiagnosed. In this study, we present a case of bizarre leiomyoma of the scrotum in a 53-year-old male. The patient presented with a painless scrotal mass that was insidious in the right side of the scrotum with no sudden increase in size. Definitive preoperative diagnosis could not be established; however, following surgical resection of the tumor, a diagnosis of bizarre leiomyoma of the scrotum was determined by pathological examination. The patient was followed up six months following resection and no problems were reported. This is the first reported case of bizarre leiomyoma of the scrotum in China. A supplementary review of previously published cases and literature is also presented

microRNA-184 functions as tumor suppressor in renal cell carcinoma

microRNAs (miRNAs) are evolutionarily conserved, endogenous, small, noncoding RNA molecules of approximately 22 nucleotides in length that function as post-transcriptional gene regulators. Their aberrant expression may be involved in human diseases, including cancer. Although miRNA-184 (miR-184) has been reported in other tumors, its function in renal cell carcinoma (RCC) is still unknown. The aim of the present study was to investigate the role of miR-184 in RCC. The impacts of miR-184 on cell migration, proliferation and apoptosis were evaluated using migration scratch, 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assay. Our studies revealed that miR-184 mimic significantly inhibits cell migration, suppresses cell proliferation and induces renal cancer cell apoptosis in vitro when compared with the negative control (P<0.05). In this study, it was observed that miR-184 played a significant role as a tumor suppressor in RCC. Therefore, miR-184 may be a promising therapeutic target for renal cancer treatment in the future

PROSTATIC SCHISTOSOMA JAPONICUM WITH ATYPICAL IMMUNOPHENOTYPING OF INDIVIDUAL GLANDULAR TUBES: A CASE REPORT AND REVIEW OF THE LITERATURE

There are few cases of prostatic schistosomiasis. Here we report a case of Schistosoma japonicum of the prostate, in which the immunophenotyping of individual glandular tubes was atypical. Whether the S. japonicum infection contributed to the lesion or not is unknown. We suspect the lesion was a sign of early precancerous hyperplasia. Follow-up of this patient may give clues about the relationship between schistosomiasis and prostate cancer. This is the first case report of prostatic S. japonicum in the English literatures. A review of the literature is carried out

Sequencing maps of the PCR product of some calli lines in the T0 generation.

<p>4–1 is a chimera, 4–3 is a homozygote, and 4–14, 4–19, and 6–5 are heterozygotes. The sequence between two arrows is the spacer cut by TALEN. The scissors indicate three-base deletions and the asterisk indicates the targeted base substitution C³¹⁷-T.</p

The segregation patterns of TALEN construct and the <i>OsEPSPS</i> genotype from T0 to T1 generation in some transgenic lines.

<p>T1 population are divided into two groups according to whether the TALEN construct exists (+) or not (–) in the genome. The number before genotype indicates the amount of T1 plants and the number after ‘d’ indicates the amount of bases that have been deleted. WT, wild-type sequence; d, base deletion.</p

Phenotypic assessments of heterozygous <i>OsEPSPS</i> mutants with either one base deletion (WTd₁) or three bases deletion (WTd₃) and wild-type plant (WTWT) in T1 generation.

<p>A. Glyphosate test (Roundup 300×dilution). Heterozygous mutants are more sensitive to glyphosate; B. Spikelets; C. Seed-setting rate. Bar = 10 cm.</p

Mutations from the calli flow to the regenerated plants.

<p>The genotype and detailed sequence variation of some typical mutants from chimera, homozygote and heterozygote are presented. Each dashed line represents a deleted nucleotide. The nucleotide in gray represents substitution. WT, wild-type sequence; d, base deletion; s, base substitution. The number after ‘d’ or ‘s’ represents the number of bases that have been deleted or substituted. The box marks the variant sequence which might result from mismatch repair in <i>E</i>. <i>coli</i> during PCR product cloning.</p

Chimeric RNA/DNA oligonucleotides (COs).

<p>Uppercase letters represent DNA residues, lowercase letters represent 2′-O-methyl-RNA residues, and the box indicates the nucleotide that should be introduced into the target sequence.</p

Gene Editing by Co-Transformation of TALEN and Chimeric RNA/DNA Oligonucleotides on the Rice <i>OsEPSPS</i> Gene and the Inheritance of Mutations

<div><p>Although several site-specific nucleases (SSNs), such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas, have emerged as powerful tools for targeted gene editing in many organisms, to date, gene targeting (GT) in plants remains a formidable challenge. In the present study, we attempted to substitute a single base <i>in situ </i>on the rice <i>OsEPSPS </i>gene by co-transformation of TALEN with chimeric RNA/DNA oligonucleotides (COs), including different strand composition such as RNA/DNA (C1) or DNA/RNA (C2) but contained the same target base to be substituted. In contrast to zero GT event obtained by the co-transformation of TALEN with homologous recombination plasmid (HRP), we obtained one mutant showing target base substitution although accompanied by undesired deletion of 12 bases downstream the target site from the co-transformation of TALEN and C1. In addition to this typical event, we also obtained 16 mutants with different length of base deletions around the target site among 105 calli lines derived from transformation of TALEN alone (4/19) as well as co-transformation of TELAN with either HRP (5/30) or C1 (2/25) or C2 (5/31). Further analysis demonstrated that the homozygous gene-edited mutants without foreign gene insertion could be obtained in one generation. The induced mutations in transgenic generation were also capable to pass to the next generation stably. However, the genotypes of mutants did not segregate normally in T1 population, probably due to lethal mutations. Phenotypic assessments in T1 generation showed that the heterozygous plants with either one or three bases deletion on target sequence, called d₁ and d₃, were more sensitive to glyphosate and the heterozygous d1 plants had significantly lower seed-setting rate than wild-type.</p></div