Assessment of the potential germ cell mutagenicity of industrial and plant protection chemicals as part of an integrated study of genotoxicity in vitro and in vivo

Cucumber mosaic virus (CMV) is a destructive plant virus with worldwide distribution and the broadest host range of any known plant virus, as well as a model plant virus for understanding plant–virus interactions. Since the discovery of RNA interference (RNAi) as a major antiviral defense, RNAi-based technologies have been developed for plant protection against viral diseases. In plants and animals, a key trigger of RNAi is double-stranded RNA (dsRNA) processed by Dicer and Dicer-like (DCL) family proteins in small interfering RNAs (siRNAs). In the present study, dsRNAs for coat protein (CP) and 2b genes of CMV were produced in vitro and in vivo and applied onto tobacco plants representing a systemic solanaceous host as well as on a local host plant Chenopodium quinoa. Both dsRNA treatments protected plants from local and systemic infection with CMV, but not against infection with unrelated viruses, confirming sequence specificity of antiviral RNAi. Antiviral RNAi was effective when dsRNAs were applied simultaneously with or four days prior to CMV inoculation, but not four days post inoculation. In vivo-produced dsRNAs were more effective than the in vitro-produced; in treatments with in vivo dsRNAs, dsRNA-CP was more effective than dsRNA-2b, while the effects were opposite with in vitro dsRNAs. Illumina sequencing of small RNAs from in vivo dsRNA-CP treated and non-treated tobacco plants revealed that interference with CMV infection in systemic leaves coincides with strongly reduced accumulation of virus-derived 21- and 22-nucleotide (nt) siRNAs, likely generated by tobacco DCL4 and DCL2, respectively. While the 21-nt class of viral siRNAs was predominant in non-treated plants, 21-nt and 22-nt classes accumulated at almost equal (but low) levels in dsRNA treated plants, suggesting that dsRNA treatment may boost DCL2 activity. Taken together, our findings confirm the efficacy of topical application of dsRNA for plant protection against viruses and shed more light on the mechanism of antiviral RNAi.

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Differentiation of human germ cell tumor cells in vivo and in vitro.

ACTA HISTOCHEMICA ET CYTOCHEMICA ◽

10.1267/ahc.25.563 ◽

1992 ◽

Vol 25 (5) ◽

pp. 563-576 ◽

Cited By ~ 20

Author(s):

JUN-ICHI HATA ◽

JUNICHIRO FUJIMOTO ◽

EIZABURO ISHII ◽

AKIHIRO UMEZAWA ◽

YASUO KOKAI ◽

...

Keyword(s):

Germ Cell ◽

Tumor Cells ◽

Germ Cell Tumor ◽

Cell Tumor ◽

Human Germ Cell Tumor

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Synergistic Antifungal Activity of Graphene Oxide and Fungicides against Fusarium Head Blight In Vitro and In Vivo

Nanomaterials ◽

10.3390/nano11092393 ◽

2021 ◽

Vol 11 (9) ◽

pp. 2393

Author(s):

Xiuping Wang ◽

Fei Peng ◽

Caihong Cheng ◽

Lina Chen ◽

Xuejuan Shi ◽

...

Keyword(s):

Graphene Oxide ◽

Plant Pathogens ◽

Antimicrobial Agents ◽

Plant Protection ◽

Disease Incidence ◽

Agricultural Science ◽

Plant Diseases ◽

Synergistic Activity

Plant pathogens constantly develop resistance to antimicrobial agents, and this poses great challenges to plant protection. Therefore, there is a pressing need to search for new antimicrobials. The combined use of antimicrobial agents with different antifungal mechanisms has been recognized as a promising approach to manage plant diseases. Graphene oxide (GO) is a newly emerging and highly promising antimicrobial agent against various plant pathogens in agricultural science. In this study, the inhibitory activity of GO combined with fungicides (Mancozeb, Cyproconazol and Difenoconazole) against Fusarium graminearum was investigated in vivo and in vitro. The results revealed that the combination of GO and fungicides has significant synergistic inhibitory effects on the mycelial growth, mycelial biomass and spore germination of F. graminearum relative to single fungicides. The magnitude of synergy was found to depend on the ratio of GO and fungicide in the composite. In field tests, GO–fungicides could significantly reduce the disease incidence and disease severity, exhibiting a significantly improved control efficacy on F. graminearum. The strong synergistic activity of GO with existing fungicides demonstrates the great application potential of GO in pest management.

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Susceptibility of endophyte-infected grasses to winter pathogens (snow molds)

Canadian Journal of Botany ◽

10.1139/b06-075 ◽

2006 ◽

Vol 84 (7) ◽

pp. 1043-1051 ◽

Cited By ~ 42

Author(s):

Piippa R. Wäli ◽

Marjo Helander ◽

Oiva Nissinen ◽

Kari Saikkonen

Keyword(s):

Field Experiment ◽

Plant Protection ◽

Inhibition Zone ◽

Snow Mold ◽

Dual Cultures ◽

Endophyte Infection ◽

Neotyphodium Endophyte ◽

Status Interaction

Neotyphodium endophytes are suggested to be mutualistic symbionts of grasses and regarded as potential biological plant protection agents. We examined the effects of the Neotyphodium endophyte of meadow ryegrass on snow molds in vitro with dual cultures of endophyte and Typhula ishikariensis , and on grass–snow mold interactions in vivo in a greenhouse and in a field experiment. In dual cultures, the endophytes formed an inhibition zone and retarded the growth of T. ishikariensis. In the field experiment, however, the endophyte-infected (E+) meadow ryegrasses were more susceptible to T. ishikariensis than the endophyte-free (E–) grasses. Endophyte infection increased the winter damage of grasses both in greenhouse and in field conditions. After winter, the growth of E+ grasses exceeded the growth of E– plants in the field experiment, indicating the marked tolerance of E+ grasses against winter damage. We detected differences in growth and pathogenesis between the different T. ishikariensis strains and found meadow ryegrass cultivar–endophyte status interaction in the growth of meadow ryegrasses, which highlight the effects of the genetic background of the participants on endophyte–grass–snow mold interactions.

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Secretion of glutathione S-transferase isoforms in the seminiferous tubular fluid, tissue distribution and sex steroid binding by rat GSTM1

Biochemical Journal ◽

10.1042/bj3400309 ◽

1999 ◽

Vol 340 (1) ◽

pp. 309-320 ◽

Cited By ~ 11

Author(s):

Sikha Bettina MUKHERJEE ◽

S. ARAVINDA ◽

B. GOPALAKRISHNAN ◽

Sushma NAGPAL ◽

Dinakar M. SALUNKE ◽

...

Keyword(s):

Cell Culture ◽

Germ Cell ◽

Sertoli Cell ◽

Sertoli Cells ◽

Culture Media ◽

Glutathione S Transferase ◽

Steroid Binding ◽

Spent Media

The seminiferous tubular fluid (STF) provides the microenvironment necessary for spermatogenesis in the adluminal compartment of the seminiferous tubule (ST), primarily through secretions of the Sertoli cell. Earlier studies from this laboratory demonstrated the presence of glutathione S-transferase (GST) in STF collected from adult rat testis and in the spent media of ST cultures. This study describes the cellular source, isoform composition and possible function of GSTs in the STF. The major GST isoforms present in STF in vivo share extensive N-terminal similarity with rat GSTM1 (rGSTM1), rGSTM2, rGSTM3 and rGST-Alpha. Molecular masses of rGSTM2, rGSTM3 and rGST-Alpha from liver and testis sources were similar, unlike STF-GSTM1, which was larger by 325 Da than its liver counterpart. Peptide digest analysis profiles on reverse-phase HPLC between liver and STF isoforms were identical, and N-terminal sequences of selected peptides obtained by digestion of the various isoforms were closely similar. The above results confirmed close structural similarity between liver and STF-GST isoforms. Active synthesis and secretion of GSTs by the STs were evident from recovery of radiolabelled GST from the spent media of ST cultures. Analysis of secreted GST isoforms showed that GST-Alpha was not secreted by the STs in vitro, whereas there was an induction of GST-Pi secretion. Detection of immunostainable GST-Mu in Sertoli cells in vitro and during different stages of the seminiferous epithelium in vivo, coupled with the recovery of radiolabelled GST from Sertoli cell-culture media, provided evidence for Sertoli cells as secretors of GST. In addition, STF of ‘Sertoli cell only’ animals showed no change in the profile of GST isoform secretion, thereby confirming Sertoli cells as prime GST secretors. Non-recovery of [35S]methionine-labelled GSTs from germ cell culture supernatants, but their presence in germ cell lysates, confirm the ability of the germ cells to synthesize, but not to release, GSTs. Functionally, STF-GSTM1 appeared to serve as a steroid-binding protein by its ability to bind to testosterone and oestradiol, two important hormones in the ST that are essential for spermatogenesis, with binding constants of < 9.8×10-7 M for testosterone and 9×10-6 M for oestradiol respectively.

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Erythropoietin production by a human testicular germ cell line

Blood ◽

10.1182/blood.v62.5.1132.1132 ◽

1983 ◽

Vol 62 (5) ◽

pp. 1132-1134 ◽

Cited By ~ 12

Author(s):

JL Ascensao ◽

F Gaylis ◽

D Bronson ◽

EE Fraley ◽

ED Zanjani

Keyword(s):

Germ Cell ◽

Cell Line ◽

Continuous Cell Line ◽

Human Testis ◽

Testicular Germ Cell ◽

Germ Cell Line ◽

Erythropoietin Production ◽

Unique Model

Abstract An established human testis germ cell line (1411-H) was found to produce significant amounts of erythropoietin (Ep), the primary regulator of erythropoiesis. Media conditioned by the 1411-H cells stimulated erythropoiesis both in vivo and in vitro. This activity was neutralized by anti-Ep. This continuous cell line provides a unique model for the study of the mechanisms of control of Ep biogenesis.

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Biocontrol of Tomato Fusarium wilt by Trichoderma Species under in vitro and in vivo Conditions

Cercetari Agronomice in Moldova ◽

10.1515/cerce-2016-0008 ◽

2016 ◽

Vol 49 (1) ◽

pp. 91-98 ◽

Cited By ~ 6

Author(s):

H. Barari

Keyword(s):

Plant Protection ◽

Disease Incidence ◽

Plant Diseases ◽

Growth And Yield ◽

Mazandaran Province ◽

Trichoderma Species ◽

Tomato Plants ◽

Geographical Regions

AbstractTrichodermaspp. have long been used as biological control agents against plant fungal diseases, but the mechanisms by which the fungi confer protection are not well understood. Our goal in this study was to isolate species ofTrichoderma, that exhibit high levels of biocontrol efficacy from natural environments and to investigate the mechanisms by which these strains confer plant protection. In this study, efficacy of the native isolates ofTrichodermaspecies to promote the growth and yield parameters of tomato and to manageFusariumwilt disease underin vitroandin vivoconditions were investigated. The dominant pathogen, which causesFusariumwilt of tomato, was isolated and identified asFusarium oxysporumf. sp.lycopersici(FOL). Twenty eight nativeTrichodermaantagonists were isolated from healthy tomato rhizosphere soil in different geographical regions of Mazandaran province, Iran. Underin vitroconditions, the results revealed thatTrichoderma harzianum, isolate N-8, was found to inhibit effectively the radial mycelial growth of the pathogen (by 68.22%). Under greenhouse conditions, the application ofT. harzianum(N-8) exhibited the least disease incidence (by 14.75%). Also, tomato plants treated withT. harzianum(N-8) isolate showed a significant stimulatory effect on plant height (by 70.13 cm) and the dry weight (by 265.42 g) of tomato plants, in comparison to untreated control (54.6 cm and 195.5 g). Therefore, the antagonistT. harzianum(N-8) is chosen to be the most promising bio-control agent forF. oxysporumf. sp.lycopersici. On the base of present study, the biocontrol agents of plant diseases might be exploited for sustainable disease management programs to save environmental risk.

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PRODUCTION OF ALPHA-FETOPROTEIN BY HUMAN GERM CELL TUMORS IN VIVO AND IN VITRO

Pathology International ◽

10.1111/j.1440-1827.1987.tb00459.x ◽

1987 ◽

Vol 37 (8) ◽

pp. 1263-1277 ◽

Cited By ~ 1

Author(s):

Teiichi Motoyama ◽

Hidenobu Watanabe ◽

Takahiko Yamamoto ◽

Morimasa Sekiguchi

Keyword(s):

Germ Cell ◽

Germ Cell Tumors ◽

Alpha Fetoprotein

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Dynein 1 supports spermatid transport and spermiation during spermatogenesis in the rat testis

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00114.2018 ◽

2018 ◽

Vol 315 (5) ◽

pp. E924-E948 ◽

Cited By ~ 15

Author(s):

Qing Wen ◽

Elizabeth I. Tang ◽

Wing-yee Lui ◽

Will M. Lee ◽

Chris K. C. Wong ◽

...

Keyword(s):

Germ Cell ◽

Heavy Chain ◽

Sertoli Cells ◽

Cytoplasmic Dynein ◽

Seminiferous Epithelium ◽

Organelle Transport ◽

Cellular Events

In the mammalian testis, spermatogenesis is dependent on the microtubule (MT)-specific motor proteins, such as dynein 1, that serve as the engine to support germ cell and organelle transport across the seminiferous epithelium at different stages of the epithelial cycle. Yet the underlying molecular mechanism(s) that support this series of cellular events remain unknown. Herein, we used RNAi to knockdown cytoplasmic dynein 1 heavy chain (Dync1h1) and an inhibitor ciliobrevin D to inactivate dynein in Sertoli cells in vitro and the testis in vivo, thereby probing the role of dynein 1 in spermatogenesis. Both treatments were shown to extensively induce disruption of MT organization across Sertoli cells in vitro and the testis in vivo. These changes also perturbed the transport of spermatids and other organelles (such as phagosomes) across the epithelium. These changes thus led to disruption of spermatogenesis. Interestingly, the knockdown of dynein 1 or its inactivation by ciliobrevin D also perturbed gross disruption of F-actin across the Sertoli cells in vitro and the seminiferous epithelium in vivo, illustrating there are cross talks between the two cytoskeletons in the testis. In summary, these findings confirm the role of cytoplasmic dynein 1 to support the transport of spermatids and organelles across the seminiferous epithelium during the epithelial cycle of spermatogenesis.

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Overexpression of Bcl-w in the Testis Disrupts Spermatogenesis: Revelation of a Role of BCL-W in Male Germ Cell Cycle Control

Molecular Endocrinology ◽

10.1210/me.2002-0389 ◽

2003 ◽

Vol 17 (9) ◽

pp. 1868-1879 ◽

Cited By ~ 22

Author(s):

Wei Yan ◽

Jun-Xing Huang ◽

Anna-Stina Lax ◽

Lauri Pelliniemi ◽

Eeva Salminen ◽

...

Keyword(s):

Cell Cycle ◽

Germ Cell ◽

Sertoli Cell ◽

Germ Cells ◽

Cell Cycle Progression ◽

Cell Cycle Control ◽

Testicular Development ◽

Dna Ladder

Abstract To explore physiological roles of BCL-W, a prosurvival member of the BCL-2 protein family, we generated transgenic (TG) mice overexpressing Bcl-w driven by a chicken β-actin promoter. Male Bcl-w TG mice developed normally but were infertile. The adult TG testes displayed disrupted spermatogenesis with various severities ranging from thin seminiferous epithelium containing less germ cells to Sertoli cell-only appearance. No overpopulation of any type of germ cells was observed during testicular development. In contrast, the developing TG testes displayed decreased number of spermatogonia, degeneration, and detachment of spermatocytes and Sertoli cell vacuolization. The proliferative activity of germ cells was significantly reduced during testicular development and spermatogenesis, as determined by in vivo and in vitro 5′-bromo-2′deoxyuridine incorporation assays. Sertoli cells were structurally and functionally normal. The degenerating germ cells were TUNEL-negative and no typical apoptotic DNA ladder was detected. Our data suggest that regulated spatial and temporal expression of BCL-W is required for normal testicular development and spermatogenesis, and overexpression of BCL-W inhibits germ cell cycle entry and/or cell cycle progression leading to disrupted spermatogenesis.

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