scholarly journals Different components of the RNAi machinery are required for conidiation, ascosporogenesis, virulence, DON production and fungal inhibition by exogenous dsRNA in the Head Blight pathogenFusarium graminearum

2019 ◽  
Author(s):  
Fatima Yousif Gaffar ◽  
Jafargholi Imani ◽  
Petr Karlovsky ◽  
Aline Koch ◽  
Karl-Heinz Kogel
Keyword(s):  
Gene Silencing ◽  
Recq Helicase ◽  
Head Blight ◽  
Low Light ◽  
Rnai Machinery ◽  
Interacting Protein ◽  
Knock Out ◽  
Fungal Inhibition ◽  
Asexual Multiplication ◽  
Wheat Kernels

AbstractGene silencing through RNA interference (RNAi) shapes many biological processes in filamentous fungi, including pathogenicity. In this study we explored the requirement of key components of fungal RNAi machinery, including DICER-like 1 and 2 (FgDCL1,FgDCL2), ARGONAUTE 1 and 2 (FgAGO1,FgAGO2), AGO-interacting proteinFgQIP (QDE2-interacting protein), RecQ helicase (FgQDE3), and four RNA-dependent RNA polymerases (FgRdRP1,FgRdRP2,FgRdRP3,FgRdRP4), in the ascomycete mycotoxin-producing fungal pathogenFusarium graminearum(Fg) for sexual and asexual multiplication, pathogenicity, and its sensitivity to double-stranded (ds)RNA. We corroborate and extend earlier findings that conidiation, ascosporogenesis and Fusarium Head Blight (FHB) symptom development require an operable RNAi machinery. The involvement of RNAi in conidiation is dependent on environmental conditions as it is detectable only under low light (< 2 µmol m−2s−1). Although both DCLs and AGOs partially share their functions, the sexual ascosporogenesis is mediated primarily byFgDCL1 andFgAGO2, whileFgDCL2 andFgAGO1 contribute to asexual conidia formation and germination.FgDCL1 andFgAGO2 also account for pathogenesis as their knock-out (KO) results in reduced FHB development. Apart from KO mutantsΔdcl2andΔago1, mutantsΔrdrp2, Δrdrp3, Δrdrp4, Δqde3andΔqipare strongly compromised for conidiation, while KO mutations in allRdPRs, QDE3andQIPstrongly affect ascosporogenesis. Analysis of trichothecenes mycotoxins in wheat kernels showed that the relative amount of deoxynivalenol (DON), calculated as [DON] per amount of fungal genomic DNA, was reduced in all spikes infected with RNAi mutants, suggesting the possibility that the fungal RNAi pathways affectFg’s DON production in wheat spikes. Moreover, gene silencing by exogenous target gene specific dsRNA (spray-induced gene silencing, SIGS) is dependent on fungal DCLs, AGOs, and QIP, but not on QDE3. Together these data show that inF. graminearumdifferent key components of the RNAi machinery are crucial in different steps of fungal development and pathogenicity.

scholarly journals Viroids as a Tool to Study RNA-Directed DNA Methylation in Plants

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1187
Author(s):  
Michael Wassenegger ◽  
Athanasios Dalakouras
Keyword(s):  
Dna Methylation ◽  
Rna Interference ◽  
Gene Silencing ◽  
Plant Cells ◽  
Transcriptional Gene Silencing ◽  
Rnai Machinery ◽  
Double Stranded Rna ◽  
Post Transcriptional Gene Silencing ◽  
Cumulative Amount

Viroids are plant pathogenic, circular, non-coding, single-stranded RNAs (ssRNAs). Members of the Pospiviroidae family replicate in the nucleus of plant cells through double-stranded RNA (dsRNA) intermediates, thus triggering the host’s RNA interference (RNAi) machinery. In plants, the two RNAi pillars are Post-Transcriptional Gene Silencing (PTGS) and RNA-directed DNA Methylation (RdDM), and the latter has the potential to trigger Transcriptional Gene Silencing (TGS). Over the last three decades, the employment of viroid-based systems has immensely contributed to our understanding of both of these RNAi facets. In this review, we highlight the role of Pospiviroidae in the discovery of RdDM, expound the gradual elucidation through the years of the diverse array of RdDM’s mechanistic details and propose a revised RdDM model based on the cumulative amount of evidence from viroid and non-viroid systems.

scholarly journals The CBL-Interacting Protein Kinase NtCIPK23 Positively Regulates Seed Germination and Early Seedling Development in Tobacco (Nicotiana tabacum L.)

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 323
Author(s):  
Sujuan Shi ◽  
Lulu An ◽  
Jingjing Mao ◽  
Oluwaseun Olayemi Aluko ◽  
Zia Ullah ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Seed Germination ◽  
Seedling Growth ◽  
Hypocotyl Elongation ◽  
Interacting Protein ◽  
Knock Out ◽  
Early Seedling Growth ◽  
Tobacco Seedlings ◽  
Early Seedling ◽  
Cotyledon Expansion

CBL-interacting protein kinase (CIPK) family is a unique group of serine/threonine protein kinase family identified in plants. Among this family, AtCIPK23 and its homologs in some plants are taken as a notable group for their importance in ions transport and stress responses. However, there are limited reports on their roles in seedling growth and development, especially in Solanaceae plants. In this study, NtCIPK23, a homolog of AtCIPK23 was cloned from Nicotiana tabacum. Expression analysis showed that NtCIPK23 is mainly expressed in the radicle, hypocotyl, and cotyledons of young tobacco seedlings. The transcriptional level of NtCIPK23 changes rapidly and spatiotemporally during seed germination and early seedling growth. To study the biological function of NtCIPK23 at these stages, the overexpressing and CRISPR/Cas9-mediated knock-out (ntcipk23) tobacco lines were generated. Phenotype analysis indicated that knock-out of NtCIPK23 significantly delays seed germination and the appearance of green cotyledon of young tobacco seedling. Overexpression of NtCIPK23 promotes cotyledon expansion and hypocotyl elongation of young tobacco seedlings. The expression of NtCIPK23 in hypocotyl is strongly upregulated by darkness and inhibited under light, suggesting that a regulatory mechanism of light might underlie. Consistently, a more obvious difference in hypocotyl length among different tobacco materials was observed in the dark, compared to that under the light, indicating that the upregulation of NtCIPK23 contributes greatly to the hypocotyl elongation. Taken together, NtCIPK23 not only enhances tobacco seed germination, but also accelerate early seedling growth by promoting cotyledon greening rate, cotyledon expansion and hypocotyl elongation of young tobacco seedlings.

Gene silencing of receptor-interacting protein 2 protects against cigarette smoke-induced acute lung injury

2019 ◽  
Vol 139 ◽  
pp. 560-568 ◽  
Author(s):  
Jinrui Dong ◽  
Wupeng Liao ◽  
Lay Hong Tan ◽  
Amy Yong ◽  
Wen Yan Peh ◽  
...  
Keyword(s):  
Acute Lung Injury ◽  
Lung Injury ◽  
Gene Silencing ◽  
Cigarette Smoke ◽  
Interacting Protein

scholarly journals Receptor-interacting Protein 140 Directly Recruits Histone Deacetylases for Gene Silencing

2000 ◽  
Vol 275 (52) ◽  
pp. 40782-40787 ◽  
Author(s):  
Li-Na Wei ◽  
Xinli Hu ◽  
Dhyan Chandra ◽  
Edward Seto ◽  
Maria Farooqui
Keyword(s):  
Gene Silencing ◽  
Histone Deacetylases ◽  
Interacting Protein

scholarly journals Novel Genes of Fusarium graminearum That Negatively Regulate Deoxynivalenol Production and Virulence

2009 ◽  
Vol 22 (12) ◽  
pp. 1588-1600 ◽  
Author(s):  
Donald M. Gardiner ◽  
Kemal Kazan ◽  
John M. Manners
Keyword(s):  
Gene Expression ◽  
Fusarium Graminearum ◽  
Global Analysis ◽  
Gene Clusters ◽  
Head Blight ◽  
Knock Out ◽  
Serious Disease ◽  
Amine Compounds ◽  
Fungal Genes ◽  
Fungal Gene Expression

Fusarium head blight of wheat, caused by Fusarium graminearum, is a serious disease resulting in both reduced yields and contamination of grain with trichothecene toxins, with severe consequences for mammalian health. Recently, we have identified several related amine compounds such as agmatine and putrescine that promote the production of high levels of trichothecene toxins, such as deoxynivalenol (DON), in culture by F. graminearum and F. sporotrichioides. Here, a global analysis of fungal gene expression using the Affymetrix Fusarium GeneChip during culture under DON-inducing conditions compared with noninducing conditions is reported. Agmatine differentially regulated a large number of fungal genes, including both known and previously uncharacterized putative secondary metabolite biosynthetic gene clusters. In silico prediction of binding sites for the transcriptional regulator (TRI6) controlling TRI gene expression and gene expression analysis in a TRI6 mutant of F. graminearum showed that three of the differentially regulated genes were under the control of TRI6. Gene knock-out mutations of two of these genes resulted in mutants with massively increased production of DON and increased aggressiveness toward wheat. Our results not only identify a novel mechanism of negative regulation of DON production and virulence in F. graminearum but also point out the potential of this pathogen to evolve with an ability to produce massively increased amounts of toxins and increased virulence.

Vesper hard red spring wheat

2013 ◽  
Vol 93 (2) ◽  
pp. 315-321 ◽  
Author(s):  
J. Thomas ◽  
S. Fox ◽  
B. McCallum ◽  
T. Fetch ◽  
J. Gilbert ◽  
...  
Keyword(s):  
Spring Wheat ◽  
High Water ◽  
Head Blight ◽  
Hard Red Spring Wheat ◽  
Wheat Midge ◽  
End Use ◽  
Sprouting Resistance ◽  
Moderately Resistant ◽  
Wheat Kernels

Thomas, J., Fox, S., McCallum, B., Fetch, T., Gilbert, J., Menzies, J., Wise, I., Smith, M., Gaudet, D., Niziol, D., Humphreys, G. and Brown, D. 2013. Vesper hard red spring wheat. Can. J. Plant Sci. 93: 315–321. Vesper is a high-yielding, hard red spring wheat that is adapted to the wheat-growing regions of Manitoba and Saskatchewan. In the Central Bread Wheat Cooperative Registration Trials of 2007, 2008 and 2009, Vesper out-yielded five check cultivars by an average of 12%, and Unity, which was the highest yielding check, by 4.6%. Lodging, height and maturity scores of Vesper were all intermediate (similar to the check mean). Pre-harvest sprouting resistance of Vesper was equivalent to the poorer checks (Katepwa and CDC Teal). Kernels of Vesper were heavier than all five checks and test weight was high (exceeded only by Unity). Vesper was resistant to leaf rust and was moderately resistant to stem rust and Fusarium head blight. Vesper was intermediately resistant to loose smut and was susceptible to common bunt. Spikes of Vesper showed two forms of resistance to wheat midge: antibiotic resistance (no larvae observed in the field) and antixenotic resistance (reduced egg numbers laid by caged ovipositing females). Over 3 yr of testing, end use quality of Vesper was rated as eligible for the Canada Western Red Spring (CWRS) market class of wheat. Kernels of Vesper were relatively hard with consequent high water absorption.

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