Global and local perturbation of the tomato microRNA pathway by a<i>trans</i>-activated<i>DICER-LIKE 1</i>mutant

Kravchik, Michael; Sunkar, Ramanjulu; Damodharan, Subha; Stav, Ran; Zohar, Matat; Isaacson, Tal; Arazi, Tzahi

doi:10.1093/jxb/ert428

Cited by 29 publications

(40 citation statements)

References 65 publications

(124 reference statements)

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“…Arabidopsis thaliana DCL1i plants showed significant phenotypic variability (Lackey et al , ), which correlates with the phenotypic stochasticity observed here, whereas Oryza sativa DCL1i plants showed severe dwarfism, curly leaves and tortuous shoots (Liu et al , ). Physcomitrella patens dcl1a null mutants presented abnormality in cell size, shape and growth rate (Khraiwesh et al , ) and, finally, trans‐activated DCL1i plants showed malformation in leaf primordia (Kravchik et al , ). NbDCL2i plants showed increased branching (Fig.…”

Section: Resultsmentioning

confidence: 99%

DCL‐suppressed Nicotiana benthamiana plants: valuable tools in research and biotechnology

Κατσαρού

Mitta

Bardani

et al. 2018

Molecular Plant Pathology

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Summary RNA silencing is a universal mechanism involved in development, epigenetic modifications and responses to biotic and abiotic stresses. The major components of this mechanism are Dicer‐like (DCL), Argonaute (AGO) and RNA‐dependent RNA polymerase (RDR) proteins. Understanding the role of each component is of great scientific and agronomic importance. Plants, including Nicotiana benthamiana , an important plant model, usually possess four DCL proteins, each of which has a specific role, namely being responsible for the production of an exclusive small RNA population. Here, we used RNA interference (RNAi) technology to target DCL proteins and produced single and combinatorial mutants for DCL. We analysed the phenotype for each DCL knockdown plant, together with the small RNA profile, by next‐generation sequencing (NGS). We also investigated transgene expression, as well as viral infections, and were able to show that DCL suppression results in distinct developmental defects, changes in small RNA populations, increases in transgene expression and, finally, higher susceptibility in certain RNA viruses. Therefore, these plants are excellent tools for the following: (i) to study the role of DCL enzymes; (ii) to overexpress proteins of interest; and (iii) to understand the complex relationship between the plant silencing mechanism and biotic or abiotic stresses.

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Section: Resultsmentioning

confidence: 99%

DCL‐suppressed Nicotiana benthamiana plants: valuable tools in research and biotechnology

Κατσαρού

Mitta

Bardani

et al. 2018

Molecular Plant Pathology

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“…Loss-of-function mutants and antisense transformants of DCL1 or AGO1 showed varying degrees of developmental defects and lethality due to decreased miRNA expression levels in different species (Park et al 2002;Schauer et al 2002;Liu et al 2005;Mi et al 2008;Khraiwesh et al 2010;Bozorov et al 2012;Kravchik et al 2014). In our degradome library, DCL1 and AGO1 were targeted by miR162 and miR168, respectively.…”

Section: Autoregulation Of Mirna Biogenesismentioning

confidence: 99%

Small RNA and degradome sequencing reveals microRNAs and their targets involved in tomato pedicel abscission

Wang

Liu

et al. 2015

Planta

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We constructed small RNA and degradome sequencing libraries to identify miRNAs and targets involved in tomato pedicel abscission, and confirmed their roles via quantitative real-time PCR. MicroRNAs (miRNAs) are endogenous small RNAs which play crucial negatively regulatory roles at both the transcriptional and post-transcriptional levels in plants; however, limited knowledge is available on the expression profiles of miRNAs and their target genes during tomato pedicel abscission. Taking advantage of small RNA (sRNA) and degradome sequencing technology, a total of 56 known and 11 novel candidate miRNAs targeting 223 mRNA genes were confirmed during pedicel abscission. Gene ontology annotation and KEGG pathway analysis showed that these target genes were significantly enriched in intracellular, membrane-bounded organelle-related biological processes as well as in metabolic, plant-pathogen interaction and hormone signaling pathways. We screened 17 miRNA/target pairs for further analysis and performed quantitative real-time PCR to identify the roles. Cluster analysis of selected miRNAs revealed that the expression profiles of miRNAs varied in different stages of abscission and could be impacted by ethylene treatment. In the present study, the correlations between miRNAs and targets suggested a complex regulatory network of miRNA-mediated target interaction during pedicel abscission. Additionally, the expression profiles of miRNAs and their targets changed by ethylene might be a considerable reason why ethylene promotes pedicel abscission. Our study provides new insights into the expression and regulatory profiles of miRNAs during tomato pedicel abscission.

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“…The MIR396a generates two mature sequences, miR396a-5p and miR396a-3p, which are complementary strands (Kozomara and Griffiths-Jones 2014). Previous study has shown that miR396a-5p can act as a guide strand and its main function is to inhibit the expression of its target genes at the mRNA level, and is described as miR396a (Kravchik et al 2014). MiR396a-3p can also act as a passenger strand, but this may be rapidly degraded in the cytoplasm.…”

Section: Introductionmentioning

confidence: 99%

Sp-miR396a-5p acts as a stress-responsive genes regulator by conferring tolerance to abiotic stresses and susceptibility to Phytophthora nicotianae infection in transgenic tobacco

2015

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Overexpression of Sp-miR396a-5p in tobacco increased tolerance to salt, drought, cold stress and susceptibility to Phytophthora nicotianae infection. MicroRNA396 (miR396) is one of the conserved microRNA families in plants, and it targeted growth-regulating factors (GRFs) family. The GRF transcription factors are associated with growth and stress responses. However, the molecular mechanisms of miR396 responding to environmental stresses are elusive. The purpose of this study was to explore the function of tomato miR396a-5p (Sp-miR396a-5p) in Solanaceae responses to abiotic and biotic stresses. We showed that Sp-miR396a-5p transcript levels were up-regulated under salt and drought stresses and down-regulated after Phytophthora infestans (P. infestans) infection. Consistently, overexpression of Sp-miR396a-5p in tobacco enhanced its tolerance to salt, drought and cold stresses. Additionally, the expression of Sp-miR396a-5p was found to be down-regulated under pathogen-related biotic stress. Tobacco plants overexpressing Sp-miR396a-5p showed increased susceptibility to Phytophthora nicotianae (P. nicotianae) infection. Physiological analysis indicated that Sp-miR396a-5p overexpression enhanced osmoregulation and decreased production of reactive oxygen species (ROS). Furthermore, four Sp-miR396a-5p target genes, NtGRF1, NtGRF3, NtGRF7 and NtGRF8, were down-regulated in these plants. Our results suggested that Sp-miR396a-5p plays critical roles in both abiotic stresses through targeting NtGRF7-regulated expression of osmotic stress-responsive genes and pathogen infection via the regulatory networks of NtGRF1 and NtGRF3.

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Global and local perturbation of the tomato microRNA pathway by atrans-activatedDICER-LIKE 1mutant

Cited by 29 publications

References 65 publications

DCL‐suppressed Nicotiana benthamiana plants: valuable tools in research and biotechnology

DCL‐suppressed Nicotiana benthamiana plants: valuable tools in research and biotechnology

Small RNA and degradome sequencing reveals microRNAs and their targets involved in tomato pedicel abscission

Sp-miR396a-5p acts as a stress-responsive genes regulator by conferring tolerance to abiotic stresses and susceptibility to Phytophthora nicotianae infection in transgenic tobacco

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