scholarly journals Overexpression of HOP2 induces developmental defects and compromises growth in Arabidopsis

2021 ◽  
Author(s):  
Ameth N. Garrido ◽  
Therese Francom ◽  
Sakina Divan ◽  
Mohamad Kesserwan ◽  
Jenya Daradur ◽  
...  
Keyword(s):  
Floral Organ ◽  
Developmental Defects ◽  
Growth Defects ◽  
Homologous Chromosome Pairing ◽  
Developmental Growth ◽  
Gfp Reporter ◽  
Growth Properties ◽  
Dna Insertion ◽  
Tail Pcr ◽  
Insertion Mutants

HOMOLOGOUS PAIRING 2 (HOP2) is a predominantly meiotic protein that plays a pivotal role in homologous chromosome pairing in organisms as diverse as yeast and mammals. While generating HOP2::GFP reporter lines, we identified two Arabidopsis T-DNA insertion mutants, stunted1(std1) and stunted2 (std2) that exhibit pleiotropic phenotypes, including fasciated stems, altered phyllotaxy, floral organ defects, reduced fecundity, and an overall reduction in growth properties. TAIL-PCR followed by sequencing revealed several insertions near genes, but genotyping showed that none of the insertions are causal. Analysis the std mutants by qRT-PCR, and analysis of dexamethasone inducible HOP2 transgenic plants demonstrated that the std phenotypes are associated with ectopic/overexpression of HOP2. Based on the postulated mechanisms of HOP2 action, we speculate on how overexpression leads to these developmental/growth defects.

scholarly journals The mitochondrial monothiol glutaredoxin S15 is essential for iron-sulfur protein maturation in Arabidopsis thaliana

2015 ◽  
Vol 112 (44) ◽  
pp. 13735-13740 ◽  
Author(s):  
Anna Moseler ◽  
Isabel Aller ◽  
Stephan Wagner ◽  
Thomas Nietzel ◽  
Jonathan Przybyla-Toscano ◽  
...  
Keyword(s):  
Amino Acid Residues ◽  
Iron Sulfur Cluster ◽  
Growth Defects ◽  
Iron Sulfur ◽  
Iron Sulfur Proteins ◽  
Dna Insertion ◽  
Sulfur Protein ◽  
Metabolic Reactions ◽  
Insertion Mutants

The iron-sulfur cluster (ISC) is an ancient and essential cofactor of many proteins involved in electron transfer and metabolic reactions. In Arabidopsis, three pathways exist for the maturation of iron-sulfur proteins in the cytosol, plastids, and mitochondria. We functionally characterized the role of mitochondrial glutaredoxin S15 (GRXS15) in biogenesis of ISC containing aconitase through a combination of genetic, physiological, and biochemical approaches. Two Arabidopsis T-DNA insertion mutants were identified as null mutants with early embryonic lethal phenotypes that could be rescued by GRXS15. Furthermore, we showed that recombinant GRXS15 is able to coordinate and transfer an ISC and that this coordination depends on reduced glutathione (GSH). We found the Arabidopsis GRXS15 able to complement growth defects based on disturbed ISC protein assembly of a yeast Δgrx5 mutant. Modeling of GRXS15 onto the crystal structures of related nonplant proteins highlighted amino acid residues that after mutation diminished GSH and subsequently ISC coordination, as well as the ability to rescue the yeast mutant. When used for plant complementation, one of these mutant variants, GRXS15K83/A, led to severe developmental delay and a pronounced decrease in aconitase activity by approximately 65%. These results indicate that mitochondrial GRXS15 is an essential protein in Arabidopsis, required for full activity of iron-sulfur proteins.

scholarly journals mRNA Surveillance Complex PELOTA-HBS1 Regulates Phosphoinositide-Dependent Protein Kinase1 and Plant Growth

2021 ◽  
Author(s):  
Wei Kong ◽  
Shutang Tan ◽  
Qing Zhao ◽  
De-Li Lin ◽  
Zhi-Hong Xu ◽  
...  
Keyword(s):  
Quality Control ◽  
Messenger Rna ◽  
Loss Of Function ◽  
Yeast Saccharomyces Cerevisiae ◽  
Developmental Defects ◽  
Mrna Surveillance ◽  
Dna Insertion ◽  
Dependent Protein ◽  
Heterodimeric Complex ◽  
Severe Growth

Abstract The quality control system for messenger RNA (mRNA) is fundamental for cellular activities in eukaryotes. To elucidate the molecular mechanism of 3’-Phosphoinositide-Dependent Protein Kinase1 (PDK1), a master regulator that is essential throughout eukaryotic growth and development, we employed a forward genetic approach to screen for suppressors of the loss-of-function T-DNA insertion double mutant pdk1.1 pdk1.2 in Arabidopsis thaliana. Notably, the severe growth attenuation of pdk1.1 pdk1.2 was rescued by sop21 (suppressor of pdk1.1 pdk1.2), which harbours a loss-of-function mutation in PELOTA1 (PEL1). PEL1 is a homologue of mammalian PELOTA and yeast (Saccharomyces cerevisiae) DOM34p, which each form a heterodimeric complex with the GTPase HBS1 (HSP70 SUBFAMILY B SUPPRESSOR1, also called SUPERKILLER PROTEIN7, SKI7), a protein that is responsible for ribosomal rescue and thereby assures the quality and fidelity of mRNA molecules during translation. Genetic analysis further revealed that a dysfunctional PEL1-HBS1 complex failed to degrade the T-DNA-disrupted PDK1 transcripts, which were truncated but functional, and thus rescued the growth and developmental defects of pdk1.1 pdk1.2. Our studies demonstrated the functionality of a homologous PELOTA-HBS1 complex and identified its essential regulatory role in plants, providing insights into the mechanism of mRNA quality control.

scholarly journals Epigenetic Suppression of T-DNA Insertion Mutants in Arabidopsis

2013 ◽  
Vol 6 (2) ◽  
pp. 539-545 ◽  
Author(s):  
Yangbin Gao ◽  
Yunde Zhao
Keyword(s):  
Dna Insertion ◽  
Epigenetic Suppression ◽  
Insertion Mutants

scholarly journals Mutation of ONAC096 Enhances Grain Yield by Increasing Panicle Number and Delaying Leaf Senescence during Grain Filling in Rice

2019 ◽  
Vol 20 (20) ◽  
pp. 5241 ◽  
Author(s):  
Kiyoon Kang ◽  
Yejin Shim ◽  
Eunji Gi ◽  
Gynheung An ◽  
Nam-Chon Paek
Keyword(s):  
Grain Yield ◽  
Leaf Senescence ◽  
Grain Filling ◽  
Pcr Analysis ◽  
Cytokinin Oxidase ◽  
Panicle Number ◽  
Reverse Transcription Quantitative Pcr ◽  
Dna Insertion ◽  
Upregulated Genes ◽  
Insertion Mutants

Exploring genetic methods to improve yield in grain crops such as rice (Oryza sativa) is essential to help meet the needs of the increasing population. Here, we report that rice ONAC096 affects grain yield by regulating leaf senescence and panicle number. ONAC096 expression increased rapidly in rice leaves upon the initiation of aging- and dark-induced senescence. Two independent T-DNA insertion mutants (onac096-1 and onac096-2) with downregulated ONAC096 expression retained their green leaf color during natural senescence in the field, thus extending their photosynthetic capacity. Reverse-transcription quantitative PCR analysis showed that ONAC096 upregulated genes controlling chlorophyll degradation and leaf senescence. Repressed OsCKX2 (encoding cytokinin oxidase/dehydrogenase) expression in the onac096 mutants led to a 15% increase in panicle number without affecting grain weight or fertility. ONAC096 mediates abscisic acid (ABA)-induced leaf senescence by upregulating the ABA signaling genes ABA INSENSITIVE5 and ENHANCED EM LEVEL. The onac096 mutants showed a 16% increase in grain yield, highlighting the potential for using this gene to increase grain production.

Characterization of T-DNA insertion mutants with decreased virulence in the entomopathogenic fungus Beauveria bassiana JEF-007

2016 ◽  
Vol 100 (20) ◽  
pp. 8889-8900 ◽  
Author(s):  
Sihyeon Kim ◽  
Se Jin Lee ◽  
Yu-Shin Nai ◽  
Jeong Seon Yu ◽  
Mi Rong Lee ◽  
...  
Keyword(s):  
Beauveria Bassiana ◽  
Entomopathogenic Fungus ◽  
Dna Insertion ◽  
Fungus Beauveria Bassiana ◽  
Insertion Mutants

scholarly journals Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene

2011 ◽  
Vol 62 (15) ◽  
pp. 5531-5545 ◽  
Author(s):  
Miloslava Fojtová ◽  
Vratislav Peška ◽  
Zuzana Dobšáková ◽  
Iva Mozgová ◽  
Jiří Fajkus ◽  
...  
Keyword(s):  
Molecular Analysis ◽  
Regulatory Elements ◽  
Dna Insertion ◽  
Insertion Mutants

The genetic and physiological analysis of late-flowering phenotype of T-DNA insertion mutants of AtCAL1 and AtCAL2 in Arabidopsis

2011 ◽  
Vol 39 (2) ◽  
pp. 1527-1535 ◽  
Author(s):  
Jihong Zhang ◽  
Xinhong Guo ◽  
Xiushan Li ◽  
Feng Xiang ◽  
Bo Zhou ◽  
...  
Keyword(s):  
Late Flowering ◽  
Physiological Analysis ◽  
Dna Insertion ◽  
Insertion Mutants

Analyses of Arabidopsis trr14 T-DNA Insertion Mutants Reveal an Essential Role in Seed Germination

2011 ◽  
Vol 30 (2) ◽  
pp. 319-329 ◽  
Author(s):  
Mahnaz Aghdasi ◽  
Fariba Fazli ◽  
Mohammad B. Bagherieh-Najjar
Keyword(s):  
Seed Germination ◽  
Essential Role ◽  
Dna Insertion ◽  
Insertion Mutants

scholarly journals Species-specific gene duplication in Arabidopsis thaliana evolved novel phenotypic effects on morphological traits under strong positive selection

2021 ◽  
Author(s):  
Yuan Huang ◽  
Jiahui Chen ◽  
Chuan Dong ◽  
Dylan Sosa ◽  
Shengqian Xia ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Morphological Traits ◽  
Morphological Changes ◽  
Specific Gene ◽  
Phenotypic Evolution ◽  
Parental Gene ◽  
Dna Insertion ◽  
New Gene ◽  
Species Specific ◽  
Insertion Mutants

Gene duplication is increasingly recognized as an important mechanism for the origination of new genes, as revealed by comparative genomic analysis. However, the ways in which new duplicate genes contribute to phenotypic evolution remain largely unknown, especially in plants, owing to a lack of experimental and phenotypic data. In this study, we identified the new gene Exov, derived from a partial gene region duplication of its parental gene Exov-L, which is a member of an exonuclease family, into a different chromosome in Arabidopsis thaliana. We experimentally investigated the phenotypic effects of Exov and Exov-L in an attempt to understand how the new gene diverged from the parental copy and contributes to phenotypic evolution. Evolutionary analysis demonstrated that Exov is a species-specific gene that originated within the last 3.5 million years and shows strong signals of positive selection. Unexpectedly, RNAseq analyses reveal that the new gene, despite its young age, has acquired a large number of novel direct and indirect interactions in which the parental gene does not engage. This is consistent with a high, selection-driven substitution rate in the protein sequence encoded by Exov in contrast to the slowly evolving Exov-L, suggesting an important role for Exov in phenotypic evolution. We analyzed phenotypic effects of exov and exov-l single T-DNA-insertion mutants; double exov, exov-l T-DNA insertion mutants; and CRISPR/Cas9-mediated exovcrp and exov-lcrp knockouts on seven morphological traits in both the new and parental genes. We detected significant segregation of morphological changes for all seven traits when assessed in terms of single mutants, as well as morphological changes for seven traits associated with segregation of double exov, exov-l mutants. Substantial divergence of phenotypic effects between new and parental genes was revealed by principal component analyses, suggesting neofunctionalization in the new gene. These results reveal a young gene that plays critical roles in biological processes that underlie morphological and developmental evolution in Arabidopsis thaliana.

Jasmonate-dependent expression of a galactinol synthase gene is involved in priming of systemic fungal resistance in Arabidopsis thaliana

Botany ◽  
2010 ◽  
Vol 88 (5) ◽  
pp. 452-461 ◽  
Author(s):  
Song Mi Cho ◽  
Eun Young Kang ◽  
Mi Seong Kim ◽  
Seung Jin Yoo ◽  
Yang Ju Im ◽  
...  
Keyword(s):  
Fungal Pathogen ◽  
Target Gene ◽  
Fungal Resistance ◽  
Systemic Resistance ◽  
Raffinose Family Oligosaccharides ◽  
Galactinol Synthase ◽  
Dna Insertion ◽  
Mutant Lines ◽  
Dependent Pathway ◽  
Insertion Mutants

Previously, root colonization by the rhizobacterium, Pseudomonas chlororaphis O6, was shown to induce expression of galactinol synthase conferring systemic resistance against a fungal pathogen in cucumber leaves. Here, the Arabidopsis – Botrytis cinerea system is introduced to better understand signal transduction of galactinol and (or) raffinose family oligosaccharides (RFO) during O6-mediated induced systemic resistance (ISR). Among the 10 Arabidopsis galactinol synthase genes, only AtGolS1 was specifically induced upon infection with the fungal pathogen B. cinerea. AtGolS1 was primed by O6 colonization against the pathogen in Arabidopsis leaves. Arabidopsis T-DNA insertion mutants at the AtGolS1 gene site compromised O6-mediated ISR against the pathogen, thereby suggesting that AtGolS1 plays an important role in ISR. O6 colonization increased AtGolS1 transcription as well as ISR in several Arabidopsis signaling mutants, but not in the jar1-1 and coi1 mutant lines. Exogenous jasmonate treatment induced transcription of AtGolS1 in wild-type Col-0 plants, but salicylic acid and 1-aminocyclopropane-1-carboxylate did not. These studies on signaling mutants and target gene expression indicate that expression of AtGolS1 in response to O6 colonization is mediated through the jasmonate-dependent pathway, stimulating ISR in Arabidopsis against B. cinerea infection.

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