Photosynthetic efficiency and mesophyll conductance are unaffected in Arabidopsis thaliana aquaporin knock-out lines

Stress granules (SGs) are dynamic RNA–protein complexes localized in the cytoplasm that rapidly form under stress conditions and disperse when normal conditions are restored. The formation of SGs depends on the Ras-GAP SH3 domain-binding protein (G3BP). Formations, interactions and functions of plant and human SGs are strikingly similar, suggesting a conserved mechanism. However, functional analyses of plant G3BPs are missing. Thus, members of the Arabidopsis thaliana G3BP (AtG3BP) protein family were investigated in a complementation assay in a human G3BP knock-out cell line. It was shown that two out of seven AtG3BPs were able to complement the function of their human homolog. GFP-AtG3BP fusion proteins co-localized with human SG marker proteins Caprin-1 and eIF4G1 and restored SG formation in G3BP double KO cells. Interaction between AtG3BP-1 and -7 and known human G3BP interaction partners such as Caprin-1 and USP10 was also demonstrated by co-immunoprecipitation. In addition, an RG/RGG domain exchange from Arabidopsis G3BP into the human G3BP background showed the ability for complementation. In summary, our results support a conserved mechanism of SG function over the kingdoms, which will help to further elucidate the biological function of the Arabidopsis G3BP protein family.

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Acclimation of mesophyll conductance and anatomy to light during leaf ageing in Arabidopsis thaliana

Physiologia Plantarum ◽

10.1111/ppl.13398 ◽

2021 ◽

Author(s):

Marc Carriquí ◽

Miquel Nadal ◽

Jaume Flexas

Keyword(s):

Arabidopsis Thaliana ◽

Mesophyll Conductance ◽

Leaf Ageing

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Ca2+-Dependent Protein Kinase 6 Enhances KAT2 Shaker Channel Activity in Arabidopsis thaliana

International Journal of Molecular Sciences ◽

10.3390/ijms22041596 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1596

Author(s):

Elsa Ronzier ◽

Claire Corratgé-Faillie ◽

Frédéric Sanchez ◽

Christian Brière ◽

Tou Cheu Xiong

Keyword(s):

Arabidopsis Thaliana ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

Physical Interaction ◽

Fluorescence Lifetime Imaging ◽

Dependent Manner ◽

In Planta ◽

Knock Out ◽

Calcium Dependent ◽

Dependent Protein

Post-translational regulations of Shaker-like voltage-gated K+ channels were reported to be essential for rapid responses to environmental stresses in plants. In particular, it has been shown that calcium-dependent protein kinases (CPKs) regulate Shaker channels in plants. Here, the focus was on KAT2, a Shaker channel cloned in the model plant Arabidopsis thaliana, where is it expressed namely in the vascular tissues of leaves. After co-expression of KAT2 with AtCPK6 in Xenopuslaevis oocytes, voltage-clamp recordings demonstrated that AtCPK6 stimulates the activity of KAT2 in a calcium-dependent manner. A physical interaction between these two proteins has also been shown by Förster resonance energy transfer by fluorescence lifetime imaging (FRET-FLIM). Peptide array assays support that AtCPK6 phosphorylates KAT2 at several positions, also in a calcium-dependent manner. Finally, K+ fluorescence imaging in planta suggests that K+ distribution is impaired in kat2 knock-out mutant leaves. We propose that the AtCPK6/KAT2 couple plays a role in the homeostasis of K+ distribution in leaves.

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The UV-B photoreceptor UVR8 promotes photosynthetic efficiency in Arabidopsis thaliana exposed to elevated levels of UV-B

Photosynthesis Research ◽

10.1007/s11120-012-9785-y ◽

2012 ◽

Vol 114 (2) ◽

pp. 121-131 ◽

Cited By ~ 36

Author(s):

Matthew P. Davey ◽

Novita I. Susanti ◽

Jason J. Wargent ◽

Jane E. Findlay ◽

W. Paul Quick ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Photosynthetic Efficiency

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A dual sgRNA approach for functional genomics in Arabidopsis thaliana[OPEN]

10.1101/172676 ◽

2017 ◽

Author(s):

Laurens Pauwels ◽

Rebecca De Clercq ◽

Jonas Goossens ◽

Sabrina Iñigo ◽

Clara Williams ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Gene Editing ◽

Reverse Genetics ◽

Gene Families ◽

Null Alleles ◽

Loss Of Function ◽

Knock Out ◽

Nonsense Mediated Decay ◽

Dna Insertion ◽

Entire Gene

AbstractReverse genetics uses loss-of-function alleles to interrogate gene function. The advent of CRISPR/Cas9-based gene editing now allows to generate knock-out alleles for any gene and entire gene families. Even in the model plant Arabidopsis thaliana, gene editing is welcomed as T-DNA insertion lines do not always generate null alleles. Here, we show efficient generation of heritable mutations in Arabidopsis using CRISPR/Cas9 with a workload similar to generating overexpression lines. We obtain Cas9 null-segregants with bi-allelic mutations in the T2 generation. Out of seven new mutant alleles we report here, one allele for GRXS17, the ortholog of human GRX3/PICOT, did not phenocopy previously characterized nulls. Notwithstanding, the mutation caused a frameshift and triggered nonsense-mediated decay. We demonstrate that our workflow is also compatible with a dual sgRNA approach in which a gene is targeted by two sgRNAs simultaneously. This paired nuclease method can result in a more reliable loss-of-function alleles that lack a large essential part of the gene. The ease in the CRISPR/Cas9 workflow should help in the eventual generation of true null alleles of every gene in the Arabidopsis genome, which will advance both basic and applied plant research.One-sentence summaryWe present a dual sgRNA approach to delete Arabidopsis gene 34 fragments in order to obtain reliable functional knock-outs.

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Evaluating Population Genomic Candidate Genes Underlying Flowering Time in Arabidopsis thaliana Using T-DNA Insertion Lines

Journal of Heredity ◽

10.1093/jhered/esz026 ◽

2019 ◽

Vol 110 (4) ◽

pp. 445-454 ◽

Cited By ~ 2

Author(s):

Veronica K Chong ◽

John R Stinchcombe

Keyword(s):

Arabidopsis Thaliana ◽

Flowering Time ◽

Candidate Genes ◽

Insertional Mutagenesis ◽

Knock Out ◽

Population Genomic ◽

Outlier Loci ◽

Genomic Studies ◽

Dna Insertion ◽

Genomic Scans

Abstract Population genomic scans have emerged as a powerful tool to detect regions of the genome that are potential targets of selection. Despite the success of genomic scans in identifying novel lists of loci potentially underlying adaptation, few studies proceed to validate the function of these candidate genes. In this study, we used transfer-DNA (T-DNA) insertion lines to evaluate the effects of 27 candidate genes on flowering time in North American accessions of Arabidopsis thaliana. We compared the flowering time of T-DNA insertion lines that knock out the function of a candidate gene obtained from population genomic studies to a wild type under long- and short-day conditions. We also did the same for a collection of randomly chosen genes that had not been identified as candidates. We validated the well-known effect of long-day conditions in accelerating flowering time and found that gene disruption caused by insertional mutagenesis tends to delay flowering. Surprisingly, we found that knockouts in random genes were just as likely to produce significant phenotypic effects as knockouts in candidate genes. T-DNA insertions at a handful of candidate genes that had previously been identified as outlier loci showed significant delays in flowering time under both long and short days, suggesting that they are promising candidates for future investigation.

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Leaf Age-Dependent Effects of Foliar-Sprayed CuZn Nanoparticles on Photosynthetic Efficiency and ROS Generation in Arabidopsis thaliana

Materials ◽

10.3390/ma12152498 ◽

2019 ◽

Vol 12 (15) ◽

pp. 2498 ◽

Cited By ~ 6

Author(s):

Sperdouli ◽

Moustaka ◽

Antonoglou ◽

Adamakis ◽

Dendrinou-Samara ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Photosynthetic Efficiency ◽

Leaf Age ◽

Foliar Spray ◽

Ros Generation ◽

H2o2 Production ◽

Mature Leaves ◽

Transmission Electron ◽

Psii Photochemistry ◽

Young Leaves

Young and mature leaves of Arabidopsis thaliana were exposed by foliar spray to 30 mg L−1 of CuZn nanoparticles (NPs). The NPs were synthesized by a microwave-assisted polyol process and characterized by dynamic light scattering (DLS), X-ray diffraction (XRD), and transmission electron microscopy (TEM). CuZn NPs effects in Arabidopsis leaves were evaluated by chlorophyll fluorescence imaging analysis that revealed spatiotemporal heterogeneity of the quantum efficiency of PSII photochemistry (ΦPSΙΙ) and the redox state of the plastoquinone (PQ) pool (qp), measured 30 min, 90 min, 180 min, and 240 min after spraying. Photosystem II (PSII) function in young leaves was observed to be negatively influenced, especially 30 min after spraying, at which point increased H2O2 generation was correlated to the lower oxidized state of the PQ pool.. Recovery of young leaves photosynthetic efficiency appeared only after 240 min of NPs spray when also the level of ROS accumulation was similar to control leaves. On the contrary, a beneficial effect on PSII function in mature leaves after 30 min of the CuZn NPs spray was observed, with increased ΦPSΙΙ, an increased electron transport rate (ETR), decreased singlet oxygen (1O2) formation, and H2O2 production at the same level of control leaves.An explanation for this differential response is suggested.

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