Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants

Sparkes, Imogen; Runions, John; Kearns, Anne; Hawes, Chris

doi:10.1038/nprot.2006.286

Cited by 1,577 publications

(1,092 citation statements)

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“…Transient expression in N. benthamiana was performed as described previously [39]. Agrobacterium strains (GV3101) harboring above-described constructs were cultured in liquid Luria-Bertani medium overnight.…”

Section: Transient Expression In Nicotiana Benthamianamentioning

confidence: 99%

Signature motif-guided identification of receptors for peptide hormones essential for root meristem growth

et al. 2016

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Peptide-mediated cell-to-cell signaling has crucial roles in coordination and definition of cellular functions in plants. Peptide-receptor matching is important for understanding the mechanisms underlying peptide-mediated signaling. Here we report the structure-guided identification of root meristem growth factor (RGF) receptors important for plant development. An assay based on a signature ligand recognition motif (Arg-x-Arg) conserved in a subfamily of leucine-rich repeat receptor kinases (LRR-RKs) identified the functionally uncharacterized LRR-RK At4g26540 as a receptor of RGF1 (RGFR1). We further solved the crystal structure of RGF1 in complex with the LRR domain of RGFR1 at a resolution of 2.6 Å, which reveals that the Arg-x-Gly-Gly (RxGG) motif is responsible for specific recognition of the sulfate group of RGF1 by RGFR1. Based on the RxGG motif, we identified additional four RGFRs. Participation of the five RGFRs in RGF-induced signaling is supported by biochemical and genetic data. We also offer evidence showing that SERKs function as co-receptors for RGFs. Taken together, our study identifies RGF receptors and co-receptors that can link RGF signals with their downstream components and provides a proof of principle for structure-based matching of LRR-RKs with their peptide ligands.

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Section: Transient Expression In Nicotiana Benthamianamentioning

confidence: 99%

Signature motif-guided identification of receptors for peptide hormones essential for root meristem growth

et al. 2016

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“…Fluorescent protein fusion constructs were transformed into tobacco (Nicotiana tabacum) leaves using agroinfiltration with a variety of A. tumefaciens cell densities and the method described by Sparkes et al (2006). Microscopy using system 1 was performed 3 to 4 d after infiltration.…”

Section: Transient Transformation Of Tobacco Leaf Cellsmentioning

confidence: 99%

FRIENDLY Regulates Mitochondrial Distribution, Fusion, and Quality Control in Arabidopsis

et al. 2014

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Mitochondria are defining components of most eukaryotes. However, higher plant mitochondria differ biochemically, morphologically, and dynamically from those in other eukaryotes. FRIENDLY, a member of the CLUSTERED MITOCHONDRIA superfamily, is conserved among eukaryotes and is required for correct distribution of mitochondria within the cell. We sought to understand how disruption of FRIENDLY function in Arabidopsis (Arabidopsis thaliana) leads to mitochondrial clustering and the effects of this aberrant chondriome on cell and whole-plant physiology. We present evidence for a role of FRIENDLY in mediating intermitochondrial association, which is a necessary prelude to mitochondrial fusion. We demonstrate that disruption of mitochondrial association, motility, and chondriome structure in friendly affects mitochondrial quality control and leads to mitochondrial stress, cell death, and strong growth phenotypes.

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“…The fusion construct and control were transformed into onion epidermis cells by particle bombardment using PDS-1000/He (BIO-RAD) [38]. This construct was also introduced into Agrobacterium tumefaciens GV3101 and then infiltrated into the leaves of 3-week-old Nicotiana benthamiana plants [39,40]. GFP was detected by a confocal laser scanning microscopy (Leica TCS SP5) 24 h after transformation of onion epidermis cells and 48 h after infiltration of N. benthamiana leaves.…”

Section: Subcellular Localization Of Dep2mentioning

confidence: 99%

Rice DENSE AND ERECT PANICLE 2 is essential for determining panicle outgrowth and elongation

et al. 2010

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The architecture of the panicle, including grain size and panicle morphology, directly determines grain yield. Panicle erectness, which is selected for achieving ideal plant architecture in the northern part of China, has drawn increasing attention of rice breeders. Here, dense and erect panicle 2 (dep2) mutant, which shows a dense and erect panicle phenotype, was identified. DEP2 encodes a plant-specific protein without any known functional domain. Expression profiling of DEP2 revealed that it is highly expressed in young tissues, with most abundance in young panicles. Morphological and expression analysis indicated that mutation in DEP2 mainly affects the rapid elongation of rachis and primary and secondary branches, but does not impair the initiation or formation of panicle primordia. Further analysis suggests that decrease of panicle length in dep2 is caused by a defect in cell proliferation during the exponential elongation of panicle. Despite a more compact plant type in the dep2 mutant, no significant alteration in grain production was found between wild type and dep2 mutant. Therefore, the study of DEP2 not only strengthens our understanding of the molecular genetic basis of panicle architecture but also has important implications for rice breeding.

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Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants

Cited by 1,577 publications

References 20 publications

Signature motif-guided identification of receptors for peptide hormones essential for root meristem growth

Signature motif-guided identification of receptors for peptide hormones essential for root meristem growth

FRIENDLY Regulates Mitochondrial Distribution, Fusion, and Quality Control in Arabidopsis

Rice DENSE AND ERECT PANICLE 2 is essential for determining panicle outgrowth and elongation

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