A New Dynamin-Like Protein, ADL6, Is Involved in Trafficking from the trans-Golgi Network to the Central Vacuole in Arabidopsis

Jin, Jing Bo; Kim, Young A; Kim, Soo Jin; Lee, Sung Hoon; Kim, Dae Heon; Cheong, Gang-Won; Hwang, Inhwan

doi:10.2307/3871383

Cited by 79 publications

(142 citation statements)

References 51 publications

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“…Interesting regulatory roles of dynamin-like ADL6 (ref. 21), small GTP binding protein Rha1 (ref. 22) and actin filaments 23 have been studied in vacuolar trafficking.…”

Section: Applicationsmentioning

confidence: 99%

Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis

2007

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The transient gene expression system using Arabidopsis mesophyll protoplasts has proven an important and versatile tool for conducting cell-based experiments using molecular, cellular, biochemical, genetic, genomic and proteomic approaches to analyze the functions of diverse signaling pathways and cellular machineries. A well-established protocol that has been extensively tested and applied in numerous experiments is presented here. The method includes protoplast isolation, PEG-calcium transfection of plasmid DNA and protoplast culture. Physiological responses and high-throughput capability enable facile and cost-effective explorations as well as hypothesis-driven tests. The protoplast isolation and DNA transfection procedures take 6-8 h, and the results can be obtained in 2-24 h. The cell system offers reliable guidelines for further comprehensive analysis of complex regulatory mechanisms in whole-plant physiology, immunity, growth and development. INTRODUCTIONPlant protoplasts without cell walls offer a versatile cell-based experimental system. Macromolecules such as DNA, RNA and proteins can be delivered into protoplasts using various methods, e.g., PEG-calcium fusion, electroporation and microinjection. Signal transduction and metabolic pathways as well as transcription and translation machineries can be transiently manipulated to investigate cell-autonomous regulation and responses. The method is especially powerful in functional genomics analysis when combined with rich genetic resources, such as insertion libraries, available in the model plant Arabidopsis thaliana. We describe here a protocol for a system of transient expression in Arabidopsis mesophyll protoplast (TEAMP) and provide troubleshooting advice and detailed information on useful reporter constructs. Applications, advantages and limitations of the TEAMP assay are also discussed. More general information about plant protoplasts can be found in a review 1 and a live performance of the TEAMP procedure can be seen at the Sheen lab website (http://genetics. mgh.harvard.edu/sheenweb/protocols_reg.html).

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“…Interesting regulatory roles of dynamin-like ADL6 (ref. 21), small GTP binding protein Rha1 (ref. 22) and actin filaments 23 have been studied in vacuolar trafficking.…”

Section: Applicationsmentioning

confidence: 99%

Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis

2007

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“…Transient transformation of A. thaliana protoplasts was done as described (Jin et al, 2001) using the following modification. Instead of 4% and 1%, respectively, we used only 1% w/v cellulase (Serva) and 0.25% w/v macerozyme (Yakult Honsha Co.).…”

Section: Gfp-constructs and Transient Transformation Of A Thaliana Pmentioning

confidence: 99%

AtToc90, a New GTP-Binding Component of the Arabidopsis Chloroplast Protein Import Machinery

et al. 2004

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AtToc159 is a GTP-binding chloroplast protein import receptor. In vivo, atToc159 is required for massive accumulation of photosynthetic proteins during chloroplast biogenesis. Yet, in mutants lacking atToc159 photosynthetic proteins still accumulate, but at strongly reduced levels whereas non-photosynthetic proteins are imported normally: This suggests a role for the homologues of atToc159 (atToc132, -120 and -90). Here, we show that atToc90 supports accumulation of photosynthetic proteins in plastids, but is not required for import of several constitutive proteins. Part of atToc90 associates with the chloroplast surface in vivo and with the Toc-complex core components (atToc75 and atToc33) in vitro suggesting a function in chloroplast protein import similar to that of atToc159. As both proteins specifically contribute to the accumulation of photosynthetic proteins in chloroplasts they may be components of the same import pathway.

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“…The localization was examined by fluorescence microscopy (Jin et al, 2001). Images were processed using Adobe Photoshop and presented in pseudocolor.…”

Section: Qrt-reverse Transcription-pcrmentioning

confidence: 99%

“…To generate the UGT71B6:GFP, UGT71B7:GFP, UGT71B8:GFP, and UGT73B1: GFP constructs, the complementary DNAs (cDNAs) of UGT71B6, UGT71B7, UGT71B8, and UGT73B1 were amplified from a Col-0 flower cDNA library by PCR using the gene-specific primers 71B6-5 and 71B6-3, 71B7-5 and 71B7-3, 71B8-5 and 71B8-3, and 73B1-5 and 73B1-3, respectively (for nucleotide sequences of the primers, see Supplemental Table S1), and fused to GFP of the 326-GFP vector driven by the 35S promoter (Jin et al, 2001). These constructs were transferred to the binary vector pCsV1300 (Invitrogen) containing the strong cassava vein mosaic virus promoter.…”

Section: Construction Of Plasmidsmentioning

confidence: 99%

“…A suitable amount of plasmids expressing effectors, reporter, and normalizer were cotransformed into protoplasts derived from wild-type plants using the polyethylene glycol transformation method (Jin et al, 2001) and incubated for 20 h at room temperature, and total RNA was extracted from the transformed protoplasts. The transcript levels of LUC were detected by qRT-PCR.…”

Section: Reporter Assaymentioning

confidence: 99%

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Abscisic Acid Uridine Diphosphate Glucosyltransferases Play a Crucial Role in Abscisic Acid Homeostasis in Arabidopsis

Park

Kim

et al. 2014

Plant Physiol.

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The phytohormone abscisic acid (ABA) is crucial for plant growth and adaptive responses to various stress conditions. Plants continuously adjust the ABA level to meet physiological needs, but how ABA homeostasis occurs is not fully understood. This study provides evidence that UGT71B6, an ABA uridine diphosphate glucosyltransferase (UGT), and its two closely related homologs, UGT71B7 and UGT71B8, play crucial roles in ABA homeostasis and in adaptation to dehydration, osmotic stress, and high-salinity stresses in Arabidopsis (Arabidopsis thaliana). UGT RNA interference plants that had low levels of these three UGT transcripts displayed hypersensitivity to exogenous ABA and high-salt conditions during germination and exhibited a defect in plant growth. However, the ectopic expression of UGT71B6 in the atbg1 (for b-glucosidase) mutant background aggravated the ABA-deficient phenotype of atbg1 mutant plants. In addition, modulation of the expression of the three UGTs affects the expression of CYP707A1 to CYP707A4, which encode ABA 89-hydroxylases; four CYP707As were expressed at higher levels in the UGT RNA interference plants but at lower levels in the UGT71B6:GFP-overexpressing plants. Based on these data, this study proposes that UGT71B6 and its two homologs play a critical role in ABA homeostasis by converting active ABA to an inactive form (abscisic acid-glucose ester) depending on intrinsic cellular and environmental conditions in plants.

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A New Dynamin-Like Protein, ADL6, Is Involved in Trafficking from the trans-Golgi Network to the Central Vacuole in Arabidopsis

Cited by 79 publications

References 51 publications

Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis

Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis

AtToc90, a New GTP-Binding Component of the Arabidopsis Chloroplast Protein Import Machinery

Abscisic Acid Uridine Diphosphate Glucosyltransferases Play a Crucial Role in Abscisic Acid Homeostasis in Arabidopsis

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