Gene Expression Pattern and Protein Localization of Arabidopsis Phospholipase D Alpha 1 Revealed by Advanced Light-Sheet and Super-Resolution Microscopy

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F18%3A73589790" target="_blank" >RIV/61989592:15310/18:73589790 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.frontiersin.org/articles/10.3389/fpls.2018.00371/full" target="_blank" >https://www.frontiersin.org/articles/10.3389/fpls.2018.00371/full</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3389/fpls.2018.00371" target="_blank" >10.3389/fpls.2018.00371</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Gene Expression Pattern and Protein Localization of Arabidopsis Phospholipase D Alpha 1 Revealed by Advanced Light-Sheet and Super-Resolution Microscopy

Popis výsledku v původním jazyce

Phospholipase D alpha 1 (PLD alpha 1, At3g15730) and its product phosphatidic acid (PA) are involved in a variety of cellular and physiological processes, such as cytoskeletal remodeling, regulation of stomatal closure and opening, as well as biotic and abiotic stress signaling. Here we aimed to study developmental expression patterns and subcellular localization of PLD alpha 1 in Arabidopsis using advanced microscopy methods such as light-sheet fluorescencemicroscopy (LSFM) and structured illumination microscopy (SIM). We complemented two knockout pld alpha 1 mutants with a YFP-tagged PLD alpha 1 expressed under the PLD alpha 1 native promoter in order to study developmental expression pattern and subcellular localization of PLD alpha 1 in Arabidopsis thaliana under natural conditions. Imaging of tissue-specific and developmentally-regulated localization of YFP-tagged PLD alpha 1 by LSFM in roots of growing seedlings showed accumulation of PLD alpha 1-YFP in the root cap and the rhizodermis. Expression of PLD alpha 1-YFP in the rhizodermis was considerably higher in trichoblasts before and during root hair formation and growth. Thus, PLD alpha 1-YFP accumulated in emerging root hairs and in the tips of growing root hairs. PLD alpha 1-YFP showed cytoplasmic subcellular localization in root cap cells and in cells of the root transition zone. In aerial parts of plants PLD alpha 1-YFP was also localized in the cytoplasm showing enhanced accumulation in the cortical cytoplasmic layer of epidermal non-dividing cells of hypocotyls, leaves, and leaf petioles. However, in dividing cells of root apical meristem and leaf petiole epidermis PLD alpha 1-YFP was enriched in mitotic spindles and phragmoplasts, as revealed by co-visualization with microtubules. Finally, super-resolution SIM imaging revealed association of PLD alpha 1-YFP with both microtubules and clathrin-coated vesicles (CCVs) and pits (CCPs). In conclusion, this study shows the developmentally-controlled expression and subcellular localization of PLD alpha 1 in dividing and non-dividing Arabidopsis cells.

Název v anglickém jazyce

Gene Expression Pattern and Protein Localization of Arabidopsis Phospholipase D Alpha 1 Revealed by Advanced Light-Sheet and Super-Resolution Microscopy

Popis výsledku anglicky

Phospholipase D alpha 1 (PLD alpha 1, At3g15730) and its product phosphatidic acid (PA) are involved in a variety of cellular and physiological processes, such as cytoskeletal remodeling, regulation of stomatal closure and opening, as well as biotic and abiotic stress signaling. Here we aimed to study developmental expression patterns and subcellular localization of PLD alpha 1 in Arabidopsis using advanced microscopy methods such as light-sheet fluorescencemicroscopy (LSFM) and structured illumination microscopy (SIM). We complemented two knockout pld alpha 1 mutants with a YFP-tagged PLD alpha 1 expressed under the PLD alpha 1 native promoter in order to study developmental expression pattern and subcellular localization of PLD alpha 1 in Arabidopsis thaliana under natural conditions. Imaging of tissue-specific and developmentally-regulated localization of YFP-tagged PLD alpha 1 by LSFM in roots of growing seedlings showed accumulation of PLD alpha 1-YFP in the root cap and the rhizodermis. Expression of PLD alpha 1-YFP in the rhizodermis was considerably higher in trichoblasts before and during root hair formation and growth. Thus, PLD alpha 1-YFP accumulated in emerging root hairs and in the tips of growing root hairs. PLD alpha 1-YFP showed cytoplasmic subcellular localization in root cap cells and in cells of the root transition zone. In aerial parts of plants PLD alpha 1-YFP was also localized in the cytoplasm showing enhanced accumulation in the cortical cytoplasmic layer of epidermal non-dividing cells of hypocotyls, leaves, and leaf petioles. However, in dividing cells of root apical meristem and leaf petiole epidermis PLD alpha 1-YFP was enriched in mitotic spindles and phragmoplasts, as revealed by co-visualization with microtubules. Finally, super-resolution SIM imaging revealed association of PLD alpha 1-YFP with both microtubules and clathrin-coated vesicles (CCVs) and pits (CCPs). In conclusion, this study shows the developmentally-controlled expression and subcellular localization of PLD alpha 1 in dividing and non-dividing Arabidopsis cells.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10605 - Developmental biology

Projekt

<a href="/cs/project/GA16-22044S" target="_blank" >GA16-22044S: Funkční regulace fosfolipasy D alfa 1 prostřednictvím MPK3-závislé fosforylace</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Frontiers in Plant Science

ISSN

1664-462X

e-ISSN

—

Svazek periodika

9

Číslo periodika v rámci svazku

MAR

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

21

Strana od-do

"371-1"-"371-21"

Kód UT WoS článku

000427928800001

EID výsledku v databázi Scopus

2-s2.0-85045444953