Manual Isolation of Living Cells from the Arabidopsis thaliana Female Gametophyte by Micromanipulation

2017 ◽  
pp. 221-234 ◽  
Author(s):  
Maria Englhart ◽  
Lucija Šoljić ◽  
Stefanie Sprunck
Keyword(s):  
Arabidopsis Thaliana ◽  
Female Gametophyte ◽  
Living Cells

scholarly journals Characterization of Suspension Cultures of Arabidopsis thaliana (L.) Heynh Cells with Altered Expression of the Gene of Alternative Mitochondrial Oxidase Aox1a and Analysis of their Frost Resistance

2021 ◽  
Vol 35 ◽  
pp. 3-18
Author(s):  
A. V. Stepanov ◽  
◽  
S. A. Kashin ◽  
N. S. Zabanova ◽  
O. A. Fedotova ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Suspension Culture ◽  
Mitochondrial Respiration ◽  
Frost Resistance ◽  
Alternative Oxidase ◽  
Uncoupling Protein ◽  
Living Cells ◽  
Subzero Temperature ◽  
Wild Type ◽  
Culture Cells

The enzyme alternative cyanide-resistant oxidase (AOX) localized in mitochondria is involved in the processes of plant adaptation to various unfavorable biotic and abiotic factors. Transfer of electrons from ubiquinone to oxygen by alternative oxidase has a nonprotonmotive character and, by bypassing two sites of H+ pumping in complexes III and IV, lowers the energy efficiency of respiration and energy of electron flow through AOX is released as heat. In this work, we characterized heterotrophic suspension cultures of Arabidopsis thaliana (L.) Heynh cells obtained from seeds of plants with altered (reduced (AS-12 line) and increased (XX-2 line)) expression of the alternative oxidase gene AOX1a and studied their viability under subzero temperature (-10 °С for 3, 6, 9 hours). Cell viability and reactive oxygen species (ROS) production were assessed using fluorescence microscopy with fluorescein diacetate (FDA) and propidium iodide (PI) for cell viability measurement and H2DCF-DA for ROS measurement. The proportion of living cells was calculated as the proportion of FDApositive and PI-negative cells. Differences between the studied lines were determined in the content of mitochondrial proteins of the respiratory chain (AOX, COXII, NDB) and uncoupling protein (UCP), as well as in the intensity of formation of ROS and frost resistance. The obtained results confirmed the higher content of the AOX protein and its high contribution to mitochondrial respiration in line XX-2. Suspension culture cells of the AS-12 line showed a decrease in the AOX protein content and its contribution to mitochondrial respiration, compared to the wild type (Col-0) and line XX-2. Simultaneously with a decrease in the AOX protein content in the AS-12 cell culture, an increase in the content of the uncoupling protein UCP and subunit II of cytochrome oxidase (COXII) was observed. ROS generation was reduced in cell cultures of both XX-2 and AS-12. The obtained results indicate that the cells of the wildtype (Col-0) suspension culture were subjected to the most significant effect of subzero temperature. Long-term exposure (for 9 h) under -10 °С revealed significant differences in the viability of wild-type culture cells and lines with altered AOX1a gene expression. Cells of line XX-2 with an increased content of AOX turned out to be more resistant to subzero temperature compared to wild-type and AS-12 cells. However, while the proportion of living cells in the culture of the AS-12 line 48 h after exposure remained at the same level as immediately after it, in the suspension culture of the wild type cell death developed over time. The obtained results indicate the importance of alternative oxidase in the development of frost resistance in plant cell.

scholarly journals Molecular movement in the Arabidopsis thaliana female gametophyte

2017 ◽  
Vol 30 (3) ◽  
pp. 141-146 ◽  
Author(s):  
Robert M. Erdmann ◽  
Anja Hoffmann ◽  
Heidi-Kristin Walter ◽  
Hans-Achim Wagenknecht ◽  
Rita Groß-Hardt ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Female Gametophyte ◽  
Molecular Movement

scholarly journals The MIDASIN and NOTCHLESS genes are essential for female gametophyte development in Arabidopsis thaliana

2010 ◽  
Vol 16 (1) ◽  
pp. 3-18 ◽  
Author(s):  
Sier-Ching Chantha ◽  
Madoka Gray-Mitsumune ◽  
Josée Houde ◽  
Daniel P. Matton
Keyword(s):  
Arabidopsis Thaliana ◽  
Female Gametophyte ◽  
Gametophyte Development ◽  
Female Gametophyte Development

scholarly journals Protein Interaction Network of Arabidopsis thaliana Female Gametophyte Development Identifies Novel Proteins and Relations

PLoS ONE ◽  
2012 ◽  
Vol 7 (12) ◽  
pp. e49931 ◽  
Author(s):  
Batool Hosseinpour ◽  
Vahid HajiHoseini ◽  
Rafieh Kashfi ◽  
Esmaeil Ebrahimie ◽  
Farhid Hemmatzadeh
Keyword(s):  
Arabidopsis Thaliana ◽  
Protein Interaction ◽  
Protein Interaction Network ◽  
Female Gametophyte ◽  
Interaction Network ◽  
Gametophyte Development ◽  
Novel Proteins ◽  
Female Gametophyte Development

scholarly journals Identification and characterization of a female gametophyte defect in sdk1-7 +/- abi3-6 +/- heterozygotes of Arabidopsis thaliana

2020 ◽  
Vol 15 (8) ◽  
pp. 1780038
Author(s):  
Subramanian Sankaranarayanan ◽  
Muhammad Jamshed ◽  
Frédéric Delmas ◽  
Edward C. Yeung ◽  
Marcus A. Samuel
Keyword(s):  
Arabidopsis Thaliana ◽  
Female Gametophyte ◽  
Identification And Characterization

scholarly journals Functional Analysis of Sporophytic Transcripts Repressed by the Female Gametophyte in the Ovule of Arabidopsis thaliana

PLoS ONE ◽  
2013 ◽  
Vol 8 (10) ◽  
pp. e76977 ◽  
Author(s):  
Alma Armenta-Medina ◽  
Wilson Huanca-Mamani ◽  
Nidia Sanchez-León ◽  
Isaac Rodríguez-Arévalo ◽  
Jean-Philippe Vielle-Calzada
Keyword(s):  
Functional Analysis ◽  
Arabidopsis Thaliana ◽  
Female Gametophyte

scholarly journals RanGAP is required for post-meiotic mitosis in female gametophyte development in Arabidopsis thaliana

2011 ◽  
Vol 62 (8) ◽  
pp. 2705-2714 ◽  
Author(s):  
Thushani Rodrigo-Peiris ◽  
Xianfeng Morgan Xu ◽  
Qiao Zhao ◽  
Horng-Jing Wang ◽  
Iris Meier
Keyword(s):  
Arabidopsis Thaliana ◽  
Female Gametophyte ◽  
Gametophyte Development ◽  
Female Gametophyte Development

scholarly journals Coalescence and directed anisotropic growth of starch granule initials in subdomains of Arabidopsis thaliana chloroplasts

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Léo Bürgy ◽  
Simona Eicke ◽  
Christophe Kopp ◽  
Camilla Jenny ◽  
Kuan Jen Lu ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Enzyme Activities ◽  
Starch Granule ◽  
Imaging Techniques ◽  
Thylakoid Membranes ◽  
Living Cells ◽  
Anisotropic Growth ◽  
Starch Granules ◽  
Glucose Storage ◽  
Granule Surface

AbstractLiving cells orchestrate enzyme activities to produce myriads of biopolymers but cell-biological understanding of such processes is scarce. Starch, a plant biopolymer forming discrete, semi-crystalline granules within plastids, plays a central role in glucose storage, which is fundamental to life. Combining complementary imaging techniques and Arabidopsis genetics we reveal that, in chloroplasts, multiple starch granules initiate in stromal pockets between thylakoid membranes. These initials coalesce, then grow anisotropically to form lenticular granules. The major starch polymer, amylopectin, is synthesized at the granule surface, while the minor amylose component is deposited internally. The non-enzymatic domain of STARCH SYNTHASE 4, which controls the protein’s localization, is required for anisotropic growth. These results present us with a conceptual framework for understanding the biosynthesis of this key nutrient.

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