Measuring Cell Ploidy Level in Arabidopsis thaliana by Flow Cytometry

Yang, Leiyun; Wang, Zhixue; Hua, Jian

doi:10.1007/978-1-4939-9458-8_11

Cited by 10 publications

(6 citation statements)

References 9 publications

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“…Flow cytometry. Nuclei were isolated as previously described with minor changes 78 . Briefly, the first pairs of true leaves of 21-day-old soil-grown plants were chopped with a razor blade in precooled buffer (10 mM MgSO 4 , 50 mM KCl, 5 mM HEPES, 10 mM DTT, 2.5% (v/v) Triton X-100) to release nuclei, and the homogenates passed through a 30 μm CellTrics filter (Sysmex Partec, Münster, DE) to remove debris.…”

Section: Methodsmentioning

confidence: 99%

Translational fidelity and growth of Arabidopsis require stress-sensitive diphthamide biosynthesis

et al. 2022

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Diphthamide, a post-translationally modified histidine residue of eukaryotic TRANSLATION ELONGATION FACTOR2 (eEF2), is the human host cell-sensitizing target of diphtheria toxin. Diphthamide biosynthesis depends on the 4Fe-4S-cluster protein Dph1 catalyzing the first committed step, as well as Dph2 to Dph7, in yeast and mammals. Here we show that diphthamide modification of eEF2 is conserved in Arabidopsis thaliana and requires AtDPH1. Ribosomal −1 frameshifting-error rates are increased in Arabidopsis dph1 mutants, similar to yeast and mice. Compared to the wild type, shorter roots and smaller rosettes of dph1 mutants result from fewer formed cells. TARGET OF RAPAMYCIN (TOR) kinase activity is attenuated, and autophagy is activated, in dph1 mutants. Under abiotic stress diphthamide-unmodified eEF2 accumulates in wild-type seedlings, most strongly upon heavy metal excess, which is conserved in human cells. In summary, our results suggest that diphthamide contributes to the functionality of the translational machinery monitored by plants to regulate growth.

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Section: Methodsmentioning

confidence: 99%

Translational fidelity and growth of Arabidopsis require stress-sensitive diphthamide biosynthesis

et al. 2022

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“…For each experiment, more than 5 000 nuclei were analyzed. The endoreduplication index was calculated as follows: (% 2 C nuclei

\times

1) + (% 4 C nuclei

\times

2) + (% 8 C nuclei

\times

3) + (% 16 C nuclei

\times

4) + (% 32 C nuclei

\times

5) (Yang et al, 2019).…”

Section: Methodsmentioning

confidence: 99%

“…For each experiment, more than 5 000 nuclei were analyzed. The endoreduplication index was calculated as follows: (Yang et al, 2019).…”

Section: Determination Of Cell Polyploidy Levelsmentioning

confidence: 99%

ERF4 interacts with and antagonizes TCP15 in regulating endoreduplication and cell growth in Arabidopsis

Ding

Xie³

et al. 2022

JIPB

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Endoreduplication is prevalent during plant growth and development, and is often correlated with large cell and organ size. Despite its prevalence, the transcriptional regulatory mechanisms underlying the transition from mitotic cell division to endoreduplication remain elusive. Here, we characterize ETHYLENE-RESPONSIVE ELEMENT BINDING FACTOR 4 (ERF4) as a positive regulator of endoreduplication through its function as a transcriptional repressor. ERF4 was specifically expressed in mature tissues in which the cells were undergoing expansion, but was rarely expressed in young organs. Plants overexpressing ERF4 exhibited much larger

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“…Ploidy analysis method was adapted from Yang et al (2019). Sixteen carpels for each genotype were dissected from stage 14 flowers at inflorescence positions 1 and 2.…”

Section: Ploidy Analysismentioning

confidence: 99%

Histone Demethylases ELF6 and JMJ13 Antagonistically Regulate Self-Fertility in Arabidopsis

et al. 2021

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The chromatin modification H3K27me3 is involved in almost every developmental stage in Arabidopsis. Much remains unknown about the dynamic regulation of this histone modification in flower development and control of self-fertility. Here we demonstrate that the H3K27me3-specific demethylases ELF6 and JMJ13 antagonistically regulate carpel and stamen growth and thus modulate self-fertility. Transcriptome and epigenome data are used to identify potential targets of ELF6 and JMJ13 responsible for these physiological functions. We find that ELF6 relieves expansin genes of epigenetic silencing to promote cell elongation in the carpel, enhancing carpel growth and therefore encouraging out-crossing. On the other hand, JMJ13 activates genes of the jasmonic acid regulatory network alongside the auxin responsive SAUR26, to inhibit carpel growth, enhance stamen growth, and overall promote self-pollination. Our evidence provides novel mechanisms of self-fertility regulation in A. thaliana demonstrating how chromatin modifying enzymes govern the equilibrium between flower self-pollination and out-crossing.

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Measuring Cell Ploidy Level in Arabidopsis thaliana by Flow Cytometry

Cited by 10 publications

References 9 publications

Translational fidelity and growth of Arabidopsis require stress-sensitive diphthamide biosynthesis

Translational fidelity and growth of Arabidopsis require stress-sensitive diphthamide biosynthesis

ERF4 interacts with and antagonizes TCP15 in regulating endoreduplication and cell growth in Arabidopsis

Histone Demethylases ELF6 and JMJ13 Antagonistically Regulate Self-Fertility in Arabidopsis

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