Advances and applications of single‐cell omics technologies in plant research

Mo, Yajin; Jiao, Yuling

doi:10.1111/tpj.15772

Cited by 41 publications

(19 citation statements)

References 163 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To further clarify sperm cell identification of cdka;1 SLCs at the molecular level, we adopted single‐cell RNA‐sequencing technology (Tang et al, 2009; Ryu et al, 2021; Mo and Jiao, 2022). We manually separated and collected 10 individual cdka;1 SLCs and 17 wild‐type sperm cells that were released from semi‐ in vivo grown pollen tubes.…”

Section: Resultsmentioning

confidence: 99%

The egg cell is preferentially fertilized in Arabidopsis double fertilization

Hou

Wang

et al. 2022

JIPB

View full text Add to dashboard Cite

In flowering plants (angiosperms), fertilization of the egg cell by one sperm cell produces an embryo, whereas fusion of a second sperm cell with the central cell generates the endosperm. In most angiosperms like Arabidopsis, a pollen grain contains two isomorphic sperm cells required for this double fertilization process. A long‐standing unsolved question is whether the two fertilization events have any preference. A tool to address this question is the usage of the cyclin‐dependent kinase a1 (cdka;1) mutant pollen, which produces a single sperm‐like cell (SLC). Here, we first adopt a complementation‐based fluorescence‐labeling method to successfully separate and collect cdka;1 mutant pollen containing a single SLC. Single‐cell RNA‐sequencing analysis revealed that cdka;1 SLCs show a gene expression profile highly similar to that of sperm cells and not to the generative cell, precursor of the two sperm cells. Pollination assays using a limited number of cdka;1 mutant pollen revealed that in 98.2% of the ovules, single fertilization of the egg cell occurred. Pollination of pistils with excessive cdka;1 mutant pollen allowed the delivery of a second SLC via fertilization recovery, which fertilized the central cell, resulting in 20.7% double‐fertilized ovules. This indicates that cdka;1 SLCs are able to fertilize both the egg and the central cell. Taken together, our findings have answered a long‐standing question and support that preferential fertilization of the egg cell is evident in Arabidopsis.

show abstract

Section: Resultsmentioning

confidence: 99%

The egg cell is preferentially fertilized in Arabidopsis double fertilization

Hou

Wang

et al. 2022

JIPB

View full text Add to dashboard Cite

show abstract

“…Technological breakthroughs in single-cell transcriptomic approaches have aided investigations of expression dynamics and biological processes at the cellular level (Mo & Jiao, 2022). In the study of plant molecular biology processes, the establishment of a suitable protocol for protoplast transformation will not only allow a detailed analysis of early signs of protoplast regeneration (e.g.…”

Section: Discussionmentioning

confidence: 99%

An Integrated Physiological, Cytology and Proteomics Reveals Network of Sugarcane Protoplasts Responses to Enzymolysis

Zhang

Wang

Xiao

et al. 2022

Preprint

View full text Add to dashboard Cite

The protoplast experimental system has been becoming a powerful tool for functional genomics and cell fusion breeding. However, the physiology and molecular mechanism during enzymolysis is not completely understood and has become a major obstacle to protoplast regeneration. Our study used physiological, cytology, iTRAQ (Isobaric Tags for Relative and Absolute Quantification) -based proteomic and RT-PCR analyses to compare the young leaves of sugarcane (ROC22) and protoplasts of more than 90% viability. We found that oxidation product MDA content increased in the protoplasts after enzymolysis and several antioxidant enzymes such as POD, CAT, APX, and O2- content significantly decreased. The cytology results showed that after enzymolysis, the cell membranes were perforated to different degrees, the nuclear activity was weakened, the nucleolus structure was not obvious, and the microtubules depolymerized and formed many short rod-like structures in protoplasts. The proteomic results showed that 1,477 differential proteins were down-regulated and 810 were up-regulated after enzymolysis of sugarcane young leaves. The GO terms, KEGG and KOG enrichment analysis revealed that differentially abundant proteins were mainly involved in bioenergetic metabolism, cellular processes, osmotic stress, and redox homeostasis of protoplasts, which would allow protein biosynthesis or / degradation. The RT-PCR analysis revealed the expression of osmotic stress resistance genes such as DREB, WRKY, MAPK4, and NAC were up-regulated. Meanwhile, the expression of key regeneration genes such as CyclinD3, CyclinA, CyclinB, Cdc2, PSK, CESA and GAUT were significantly down-regulated in the protoplasts. Hierarchical clustering, identification of redox proteins and oxidation products showed that these proteins were involved in dynamic networks in response to oxidative stress after enzymolysis. We used a variety of methods to figure out how young sugarcane leaves react to enzymes.

show abstract

“…Furthermore, recent successes in rapid screening CRISPR/Cas9 constructs for gene edition after protoplast transformation show another application for protoplasts in synthetic biology ( Lin et al, 2018 ; Toda et al, 2019 ; Rather et al, 2022 ). Additionally, with the recent appearance of practical methods for single cell–based -omics research using protoplasts ( Chen et al, 2021; Farmer et al, 2021; Liu et al, 2021; Seyfferth et al, 2021; Liu et al, 2022; Mo and Jiao, 2022 ), the potential applications are continuing to grow in advanced research fields.…”

Section: Introductionmentioning

confidence: 99%

Synthetic Promoter Screening Using Poplar Mesophyll Protoplast Transformation

Yang¹,

Shao²,

Chaffin³

et al. 2023

BIO-PROTOCOL

View full text Add to dashboard Cite

Plant protoplasts are useful to study both transcriptional regulation and protein subcellular localization in rapid screens. Protoplast transformation can be used in automated platforms for design-build-test cycles of plant promoters, including synthetic promoters. A notable application of protoplasts comes from recent successes in dissecting synthetic promoter activity with poplar mesophyll protoplasts. For this purpose, we constructed plasmids with TurboGFP driven by a synthetic promoter together with TurboRFP constitutively controlled by a 35S promoter, to monitor transformation efficiency, allowing versatile screening of high numbers of cells by monitoring green fluorescent protein expression in transformed protoplasts. Herein, we introduce a protocol for poplar mesophyll protoplast isolation followed by protoplast transformation and image analysis for the selection of valuable synthetic promoters. Graphical overview

show abstract

Advances and applications of single‐cell omics technologies in plant research

Cited by 41 publications

References 163 publications

The egg cell is preferentially fertilized in Arabidopsis double fertilization

The egg cell is preferentially fertilized in Arabidopsis double fertilization

An Integrated Physiological, Cytology and Proteomics Reveals Network of Sugarcane Protoplasts Responses to Enzymolysis

Synthetic Promoter Screening Using Poplar Mesophyll Protoplast Transformation

Contact Info

Product

Resources

About