Abstract
Metabolic pathway reconstruction and gene edits for native natural product synthesis in single plant cells are considered to be less complicated when compared to the production of non-native metabolites. Being an efficient eukaryotic system, plants encompass suitable post-translational modifications. However, slow cell division rate and heterogeneous nature is an impediment for consistent product retrieval from plant cells. Plant cell synchrony can be attained in cultures developed in vitro. Isolated plant protoplasts capable of division, can potentially enhance the unimpaired yield of target bioactives, similar to microbes and unicellular eukaryotes. Evidence from yeast experiments suggests that ‘critical cell size’ and division rates for enhancement machinery, primarily depend on culture conditions and nutrient availability. The cell size control mechanisms in Arabidopsis shoot apical meristem is analogous to yeast notably, fission yeast. If protoplasts isolated from plants are subjected to cell size studies and cell cycle progression in culture, it will answer the underlying molecular mechanisms such as, unicellular to multicellular transition states, longevity, senescence, ‘cell-size resetting’ during organogenesis, and adaptation to external cues.
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Abbreviations
- CDKB:
-
Cyclin-dependent kinase B
- CRISPR-Cas9:
-
Clustered regularly interspaced short palindromic repeats-associated protein 9
- HDR:
-
Homology-directed repair
- PEPC:
-
Phosphoenolpuruvate carboxykinase
- PVP:
-
Poly Vinyl pyrrolidone
- RNP:
-
Ribonucleoprotein
- ROS:
-
Reactive oxygen species
- RuBisCO:
-
Ribulose bisphosphate carboxylase oxygenase
- SAM:
-
Shoot apical meristem
- sgRNAs:
-
Single guide RNAs
- TOR:
-
Target of rapamycin
- vir :
-
Virulence gene of tumour-inducing plasmid
References
Adedeji OS, Naing AH, Kim CK (2020) Protoplast isolation and shoot regeneration from protoplast-derived calli of Chrysanthemum cv. White ND. Plant Cell Tissue Organ Cult 141:571–581. https://doi.org/10.1007/s11240-020-01816-3
Asfaw KG, Liu Q, Xu X et al (2020) A mitochondria-targeted coenzyme Q peptoid induces superoxide dismutase and alleviates salinity stress in plant cells. Sci Rep 10:11563. https://doi.org/10.1038/s41598-020-68491-4
Barber F, Amir A, Murray AW (2020) Cell size regulation in budding yeast does not depend on linear accumulation of Whi5. bioRxiv. https://doi.org/10.1101/2020.01.20.912832
Bertoni G (2020) Phosphorus sensing by LST8 acts as a TOR guide for cell growth in Chlamydomonas. Plant Cell 32:7. https://doi.org/10.1105/tpc.19.00888
Chebli Y, Geitmann A (2011) Gravity research on plants: use of single-cell experimental models. Front Plant Sci 2:56. https://doi.org/10.3389/fpls.2011.00056
Chen K, Wang Y, Zhang R et al (2019) CRISPR/Cas genome editing and precision plant breeding in agriculture. Annu Rev Plant Biol 70:667–697. https://doi.org/10.1146/annurev-arplant-050718-100049
Christey MC (2004) Brassica protoplast culture and somatic hybridization. Biotechnology in agriculture and forestry. Springer, Berlin, pp 119–148
Constabel F, Kurz WGW, Chatson B, Gamborg OL (1974) Induction of partial synchrony in soybean cell cultures. Exp Cell Res 85:105–110. https://doi.org/10.1016/0014-4827(74)90218-3
Davey MR, Anthony P, Power JB, Lowe KC (2005) Plant protoplasts: status and biotechnological perspectives. Biotechnol Adv 23:131–171
Davis HR, Maddison AL, Phillips DW, Jones HD (2020) Genetic transformation of protoplasts isolated from leaves of Lolium temulentum and Lolium perenne. Methods in molecular biology. Humana Press Inc., Totowa, pp 199–205
Du J, Liao W, Liu W, Bissonnette M, He C, Li YC (2020) N6 -Adenosine methylation of Socs1 mRNA is required to sustain the negative feedback control of macrophage activation. Dev Cell. https://doi.org/10.1016/j.devcel.2020.10.023
Ermakova M, Danila FR, Furbank RT, von Caemmerer S (2020) On the road to C4 rice: advances and perspectives. Plant J 101:940–950. https://doi.org/10.1111/tpj.14562
Fan Y, Xin S, Dai X et al (2020) Efficient genome editing of rubber tree (Hevea brasiliensis) protoplasts using CRISPR/Cas9 ribonucleoproteins. Ind Crops Prod 146:112146. https://doi.org/10.1016/j.indcrop.2020.112146
Fowke, Fowke LC (2018) Protoplasts for studies of cell organelles. Plant protoplasts. CRC Press, Boca Raton, pp 39–52
Fowke LC, Griffing LR, Mersey BG, Valk P (1983) Protoplasts for studies of the plasma membrane and associated cell organelles. Protoplasts 1983. Birkhäuser, Basel, pp 101–110
Gao J, Wang G, Ma S et al (2015) CRISPR/Cas9-mediated targeted mutagenesis in Nicotiana tabacum. Plant Mol Biol 87:99–110. https://doi.org/10.1007/s11103-014-0263-0
García-Gómez ML, Ornelas-Ayala D, Garay-Arroyo A et al (2020) A system-level mechanistic explanation for asymmetric stem cell fates: Arabidopsis thaliana root niche as a study system. Sci Rep 10:1–16. https://doi.org/10.1038/s41598-020-60251-8
González A, Hall MN, Lin SC, Hardie DG (2020) AMPK and TOR: the Yin and Yang of cellular nutrient sensing and growth control. Cell Metab 31:472–492. https://doi.org/10.1016/j.cmet.2020.01.015
Gupta S, Gadgil VN (1972) Effect of light on growth and mitotic periodicity in tissue cultures. Indian J Exp Biol 10:62–64
Heldt FS, Tyson JJ, Cross FR, Novák B (2020) A single light-responsive sizer can control multiple-fission cycles in Chlamydomonas. Curr Biol 30:634-644.e7. https://doi.org/10.1016/j.cub.2019.12.026
Hlavová M, Vítová M, Bišová K (2016) Synchronization of green algae by light and dark regimes for cell cycle and cell division studies. Methods in molecular biology. Humana Press Inc., Totowa, pp 3–16
Ho P-Y, Lin J, Amir A (2018) Modeling cell size regulation: from single-cell-level statistics to molecular mechanisms and population-level effects. Annu Rev Biophys 47:251–271. https://doi.org/10.1146/annurev-biophys-070317-032955
John F, Roffler S, Wicker T, Ringli C (2011) Plant tor signaling components. Plant Signal Behav 6:1700–1705. https://doi.org/10.4161/psb.6.11.17662
Jones AMP, Chattopadhyay A, Shukla M et al (2012) Inhibition of phenylpropanoid biosynthesis increases cell wall digestibility, protoplast isolation, and facilitates sustained cell division in American elm (Ulmus americana). BMC Plant Biol 12:75. https://doi.org/10.1186/1471-2229-12-75
Jones AR, Forero-Vargas M, Withers SP et al (2017) Cell-size dependent progression of the cell cycle creates homeostasis and flexibility of plant cell size. Nat Commun 8:15060. https://doi.org/10.1038/ncomms15060
Kim H, Kim ST, Ryu J et al (2017) CRISPR/Cpf1-mediated DNA-free plant genome editing. Nat Commun 8:14406. https://doi.org/10.1038/ncomms14406
King PJ, Mansfield KJ, Street HE (1973) Control of growth and metabolism of cultured plant cells. Can J Bot 51:1807
Kozgunova E, Yoshida MW, Goshima G (2020) Spindle position dictates division site during asymmetric cell division in moss. bioRxiv. https://doi.org/10.1101/2020.03.03.975557
Lee ME, Rusin SF, Jenkins N et al (2018) Mechanisms connecting the conserved protein kinases Ssp1, Kin1, and Pom1 in fission yeast cell polarity and division. Curr Biol 28:84-92.e4. https://doi.org/10.1016/j.cub.2017.11.034
Lin CS, Hsu CT, Yang LH et al (2018) Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration. Plant Biotechnol J 16:1295–1310. https://doi.org/10.1111/pbi.12870
Liu S, Lin YH, Murphy A et al (2020) Mapping reaction-diffusion networks at the plant wound site with pathogens. Front Plant Sci 11:1074. https://doi.org/10.3389/fpls.2020.01074
Mahmoud LM, Grosser JW, Dutt M (2020) Silver compounds regulate leaf drop and improve in vitro regeneration from mature tissues of Australian finger lime (Citrus australasica). Plant Cell Tissue Organ Cult 141:455–464. https://doi.org/10.1007/s11240-020-01803-8
Muranaka T, Oyama T (2020) Application of single-cell bioluminescent imaging to monitor circadian rhythms of individual plant cells. Methods in molecular biology. Humana Press Inc., Totowa, pp 231–242
Mustafa NS, Taha RA, Hassan SAM et al (2013) Overcoming phenolic accumulation of date palm in vitro culture using α-tocopherol and cold pre-treatment. Middle East J Sci Res 15:344–350. https://doi.org/10.5829/idosi.mejsr.2013.15.3.11080
Orr RG, Cheng X, Vidali L, Bezanilla M (2020) Orchestrating cell morphology from the inside out—using polarized cell expansion in plants as a model. Curr Opin Cell Biol 62:46–53. https://doi.org/10.1016/j.ceb.2019.08.004
Paksa A, Bandemer J, Hoeckendorf B et al (2016) Repulsive cues combined with physical barriers and cell-cell adhesion determine progenitor cell positioning during organogenesis. Nat Commun. https://doi.org/10.1038/ncomms11288
Pasternak T, Lystvan K, Betekhtin A, Hasterok R (2020) From single cell to plants: mesophyll protoplasts as a versatile system for investigating plant cell reprogramming. Int J Mol Sci 21:1–15
Saravana Kumar RM, Wang Y, Zhang X et al (2020) Redox components: key regulators of epigenetic modifications in plants. Int J Mol Sci 21. https://www.mdpi.com/1422-0067/21/4/1419
Satterlee JW, Strable J, Scanlon MJ (2020) Plant stem cell organization and differentiation at single-cell resolution 2 Plants maintain populations of pluripotent stem cells in shoot apical meristems (SAMs). bioRxiv. https://doi.org/10.1101/2020.08.25.267427
Schaeffer SM, Nakata PA (2015) CRISPR/Cas9-mediated genome editing and gene replacement in plants: transitioning from lab to field. Plant Sci 240:130–142
Si X, Zhang H, Wang Y et al (2020) Manipulating gene translation in plants by CRISPR–Cas9-mediated genome editing of upstream open reading frames. Nat Protoc 15:338–363. https://doi.org/10.1038/s41596-019-0238-3
Simmonds DH (1992) Plant cell wall removal: cause for microtubule instability and division abnormalities in protoplast cultures? Physiol Plant 85:387–390. https://doi.org/10.1034/j.1399-3054.1992.850237.x
Simpson KJ, Bennett C, Atkinson RRL et al (2020) C4 photosynthesis and the economic spectra of leaf and root traits independently influence growth rates in grasses. J Ecol. https://doi.org/10.1111/1365-2745.13412
Slovak R, Setzer C, Roiuk M et al (2020) Ribosome assembly factor Adenylate Kinase 6 maintains cell proliferation and cell size homeostasis during root growth. New Phytol 225:2064–2076. https://doi.org/10.1111/nph.16291
Son O, Kim S, Kim D et al (2018) Involvement of TOR signaling motif in the regulation of plant autophagy. Biochem Biophys Res Commun 501:643–647. https://doi.org/10.1016/J.BBRC.2018.05.027
Suzuki K, Itoh T, Sasamoto H (1998) Cell wall architecture prerequisite for the cell division in the protoplasts of white poplar, Populus alba L. Plant Cell Physiol 39:632–638. https://doi.org/10.1093/oxfordjournals.pcp.a029415
Thomas A, Pujari I, Shetty V et al (2017) Dendrobium protoplast co-culture promotes phytochemical assemblage in vitro. Protoplasma 254:1517–1528. https://doi.org/10.1007/s00709-016-1043-2
van Ark HF, Hall RD, Creemers-Molenaar J, Krens FA (1992) High yields of cytoplasts from protoplasts of Lolium perenne and Beta vulgaris using gradient centrifugation. Plant Cell Tissue Organ Cult 31:223–232. https://doi.org/10.1007/BF00036228
van Es SW, Silveira SR, Rocha DI et al (2018) Novel functions of the Arabidopsis transcription factor TCP5 in petal development and ethylene biosynthesis. Plant J 94:867–879. https://doi.org/10.1111/tpj.13904
Wang Q, Yu G, Chen Z et al (2020) Optimization of protoplast isolation, transformation and its application in sugarcane (Saccharum spontaneum L.). Crop J. https://doi.org/10.1016/j.cj.2020.05.006
Willis L, Refahi Y, Wightman R et al (2016) Cell size and growth regulation in the Arabidopsis thaliana apical stem cell niche. Proc Natl Acad Sci 113:E8238–E8246. https://doi.org/10.1073/pnas.1616768113
Winnicki K (2020) The winner takes it all: auxin-the main player during plant embryogenesis. Cells 9:606. https://doi.org/10.3390/cells9030606
Woo JW, Kim J, Il KS et al (2015) DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins. Nat Biotechnol 33:1162–1164. https://doi.org/10.1038/nbt.3389
Wu Q, Xu F, Jackson D (2018) All together now, a magical mystery tour of the maize shoot meristem. Curr Opin Plant Biol 45:26–35. https://doi.org/10.1016/j.pbi.2018.04.010
Xiao L, Zhang L, Yang G et al (2012) Transcriptome of protoplasts reprogrammed into stem cells in Physcomitrella patens. PLoS ONE 7:e35961. https://doi.org/10.1371/journal.pone.0035961
Yadav V, Wang Z, Wei C et al (2020) Phenylpropanoid pathway engineering: An emerging approach towards plant defense. Pathogens 9:312
Zhang Y, Liang Z, Zong Y et al (2016) Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA. Nat Commun 7:12617. https://doi.org/10.1038/ncomms12617
Zhang Y, Ran Y, Nagy I et al (2020) Targeted mutagenesis in Ryegrass (Lolium spp.) using the CRISPR/Cas9 system. Plant Biotechnol J. https://doi.org/10.1111/pbi.13359
Zhu M, Roeder AH (2020) Plants are better engineers: the complexity of plant organ morphogenesis. Curr Opin Genet Dev 63:16–23. https://doi.org/10.1016/j.gde.2020.02.008
Acknowledgements
The authors thank the Science and Engineering Research Board—Extra Mural Research (SERB-EMR) (presently called Core Research Grant [CRG]), Government of India, File No. EMR/2015/001816 for funding the research project. IP thank Manipal Academy of Higher Education, Manipal, India, for providing the prestigious Dr. T. M. A. Pai Ph.D. Scholarship. IP acknowledges SERB for granting the Junior Research Fellowship since May 2018.
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VSB conceived the idea. VSB designed the illustration. IP, AT and VSB wrote the manuscript. PSR and KS reviewed and gave suggestions to improve the document. All the authors read and approved the manuscript.
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IP, AT, PSR, KS and VSB declare that they have no conflicts of interest relevant to this article.
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Pujari, I., Thomas, A., Rai, P.S. et al. Cell size: a key determinant of meristematic potential in plant protoplasts. aBIOTECH 2, 96–104 (2021). https://doi.org/10.1007/s42994-020-00033-y
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DOI: https://doi.org/10.1007/s42994-020-00033-y