The Cell Wall Regeneration of Tobacco Protoplasts Based on Microfluidic System

The cell wall, serving as the exoskeleton of plants, is naturally a barrier to resist external stresses. Protoplasts can be obtained by dissolving the cell walls of plant cells without damaging the cell membrane, and are widely used in the rapid propagation, transgenic breeding, and somatic hybridization of plants. However, to regenerate the cell wall is a precondition for cell division. Therefore, to study the culture condition and influencing factors during the cell wall regeneration of protoplasts is vital. Traditionally, culture medium is used to cultivate protoplasts, but it has some disadvantages. Herein, a microfluidic system with crossed channels was constructed to isolate and cultivate the protoplasts of tobacco. Then, the cell wall regeneration of the tobacco protoplasts was also studied based on this microfluidic system. It was found that, compared with the control, benzo-(1, 2, 3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH) could accelerate the regeneration of the cell wall, while Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) could inhibit the regeneration of the cell wall within 24 h. To conclude, this study demonstrated that a crossed microfluidic chip could be an effective tool to study cell wall regeneration or other behavior of plant cells in situ with high resolution. In addition, this study revealed the rate of cell wall regeneration under BTH and Pst DC3000 treatment.

[1]  Zhenbiao Yang,et al.  Mechano-transduction via the pectin-FERONIA complex activates ROP6 GTPase signaling in Arabidopsis pavement cell morphogenesis , 2021, Current Biology.

[2]  Hai Liu,et al.  Cellulose synthase‐like protein OsCSLD4 plays an important role in the response of rice to salt stress by mediating abscisic acid biosynthesis to regulate osmotic stress tolerance , 2021, Plant biotechnology journal.

[3]  A. Geitmann,et al.  Microfluidics-Based Bioassays and Imaging of Plant Cells , 2021, Plant & cell physiology.

[4]  Hang Gao,et al.  Signals in systemic acquired resistance of plants against microbial pathogens , 2021, Molecular Biology Reports.

[5]  Yihua Zhou,et al.  The Plant Cell Wall: Biosynthesis, construction, and functions. , 2020, Journal of integrative plant biology.

[6]  Haibing Yang,et al.  Microfluidic devices for monitoring the root morphology of Arabidopsis thaliana in situ. , 2020, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[7]  B. Cooper,et al.  Benzothiadiazole conditions the bean proteome for immunity to bean rust. , 2020, Molecular plant-microbe interactions : MPMI.

[8]  Lingling Wu,et al.  Microfluidic Single-Cell Omics Analysis. , 2020, Small.

[9]  L. Vaahtera,et al.  Cell wall integrity maintenance during plant development and interaction with the environment , 2019, Nature Plants.

[10]  H. Kang,et al.  Single-Cell RNA Sequencing Resolves Molecular Relationships Among Individual Plant Cells1[OPEN] , 2019, Plant Physiology.

[11]  G. Spangenberg,et al.  Mesophyll Protoplasts and PEG-Mediated Transfections: Transient Assays and Generation of Stable Transgenic Canola Plants. , 2018, Methods in molecular biology.

[12]  P. Kachroo,et al.  Signaling mechanisms underlying systemic acquired resistance to microbial pathogens. , 2019, Plant science : an international journal of experimental plant biology.

[13]  Yolanda Ruiz-García,et al.  Preharvest Application of Elicitors to Monastrell Grapes: Impact on Wine Polysaccharide and Oligosaccharide Composition. , 2018, Journal of agricultural and food chemistry.

[14]  A. Wiszniewska,et al.  Studies on Cell Wall Regeneration in Protoplast Culture of Legumes - the Effect of Organic Medium Additives on Cell Wall Components , 2018 .

[15]  A. Molina,et al.  Plant cell wall-mediated immunity: cell wall changes trigger disease resistance responses. , 2018, The Plant journal : for cell and molecular biology.

[16]  A. Lee,et al.  High-throughput microfluidic single-cell trapping arrays for biomolecular and imaging analysis. , 2018, Methods in cell biology.

[17]  Christian R. Boehm,et al.  Droplet-based microfluidic analysis and screening of single plant cells , 2017, bioRxiv.

[18]  X. Wang,et al.  IDL6‐HAE/HSL2 impacts pectin degradation and resistance to Pseudomonas syringae pv tomato DC3000 in Arabidopsis leaves , 2017, The Plant journal : for cell and molecular biology.

[19]  E. Prats,et al.  BTH and BABA induce resistance in pea against rust (Uromyces pisi) involving differential phytoalexin accumulation , 2015, Planta.

[20]  Y. Bi,et al.  BTH treatment caused physiological, biochemical and proteomic changes of muskmelon (Cucumis melo L.) fruit during ripening. , 2015, Journal of proteomics.

[21]  A. Sanati Nezhad.  Microfluidic platforms for plant cells studies. , 2014, Lab on a chip.

[22]  S. He,et al.  Pseudomonas syringae pv. tomato DC3000: a model pathogen for probing disease susceptibility and hormone signaling in plants. , 2013, Annual review of phytopathology.

[23]  De-quan Li,et al.  Response of tobacco to the Pseudomonas syringae pv. Tomato DC3000 is mainly dependent on salicylic acid signaling pathway. , 2013, FEMS microbiology letters.

[24]  M. Packirisamy,et al.  TipChip: a modular, MEMS-based platform for experimentation and phenotyping of tip-growing cells. , 2013, The Plant journal : for cell and molecular biology.

[25]  S. Quake,et al.  The RootChip: An Integrated Microfluidic Chip for Plant Science[W][OA] , 2011, Plant Cell.

[26]  M. Iriti,et al.  Benzothiadiazole (BTH) activates sterol pathway and affects vitamin D3 metabolism in Solanum malacoxylon cell cultures , 2011, Plant Cell Reports.

[27]  Wenming Liu,et al.  Culture and chemical-induced fusion of tobacco mesophyll protoplasts in a microfluidic device , 2011 .

[28]  C. Goulet,et al.  On-chip detection of low-molecular-weight recombinant proteins in plant crude extracts by SELDI-TOF MS. , 2009, Methods in molecular biology.

[29]  A. Furini,et al.  Plant regeneration from leaf protoplasts of Solanum virginianum L. (Solanaceae) , 2007, Plant Cell, Tissue and Organ Culture.

[30]  Rustem F. Ismagilov,et al.  Dynamics of Drosophila embryonic patterning network perturbed in space and time using microfluidics , 2005, Nature.

[31]  Shaukat Ali,et al.  Somatic cybridization between Nicotiana tabacum and N. repanda based on a single inactivation procedure of nuclear donor parental protoplasts , 2005 .

[32]  J. Ryals,et al.  Benzothiadiazole induces disease resistance in Arabidopsis by activation of the systemic acquired resistance signal transduction pathway. , 1996, The Plant journal : for cell and molecular biology.