Waiting
Elaborazione accesso...

Trial ends in Request Full Access Tell Your Colleague About Jove
Click here for the English version

Biology

可视化Stromule频率与荧光显微镜

Published: November 23, 2016 doi: 10.3791/54692

Materials

Name Company Catalog Number Comments
Hepes Sigma-Aldrich H3375
NaOH Fischer-Scientific S320-1
Sorbitol Sigma-Aldrich S1876
EDTA Fischer-Biotech BP121
MnCl2 Sigma-Aldrich 221279
MgCl2 Sigma-Aldrich M0250
KCl Sigma-Aldrich P3911
NaCl Sigma-Aldrich S9625
Laser Scanning Confocal Microscope  Carl Zeiss Inc Model: LSM710
Carboxyfluorescein diacetate (CFDA) Sigma-Aldrich 21879 
Dimethyl sulfoxide (DMSO) EMD MX1458-6
Waring blender Waring  Model: 31BL92
Fiji fiji.sc Open-source software for analyzing biological images

DOWNLOAD MATERIALS LIST

References

  1. Koussevitzky, S., et al. Signals from chloroplasts converge to regulate nuclear gene expression. Science. 316 (5825), 715-719 (2007).
  2. Burch-Smith, T. M., Brunkard, J. O., Choi, Y. G., Zambryski, P. C. PNAS Plus: Organelle-nucleus cross-talk regulates plant intercellular communication via plasmodesmata. Proc. Natl. Acad. Sci. USA. 108 (51), E1451-E1460 (2011).
  3. Stonebloom, S., Brunkard, J. O., Cheung, A. C., Jiang, K., Feldman, L., Zambryski, P. Redox states of plastids and mitochondria differentially regulate intercellular transport via plasmodesmata. Plant Physiol. 158 (1), 190-199 (2012).
  4. Nomura, H., et al. Chloroplast-mediated activation of plant immune signalling in Arabidopsis. Nat. Commun. 3, 926 (2012).
  5. Avendaño-Vázquez, A. -O., et al. An uncharacterized apocarotenoid-derived signal generated in ζ-carotene desaturase mutants regulates leaf development and the expression of chloroplast and nuclear genes in Arabidopsis. Plant Cell. 26 (June), 2524-2537 (2014).
  6. Caplan, J. L., et al. Chloroplast stromules runction during innate immunity. Dev. Cell. 34 (1), 45-57 (2015).
  7. Hanson, M. R., Sattarzadeh, A. Stromules: recent insights into a long neglected feature of plastid morphology and function. Plant Physiol. 155 (4), 1486-1492 (2011).
  8. Gray, J. C., et al. Plastid stromules are induced by stress treatments acting through abscisic acid. Plant J. 69 (3), 387-398 (2012).
  9. Brunkard, J., Runkel, A., Zambryski, P. Chloroplasts extend stromules independently and in response to internal redox signals. Proc. Natl. Acad. Sci. USA. 112 (32), 10044-10049 (2015).
  10. Dupree, P., Pwee, K. -H., Gray, J. C. Expression of photosynthesis gene-promoter fusions in leaf epidermal cells of transgenic tobacco plants. Plant J. 1 (1), 115-120 (1991).
  11. Charuvi, D., Kiss, V., Nevo, R., Shimoni, E., Adam, Z., Reich, Z. Gain and loss of photosynthetic membranes during plastid differentiation in the shoot apex of Arabidopsis. Plant Cell. 24 (3), 1143-1157 (2012).
  12. Chiang, Y. -H., et al. Functional characterization of the GATA transcription factors GNC and CGA1 reveals their key role in chloroplast development, growth, and division in Arabidopsis. Plant Physiol. 160 (1), 332-348 (2012).
  13. Schattat, M., Barton, K., Baudisch, B., Klösgen, R. B., Mathur, J. Plastid stromule branching coincides with contiguous endoplasmic reticulum dynamics. Plant Physiol. 155 (4), 1667-1677 (2011).
  14. Erickson, J. L., et al. Agrobacterium-derived cytokinin influences plastid morphology and starch accumulation in Nicotiana benthamiana during transient assays. BMC Plant Biol. 14 (1), 127 (2014).
  15. Ho, J., Theg, S. M. The formation of stromules in vitro from chloroplasts isolated from Nicotiana benthamiana. PLoS One. 11 (2), e0146489 (2016).
  16. Brunkard, J. O., Burch-Smith, T. M., Runkel, A. M., Zambryski, P. Investigating plasmodesmata genetics with virus-induced gene silencing and an Agrobacterium-mediated GFP movement assay. Method. Mol. Biol. , 185-198 (2015).
  17. Schattat, M. H., Klösgen, R. B. Induction of stromule formation by extracellular sucrose and glucose in epidermal leaf tissue of Arabidopsis thaliana. BMC Plant Biol. 11, 115 (2011).
  18. Joly, D., Carpentier, R. Rapid isolation of intact chloroplasts from spinach leaves. Method. Mol. Biol. 684, 321-325 (2011).
  19. Waters, M. T., Fray, R. G., Pyke, K. A. Stromule formation is dependent upon plastid size, plastid differentiation status and the density of plastids within the cell. Plant J. 39 (4), 655-667 (2004).
可视化Stromule频率与荧光显微镜
Play Video
PDF DOI DOWNLOAD MATERIALS LIST

Cite this Article

Brunkard, J. O., Runkel, A. M.,More

Brunkard, J. O., Runkel, A. M., Zambryski, P. Visualizing Stromule Frequency with Fluorescence Microscopy. J. Vis. Exp. (117), e54692, doi:10.3791/54692 (2016).

Less
Copy Citation Download Citation Reprints and Permissions
View Video

Get cutting-edge science videos from JoVE sent straight to your inbox every month.

Waiting X
Simple Hit Counter