Comparison of Microtubule Organization in Arabidopsis thaliana TUBGFP and MBD-GFP Mutants Exposed to UV-B Radiation

Abstract

Microtubule organization was compared between the Arabidopsis thaliana TUB-GFP and MBD-GFP mutants. Plant height and primary root length were measured, and microtubule dynamics were examined by confocal laser scanning microscopy after UV-B radiation to reveal changes in microtubules. Damage caused by UV-B was comparable between transgenic lines and wild-type plants, although transgenic lines were more sensitive to UV-B than the wild-type. Spots and depolymerization of microtubules were detected in both TUB-GFP and MBD-GFP plants; however, MBD-GFP showed better adaptation of changes induced by UV-B treatment. These results indicated that UV-B inhibits the growth and development of transgenic lines, and the inhibitory effects might result from changes in microtubules, as determined by comparison between the TUB-GFP and MBD-GFP lines. 

Keywords

• Microtubules; UV-B radiation; Arabidopsis thaliana TUB-GFP; Arabidopsis thaliana MBD-GFP

References

1. Bisgrove S R (2008). The roles of microtubules in tropisms. Plant Science 175(6): 747-755
2. Chen H & Han R (2015). F-actin participates in the process of the “partition-bundle division”. Russian Journal of Plant Physiology 62(2): 187-194
3. Cleary A L & Hardham A R (1993). Pressure induced reorientation of cortical microtubules in epidermal cells of Lolium rigidum leaves. Plant and Cell Physiology 34(7): 1003-1008
4. Eisinger W, Ehrhardt D & Briggs W (2012). Microtubules are essential for guard-cell function in Vicia and Arabidopsis. Molecular Plant 5(3): 601-610
5. Granger C L & Cyr R J (2000). Microtubule reorganization in tobacco BY-2 cells stably expressing GFP-MBD. Planta 210: 502-509
6. Granger C L & Cyr R J (2001). Spatiotemporal relationships between growth and microtubule orientation as revealed in living root cells of Arabidopsis thaliana transformed with green-fluorescent-protein gene construct GFP-MBD. Protoplasma 216(3-4): 201-214
7. Guo A H, Gao L M, Li Y F & Han R (2010). Influence on microtubule in wheat mesophyll cell exposed to enhanced ultraviolet-B radiation and He-Ne laser irradiation. Guihaia 30(2): 250-255
8. Han R, Wang X, Yue M & Qi Z (2002). Effects of the enhanced UV-B radiation on the body cell mitosis of the wheat. Acta Genetica Sinica 29(6): 537-541

9. Hardham A R, Takemoto D & White R G (2008). Rapid and dynamic subcellular reorganization following mechanical stimulation of Arabidopsis epidermal cells mimics responses to fungal and oomycete attack. BMC Plant Biology 8(63): 1-14
10. Hectors K, Jacques E, Prinsen E, Guisez Y, Verbelen J P, Jansen M A & Vissenberg K (2010). UV radiation reduces epidermal cell expansion in leaves of Arabidopsis thaliana. Journal of Experimental Botany 61(15): 4339-4349
11. Hong Y Y, Zhang W H & Wang X M (2010). Phospholipase D and phosphatidic acid signalling in plant response to drought and salinity. Plant, Cell & Environment 33(4): 627-635
12. Iwata K, Tazawa M & Itoh T (2001). Turgor pressure regulation and the orientation of cortical microtubules in Spirogyra cells. Plant and Cell Physiology 42(6): 594-598
13. Jansen M A (2002). Ultraviolet-B radiation effects on plants: İnduction of morphogenic responses. Physiologia Plantarum 116(3): 423-429
14. Kawamura E & Wasteneys G O (2008). MOR1, the Arabidopsis thaliana homologue of Xenopus MAP215, promotes rapid growth and shrinkage, and suppresses the pausing of microtubules in vivo. Journal of Cell Science 121: 4114-4123
15. Krasylenko Y A, Yemets A I, Sheremet Y A & Blume Y B (2012). Nitric oxide as a critical factor for perception of UV-B irradiation by microtubules in Arabidopsis. Physiologia Plantarum 145(4): 505-515
16. Krasylenko Y A, Yemets A I & Blume Y B (2013). Plant microtubules reorganization under the indirect UV-B exposure and during UV-B-induced programmed cell death. Plant Signaling & Behavior 8(5): e24031, doi: 10.4161/psb.24031
17. Liu J Y, Wang B C, Zhang Y G, Wang Y C, Kong J, Zhu L Q, Yang X Y & Zha G D (2014). Microtubule dynamics is required for root elongation growth under osmotic stress in Arabidopsis. Plant Growth Regulation 74: 187-192
18. Lucas J R, Nadeau J A & Sack F D (2006). Microtubule arrays and Arabidopsis stomatal development. Journal of Experimental Botany 57(1): 71-79
19. Marc J, Granger C L, Brincat J, Fisher D D, Kao T, McCubbin A G & Cyr R J (1998). A GFP-MAP4 reporter gene for visualizing cortical microtubule rearrangements in living epidermal cells. The Plant Cell 10(11): 1927-1939
20. Nakamura M, Naoi K, Shoji T & Hashimoto T (2004). Low concentrations of propyzamide and oryzalin alter microtubule dynamics in Arabidopsis epidermal cells. Plant and Cell Physiology 45(9): 1330-1334
21. Robson T M & Aphalo P J (2012). Species-specific effect of UV-B radiation on the temporal pattern of leaf growth. Physiologia Plantarum 144(2): 146-160
22. Robson T M, Klem K, Urban O & Jansen M A (2015). Reinterpreting plant morphological responses to UV-B radiation. Plant, Cell & Environment 38(5): 856-866
23. Rozema J, Staaij J V, Björn L O & Caldwell M (1997). UV-B as an environmental factor in plant life: stress and regulation. Trends in Ecology & Evolution 12(1): 22-28
24. Sedbrook J C (2004). MAPs in plant cells: Delineating microtubule growth dynamics and organization. Current Opinion in Plant Biology 7(6): 632-640
25. Shi L C, Wang B C, Gong W, Zhang Y G, Zhu L Q & Yang X Y (2011). Actin filaments and microtubules of Arabidopsis suspension cells show different responses to changing turgor pressure. Biochemical and Biophysical Research Communications 405(4): 632-637
26. Voigt B, Timmers A C, Šamaj J, Müller J, Baluška F & Menzel D (2005). GFP-FABD2 fusion construct allows in vivo visualization of the dynamic actin cytoskeleton in all cells of Arabidopsis seedlings. European Journal of Cell Biology 84(6): 595-608
27. Wasteneys G O & Yang Z B (2004). New views on the plant cytoskeleton. Plant Physiology 136(4): 38843891
28. Yu R, Huang R F, Wang X C & Yuan M (2001). Microtubule dynamics are involved in stomatal movement of Vicia faba L. Protoplasma 216: 113-118
29. Zhang J W & Zhou Q (2009). Effect of UV-B radiation on water metabolism in plants (in Chinese). Chinese Journal of Eco-Agriculture 17(4): 829-833