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Analysis of CESA complexes dynamics in plant cytoskeletal mutants
Dubenecká, Kamila ; Schwarzerová, Kateřina (advisor) ; Malínská, Kateřina (referee)
The basis of this study are mutant plants with ARP2/3 complex lacking in one of its subunits (arpc5 and arp2). These plants also express CSC subunit CESA6 of primary cell wall tagged by YFP. Thanks to modern imaging technologies, it is possible to observe the movement of tagged cellulose synthase complexes in vivo at plasmatic membrane. Kymograph analyses was used to measure the velocity of CESA complexes. In addition to observing CESA complexes directly on the plasma membrane, experiments were made to regenerate cell walls of protoplasts of Arabidopsis thaliana plants arpc5 and WT. It was found, that observed mutants arpc5 and arp2 have reduced velocity of CESA complexes in comparison to WT and arpc5 protoplasts regenerate cellulose mesh of cell wall slower. Keywords: Cellulose synthesis, ARP2/3 complex, CESA, CSC velocity, arpc5, arp2, Arabidopsis thaliana.
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The role of PLD in early phases of aluminium toxicity
Poláková, Lucie ; Schwarzerová, Kateřina (advisor) ; Malínská, Kateřina (referee)
Aluminium toxicity is the main limiting factor in crop production on acid soils. The main symptom of aluminium toxicity is a rapid inhibition of root growth, but the mechanism of root growth cessation remains unclear. In this diploma thesis we deal with the question of whether phospholipases PLDα1 and PLDδ may play a role in the mechanism of aluminium toxicity. We compared the responses of plants lacking PLDα and PLDδ with WT plants. Growth analysis of roots was performed in hydroponic conditions. The most sensitive part of roots was transient zone in which cells were dying earlier. It was further found that pldα1 plants were less sensitive on aluminium toxicity because their roots showed less growth inhibition than WT. Pldδ plants did not differ from WT plants in their response to aluminum. During further analysis of the pldα1 reactions, it was found that the root cells were capable of cell expansion during aluminum toxicity, and the cellular malformations were formed on the roots. This phenomenon was associated with faster reorientation and even depolymerization of cortical microtubules in response to toxic aluminium in pldα plants compared to WT plants. The results indicated that PLDα1 molecule affects the stability of cortical microtubules. Microtubules were less stable and they depolymerized...
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The role of the cytoskeleton in morphogenesis of plant cells
Havelková, Lenka ; Schwarzerová, Kateřina (advisor) ; Hašek, Jiří (referee) ; Malínská, Kateřina (referee)
6. Conclusions ► The capability of ARPC2 protein (subunit of ARP2/3 complex) to bind microtubules was described in this study. Our results indicated that ARP2/3 complex may mediate direct interaction between microfilaments and microtubules in processes where the coordination of microtubule reorganization and actin mediated growth was required. ► We showed that cell-malforming effect of growth retardant ancymidol was based on its inhibitory action on cellulose synthesis. Microtubules played a passive role in this process and were not directly responsible for changes of the cell shape induced by ancymidol. ► Meristematic and suspensors cells in maturating somatic embryos differed in sensitivity to latrunculin B, probably due to different composition of actin isoforms in these cells. Application of low doses of latrunculin B resulted in selective death of suspensor cells and thus contributed to the development of high-quality embryos. ► Aluminium caused rapid cessation of root growth of Arabidopsis thaliana seedlings. We showed that the immediate reason of root growth cessation was plasma membrane rigidization and loss of endocytosis in root cells. Aluminium stabilized cortical microtubules within minutes and induced their loss within hours of treatment. However, the effect of aluminium on microtubules was not...
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