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Master's Thesis
Author of thesis: Bc. Petr Haleš
Acad. year: 2025/2026
Supervisor: Ing. Michal Kalina, Ph.D.
Reviewer: doc. Ing. Vojtěch Enev, Ph.D.
Biochar is an important soil conditioner in agriculture, serving as a long-term source of organic matter and a short-term source of minerals. However, due to the high temperatures used during its production by pyrolysis, a non-living porous matrix is often formed, which limits its direct impact on biological processes in the soil. Enriching biochar with plant growth-promoting rhizobacteria (PGPR) therefore offers a way to utilize its physicochemical and structural properties for microbial colonization and to supplement this soil conditioner with a living microbial component. Four types of commercial biochar samples (Carbohran, Novocarbo, Sonnenerde, and Zera) produced from various biomass feedstocks at different pyrolysis temperatures and the bacterial strain Azotobacter vinelandii CCM 289 were used in this study. In the introductory section, the biochars were characterized using thermogravimetric analysis (TGA), and the physicochemical properties of aqueous extracts (pH, conductivity) and their elemental composition were determined using ICP-OES. Prior to colonization, the effects of pH, ionic strength, and biochar extract on bacterial growth and its capability to colonize biochar structure were simulated in preliminary cultivation experiments. The aim was to assess separately the influence of these parameters on cell viability and concentration and the capability of bacteria to produce alginate extracellularly. These bacterial suspensions were cross-linked with a 2 wt.% CaCl₂ solution into the alginate hydrogel, which was characterized using rheometry. Subsequently, two colonization methods were compared: post-cultivation (mixing the prepared bacterial suspension with biochar) and co-cultivation (direct bacterial growth in a biochar suspension). For both approaches, the effect of biochar presence on cell concentration and viability was evaluated using flow cytometry, and the viscoelastic properties of the resulting alginate hydrogels with entrapped bacteria were monitored using rheometry. These prepared systems were freeze-dried to ensure stability for subsequent soil applications. The core of the work consisted of a 92-day cultivation experiment with the presence of a model plant, maize (Zea mays) under controlled conditions (temperature, moisture, irrigation, illumination). The post-cultivation colonization method of biochars was chosen for these soil applications. During the experiment, soil pH dynamics and agronomic parameters of the plants were monitored. After completion of the cultivation experiment, a comprehensive analysis of the soil, plant biomass, and microbial activity in the rhizosphere using the BIOLOG EcoPlate™ kit was performed. Specific PGP effects were also tested, including nitrogen fixation, phosphate solubilization, and the production of siderophores, 1-aminocyclopropane-1-carboxylate (ACC) deaminase, and indole-3-acetic acid (IAA). The obtained results indicate that the combination of biochar with PGPR has the potential to enhance the effects of biochar in soil applications, particularly in terms of pH stabilization and support for microbial activity in the rhizosphere. However, since these are preliminary experiments conducted within complex matrices, it would be necessary to repeat the measurements with a higher number of replicates to obtain statistically more significant results and subsequently verify the obtained outcomes in semi-field experiments under real-soil conditions.
Biochar, soil, plant growth-promoting rhizobacteria (PGPR), Azotobacter vinelandii, PGP effects, cultivation experiment, Zea mays
Date of defence
22.05.2026
Result of the defence
Defended (thesis was successfully defended)
Grading
A
Process of defence
Obhajoba diplomové práce proběhla podle následujícího schématu: prezentace studenta-vyjádření vedoucí/ho-oponentský posudek-reakce na posudek-diskuse s komisí. Student přednesl výborný výtah výsledků své práce, řádně zodpověděl všechny dotazy oponentské i členů komise, pohotově reagoval na připomínky. V diskusi tak student prokázal výbornou schopnost orientace v teoretických i praktických základech problematiky práce. Komise zhodnotila jeho práci celkově jako výbornou. Pekař: Kolik je potřeba biouhlu na polní experiment? Dzik: Jaký je zdroj uhlíku v čistírenských kalech?
Language of thesis
Czech
Faculty
Fakulta chemická
Department
Institute of Physical and Applied Chemistry
Study programme
Chemistry for Medical Application (NPCP_CHMA)
Specialization
Processes and Materials of Medical Applications (BF)
Composition of Committee
doc. PharmDr. Ing. Radka Ješinová, Ph.D. (člen) doc. Ing. Zdenka Kozáková, Ph.D. (člen) prof. Ing. Stanislav Obruča, Ph.D. (místopředseda) prof. Ing. Miloslav Pekař, CSc. (předseda) doc. Ing. Petr Dzik, Ph.D. (člen)
Supervisor’s reportIng. Michal Kalina, Ph.D.
Grade proposed by supervisor: A
Reviewer’s reportdoc. Ing. Vojtěch Enev, Ph.D.
Grade proposed by reviewer: A
Responsibility: Mgr. et Mgr. Hana Odstrčilová