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Resynthesis of oilseed rape from Brassica rapa and Brassica oleracea
Klíma, Miroslav ; Hilgert-Delgado, Alois ; Ulvrová, Tereza ; Bryxová, Pavla ; Vítámvás, Pavel ; Prášil, Ilja Tom ; Kosová, Klára
In order to extend the variability of the genofond, so-called resynthesis can be used, when new genotypes are created by crossing selected Brassica rapa and B. oleracea, by early isolation of immature embryos from young eggs and their in vitro fertilization (Hilgert-Delgado et al., 2015). It has been shown that resyntetized genotypes are distinct from rape materials grown at present (Sosnowska et al., 2010), so that they can serve as a source of new variability in breeding programs focusing on cytoplasmic and pollen sterility, resistance to pathogens and tolerance to stresses (Warwick et al., 2009). Representatives of the initial components are characterized by wide variability in terms of morphology and agronomic characteristics, and are a prospective source of heterosis and other traits (Wu et al. The use of resyntetized genotypes for the direct production of rapeseed varieties is not possible due to the poor quality of the seed of these materials. It is mainly the high content of glucosinolates and erucic acid, typical of most of the initial components (Jesske et al., 2013). However, the required properties can be transferred by crossing to conventional breeding materials (Hilgert-Delgado et al., 2017).
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Funkce genů rodiny TTL a jejich efekt na tvorbu postranních kořenů u Arabidopsis
Hilgert Delgado, Alois Antonín ; Soukup, Aleš (advisor) ; Cvrčková, Fatima (referee)
The work is focused on the lateral roots, which are an indispensable part of the root system and play a key role in communication with rhizosphere and the efficient exploitation of natural resources. Tetratricopeptide-repeat Thioredoxin-like 3 gene (TTL3) was identified by forward screening based on its gene expression pattern in the search for genes associated with the development of lateral roots in Arabidopsis thaliana. TTL3 also known as VIT (VH1 interacting TPR containing protein), belongs to a family of four paralogues in Arabidopsis showing a very similar arrangement of domains. It seems that this type of proteins is widespread in plants and provides protein-protein signaling as an important component of phytohormone response pathways. The presented diploma thesis is focused on characterization and comparison of expression patterns of the four members of TTL family, through creation of pTTL::uidA (GUS) transcriptional fusions. The promoter area was estimated to be contained in about 2000bp upwards the TTLs coding sequences and transcriptional fusions with uidA gene were individually constructed and transformed into Arabidopsis via Agrobacterium tumefaciens. The expression patterns of analyzed genes were microscopically confirmed and allowed comparison among members of TTL gene family with regard...
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Optimization and application of in vitro techniques in selected members of the family Brassicaceae
Hilgert-Delgado, Alois Antonín ; Fernández Cusimamani, Eloy (advisor) ; Jana, Jana (referee)
This thesis is focused on the application and optimization of biotechnological methods within the family Brassicaceae. Most of this thesis is focused specifically the genus Brassica. The main objective of this dissertation thesis was to optimize and apply selected biotechnological methods in the resynthesis of oilseed rape (B. napus) from its baseline species (B. oleracea, CC and B. rapa, AA) and subsequently create valuable genetic plant materials.
The optimized procedure implemented in my work have exhibited better results of hybrid production (resynthesis) than in similar published experiments and provides a simplified and less laborious method via simple ovule culture and early hybrid verification.
The next accomplished step was to work with a wider range of materials using a broader source of different and contrasting genotypes from Brassica rapa (spring and winter oilseed and vegetable turnips) and B. oleracea (green and purple curly kales and cabbages) for new combinations and wider genetic diversity.
It was concluded that the optimized ovule culture protocol with early verification, developed in the framework of this thesis, is satisfactorily sufficient enough to be applicable in breeding programmes, aimed at diversity expansion of winter oilseed rape gene pool, as the resynthesized embryos were derived in most combinations.
New resynthesized verified lines were colchicine treated in order to obtain diploid regenerants and the fertile plants were self-pollinized and crossed with elite oilseed rape lines for further research and breeding.
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