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• W2586151330 abstract "Sugar beet (Beta vulgaris subsp. vulgaris), a biennial long day (LD) plant belonging to the family Chenopodiaceae, stores sucrose in a thickened taproot and is the only crop cultivated for sugar production in Europe. Full bolting time control is of substantial agronomic importance to prevent stem elongation (bolting) and flowering which result in decreasing root yield. The regulation of flowering time is well studied in the model species Arabidopsis thaliana and the induction of flowering through the zinc finger transcriptional regulator CONSTANS (CO) and the FLOWERING LOCUS T (FT) gene appears to be conserved in various plant species. In B. vulgaris, a new mechanism of flowering time regulation has been observed. Two FT paralogs BvFT1 and BvFT2 have evolved antagonistic functions, in which BvFT1 is a floral repressor and BvFT2 acts as a floral inducer. Annuals that flower under long days (LD) within one season carry the dominant pseudo response regulator (PRR) gene BOLTING TIME CONTROL 1 (BTC1), which shows homology to the PRR7 gene in A. thaliana. BTC1 promotes bolting and flowering through repression of BvFT1 and activation of BvFT2. By contrast, in biennials carrying the recessive btc1 gene, BvFT1 is not repressed and plants require a period of prolonged cold temperature over winter (vernalization) followed by LD for bolting induction. Protein sequences of the dominant and recessive BTC1 genes differ by several amino acid substitutions. Besides BTC1, three additional bolting loci B2, B3 and B4 have been previously identified after EMS mutagenesis of an annual genotype. So far, the two loci B2 and B4 were mapped to chromosome 9 and 2, respectively. A B2 mutant displays a biennial phenotype and thus requires vernalization for bolting induction. The aims of my study were (1) to fine-map the bolting locus B2 and to identify putative candidate genes, and (2) to investigated the function of BTC1 in transgenic A. thaliana and B. vulgaris plants to clarify whether the polypeptides encoded by the dominant and recessive BTC1 gene have the same function in beet and whether BTC1 and PRR7 are orthologs.In order to fine-map the B2 locus, 5457 F2 plants from a cross of a biennial B2 mutant and an annual wild beet and 2549 F2:3 families were phenotyped for bolting behavior. Finally, 1301 F2 plants were genotyped with molecular markers. I used the draft assembly of the sugar beet reference sequence to develop molecular markers and identified two markers flanking the B2 locus in a genetic distance of 1.9 cM. I screened this region for candidate genes using a collection of predicted gene models of the sugar beet reference sequence. This enabled the identification of a candidate gene which shows an overall sequence identity of 55% on amino acid level to BBX19, a B-box zinc finger family protein of A. thaliana, comprising two B-box domains. Accordingly, I termed the newly detected candidate gene BvBBX19. A molecular marker located in the 3`-UTR of BvBBX19 completely co-segregates with the F2 phenotypes (1295 F2 plants). I identified a single transition in each of two independent mutants that occurred at different positions within the candidate gene. Both mutations are likely to be induced through the EMS treatment and alter the amino acid sequence of the BvBBX19 protein. These results give strong evidence that BvBBX19 is likely to be the sought gene encoding the B2 phenotype.The function of the different BTC1 alleles was first verified by constitutive expression of both BTC1 coding sequences in A. thaliana wildtype and late flowering prr7 mutant plants. I observed no significant differences for bolting and flowering in segregating T2 families. BTC1 neither accelerated flowering in the wildtype nor complemented the late flowering prr7 mutant. These results suggest that BTC1 is not an ortholog of PRR7 and has a different function in B. vulgaris compared to A. thaliana. Second, I investigated the BTC1 function in B. vulgaris plants (T1 generation). I observed no phenotypic differences in plants carrying the dominant BTC1 transgene compared to those carrying the recessive btc1 transgene, before as well as after cold treatment. However, after cold treatment I observed bolting and never bolting (non bolting after cold treatment) individuals. Expression analysis and copy number determination revealed an upregulation of BTC1 and a downregulation of BvFT1 in bolting plants that carry a single copy of the transgene. By contrast, BTC1 is completely downregulated and BvFT1 is highly upregulated in never bolting plants carrying multiple integrations of the transgene. These results demonstrate for the first time transgene-mediated co-suppression in transgenic sugar beet plants and thus confirm also that both BTC1 genes have the same function.Beyond the aims of my study, I further investigated the expression of BvBBX19 in bolting and never bolting transgenic beets as well as in the parental accessions of the F2 population, in comparison with BTC1, BvFT1 and BvFT2. The results of this analysis indicate that BvBBX19 acts in conjunction with BTC1 upstream of BvFT1 and BvFT2 to induce bolting and flowering in beet. Under the assumption that BvBBX19 and BTC1 proteins interact with each other, the resulting protein complex would have i.a. two B-box and a CCT domain and thus resemble the domain structure of CO. Accordingly I hypothesize that in beet a CO function is achieved through interaction of BvBBX19 and BTC1. The results of my work largely extended our knowledge about bolting and flowering time regulation in beet which is of substantial importance for breeding winter beets with controlled bolting behavior." @default.
• W2586151330 created "2017-02-17" @default.
• W2586151330 creator A5046424101 @default.
• W2586151330 date "2014-07-08" @default.
• W2586151330 modified "2023-09-23" @default.
• W2586151330 title "Map based cloning and functional analysis of two bolting time loci BTC1 and BvBBX19 in sugar beet (Beta vulgaris L.)" @default.
• W2586151330 hasPublicationYear "2014" @default.
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