• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.438-447
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• 2017

Flowering is one of the most important development traits related to the production of Brassica rapa crops. After planting, a sudden low temperature triggers premature flowering, which leads to a reduction in the yield and quality of harvested production. Therefore, understanding the mechanism of flowering control is important in the agricultural productivity for preventing Brassica rapa crops. Vernalization is generally known as the main factor of flowering in the Brassica plant. However, in the subspecies of Brassica rapa, some accession such as Yellow sarson and Komatsuna display the flowering phenotype without vernalization. Circadian genes, which diurnally regulate plant physiology, have a role for photoperiodic flowering but are related to the regulation of the vernalizarion mechanism. In this report, the 22 B. rapa accession were divided into two groups, vernalization and non-vernalization, and the sequenced circadian gene, BrPRR1s. Among them, the BrPRR1b gene was found to have deletion regions, which could classify the two groups. The PCR primer was designed to amplify a short band of 422bp in the vernalization type and a long band of 451bp in the non-vernalization type. This primer set was applied to distinguish the flowering types in the 43 B. rapa accession and 4 Brassica genus crop, Broccoli, cabbage, mustard, and rape. The PCR analysis results and flowering time information of each crop demonstrated that the primer set can be used as marker to discern the flowering type in Brassica crops. This marker system can be applied to the B. rapa breeding when selecting the flowering character of new progenies or introducing varieties at an early stage. In addition, these results displayed that the circadian clock genes can be a good strategy for the flowering control of B. rapa crops.

• Molecules and Cells
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• v.23 no.2
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• pp.145-153
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• 2007

Elucidation of the roles of circadian associated factors requires a better understanding of the molecular mechanisms of circadian rhythms, control of flowering time through photoperiodic pathways, and photosensory signal transduction. In Arabidopsis, the APRR1 quintet, APRRs 1, 3, 5, 7, and 9, are known as central oscillator genes. Other plants may share the molecular mechanism underlying the circadian rhythm. To identify and characterize these circadian response genes in Brassica crops whose genome was triplicated after divergence from Arabidopsis, we identified B. rapa BAC clones containing these genes by BLAST analysis of B. rapa BAC end sequences against the five corresponding Arabidopsis regions. Subsequent fingerprinting, Southern hybridization, and PCR allowed identification of five BAC clones, one for each of the five circadian-related genes. By draft shotgun sequencing of the BAC clones, we identified the complete gene sequences and cloned the five expressed B. rapa circadian-associated gene members, BrPRRs 1, 3, 5, 7, and 9. Phylogenetic analysis revealed that each BrPRR was orthologous to the corresponding APRR at the sequence level. Northern hybridization revealed that the five genes were transcribed at distinct points in the 24 hour period, and Southern hybridization revealed that they are present in 2, 1, 2, 2, and 1 copies, respectively in the B. rapa genome, which was triplicated and then diploidized during the last 15 million years.