Cloning and Expression of CYP79B2 Homologous Genes in Brassica rapa ssp. chinensis

doi:10.13560/j.cnki.biotech.bull.1985.2022-0341

Abstract

Abstract:

CYP79B is a key gene involved in auxin synthesis in Arabidopsis thaliana. Cloning the homologous gene BrcCYP79B2-2 of CYP79B2 and analyzing the tissue expression of this gene may provide a theoretical basis for the functional verification of genes regulating flowering in pak choi（Brassica rapa ssp. chinensis）. Here，RT-PCR was used to clone the BrcCYP79B2-2 of pakchoi’s CYP79B2 and the gene was analyzed by bioinformatics. RT-qPCR was employed to study the expression of BrcCYP79B2-2 in different tissues of pakchoi. The results showed that the CDS of BrcCYP79B2-2 was 1 554 bp in length，encoding 517 amino acids. The amino acid sequence of BrcCYP79B2-2 protein was of 99.42% homology with that of Brassica pekinensis，suggesting that BrcCYP79B2-2 was highly conserved in different varieties of B. pekinensis. RT-qPCR analysis revealed that the gene was expressed in different tissues，with the highest expression in the roots，lower in the buds，and lowest in stems. The expression of BrcCYP79B2-2 in the stem tip after low-temperature treatment decreased. Besides，the expressions of BrcCYP79B2-2 varied among different growth stages，higher in the vegetative growth stage but lower in the critical stage of flower bud differentiation. The expression of BrcCYP79B2-2 responded to low temperature and significantly varied in different tissues. It is speculated that the gene may affect flower bud differentiation.

Key words: Brassica rapa ssp. chinensis, auxin, BrcCYP79B2-2, gene cloning, expression analysis

HU Qi, HOU Yu-xiang, LI Rui, LI Mei-lan. Cloning and Expression of CYP79B2 Homologous Genes in Brassica rapa ssp. chinensis[J]. Biotechnology Bulletin, 2022, 38(12): 168-174.

Figures/Tables 5

Fig. 1 Amplified electrophoresis of BrcCYP79B2-2

Fig. 2 Multiple alignment of CYP79B2-2 amino acid sequences from different species

Fig. 3 Phylogenetic tree of CYP79B2-2 homologous prote-ins from different species

Fig. 4 Protein tertiary structure prediction of BrcCYP-79B2-2

Fig. 5 Expressions of BrcCYP79B2-2 in different tissues and in stem tip in different stages after low te-mperature treatment V0：0 d after vernalization（DAV0）；V10：10 d after vernalization（DAV10）；V15：critical stage of flower bud differentiation（S0）；V16：flower bud differentiation stage（S1）

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