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

doi:10.7668/hbnxb.20193182

Abstract

Abstract:

CYP79B2 is a related gene that regulates auxin synthesis in Arabidopsis.By cloning the homologous gene BrcCYP79B2-1 of CYP79B2 in B.rapa ssp. cninensis,analyzing its expression in different tissues and periods,and studying its regulatory mechanism on vernal flowering in B.rapa ssp. cninensis,to lay a theoretical foundation for the subsequent functional verification of the auxin-encoding gene in B.rapa ssp. cninensis.The homologous gene of CYP79B2 was cloned from B.rapa ssp. cninensis by qRT-PCR and named as BrcCYP79B2-1.The structure,physicochemical properties and relationship of its protein were analyzed by bioinformatics method,and its protein was analyzed by qRT-PCR method.Expression levels in different tissues and growth stages in B.rapa ssp. cninensis.And its expression in different tissues and growth stages of B.rapa ssp. cninensis was analyzed by qRT-PCR method.The results showed that the full-length coding sequence of BrcCYP79B2-1 gene was 1 623 bp,encoding 540 amino acids.The physicochemical analysis of the protein showed that the molecular mass of the protein was 60.849 73 ku,and the theoretical isoelectric point was 8.71.Compared with the amino acid sequences of other species,it was found that BrcCYP79B2-1 was highly conserved in cruciferous plants,and had the highest homology with turnip,up to 99.52%;the expression levels of different organs of B.rapa ssp. cninensis were analyzed by qRT-PCR, and it was found that the expression level of BrcCYP79B2-1 was the highest in roots, followed by leaves, and the lowest in flower buds.The expression of BrcCYP79B2-1 in seedlings after 0,10,15 and 16 days of low temperature treatment showed that the expresion of BrcCYP79B2-1 reached its peak on the 10th day of low temperature treatment, that is, the vegetative growth period, while the expression of BrcCYP79B2-1 decreased during flower bud differentiation, which indicated that the expression of BrcCYP79B2-1 was related to low temperature vernalization and could affect flower bud differentiation.

Key words: B.rapa ssp. chinensis Makino, BrcCYP79B2-1, Bioinformatics, Gene cloning, Expression analysis, Flowering

HOU Ruize, HOU Yuxiang, XU Xiaoyong, LI Xuan, LI Meilan. Cloning and Expression Analysis of CYP Homologous Genes in Brassica rapa ssp. chinensis[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 45-51. doi: 10.7668/hbnxb.20193182.

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Figures/Tables 8

Fig.1 Agarose gel electrophoresis of the PCR products of BrcCYP79B2-1 M.DNA Marker;1—3.BrcCYP79B2-1。

Fig.2 Gene and amino acid sequence of BrcCYP79B2-1

Fig.3 Multiple alignment results of the amino acid sequence of BrcCYP79B2-1

Fig.4 Phylogenetic tree of BrcCYP79B2-1 homologous protein

Fig.5 Protein secondary structure of BrcCYP79B2-1

Fig.6 Protein tertiary structure prediction of BrcCYP79B2-1

Fig.7 Expression analysis BrcCYP79B2-1 in different organs Different lowercase letters represent significant differences in relative expression at the 0.05 level.

Fig.8 Expression of BrcCYP79B2-1 in shoot apex at different developmental

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