Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (12): 168-174.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0341

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

HU Qi(), HOU Yu-xiang, LI Rui, LI Mei-lan()   

  1. College of Horticulture,Shanxi Agricultural University,Jinzhong 030800
  • Received:2022-03-21 Online:2022-12-26 Published:2022-12-29
  • Contact: LI Mei-lan E-mail:15231255323@163.com;15935485975@163.com

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