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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (08): 1371-1379.doi: 10.3724/SP.J.1006.2014.01371


Cloning and Expression Analysis of BoLH01and BoLH02 Genes in Cabbage

ZHANG Lin-Cheng1,GAO Qi-Guo1,*,PU Quan-Ming1,REN Xue-Song1,LIU Yu-Dong1,ZHU Li-Quan2,WANG Xiao-Jia1,*   

  1. 1 College of Horticulture and Landscape Architecture, Southwest University / Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Chongqing 400716, China; 2 College of Agronomy and Biotechnology, Southwest University, Chongqing 400716, China
  • Received:2013-12-02 Revised:2014-06-16 Online:2014-08-12 Published:2014-06-10


Basic helix-loop-helix (bHLH) transcription factors play an important role in the leaf morphogenesis and leaf development. In this study, through transcriptome analyses, two bHLH transcription factor genes, differentially expressed in leaves and shoot tips between cabbage rosette stage and heading stage, were screened, named BoLH01 and BoLH02 genes. The cDNA sequences of BoLH01and BoLH02 were cloned from cabbage (Brasscia oleracea var. Capitata L). The CDS of BoLH01 was 966 bp, which encoded 321 amino acids, and the CDS of BoLH02 was 870 bp, which encoded 289 amino acids. Sequence analysis showed that the similarity of BoLH01 and BoLH02 with AtbHLH18 and AtbHLH19 was 81% and 74% respectively. They contained the typical bHLH domains, conservative 13E-16R, 9E-13R-17H and Leu23 corresponding key amino acid sites. Phylogenetic tree analysis displayed that BoLH01and BoLH02 were closest to AtTCP2/3/4/10/24, and had no close genetic relationship with AtTCP9/11/14/15. Moreover, the transcriptome and real-time fluorescence quantitative PCR analysis indicated that the relative expression levels of BoLH01 and BoLH02 were low in rosette leaves, and high in heading stage. Sequence analysis showed that the ATG upstream of BoHB7, BoHB12 and BoILL6 contained the E-box motif, and those motifs could been recognized by bHLH functional domains. The real-time fluorescence quantitative PCR analysis indicated that the variation in expression levels of BoHB7, BoHB12 and BoILL6 was similar with that of BoLH01 and BoLH02 in leaves (1 cm). These results meant that BoLH01 and BoLH02 maybe participate in the regulation of cabbage leaf curl by positively regulating BoHB7, BoHB12, BoILL6.

Key words: Brassica oleracea L., Leaf curl, Gene cloning, Expression analysis

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