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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (03): 397-404.doi: 10.3724/SP.J.1006.2018.00397


Cloning of BoLH27 Gene from Cabbage and Phenotype Analysis of Transgenic Cabbage

Yun-Fei LIANG1,**, Lin-Cheng ZHANG1,**, Quan-Ming PU2, Zhen-Ze LEI1, Song-Mei SHI1, Yu-Peng JIANG1, Xue-Song REN1, Qi-Guo GAO1,*()   

  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 Institute of Vegetables, Nanchong Academy of Agricultural Sciences, Nanchong 637000, Sichuan, China
  • Received:2017-06-06 Accepted:2017-11-21 Online:2018-03-12 Published:2017-12-11
  • Contact: Yun-Fei LIANG,Lin-Cheng ZHANG,Qi-Guo GAO E-mail:gaoqg031@swu.edu.cn
  • Supported by:
    This study was supported by the National Program on Key Basic Research Project (973 program)(2012CB113900) and the National Natural Science Foundation of China (30900986).


bHLH transcription factor plays an important role in plant growth, development and morphological control. In order to explore BoLH27 gene regulatory function for leaf development and morphologic formation, the BoLH27 gene was cloned from cabbage (Brassica oleracea L.) variety 519. Sequence analysis indicated that the length of BoLH27 gene was 795 bp, which encoded 264 amino acids. The BoLH27 protein contained conservative structure domains of the bHLH family. The sense BoLH27 gene was transformed into cabbage variety 519 by Agrobacterium mediated method, PCR analysis exhibited that BoLH27 was genetically transformed into nine individual plants, qRT-PCR analysis revealed that BoLH27 had higher expression level in T2 transgenic cabbage than in wild type. The phenotype at rosette stage of transgenic cabbage grown in the test base was obvious, showing elongated the stems and leaves, elongated petiole, purple stems and petiole, and flat leaves without upward inward curling trend. It suggested that BoLH27 may play important roles in the control of leaves development.

Key words: Brassica oleracea, BoLH27, gene cloning, transgene, phenotype

Table 1

Primers used in this study"

Primer name and sequence (5°→3°)
Annealing temperature (°C)

Fig. 1

cDNA sequence of Brassica oleracea L. BoLH27 and its deduced amino acid sequence The black shadow shows bHLH domain; the full line box shows N-glycosylation site; the dotted line box shows cAMP phosphorylation site; the underline shows cGMP-dependent protein kinase phosphorylation site; the wavy line shows casein kinase II phosphorylation site; the oval box shows N-glycosylation site."

Fig. 2

bHLH domain alignment among BoLH27 and other homologous proteins Stars indicate the conserved amino acids of bHLH domain; triangle indicates amino acid involved in dimer formation."

Fig. 3

Phylogenetic relationship of amino acid sequences between Brassica oleracea L. BoLH27 and its homologous proteins"

Fig. 4

The process of obtaining pC35S-BoLH27 transgenic cabbage plants A: the growth of resistant bud; B, C: the rooting of resistant seedling; D: the seedling of transgenic cabbage."

Fig. 5

PCR analysis of transgenic plants M: DL2000; 1-10: transgenic cabbage; +: positive plamid; -: non-transgenic cabbage."

Fig. 6

Relative expression of BoLH27 in wild-type and transgenic cabbage lines WT: non-transgenic lines; S13, S14, S33: transgenic lines."

Fig. 7

Phenotypes of non-transgenic lines and BoLH27 overexpressed lines A: seedling stage with ten leaves; B: side view of rosette stage with twenty-four leaves; C: top view of rosette stage with twenty-four leaves; WT: non-transgenic lines; BoLH27: overexpressed lines; partial: partial enlarged drawing of overexpressed lines."

Table 2

Average petiole length and average leaf spacing in overexpressed lines"

Overexpressed lines
Average petiole length (cm)
Average leaf
spacing (cm)
1 16.8 1.7
2 15.2 1.8
3 14.5 1.6
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