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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (12): 1793-1801.doi: 10.3724/SP.J.1006.2018.01793


Cloning and Characterization of Brassinazole-resistant (BnaBZR1 and BnaBES1) CDS from Brassica napus L.

Tao FENG,Chun-Yun GUAN()   

  1. College of Agronomy, Hunan Agricultural University / National Oilseed Crops Improvement Center in Hunan, Changsha 410128, Hunan, China
  • Received:2018-05-03 Accepted:2018-08-20 Online:2018-12-12 Published:2018-09-19
  • Contact: Chun-Yun GUAN E-mail:guancy2011@aliyun.com
  • Supported by:
    This study was supported by the National Basic Research Program (973 Program).(2015CB150206)


Brassinazole-resistant (BZR) is a key transcription factor in brassinosteroid signaling pathway of plants, containing brassinazole-resistant 1 (BZR1) and brassinazole-resistant 2 (BES1). In this study, three novel BZR1 coding sequences (CDSs) were isolated from cDNA of Brassica napus L. cv. Xiangyou 15 leaves, which were mapped on the chromosomes A07, A06, and A06, and designated as BnaBZR1_A07, BnaBES1_A06F, and BnaBES1_A06R, respectively. These three BnaBZR CDSs were 996, 993, and 996 bp in length and encoded predicted proteins with 331, 330, and 331 amino acid residues, respectively. BnaBZR proteins were predicted to be located on the cell nucleus and have a typical plant BZR/BES conserved domain. Multiple sequence alignments and phylogenetic analysis showed that the deduced amino acid sequences of BnaBZR were highly homologous to previously reported BZR/BES of Brassica oleracea, Arabidopsis thalian, and Eruca sativa. And the similarity of BZR1 or BES1 among different related species was higher than the similarity between BZR1 and BES1 in the same species or related species, indicating that BZR1 and BES1 differentiation is an early evolutionary event. The expression patterns of BnaBZR1_A07, BnaBES1_A06F, and BnaBES1_A06R in Xiangyou 15 were similar, whit high expression level at the seedling stage and flowering stage, while slightly lower level at the stage of bolting and podgrain ripening. The expression level of BnaBZR in the aerial parts such as leaves, stems, flowers and pods was higher than that in underground parts.

Key words: Brassica napus L., BZR, gene clone, gene expression, bionformatic analysis

Fig. 1

Cloning of BnaBZR1_A07, BnaBES1_A06F, and BnaBES1_A06R M: 2K DNA marker; 1: BnaBZR1_A07CDS; 2: BnaBES1_A06F CDS; 3: BnaBES1_A06R CDS."

Table 1

Summary of the deduced BnaBZR1, BnaBES1_F, and BnaBES1_R proteins"

Serial number
Base of amino acid residues
Molar mass (Da)
Isoelectric point
BnaBZR1_A07 331 35 895.131 9.145
BnaBES1_A06F 330 36 071.176 9.745
BnaBES1_A06R 331 36 177.322 9.745

Fig. 2

Secondary structure of BnaBZR1_A07, BnaBES1_A06F, and BnaBES1_A06R"

Table 2

Summary of specific region of BnaBZR1_A07, BnaBES1_A06F, and BnaBES1_A06R"

Serial number
Domains type
Sequence motif
BnaBZR1_A07 PS00001 2 142-145, 143-146 NNSS, NSST N-{P}-[ST]-{P}
PS00004 1 21-24 RKPS [RK](2)-x-[ST]
PS00005 5 24-26, 84-86, 162-164, 179-181, 221-223 SWR, TYR, SLR, TSK, THR [ST]-x-[RK]
PS00006 4 24-27, 236-239, 230-233, 240-243 SWRE, SRGE, TIPE, STVD [ST]-x(2)-[DE]
PS00008 4 5-10, 154-159, 155-160, 325-330 GATSTS, GGIPSS, GIPSSSL, GNGKAR G-{EDRKHPFYW}-x(2)-[STAGCN]-{P}
BnaBES1_A06F PS00001 1 138-141 NIST N-{P}-[ST]-{P}
PS00004 1 20-23 RKPS [RK](2)-x-[ST]
Serial number
Domains type
Sequence motif
BnaBES1_A06R PS00005 5 18-20, 23-25, 83-85, 99-101, 174-176 TRR, SWR, TYR, SSR, TNK [ST]-x-[RK]
PS00006 6 23-26, 112-115, 134 -137, 227 -230, 237-240, 305-308 SWRE, SPFE, SRGD, TIPE, STVD, TPWE [ST]-x(2)-[DE]
PS00008 5 5-10, 149-154, 150-155, 259-264, 325-330 GATSTS, GGIPSS, GIPSSL, GVSSAV, GNAKGR G-{EDRKHPFYW}-x(2)-[STAGCN]-{P}
PS00016 1 135-137 RGD R-G-D
PS00001 1 139-142 NIST N-{P}-[ST]-{P}
PS00004 1 21-24 RKPS [RK](2)-x-[ST]
PS00005 5 18-20, 24-26, 84-86, 100-102, 175-177 TRR, SWR, TYR, SSR, TSK [ST]-x-[RK]
PS00006 6 24-27, 113-116, 135-138, 228-231, 238-241, 306-309 SWRE, SPFE, SRGD, TIPE, STVD, TPWE [ST]-x(2)-[DE]
PS00008 5 5-10, 150-155, 151-156, 260-265, 326-331 GATSTS, GGIPSS, GIPSSL, GVSSAV, GNAKGR G-{EDRKHPFYW}-x(2)-[STAGCN]-{P}
PS00016 1 136-138 RGD R-G-D

Fig. 3

Comparison of the amino acid sequences of BnaBZR"

Fig. 4

Phylogenetic analysis of BZR/BES in different plant species"

Fig. 5

BnaBZR1_A07, BnaBES1_A06F, and BnaBES1_A06R expression in root, stem, leaf, flower, and silique DAG means days after seed germination."

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