Functional Identification and Expression Analysis of BjuSPL10 in Regulating Brassica juncea Bolting and Flowering

doi:10.16420/j.issn.0513-353x.2024-0835

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (5): 1341-1350.doi: 10.16420/j.issn.0513-353x.2024-0835

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Functional Identification and Expression Analysis of BjuSPL10 in Regulating Brassica juncea Bolting and Flowering

ZHAO Yiman, LI Jingfei, TANG Zhuoran, GE Jiaxing, WEI Dayong, WANG Zhimin, ZOU Jiaqi, TANG Qinglin^**()

Chongqing Key Laboratory of Olericulture，College of Horticulture and Landscape Architecture，Southwest University，Chongqing 400715，China

Received:2025-01-14 Revised:2025-03-17 Online:2025-05-23 Published:2025-05-21
Contact: TANG Qinglin

Abstract

Abstract:

In this study，the BjuSPL10 gene was cloned from Brassica juncea. qRT-PCR analysis showed that the expression of BjuSPL10 gene was different in different parts of Brassica juncea，and the expression level was the highest in flowers. Subcellular localization revealed that BjuSPL10 protein was localized in the nucleus. Subsequently，the BjuSPL10 overexpression vector was constructed and transformed into Brassica juncea. The results showed that BjuSPL10 could significantly promote bolting and flowering in transgenic lines，and increase the expression level of flowering integrator BjuSOC1 gene. Furthermore，yeast one-hybrid and luciferase assays showed that the BjuSPL10 protein directly interacted with the promoter of BjuSOC1 gene，then enhanced its expression level.

Key words: Brassica juncea, SPL10, flowering, regulation, BjuSOC1, transcriptional activation, yeast one-hybrid, luciferase assay

ZHAO Yiman, LI Jingfei, TANG Zhuoran, GE Jiaxing, WEI Dayong, WANG Zhimin, ZOU Jiaqi, TANG Qinglin. Functional Identification and Expression Analysis of BjuSPL10 in Regulating Brassica juncea Bolting and Flowering[J]. Acta Horticulturae Sinica, 2025, 52(5): 1341-1350.

Figures/Tables 8

Table 1 Primers used in this study

用途 Purpose	基因名称 Gene name	引物序列（5′-3′） Sequences
基因克隆 Gene cloning	BjuSPL10	F：ATGGAATGCAACATGAGATC； R：TCATATAAAATGGGTAGGAG
实时荧光定量PCR qRT-PCR	qPCR-BjuSPL10	F：TGTGGGACTGGGAGCATTTG； R：ACAGGAGTGTGCAGGAAACC
实时荧光定量PCR qRT-PCR	BjuACTIN	F：GCTGACCGTATGAGCAAAGA； R：GTTGGAAAGTGCTGAGGGAT
亚细胞定位 Subcellular Localization	BjuSPL10-GFP	F：atacaccaaatcgactctagaATGGAATGCAACATGAGATCTCC； R：gcccttgctcaccatggtaccTATAAAATGGGTAGGAGGGAAA
过表达载体构建 Overexpressing vector	BjuSPL10-Bar	F：tttcatttggagaggagatctATGGAATGCAACATGAGATCTCC； R：caggacgtaacatggccatggTCATATAAAATGGGTAGGAGGGAAA
酵母单杂交 Yeast one-hybrid	BjuSPL10-AD	F：CCGGAATTCATGGAATGCAACATGAGATC； R：CGCGGATCCTCATATAAAATGGGTAGGAG
萤火虫素酶活性实验 LUC	BjuSPL10-SK	F：CGCGGATCCATGGAATGCAACATGAGATC； R：CCGGAATTCTCATATAAAATGGGTAGGAG

Fig. 1 PCR products electropherogram of BjuSPL10 in Brassica juncea

Fig. 2 Phylogenetic evolutionary tree of Brassica juncea and Arabidopsis thaliana SPL family members Ⅰ-Ⅳ are subfamilies，data indicate self-expansion values

Fig. 3 Relative expression of BjuSPL10 gene in different tissues of Brassica juncea（A）and its subcellular localization in tobacco leaves（B） Significance of differences was analyzed using the Duncan’s method，and different letters indicate significant differences in expression in different tissues（P < 0.05）

Fig. 4 Flowering phenotype of the transgenic overexpressing BjuSPL10（A）and its expression level detection in Brassica juncea（B） WT：Wild type plant；OE-1，OE-2：The transgenic overexpressing BjuSPL10 lines. Significance of differences was analyzed using the Duncan’s method，*，** and *** respectively mean significant difference at the P < 0.05，P < 0.01 and P < 0.001. The same below

Fig. 5 Expression levels of BjuSOC1 and BjuFT in BjuSPL10-overexpressing transgenic plants

Fig. 6 Identification of the BjuSPL10 protein interacting with the BjuSOC1 promoter by yeast one-hybrid NC：Negative control；PC：Positive control

Fig. 7 Identification of the BjuSPL10 protein interacting with the BjuSOC1 promoter by luciferase assays

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Functional Identification and Expression Analysis of BjuSPL10 in Regulating Brassica juncea Bolting and Flowering

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