Function Analysis of BcMAX2 in Regulating Axillary Bud Growth in Brassica campestris ssp. chinensis

doi:10.16420/j.issn.0513-353x.2022-0329

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (5): 972-984.doi: 10.16420/j.issn.0513-353x.2022-0329

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

Function Analysis of BcMAX2 in Regulating Axillary Bud Growth in Brassica campestris ssp. chinensis

XU Lanlan, ZHAO Tianzi, ZHANG Wei, ZONG Chen, HUANG Feiyi, WANG Jianjun, HOU Xilin, LI Ying()

National Key Laboratory of Crop Genetics & Germplasm Enhancement and Utilization，Key Laboratory of Biology and Genetic Improvement of Horticultural Crops（East China），Ministry of Agriculture and Rural Affairs，College of Horticulture，Nanjing Agricultural University，Nanjing 210095，China

Received:2022-11-28 Revised:2023-02-27 Online:2023-05-25 Published:2023-05-31
Contact: LI Ying E-mail:yingli@njau.edu.cn

Abstract

Abstract:

MAX2（MORE AXILLARY GROWTH 2）is a gene associated with strigolactone signal transduction. Its function is related to the growth of axillary buds. The effects of MAX2 on the growth of axillary buds were investigated by using Brassica campestris ssp. chinensis the multiple shoot branching cultivar‘Maertou’and common cultivar‘Suzhouqing’. The results showed that there were no differences in the ORF（open reading frame）sequence of BcMAX2 gene obtained from the two materials，but the BcMAX2 promoter sequence of‘Maertou’has a two-base deletion. Subcellular localization analysis indicated BcMAX2 localization in the nucleus. Overexpression of BcMAX2 in Arabidopsis increased BRC1 expression and inhibited axillary bud growth. Silencing of BcMAX2 in‘Maertou’caused the decrease of BcBRC1 expression and promoted the growth of axillary buds. Total of 197 candidate proteins interacting with BcMAX2 were obtained from the cDNA library of Brassica campestris ssp. chinensis. Most of the candidate proteins had binding and catalytic activities，and some of them were involved in signal transduction and stress response of plants. Two interacting proteins were verified by yeast two-hybrid system and bimolecular fluorescence complementation technique. The study showed that BcMAX2 may negatively regulate axillary bud growth by promoting BcBRC1 expression.

Key words: Brassica campestris ssp. chinensis, axillary bud growth, BcMAX2, BcBRC1

CLC Number:

S634.1

XU Lanlan, ZHAO Tianzi, ZHANG Wei, ZONG Chen, HUANG Feiyi, WANG Jianjun, HOU Xilin, LI Ying. Function Analysis of BcMAX2 in Regulating Axillary Bud Growth in Brassica campestris ssp. chinensis[J]. Acta Horticulturae Sinica, 2023, 50(5): 972-984.

Figures/Tables 11

References 36

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引物名称	引物序列（5′-3′）	用途
Primer name	Primer sequence	Usage
BcMAX2	F：ATGGCTTCCACCACTCTCTGC	ORF扩增
BcMAX2	R：TCAGTCAATGATGATGCGGCT	Complete ORF amplification
BcMAX2-P	F：TTCAACCGAATCAATCCACTATTCT	启动子扩增
BcMAX2-P	R：CTCCAAGATTTGCTCTCATGGCTTCCAC	Complete promoter amplification
pRI101-BcMAX2	F：TTCTTCACTGTTGATACATATGATGGCTTCCACCACTCTCTGC	载体构建
pRI101-BcMAX2	R：TCGCCCTTGCTCACCATGGATCCTCAGTCAATGATGATGCGGCT	Construct of vector
RT-BcMAX2	F：AGGAGCTTGTGCTTGACGTT	RT-PCR
RT-BcMAX2	R：AAGACAGCGACAACAACCCT
Actin1	F：GGAGCTGAGAGATTCCGTTG	白菜内参基因
Actin1	R：GAACCACCACTGAGGACGAT	Internal reference gene in Brassica
Actin2	F：TTGACAATTGATGCAAACAATGACG	拟南芥内参基因
Actin2	R：CCATTGCTTAATTCCACGGACAAAC	Internal reference genein Arabidopsis
BcMAX2-BD	F：GGAATTCCATATGATGGCTTCCACCACTCTCTGC	诱饵载体构建
BcMAX2-BD	R：GGAATTCGTCAATGATGATGCGGCT	Construct of bait vector
BcNF-YC4-AD	F：CATATGGCCATGGAGGCCAGTGAATTCATGGACAACAACAACCAGCA	载体构建
BcNF-YC4-AD	R：ATCTGCAGCTCGAGCTCGATGGATCCGGAAGCAAGCTCATTACTAGT	Construct of vector
BcCEBiP-AD	F：CATATGGCCATGGAGGCCAGTGAATTCATGGAAACTTCCCGTTTTACCC	载体构建
BcCEBiP-AD	R：ATCTGCAGCTCGAGCTCGATGGATCCGGGAGAAGGCATGCACAGAAC	Construct of vector
BcBOLA-AD	F：CATATGGCCATGGAGGCCAGTGAATTCATGGAGGAGACAGATCGTTCA GATCGTTC	载体构建 Construct of vector
BcBOLA-AD	R：ATCTGCAGCTCGAGCTCGATGGATCCGGGATGGGAATGTCTTGATGGG
AD	F：CTATTCGATGATGAAGATACCCCACCAAACCC	cDNA插入片段扩增
AD	R：GTGAACTTGCGGGGTTTTTCAGTATCTACGATT	Amplification of cDNA fragments
pCAMBIA1300-35S -NYFP-BcNF-YC4	F：GAATTCATGGACAACAACAACCAGCA	BiFC载体构建
pCAMBIA1300-35S -NYFP-BcNF-YC4	R：GTCGACAAGCAAGCTCATTACTAGT	Construct of vector BiFC
pCAMBIA1300-35S -NYFP-BcCEBiP	F：GGATCCATGGAAACTTCCCGTTTTACCC	BiFC载体构建
pCAMBIA1300-35S -NYFP-BcCEBiP	R：ACTAGTGAGAAGGCATGCACAGAAC	Construct of vector BiFC
pCAMBIA1300-35S -NYFP-BcBOLA	F：GGATCCATGGAGGAGACAGATCGTTC	BiFC载体构建
pCAMBIA1300-35S -NYFP-BcBOLA	R：ACTAGTGATGGGAATGTCTTGATGGG	Construct of vector BiFC
pCAMBIA1300-35S -CYFP-BcMAX2	F：GAATTCATGGCTTCCACCACTCTCTGC	BiFC载体构建
pCAMBIA1300-35S -CYFP-BcMAX2	R：GTCGACGGTCAATGATGATGCGGCT	Construct of vector BiFC

引物名称	引物序列（5′-3′）	用途
Primer name	Primer sequence	Usage
BcMAX2	F：ATGGCTTCCACCACTCTCTGC	ORF扩增
BcMAX2	R：TCAGTCAATGATGATGCGGCT	Complete ORF amplification
BcMAX2-P	F：TTCAACCGAATCAATCCACTATTCT	启动子扩增
BcMAX2-P	R：CTCCAAGATTTGCTCTCATGGCTTCCAC	Complete promoter amplification
pRI101-BcMAX2	F：TTCTTCACTGTTGATACATATGATGGCTTCCACCACTCTCTGC	载体构建
pRI101-BcMAX2	R：TCGCCCTTGCTCACCATGGATCCTCAGTCAATGATGATGCGGCT	Construct of vector
RT-BcMAX2	F：AGGAGCTTGTGCTTGACGTT	RT-PCR
RT-BcMAX2	R：AAGACAGCGACAACAACCCT
Actin1	F：GGAGCTGAGAGATTCCGTTG	白菜内参基因
Actin1	R：GAACCACCACTGAGGACGAT	Internal reference gene in Brassica
Actin2	F：TTGACAATTGATGCAAACAATGACG	拟南芥内参基因
Actin2	R：CCATTGCTTAATTCCACGGACAAAC	Internal reference genein Arabidopsis
BcMAX2-BD	F：GGAATTCCATATGATGGCTTCCACCACTCTCTGC	诱饵载体构建
BcMAX2-BD	R：GGAATTCGTCAATGATGATGCGGCT	Construct of bait vector
BcNF-YC4-AD	F：CATATGGCCATGGAGGCCAGTGAATTCATGGACAACAACAACCAGCA	载体构建
BcNF-YC4-AD	R：ATCTGCAGCTCGAGCTCGATGGATCCGGAAGCAAGCTCATTACTAGT	Construct of vector
BcCEBiP-AD	F：CATATGGCCATGGAGGCCAGTGAATTCATGGAAACTTCCCGTTTTACCC	载体构建
BcCEBiP-AD	R：ATCTGCAGCTCGAGCTCGATGGATCCGGGAGAAGGCATGCACAGAAC	Construct of vector
BcBOLA-AD	F：CATATGGCCATGGAGGCCAGTGAATTCATGGAGGAGACAGATCGTTCA GATCGTTC	载体构建 Construct of vector
BcBOLA-AD	R：ATCTGCAGCTCGAGCTCGATGGATCCGGGATGGGAATGTCTTGATGGG
AD	F：CTATTCGATGATGAAGATACCCCACCAAACCC	cDNA插入片段扩增
AD	R：GTGAACTTGCGGGGTTTTTCAGTATCTACGATT	Amplification of cDNA fragments
pCAMBIA1300-35S -NYFP-BcNF-YC4	F：GAATTCATGGACAACAACAACCAGCA	BiFC载体构建
pCAMBIA1300-35S -NYFP-BcNF-YC4	R：GTCGACAAGCAAGCTCATTACTAGT	Construct of vector BiFC
pCAMBIA1300-35S -NYFP-BcCEBiP	F：GGATCCATGGAAACTTCCCGTTTTACCC	BiFC载体构建
pCAMBIA1300-35S -NYFP-BcCEBiP	R：ACTAGTGAGAAGGCATGCACAGAAC	Construct of vector BiFC
pCAMBIA1300-35S -NYFP-BcBOLA	F：GGATCCATGGAGGAGACAGATCGTTC	BiFC载体构建
pCAMBIA1300-35S -NYFP-BcBOLA	R：ACTAGTGATGGGAATGTCTTGATGGG	Construct of vector BiFC
pCAMBIA1300-35S -CYFP-BcMAX2	F：GAATTCATGGCTTCCACCACTCTCTGC	BiFC载体构建
pCAMBIA1300-35S -CYFP-BcMAX2	R：GTCGACGGTCAATGATGATGCGGCT	Construct of vector BiFC

Function Analysis of BcMAX2 in Regulating Axillary Bud Growth in Brassica campestris ssp. chinensis

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