Gene Family Function and Expression Analysis of BnEXO in Brassica napus

doi:10.7668/hbnxb.20194909

Abstract

Abstract:

The EXORDIUM(EXO) gene was identified in Arabidopsis thaliana as an brassinosteroid(BR)-responsive gene that promotes plant growth by mediating cell expansion.In order to investigate the function of EXO gene in Brassica napus and its expression pattern in different tissues at flowering stage,we used B.napus Zhongshuang 6 as the material,cloned the sequence of the coding region of the EXO gene named BnEXO,and carried out bioinformatics analysis,and used Real-time Fluorescence Quantification to determine the relative expression of BnEXO gene in B.napus in roots,stems,leaves,petals,buds,and pericarps at flowering stage.The results showed that the CDS sequence of BnEXO gene was 945 bp,BnEXO was a stable hydrophilic non-transmembrane protein,which belonged to secreted proteins and was expressed extracellularly,and the secondary structure of the protein was dominated by the random coil.The results of expression analysis in different tissues showed that the expression of BnEXO gene in different tissues was in the order of petals,pericarps,buds,stems,roots and leaves,and the highest expression was found in petals.In addition,20 B.napus EXO genes(BnaEXO),11 B.rapa EXO genes (BraEXO),and 11 B.oleracea EXO genes (BoEXO)were identified in this study based on the protein sequences of eight EXO gene family members in A.thaliana.Most proteins of gene family members were stable proteins,localized extracellularly,with amino acid lengths ranging from 271 to 411 aa,isoelectric point predictions ranging from 5.76 to 9.60,and molecular masses ranging from 28.76 to 46.21 ku.Phylogenetic analysis classified the EXO genes into five subgroups,EXOA,EXOB,EXOC,EXOD,and EXOE,with the least number of members in the EXOB subgroup.Gene structure analyses showed that most members contained only one exon and no intron,and the sequences of EXO gene family members were highly conserved.The results of cis-element analysis of the promoter region of the members in B.napus indicated that the BnaEXO genes play important roles in plant growth and development and in adversity stress.

Key words: Brassica napus, EXORDIUM, Cloning, Gene family, Brassinosteroid

CHEN Yudie, ZHANG Zerong, LI Hengxiang, LI Tianle, ZENG Sijie, WU Xianmeng, XIONG Xinghua, XIAO Gang. Gene Family Function and Expression Analysis of BnEXO in Brassica napus[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(5): 24-32. doi: 10.7668/hbnxb.20194909.

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References 22

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引物 Primer	引物序列(5'-3') Primer sequence(5'-3')
BnEXOqF	CGGCGAATTCAAACCAGCTC
BnEXOqR	CCTGCGGCGAGTTTGTAGTA
ActinF	GGTTGGGATGGACCAGAAGG
ActinR	TCAGGAGCAATACGGAGC

引物 Primer	引物序列(5'-3') Primer sequence(5'-3')
BnEXOqF	CGGCGAATTCAAACCAGCTC
BnEXOqR	CCTGCGGCGAGTTTGTAGTA
ActinF	GGTTGGGATGGACCAGAAGG
ActinR	TCAGGAGCAATACGGAGC

物种 Species	基因名称 Gene name	基因ID Gene ID	氨基酸长度/个 Length of amino acids	等电点 pI	分子质量 /ku Molecular weight	亚细胞定位 Predicted subcellular localization	不稳定指数 Instability index	疏水指数 Hydrophobicity index
甘蓝型油菜	BnaEXO-A03	BnaA03g23810D	311	9.04	33.25	细胞外	25.88	-0.162
Brassica napus	BnaEXO-C03	BnaC03g28310D	314	9.04	33.63	细胞外	21.61	-0.198
	BnaEXL2-A02	BnaA02g34120D	304	9.48	32.45	细胞外	25.72	0.006
	BnaEXL2-A06	BnaA06g23380D	304	9.43	32.53	细胞外	27.69	0.002
	BnaEXL2-C02	BnaC02g48820D	304	9.54	32.42	细胞外	26.78	0.014
	BnaEXL2-C03	BnaC03g49920D	304	9.43	32.62	细胞外	29.89	0.011
	BnaEXL3-Ann	BnaAnng08160D	337	9.00	36.98	细胞外	35.28	-0.050
	BnaEXL3-C02	BnaC02g15670D	337	9.00	36.98	细胞外	35.28	-0.050
	BnaEXL4-Ann	BnaAnng05850D	276	9.23	29.34	细胞外	27.08	0.055
	BnaEXL4-A10	BnaA10g22530D	297	9.50	31.63	细胞外	32.02	0.089
	BnaEXL4a-C02	BnaC02g00880D	280	8.81	29.95	细胞外	34.00	0.010
	BnaEXL4b-C02	BnaC02g00890D	271	9.42	28.76	细胞外	20.83	0.094
	BnaEXL4-C09	BnaC09g47090D	297	9.60	31.64	细胞外	28.31	0.093
	BnaEXL5-A07	BnaA07g02490D	358	7.66	38.89	细胞外	47.84	-0.147
	BnaEXL5-C07	BnaC07g06340D	360	8.14	39.05	细胞外	47.12	-0.128
	BnaEXL6-A03	BnaA03g28180D	328	8.07	36.19	细胞外	33.47	-0.197
	BnaEXL6-Cnn	BnaCnng06140D	411	8.75	46.21	细胞外	38.09	-0.284
	BnaEXL6-C03	BnaC03g33230D	327	8.07	36.13	细胞外	32.99	-0.205
	BnaEXL7-A05	BnaA05g08790D	321	6.38	36.08	细胞外	37.80	-0.147
	BnaEXL7-C04	BnaC04g10190D	321	6.31	36.12	细胞外	41.82	-0.166
白菜	BraEXO-A03	Bra000660	314	9.21	33.53	细胞外	26.89	-0.175
Brassica rapa	BraEXL2-A02	Bra031927	304	9.48	32.45	细胞外	25.72	0.006
	BraEXL2-A06	Bra024280	304	9.43	32.53	细胞外	27.69	0.002
	BraEXL3-A02	Bra022553	337	9.00	36.98	细胞外	35.28	-0.050
	BraEXL4a-A02	Bra028634	302	9.11	32.28	细胞外	33.29	0.000
	BraEXL4b-A02	Bra028635	295	9.29	31.41	细胞外	25.27	0.060
	BraEXL4-A10	Bra009384	297	9.50	31.66	细胞外	30.18	0.101
	BraEXL5-A10	Bra002034	358	7.66	38.89	细胞外	47.84	-0.147
	BraEXL6-A02	Bra032025	324	5.76	35.92	细胞核	38.48	-0.206
	BraEXL6-A03	Bra001070	327	8.07	36.13	细胞核	33.55	-0.205
	BraEXL7-A05	Bra005364	321	6.38	36.07	细胞外	37.80	-0.139
甘蓝	BoEXO-C3	Bo3g047100	314	9.04	33.61	细胞外	21.61	-0.206
Brassica	BoEXL2-C2	Bo2g166140	304	9.54	32.38	细胞外	26.51	0.023
oleracea	BoEXL2-C3	Bo3g101490	304	9.43	32.62	细胞外	29.89	0.011
	BoEXL3-C2	Bo2g049350	337	9.00	36.96	细胞外	35.39	-0.059
	BoEXL4a-C2	Bo2g009340	290	9.51	30.82	细胞外	20.21	0.066
	BoEXL4b-C2	Bo2g009350	303	9.01	32.50	细胞外	33.26	-0.010
	BoEXL4-C9	Bo9g174010	293	9.54	31.17	细胞外	28.56	0.102
	BoEXL5-C7	Bo7g014780	360	8.14	39.05	细胞外	47.12	-0.128
	BoEXL6-C3	Bo3g055920	327	8.07	36.13	细胞核	32.99	-0.205
	BoEXL6-C5	Bo5g150320	323	7.56	35.92	细胞核	38.78	-0.253
	BoEXL7-C4	Bo4g038980	321	6.31	36.12	细胞外	41.82	-0.166

物种 Species	基因名称 Gene name	基因ID Gene ID	氨基酸长度/个 Length of amino acids	等电点 pI	分子质量 /ku Molecular weight	亚细胞定位 Predicted subcellular localization	不稳定指数 Instability index	疏水指数 Hydrophobicity index
甘蓝型油菜	BnaEXO-A03	BnaA03g23810D	311	9.04	33.25	细胞外	25.88	-0.162
Brassica napus	BnaEXO-C03	BnaC03g28310D	314	9.04	33.63	细胞外	21.61	-0.198
	BnaEXL2-A02	BnaA02g34120D	304	9.48	32.45	细胞外	25.72	0.006
	BnaEXL2-A06	BnaA06g23380D	304	9.43	32.53	细胞外	27.69	0.002
	BnaEXL2-C02	BnaC02g48820D	304	9.54	32.42	细胞外	26.78	0.014
	BnaEXL2-C03	BnaC03g49920D	304	9.43	32.62	细胞外	29.89	0.011
	BnaEXL3-Ann	BnaAnng08160D	337	9.00	36.98	细胞外	35.28	-0.050
	BnaEXL3-C02	BnaC02g15670D	337	9.00	36.98	细胞外	35.28	-0.050
	BnaEXL4-Ann	BnaAnng05850D	276	9.23	29.34	细胞外	27.08	0.055
	BnaEXL4-A10	BnaA10g22530D	297	9.50	31.63	细胞外	32.02	0.089
	BnaEXL4a-C02	BnaC02g00880D	280	8.81	29.95	细胞外	34.00	0.010
	BnaEXL4b-C02	BnaC02g00890D	271	9.42	28.76	细胞外	20.83	0.094
	BnaEXL4-C09	BnaC09g47090D	297	9.60	31.64	细胞外	28.31	0.093
	BnaEXL5-A07	BnaA07g02490D	358	7.66	38.89	细胞外	47.84	-0.147
	BnaEXL5-C07	BnaC07g06340D	360	8.14	39.05	细胞外	47.12	-0.128
	BnaEXL6-A03	BnaA03g28180D	328	8.07	36.19	细胞外	33.47	-0.197
	BnaEXL6-Cnn	BnaCnng06140D	411	8.75	46.21	细胞外	38.09	-0.284
	BnaEXL6-C03	BnaC03g33230D	327	8.07	36.13	细胞外	32.99	-0.205
	BnaEXL7-A05	BnaA05g08790D	321	6.38	36.08	细胞外	37.80	-0.147
	BnaEXL7-C04	BnaC04g10190D	321	6.31	36.12	细胞外	41.82	-0.166
白菜	BraEXO-A03	Bra000660	314	9.21	33.53	细胞外	26.89	-0.175
Brassica rapa	BraEXL2-A02	Bra031927	304	9.48	32.45	细胞外	25.72	0.006
	BraEXL2-A06	Bra024280	304	9.43	32.53	细胞外	27.69	0.002
	BraEXL3-A02	Bra022553	337	9.00	36.98	细胞外	35.28	-0.050
	BraEXL4a-A02	Bra028634	302	9.11	32.28	细胞外	33.29	0.000
	BraEXL4b-A02	Bra028635	295	9.29	31.41	细胞外	25.27	0.060
	BraEXL4-A10	Bra009384	297	9.50	31.66	细胞外	30.18	0.101
	BraEXL5-A10	Bra002034	358	7.66	38.89	细胞外	47.84	-0.147
	BraEXL6-A02	Bra032025	324	5.76	35.92	细胞核	38.48	-0.206
	BraEXL6-A03	Bra001070	327	8.07	36.13	细胞核	33.55	-0.205
	BraEXL7-A05	Bra005364	321	6.38	36.07	细胞外	37.80	-0.139
甘蓝	BoEXO-C3	Bo3g047100	314	9.04	33.61	细胞外	21.61	-0.206
Brassica	BoEXL2-C2	Bo2g166140	304	9.54	32.38	细胞外	26.51	0.023
oleracea	BoEXL2-C3	Bo3g101490	304	9.43	32.62	细胞外	29.89	0.011
	BoEXL3-C2	Bo2g049350	337	9.00	36.96	细胞外	35.39	-0.059
	BoEXL4a-C2	Bo2g009340	290	9.51	30.82	细胞外	20.21	0.066
	BoEXL4b-C2	Bo2g009350	303	9.01	32.50	细胞外	33.26	-0.010
	BoEXL4-C9	Bo9g174010	293	9.54	31.17	细胞外	28.56	0.102
	BoEXL5-C7	Bo7g014780	360	8.14	39.05	细胞外	47.12	-0.128
	BoEXL6-C3	Bo3g055920	327	8.07	36.13	细胞核	32.99	-0.205
	BoEXL6-C5	Bo5g150320	323	7.56	35.92	细胞核	38.78	-0.253
	BoEXL7-C4	Bo4g038980	321	6.31	36.12	细胞外	41.82	-0.166

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