Identification of the EfBBX Gene Family in Erianthus fulvus and Analysis of Its Expression Patterns Under Cold Stress

doi:10.13560/j.cnki.biotech.bull.1985.2025-0908

Abstract

Abstract:

Objective B-box zinc finger proteins (BBX proteins) play a crucial role in defending plants against abiotic stress. Erianthus fulvus has excellent cold tolerance and serves as a valuable genetic resource for improving cold resistance in sugarcane. We identified and characterized members of the EfBBX gene family in E. fulvus, and explored their biological functions under cold stress. Method Members of the EfBBX gene family were identified from the whole genome of E. fulvus. Bioinformatics methods were employed to analyze their physicochemical properties, phylogenetic relationships, chromosomal localization, synteny analysis, gene structure, conserved domains, and promoter cis-elements. RT-qPCR experiments were conducted to examine the expression patterns of EfBBX family members under cold stress. Using the E. fulvus clonal line E2 as material, the coding sequences (CDS) of EfBBX4, EfBBX10, and EfBBX18 were cloned. The EfBBX18 gene was then transiently expressed in Nicotiana benthamiana. Result Nineteen members of the EfBBX gene family were identified in the whole genome of E. fulvus, classified into five subfamilies and unevenly distributed across six of its ten chromosomes. Subcellular localization predictions indicated that EfBBX proteins were primarily localized in the nucleus and chloroplasts. The promoter regions of the EfBBX gene family contain numerous elements associated with hormone, abiotic stress response, and light response pathways. RT-qPCR experiments confirmed that ten genes were induced by cold stress. The EfBBX4, EfBBX10, and EfBBX18 genes were successfully cloned, and transient expression of E. fulvusEfBBX18 in Nicotiana benthamiana enhanced the plant’s cold tolerance. Conclusion Nineteen EfBBX genes are identified from the E. fulvus genome, with ten genes inducible by cold stress. Three of these genes are successfully cloned, and transient expression of EfBBX18 in N. benthamiana enhances the plant’s tolerance to cold.

Key words: Erianthusfulvus, BBX gene family, identification gene family, bioinformatics, cold stress, expression pattern, transient expression, cold resistance

NONG Wei-you, ZHAO Chang-zu, QIAN Zhen-feng, DING Qian, WANG Yu-jie, CHEN Shu-ying, HE Li-lian, LI Fu-sheng. Identification of the EfBBX Gene Family in Erianthus fulvus and Analysis of Its Expression Patterns Under Cold Stress[J]. Biotechnology Bulletin, 2026, 42(2): 1-11.

Figures/Tables 8

References 52

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基因 Gene	基因IDGene ID	氨基酸数量 Number of amino Acids （aa）	相对分子量 Molecular Weight （Da）	等电点PI	不稳定指数 Instability index	脂肪指数 Aliphatic index	亚细胞定位 Subcellular localization
EfBBX1	Erufi.01G0010670.t1	420	45 969.84	5.88	55.68	64.21	叶绿体 Chloroplast
EfBBX2	Erufi.03G0003170.t1	358	37 106.31	5.42	51.18	65.67	细胞核 Nuclear
EfBBX3	Erufi.04G0006700.t1	373	38 846.33	5.80	45.40	65.39	叶绿体 Chloroplast
EfBBX4	Erufi.04G0019680.t1	257	27 606.01	4.95	43.19	76.54	细胞质 Cytoplasmic
EfBBX5	Erufi.04G0020000.t1	328	34 599.45	5.12	44.90	66.10	细胞质 Cytoplasmic
EfBBX6	Erufi.04G0022180.t1	276	28 816.93	5.38	61.93	63.91	细胞核 Nuclear
EfBBX7	Erufi.04G0026320.t1	405	44 398.46	5.30	50.12	61.73	细胞核 Nuclear
EfBBX8	Erufi.04G0026680.t1	374	41 614.82	6.42	56.63	70.16	叶绿体 Chloroplast
EfBBX9	Erufi.04G0026930.t1	476	51 626.87	5.68	65.17	63.87	叶绿体 Chloroplast
EfBBX10	Erufi.06G0013060.t1	255	27 181.54	4.80	49.61	69.76	细胞核 Nuclear
EfBBX11	Erufi.06G0013390.t1	354	37 454.85	5.46	38.07	63.73	叶绿体 Chloroplast
EfBBX12	Erufi.06G0016230.t1	261	28 309.31	4.89	70.95	61.07	细胞核 Nuclear
EfBBX13	Erufi.09G0008130.t1	327	34 135.01	5.07	56.04	63.15	细胞核 Nuclear
EfBBX14	Erufi.10G0004850.t1	481	52 067.46	6.60	61.19	67.53	叶绿体 Chloroplast
EfBBX15	Erufi.10G0011520.t1	459	49 201.79	5.42	46.14	68.82	细胞质 Cytoplasmic
EfBBX16	Erufi.10G0012110.t1	403	43 490.53	5.31	48.03	69.58	细胞质 Cytoplasmic
EfBBX17	Erufi.10G0012650.t1	404	43 489.41	5.30	46.75	65.50	细胞核 Nuclear
EfBBX18	Erufi.10G0020560.t1	366	39 089.78	5.76	43.58	69.48	细胞质 Cytoplasmic
EfBBX19	Erufi.10G0025500.t1	303	31 346.08	4.83	57.21	70.99	叶绿体 Chloroplast

基因 Gene	基因IDGene ID	氨基酸数量 Number of amino Acids （aa）	相对分子量 Molecular Weight （Da）	等电点PI	不稳定指数 Instability index	脂肪指数 Aliphatic index	亚细胞定位 Subcellular localization
EfBBX1	Erufi.01G0010670.t1	420	45 969.84	5.88	55.68	64.21	叶绿体 Chloroplast
EfBBX2	Erufi.03G0003170.t1	358	37 106.31	5.42	51.18	65.67	细胞核 Nuclear
EfBBX3	Erufi.04G0006700.t1	373	38 846.33	5.80	45.40	65.39	叶绿体 Chloroplast
EfBBX4	Erufi.04G0019680.t1	257	27 606.01	4.95	43.19	76.54	细胞质 Cytoplasmic
EfBBX5	Erufi.04G0020000.t1	328	34 599.45	5.12	44.90	66.10	细胞质 Cytoplasmic
EfBBX6	Erufi.04G0022180.t1	276	28 816.93	5.38	61.93	63.91	细胞核 Nuclear
EfBBX7	Erufi.04G0026320.t1	405	44 398.46	5.30	50.12	61.73	细胞核 Nuclear
EfBBX8	Erufi.04G0026680.t1	374	41 614.82	6.42	56.63	70.16	叶绿体 Chloroplast
EfBBX9	Erufi.04G0026930.t1	476	51 626.87	5.68	65.17	63.87	叶绿体 Chloroplast
EfBBX10	Erufi.06G0013060.t1	255	27 181.54	4.80	49.61	69.76	细胞核 Nuclear
EfBBX11	Erufi.06G0013390.t1	354	37 454.85	5.46	38.07	63.73	叶绿体 Chloroplast
EfBBX12	Erufi.06G0016230.t1	261	28 309.31	4.89	70.95	61.07	细胞核 Nuclear
EfBBX13	Erufi.09G0008130.t1	327	34 135.01	5.07	56.04	63.15	细胞核 Nuclear
EfBBX14	Erufi.10G0004850.t1	481	52 067.46	6.60	61.19	67.53	叶绿体 Chloroplast
EfBBX15	Erufi.10G0011520.t1	459	49 201.79	5.42	46.14	68.82	细胞质 Cytoplasmic
EfBBX16	Erufi.10G0012110.t1	403	43 490.53	5.31	48.03	69.58	细胞质 Cytoplasmic
EfBBX17	Erufi.10G0012650.t1	404	43 489.41	5.30	46.75	65.50	细胞核 Nuclear
EfBBX18	Erufi.10G0020560.t1	366	39 089.78	5.76	43.58	69.48	细胞质 Cytoplasmic
EfBBX19	Erufi.10G0025500.t1	303	31 346.08	4.83	57.21	70.99	叶绿体 Chloroplast