Genome-wide Identification and Expression Analysis of the BOI Gene Family in Nicotiana benthamiana

doi:10.13560/j.cnki.biotech.bull.1985.2025-0789

Abstract

Abstract:

Objective To clarify the roles of RING-type ubiquitin E3 ligase Botrytis Susceptible1 Interactor (BOI) in gibberellin (GA) signal transduction and cucumber green mottle mosaic virus (CGMMV) infection, as well as the characteristics of its gene family, thus to provide a theoretical basis for understanding the molecular mechanisms of antiviral defense in Solanaceae crops and their chemical regulation. Method Bioinformatics approaches were used to analyze the phylogenetic relationships, gene structure, conserved motifs, promoter cis-acting elements, transcription factors, and tissue expression characteristics of NbBOI, and fluorescence quantitative PCR was applied to verify the expression changes of NbBOI family members under biotic stress and hormone treatments, and yeast two-hybrid assay was employed to screen out the BOI family genes that interacted with CGMMV encoded proteins. Result A total of thirteen BOI genes were identified in Nicotiana benthamiana, and they were divided into four subfamilies. The Motifs and gene structures of BOI family members in the same evolutionary branch were highly consistent. Six genes related to the gibberellin pathway were identified through prediction and promoter cis-acting elements analysis. Fluorescence quantitative PCR detection showed that members of the NbBOI family had differences in tissue-specific expression patterns and revealed different response patterns to virus infection and gibberellin treatment. The yeast two-hybrid assay verified that NbBRG2.1 interacted with the coat protein (CP) and movement protein encoded by CGMMV. Conclusion The BOI gene family in N. benthamiana shows high conservation and is involved in plant biotic stress defense and hormone signal transduction. Among its members, NbBRG1.2 plays a crucial role in the gibberellin (GA) biosynthesis pathway, while NbBOI.1, NbBOI.2, NbBOI.3, NbBRG1.2, NbBRG2.1, and NbBRG2.3 play a certain role in the defense mechanism of plants against CGMMV infection. During the viral infection process, viral proteins hijack the NbBRG2.1, thereby disrupting the hormonal balance in the host.

Key words: Nicotiana benthamiana, botrytis susceptible1 interactor, tissue specificity, virus infection, gibberellin signal transduction

FU Han, SUN Shu-hao, ZHANG Si-qing, AI Niu, YU Yang, YU Lian-wei, WANG Qiong-qiong, HAN Xiao-yu, SHI Yan, HAN Wei-li, YANG Xue. Genome-wide Identification and Expression Analysis of the BOI Gene Family in Nicotiana benthamiana[J]. Biotechnology Bulletin, 2026, 42(2): 207-217.

Figures/Tables 11

References 31

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基因名称 Gene name	上游引物 Forward primer （5'-3'）	下游引物 Reverse primer （5'-3'）
NbBOI.1	CGGGTGTAACGAGAGAGACG	AACACACAAATGGCGACACG
NbBOI.2	CTAGTCAATCATCGGCTTTCCA	TAGGCGTGCCATTGTTTCCT
NbBOI.3	TTGACTTAACCCAGCTCCAC	AGAATGATGTTGTTTTTGTTGGTGC
NbBRG1.2	CAACAGCAGTTGCAAAAAGACAA	GCGTCAATCTCAATCGCTCC
NbBRG2.1	TAGCTCCAGGTCCAGCTGAT	ACCCAAACGTCCATTATCCA
NbBRG2.3	ACTATTACACCACCGCCACTT	GGCATCCAAATTGCAGCTGAG

基因名称 Gene name	上游引物 Forward primer （5'-3'）	下游引物 Reverse primer （5'-3'）
NbBOI.1	CGGGTGTAACGAGAGAGACG	AACACACAAATGGCGACACG
NbBOI.2	CTAGTCAATCATCGGCTTTCCA	TAGGCGTGCCATTGTTTCCT
NbBOI.3	TTGACTTAACCCAGCTCCAC	AGAATGATGTTGTTTTTGTTGGTGC
NbBRG1.2	CAACAGCAGTTGCAAAAAGACAA	GCGTCAATCTCAATCGCTCC
NbBRG2.1	TAGCTCCAGGTCCAGCTGAT	ACCCAAACGTCCATTATCCA
NbBRG2.3	ACTATTACACCACCGCCACTT	GGCATCCAAATTGCAGCTGAG

基因名 Gene name	基因ID Gene ID	氨基酸数 Protein length （aa）	分子量 Molecular weight （kD）	等电点 pI	不稳定指数Instability index	总平均亲水系数Grand average of hydropathicity	脂肪指数 Aliphatic index	亚细胞定位 Subcellular location
NbBRG2.1	Niben101Scf04995g03009.1	337	38 311.27	5.23	48.590	-0.597	83.89	Nucleus
NbBRG2.2	Niben101Scf05551g00006.1	579	65 546.86	6.53	43.220	-0.643	75.82	Nucleus
NbBRG2.3	Niben101Scf09599g00015.1	361	40 133.59	7.49	36.470	-0.487	81.52	Nucleus
NbBRG2.4	Niben101Scf00468g01013.1	347	38 916.32	7.88	39.380	-0.493	74.21	Nucleus
NbBRG3.1	Niben101Scf25562g00007.1	327	35 967.52	5.71	49.640	-0.343	84.16	Nucleus
NbBRG3.2	Niben101Scf02156g11001.1	349	38 832.05	5.89	43.620	-0.464	77.91	Nucleus
NbBRG3.3	Niben101Scf04499g00008.1	350	39 076.28	5.9	47.580	-0.476	78.26	Nucleus
NbBRG3.4	Niben101Scf01455g00005.1	431	47 614.25	5.09	41.520	-0.472	77.42	Nucleus
NbBRG1.1	Niben101Scf06106g01018.1	283	32 891.4	5.92	63.690	-0.632	77.53	Nucleus
NbBRG1.2	Niben101Scf07614g02001.1	269	31 231.47	7.92	72.720	-0.715	65.32	Nucleus
NbBOI.1	Niben101Scf04159g01016.1	316	34 681.7	6.52	68.610	-0.671	66.2	Nucleus
NbBOI.2	Niben101Scf01526g12008.1	338	37 295.86	6.24	64.790	-0.501	79.44	Nucleus
NbBOI.3	Niben101Scf06847g02001.1	339	37 640.29	7.2	67.220	-0.561	79.23	Nucleus

基因名 Gene name	基因ID Gene ID	氨基酸数 Protein length （aa）	分子量 Molecular weight （kD）	等电点 pI	不稳定指数Instability index	总平均亲水系数Grand average of hydropathicity	脂肪指数 Aliphatic index	亚细胞定位 Subcellular location
NbBRG2.1	Niben101Scf04995g03009.1	337	38 311.27	5.23	48.590	-0.597	83.89	Nucleus
NbBRG2.2	Niben101Scf05551g00006.1	579	65 546.86	6.53	43.220	-0.643	75.82	Nucleus
NbBRG2.3	Niben101Scf09599g00015.1	361	40 133.59	7.49	36.470	-0.487	81.52	Nucleus
NbBRG2.4	Niben101Scf00468g01013.1	347	38 916.32	7.88	39.380	-0.493	74.21	Nucleus
NbBRG3.1	Niben101Scf25562g00007.1	327	35 967.52	5.71	49.640	-0.343	84.16	Nucleus
NbBRG3.2	Niben101Scf02156g11001.1	349	38 832.05	5.89	43.620	-0.464	77.91	Nucleus
NbBRG3.3	Niben101Scf04499g00008.1	350	39 076.28	5.9	47.580	-0.476	78.26	Nucleus
NbBRG3.4	Niben101Scf01455g00005.1	431	47 614.25	5.09	41.520	-0.472	77.42	Nucleus
NbBRG1.1	Niben101Scf06106g01018.1	283	32 891.4	5.92	63.690	-0.632	77.53	Nucleus
NbBRG1.2	Niben101Scf07614g02001.1	269	31 231.47	7.92	72.720	-0.715	65.32	Nucleus
NbBOI.1	Niben101Scf04159g01016.1	316	34 681.7	6.52	68.610	-0.671	66.2	Nucleus
NbBOI.2	Niben101Scf01526g12008.1	338	37 295.86	6.24	64.790	-0.501	79.44	Nucleus
NbBOI.3	Niben101Scf06847g02001.1	339	37 640.29	7.2	67.220	-0.561	79.23	Nucleus