白桦BpDMPs基因家族生物信息学及BpDMP7功能分析

doi:10.7525/j.issn.1673-5102.2025.05.008

摘要/Abstract

摘要：

白桦（Betula platyphylla）作为多年生木本植物因生长周期较长、基因型复杂等问题，育种过程中难以获得纯合植株和家系，进而导致育种工作进展较慢。双单倍体（doubled haploid，DH）育种技术是近年来兴起的一种育种方法，只需要两代即可得到纯合的植物家系。因此，DH育种技术可以大大缩短林木育种周期并获得纯合家系，而DMP基因的发现与应用为双单倍育种开辟了一个新的研究方向。该研究旨在通过生物信息学分析揭示白桦BpDMPs基因家族的理化性质、蛋白结构、系统进化、基因结构、染色体分布及其在白桦生长发育过程中潜在的功能；通过表达模式分析鉴定该基因家族中具有单倍体诱导功能的白桦DMP基因，并通过拟南芥（Arabidopsis thaliana）回补试验验证基因功能。生物信息学分析结果显示：在白桦中，DMP基因包含14个基因家族成员，分布在6条染色体上；系统发育分析表明，BpDMPs基因家族中14个基因分属5个分支；对启动子顺式作用元件的预测分析表明，BpDMPs基因家族的启动子中包含生长发育、激素代谢、胁迫响应等相关的反应元件。qRT-PCR结果显示，BpDMPs在白桦各组织中表达水平差异较大，其中，BpDMP2和BpDMP7在雄蕊中表达量较高，拟南芥回补试验证明，BpDMP7基因可以回补拟南芥dmp8dmp9突变体籽粒败育的表型。为进一步验证BpDMP7诱导白桦产生单倍体，用CRISPER/Cas9基因编辑技术将该基因进行靶向敲除并获得突变体植株。该研究为进一步研究白桦BpDMPs基因家族的功能和调控机制提供了基础信息，并为理解这些基因在植物生长发育及环境适应过程中扮演的角色提供了新的视角。

关键词: 白桦, 单倍体诱导系, DMP基因, 生物信息学

Abstract:

Birch（Betula platyphylla）， as a perennial woody plant， is difficult to obtain homozygous plants and families due to its long growth cycle and complex genotype， which leads to slow progress in its breeding work. Double haploid breeding technology is a breeding method that has emerged in recent years. It only requires two generations to obtain homozygous plant families. Therefore， double haploid breeding technology can greatly shorten the process of forest breeding and obtain homozygous families， while the discovery and application of the DMP gene has opened up a new research direction in doubled haploid breeding. This article aimed to study the BpDMPs gene family in birch， and revealed its physicochemical properties， protein structure， systematic evolution， gene structure， chromosome distribution， and potential functions in the growth and development process of birch through bioinformatics analysis； to identify the birch DMP gene with haploid induction function in the gene family through expression pattern analysis， and verify the gene function through Arabidopsis complementation experiments. The bioinformatics analysis results showed that birch contained 14 gene family members， distributed on six chromosomes. Phylogenetic analysis showed that 14 genes in the BpDMPs gene family belonged to five branches. The predictive analysis of cis-acting elements in the promoter of the BpDMPs gene family showed that the promoter contained response elements related to growth and development， hormone metabolism， stress response， and so on. The qRT-PCR results showed that there were significant differences in the expression levels of BpDMPs in various tissues of birch， with BpDMP2 and BpDMP7 having higher expression levels in the stamens. Through Arabidopsis complementation experiments， it was demonstrated that the BpDMP7 gene can replenish the phenotype of Arabidopsisdmp8dmp9 mutant grain sterility. This article provided basic information for further studying the functions and regulatory mechanisms of the BpDMPs gene family， and offered a new perspective for understanding the roles these genes play in plant growth， development， and environmental adaptation processes.

Key words: birch, haploid induction system, DMP gene, bioinformatics

中图分类号:

S792.153

武思尧, 王新宇, 孙子腾, 郑志民. 白桦BpDMPs基因家族生物信息学及BpDMP7功能分析[J]. 植物研究, 2025, 45(5): 731-744.

Siyao WU, Xinyu WANG, Ziteng SUN, Zhimin ZHENG. Bioinformatics Analysis of the BpDMPs Gene Family in Betula platyphylla and Exploration of the Function of BpDMP7 gene[J]. Bulletin of Botanical Research, 2025, 45(5): 731-744.

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基因名称 Gene name	基因ID Gene ID	所在染色体 Chromosome	基因位置 Gene location	氨基酸数目 Number of amino acids	分子质量 Molecular mass/Da	等电点 Isoelectric point	总平均亲水性 Total average hydrophilicity	亚细胞定位 Subcellular location
BpDMP1	BPChr08G17023	8	38650991-38651641	216	23 342.28	6.50	0.292	CP
BpDMP2	BPChr07G18894	7	22585554-22586180	208	22 727.21	5.61	0.279	CM
BpDMP3	BPChr11G17728	11	3219693-3220379	228	24 285.32	9.18	0.347	CM
BpDMP4	BPChr11G13419	11	25274063-25274683	206	22 923.87	9.03	0.264	CP，Mito，Nucleus
BpDMP5	BPChr08G05126	8	10638147-10638761	204	21 735.80	8.54	0.093	CM，CW
BpDMP6	BPChr01G23597	1	6378106-6378753	215	24 248.03	8.15	0.191	CM
BpDMP7	BPChr06G30721	6	19021326-19021925	199	21 589.71	5.84	0.232	CM
BpDMP8	BPChr08G05134	8	10612115-10612687	190	20 441.58	8.66	0.357	CM
BpDMP9	BPChr12G26817	12	3237953-3238363	156	17 233.41	9.13	0.160	CP
BpDMP10	BPChr06G30778	6	19025688-19026779	363	39 524.96	5.56	-0.182	CM
BpDMP11	BPChr10G03440	10	10633218-10633790	190	20 246.45	8.79	0.297	CM
BpDMP12	BPChr10G03567	10	10667452-10668458	198	21 427.86	9.08	0.360	CM
BpDMP13	BPChr10G03436	10	10671858-10672481	207	22 453.96	6.25	0.561	CM
BpDMP14	BPChr10G03476	10	10661594-10661917	107	11 381.06	9.64	-0.040	CM

载体名称 Vector name	外植体数量 Number of explants	阳性植株数量 Number of positive plants	转化效率 Transformation efficiency/%	基因编辑数量（Indel或有意义突变） Number of gene edits （Indel or significant mutation）	基因编辑效率（Indel或有意义突变） Gene editing efficiency （Indel or significant mutation）/%
合计	175	131	74.06	4	3.05
pSC1-CRISPR-BpU6-2-sgRNA1	76	56	73.68	2	3.57
pSC1-CRISPR-BpU6-2-sgRNA2	66	52	78.79	2	3.85
pSC1-CRISPR-BpU6-2-sgRNA3	33	23	69.70	—	—