基于BSA-seq的高粱开花时间基因定位及候选基因预测

doi:10.13560/j.cnki.biotech.bull.1985.2025-0890

生物技术通报 ›› 2026, Vol. 42 ›› Issue (2): 197-206.doi: 10.13560/j.cnki.biotech.bull.1985.2025-0890

基于BSA-seq的高粱开花时间基因定位及候选基因预测

王尚封¹(), 程斌²^,³, 王若若³^,⁴, 丁延庆²^,³, 徐建霞²^,³, 曹宁²^,³, 高旭²^,³, 李文贞²^,³, 张立异²^,³()

^1.贵州大学农学院，贵阳 550025
^2.贵州省农业科学院旱粮研究所，贵阳 550006
^3.贵州省特色杂粮分子育种重点实验室，贵阳 550006
^4.贵州省农业科学院生物技术研究所，贵阳 550006

收稿日期:2025-08-18 出版日期:2026-02-26 发布日期:2026-03-17
通讯作者: 张立异，女，博士，研究员，研究方向：作物分子遗传育种；E-mail: lyzhang1997@hotmail.com
作者简介:王尚封，女，硕士研究生，研究方向：作物分子遗传育种；E-mail: 619684541@qq.com
第一联系人：（王尚封、程斌并列第一作者）
基金资助:
贵州省育种科研基础平台创新能力建设(黔科合服企［2022］014);特色杂粮分子育种贵州省重点实验室(黔科合平台ZSYS［2025］026);国家自然科学基金项目(32460493);贵州省基础研究计划（自然科学）面上项目（黔科合基础-zk［2025］面上299），贵州省农业科学院青年科技基金（自然科学）黔农科博士基金(［2025］02)

Gene Mapping of Sorghum Flowering Time and Prediction of Candidate Genes Based on BSA-seq

WANG Shang-feng¹(), CHENG Bin²^,³, WANG Ruo-ruo³^,⁴, DING Yan-qing²^,³, XU Jian-xia²^,³, CAO Ning²^,³, GAO Xu²^,³, LI Wen-zhen²^,³, ZHANG Li-yi²^,³()

^1.College of Agriculture, Guizhou University, Guiyang 550025
^2.Institute of Upland Food Crops, Guizhou Academy of Agricultural Sciences, Guiyang 550006
^3.Guizhou Key Laboratory of Biology and Breeding for Specialty Crops, Guiyang 550006
^4.Institute of Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang 550006

Received:2025-08-18 Published:2026-02-26 Online:2026-03-17

摘要/Abstract

摘要：

目的开花时间作为高粱品种选育过程中的关键目标性状之一，对其生态适应性和高产潜力具有重要影响。方法以晚开花品种红缨子与早开花品种SAP001杂交构建的F₂群体为材料进行开花时间遗传定位分析。结果表型调查显示，F₂群体中晚花和早花单株的分离比符合3∶1（χ²=0.225，P>0.05），表明该性状受到一对主效基因的控制。通过对极端早花和晚花株系的BSA-Seq分析，在第3染色体59.73‒60.44 Mb区间检测到一个与开花时间显著关联的遗传位点。结合双亲重测序和转录组数据，鉴定到一个编码光系统Ⅱ叶绿素结合蛋白的关键候选基因（SbiHYZ.03G218800）。该基因启动子区存在Myb/SANT、TATA结合蛋白（TBP）结合位点突变，在亲本间表达差异显著，可能为高粱中调控开花时间的新基因。基于该候选区段中的SNP和InDel变异，开发了一个可有效区分早花与晚花植株连锁分子标记。结论揭示了中国酒用高粱开花时间的遗传基础，为开展高粱生育期分子育种提供了重要基因资源和分子标记。

关键词: 高粱, 开花时间, BSA-seq, 转录组测序, 候选基因

Abstract:

Objective Flowering time is a key target trait in sorghum breeding, significantly influencing its ecological adaptability and high-yield potential. Method An F₂ population derived from a cross between the late-flowering variety Hongyingzi and the early-flowering variety SAP001 was used for genetic mapping of flowering time. Result Phenotypic investigation revealed that the segregation ratio of late-flowering and early-flowering plants was consistent with 3∶1(χ²=0.225, P>0.05), indicating that the trait was controlled by a pair of major genes. BSA-Seq analysis of extreme early-and late-flowering lines detected a genetic locus significantly associated with flowering time within the 59.73‒60.44 Mb interval on chromosome 3. By integrating whole-genome resequencing of the parents and transcriptome data, a key candidate gene (SbiHYZ.03G218800), encoding a light-harvesting complex Ⅱ chlorophyll-binding protein, was identified as a putative novel regulator of flowering time in sorghum (Sorghum bicolor (L.)Moench). There were mutations in the binding sites of Myb/SANT and TATA-binding protein (TBP) within its promoter region, and the expression of this gene differed significantly between the parents, possibly it was a new gene regulating flowering time of sorghum. Based on SNP and InDel variations in this candidate region, a closely linked molecular marker effectively distinguishing between early- and late-flowering plants was developed. Conclusion This study preliminarily reveals the genetic basis of flowering time in Chinese liquor-brewing sorghum germplasm and provides valuable genetic resources and molecular markers for molecular breeding of sorghum maturity.

Key words: sorghum, flowering time, BSA-seq, transcriptome sequencing, candidate genes

王尚封, 程斌, 王若若, 丁延庆, 徐建霞, 曹宁, 高旭, 李文贞, 张立异. 基于BSA-seq的高粱开花时间基因定位及候选基因预测[J]. 生物技术通报, 2026, 42(2): 197-206.

WANG Shang-feng, CHENG Bin, WANG Ruo-ruo, DING Yan-qing, XU Jian-xia, CAO Ning, GAO Xu, LI Wen-zhen, ZHANG Li-yi. Gene Mapping of Sorghum Flowering Time and Prediction of Candidate Genes Based on BSA-seq[J]. Biotechnology Bulletin, 2026, 42(2): 197-206.

图/表 9

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引物名称 Primer name	orward primer （5′‒3′）正向引物 F	everse primer （5′‒3′）反向引物 R
V1f1	CAACTTCAGTGCAACGGTCG	AGCAAGGATTGTGTGGTGCT
V423	TCAGTGCAACGGTCGACAAA	TGAATGGTGAACAGCCGACC
Hd8800	CACATCAGTGTGTGCCGTTG	TTAGGGGCGTTCTTCTGCTC
V5f1	CATCCACGGGGTTGAAGTCT	AGCGGTCCATTGCTCAAAGT

引物名称 Primer name	orward primer （5′‒3′）正向引物 F	everse primer （5′‒3′）反向引物 R
V1f1	CAACTTCAGTGCAACGGTCG	AGCAAGGATTGTGTGGTGCT
V423	TCAGTGCAACGGTCGACAAA	TGAATGGTGAACAGCCGACC
Hd8800	CACATCAGTGTGTGCCGTTG	TTAGGGGCGTTCTTCTGCTC
V5f1	CATCCACGGGGTTGAAGTCT	AGCGGTCCATTGCTCAAAGT

样本Sample	平均测序深度Mean depth （X）	短片段总数Number of total Reads	总碱基读数Clean bases （bp）	GC （%）	Q30 （%）	比对Mapped reads （%）
红缨子	67.04	369 858 258	55 478 738 700	44.63	91.96	99.41
SAP001	70.27	355 144 466	53 271 669 900	43.54	96.28	98.88
晚熟池	77.31	387 856 226	58 178 433 900	43.67	97.29	99.34
早熟池	86.91	443 492 672	66 523 900 800	43.53	97.36	98.95

样本Sample	平均测序深度Mean depth （X）	短片段总数Number of total Reads	总碱基读数Clean bases （bp）	GC （%）	Q30 （%）	比对Mapped reads （%）
红缨子	67.04	369 858 258	55 478 738 700	44.63	91.96	99.41
SAP001	70.27	355 144 466	53 271 669 900	43.54	96.28	98.88
晚熟池	77.31	387 856 226	58 178 433 900	43.67	97.29	99.34
早熟池	86.91	443 492 672	66 523 900 800	43.53	97.36	98.95

基因 Gene	变异位置 Variation position （bp）	参考基因组 Reference	等位变异Alternative	CDS突变类型Type of CDS mutation	启动子突变Promoter mutation
SbiHYZ.03G216400	59 930 660	A	-	-	碱基缺失
SbiHYZ.03G216700	59 972 784	C	A	非同义突变
SbiHYZ.03G216800	59 974 642	C	A	-	碱基突变
SbiHYZ.03G216900	59 980 499	A	-	缺失突变/移码
	59 980 657	G	A	非同义突变
	59 980 792	G	T	非同义突变
	59 980 816	G	C	非同义突变
SbiHYZ.03G217000	60 060 314	A	-	-	碱基缺失
SbiHYZ.03G217200	60 093 586	A	C	非同义突变
SbiHYZ.03G217700	60 140 432	G	A	非同义突变
SbiHYZ.03G218800	60 365 936	C	T	-	碱基突变
SbiHYZ.03G218900	60 375 442	C	T	非同义突变

基于BSA-seq的高粱开花时间基因定位及候选基因预测

Gene Mapping of Sorghum Flowering Time and Prediction of Candidate Genes Based on BSA-seq

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基因 Gene	物理位置 Physical position （bp）	同源基因 Paralogous gene	功能注释 Functional annotation	相似率 Similarity （%）
SbiHYZ.03G216400	59 931 181‒59 935 674	LOC_Os01g41250	F-box/FBD结构域 F-box/FBD domain	66
SbiHYZ.03G216700	59 971 690‒59 974 228	LOC_Os01g41260	F-box/FBD结构域 F-box/FBD domain	63
SbiHYZ.03G216800	59 974 820‒59 977 137	Zm00001d011276	F-box/FBD结构域 F-box/FBD domain	80
SbiHYZ.03G216900	59 978 600‒59 981 036	LOC_Os01g41310	F-box/FBD结构域 F-box/FBD domain	51
SbiHYZ.03G217000	60 061 333‒60 063 755	Zm00001d044385	F-box/FBD结构域 F-box/FBD domain	83
SbiHYZ.03G217200	60 090 312‒60 093 766	Zm00001d044386	F-box/FBD结构域 F-box/FBD domain	75
SbiHYZ.03G217700	60 139 306‒60 141 819	LOC_Os01g41530	F-box/FBD结构域 F-box/FBD domain	50
SbiHYZ.03G218800	60 366 296‒60 367 982	ZM00001d011285	光捕获复合物Ⅱ叶绿素a/b结合蛋白 Light-harvesting complexⅡchlorophyll a/b-binding protein	97
SbiHYZ.03G218900	60 374 509‒60 375 642	Zm00001d011285	光捕获复合物Ⅱ叶绿素a/b结合蛋白 Light-harvesting complexⅡ chlorophyll a/b-binding protein	98

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