重齿毛当归核心种质构建及其有效成分关联分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0582

摘要/Abstract

摘要：

目的分析重齿毛当归种质资源遗传多样性，构建核心种质，发掘与有效成分含量关联的分子标记，为重齿毛当归种质保护及品种选育奠定基础。方法对96份重齿毛当归种质表型性状与有效成分含量进行差异分析，并利用12条ISSR核心引物对供试种质进行遗传结构分析及核心种质构建，采用Talssel软件进行ISSR标记与有效成分的关联分析。结果重齿毛当归表型性状与有效成分含量变异较丰富，群体结构分析将供试材料划分为3个亚群，Gen Al Ex软件分析表明，重齿毛当归3个亚群的遗传多样性水平均较高，种群变异主要发生在亚群内。通过15.6%‒58.3%的抽样比例法，构建了37份代表原始群体的核心种质。关联分析的FarmCPU、GLM和MLM模型共同鉴定了CBD含量与3个标记（UBC 812、UBC 816、UBC 819）呈显著正相关（P<0.05）；CBT含量与UBC 816、UBC 817呈显著正相关（P<0.05）；IPT含量与UBC 819呈极显著正相关（P<0.01）；OST含量与UBC 819呈显著正相关（P<0.05）；UMB含量均与UBC 810、UBC 812标记呈显著正相关（P<0.05）。OHN含量仅在GLM模型中发现与UBC 816呈显著正相关（P<0.05）。结论供试重齿毛当归种质被划分为3个亚群，构建的37份核心种质能代表原始群体的遗传多样性水平，筛选出6个与有效成分含量关联的ISSR分子标记。

关键词: 重齿毛当归, 香豆素, 分子标记, 核心种质, 群体结构, 关联分析

Abstract:

Objective The genetic diversity of Angelica pubescens germplasm resources was analyzed, a core collection was constructed, and molecular markers associated with the contents of active ingredients were screened, so as to lay the foundation for the breeding of A. pubescens varieties. Method A differential analysis of the contents of active ingredients in 96 germplasms of A. pubescens was conducted, and 8 ISSR core primers was used to analyze the genetic structure and core collection construction. Talssel software was used to perform association analysis between ISSR markers and active ingredients. Result The variation in the phenotype and the content of active ingredients of A. pubescens is rich, and the population structure analysis can divide the materials into 3 subgroups. Gen Al Ex software analysis shows that the genetic diversity of the three subgroups of A. pubescens is relatively high, and the variation occurs mainly within the subgroups. By using a sampling ratio ranging from 15.6% to 58.3%, we constructed a core collection of 37 samples representing the original population. The association analyses of FarmCPU, GLM, and MLM models indicate that the content of CBD is significantly positively correlated with 3 markers (UBC 812, UBC 816, UBC 819) (P<0.05); CBT content is significantly correlated with UBC 816, UBC 817; IPT content is extremely significantly positively correlated with UBC 819 (P<0.01); OST content is significantly correlated with UBC 819 (P<0.05); UMB content is positively correlated with UBC 810, UBC 812 markers. OHN content is only found to be significantly correlated with UBC 816 in the GLM model. Conclusion The germplasms of A. pubescens can be divided into 3 subgroups, and the constructed core collection of 37 samples can represent the genetic diversity level of the original population, and 6 ISSR molecular markers associated with the content of active ingredients are screened.

Key words: Angelica pubescens, coumarin, molecular marker, core germplasm, population structure, association analysis

蒋小刚, 余凯迪, 郭晓亮. 重齿毛当归核心种质构建及其有效成分关联分析[J]. 生物技术通报, 2025, 41(11): 236-246.

JIANG Xiao-gang, YU Kai-di, GUO Xiao-liang. Construction of Angelica pubescens Core Germplasm and Its Association Analysis of Active Ingredients[J]. Biotechnology Bulletin, 2025, 41(11): 236-246.

图/表 14

参考文献 25

[1]	国家药典委员会. 中华人民共和国药典[M]. 一部. 北京: 中国医药科技出版社, 2020: 274.
	National Pharmacopoeia Committee. Pharmacopoeia of the People’s Republic of China (Part One) [M]. Beijing: China Medical Science and Technology Press, 2020:274.
[2]	郭晓亮, 郭杰, 段媛媛, 等. 重齿毛当归开花与授粉特性初探 [J]. 分子植物育种, 2025, 23(6): 1993-1999.
	Guo XL, Guo J, Duan YY, et al. Preliminary study on flowering and pollination characteristics of Angelica pubescens maxim [J]. Mol Plant Breed, 2025, 23(6): 1993-1999.
[3]	袁媛, 黄璐琦. 论基于基因组学的中药材定向培育策略与展望 [J]. 科学通报, 2024, 69(S1): 499-509.
	Yuan Y, Huang LQ. Research strategy and prospects for the genomics-driven targeted breeding of Chinese herbal medicine varieties [J]. Chin Sci Bull, 2024, 69(S1): 499-509.
[4]	黄璐琦, 吕冬梅, 杨滨, 等. 药用植物种质资源研究的发展‒‒核心种质的构建 [J]. 中国中药杂志, 2005, 30(20): 1565-1568, 1586.
	Huang LQ, Lv DM, Yang B, et al. Development of the study on germplasm resources of medicinal plants: construction of core collection [J]. China J Chin Mater Med, 2005, 30(20): 1565-1568, 1586.
[5]	赵慧, 马芹, 刘振华, 等. 药用植物核心种质研究进展 [J]. 山东农业科学, 2023, 55(7): 173-180.
	Zhao H, Ma Q, Liu ZH, et al. Research progress of core collection of medicinal plants [J]. Shandong Agric Sci, 2023, 55(7): 173-180.
[6]	Liu M, Hu X, Wang X, et al. Constructing a core collection of the medicinal plant Angelica biserrata using genetic and metabolic data [J]. Front Plant Sci, 2020, 11: 600249.
[7]	李嘉惠. 何首乌种质研究及核心种质初步构建 [D]. 广州: 广东药科大学, 2020.
	Li JH. Studies of germplasm and preliminaryconstruction of core collection of Fallopia multiflora (Thunb.) Harald [D]. Guangzhou: Guangdong Pharmaceutical University, 2020.
[8]	莫长明, 谢文娟, 郭文锋, 等. 罗汉果遗传多样性与群体结构及核心种质研究 [J]. 中草药, 2023, 54(18): 6040-6054.
	Mo CM, Xie WJ, Guo WF, et al. Study on genetic diversity and population structure and core collection of Siraitia grosvenorii [J]. Chin Tradit Herbal Drugs, 2023, 54(18): 6040-6054.
[9]	黄梅, 李鑫, 王凯, 等. 裸花紫珠群体遗传结构及其有效成分与SSR标记的关联分析 [J]. 中草药, 2024, 55(23): 8162-8170.
	Huang M, Li X, Wang K, et al. Analysis of population genetics and effective components associations with SSR markers in Callicarpa nudiflora [J]. Chin Tradit Herbal Drugs, 2024, 55(23): 8162-8170.
[10]	刘容秀. 黄芩核心种质构建与优良种质筛选研究 [D]. 北京: 北京中医药大学, 2016.
	Liu RX. Research on the construction of core germplasm and the screening of excellent germplasm of Scutellaria [D]. Beijing: Beijing University of Chinese Medicine, 2016.
[11]	潘妍, 范丽莉, 孙海霞, 等. 独活属植物香豆素类化合物及其生物活性研究进展 [J]. 中国农学通报, 2023, 39(29): 14-23.
	Pan Y, Fan LL, Sun HX, et al. Coumarin compounds of genus Heracleum L. and their bioactivity: a review [J]. Chin Agric Sci Bull, 2023, 39(29): 14-23.
[12]	张玉莲, 刘聪, 赵少迪, 等. 远志种质资源ISSR遗传多样性及其与有效成分关联分析 [J]. 中药材, 2024, 47(11): 2694-2701.
	Zhang YL, Liu C, Zhao SD, et al. Analysis of ISSR genetic diversity of Polygala tenuifolia germplasm resources and its association with active components [J]. J Chin Med Mater, 2024, 47(11): 2694-2701.
[13]	孟祥善, 周玉梅, 代晓华, 等. 黄芪ISSR标记遗传多样性及其与主要药用成分的关联分析 [J]. 中国药学杂志, 2019, 54(24): 2060-2070.
	Meng XS, Zhou YM, Dai XH, et al. Genetic diversity and association analysis between medicinal components and ISSR markers in astragali Radix [J]. Chin Pharm J, 2019, 54(24): 2060-2070.
[14]	吴怡超, 李晓瑜, 何星星, 等. 苦荞种质资源芦丁含量与SSR标记关联分析 [J]. 西南大学学报: 自然科学版, 2023, 45(9): 2-11.
	Wu YC, Li XY, He XX, et al. Association analysis of rutin content with SSR markers in Tartary buckwheat germplasm resources [J]. J Southwest Univ Nat Sci Ed, 2023, 45(9): 2-11.
[15]	郭晓亮, 段媛媛, 郭杰, 等. 重齿毛当归ISSR-PCR反应体系的建立与优化 [J]. 分子植物育种, 2022, 20(8): 2682-2688.
	Guo XL, Duan YY, Guo J, et al. Establishment and optimization of ISSR-PCR reaction system of Angelica pubescence maxim [J]. Mol Plant Breed, 2022, 20(8): 2682-2688.
[16]	张元萍, 仇锦林. 不同炮制工艺独活中4种香豆素类成分的定量分析 [J]. 中国药师, 2020, 23(8): 1659-1662.
	Zhang YP, Qiu JL. Quantitative analysis of four coumarins in Angelica pubescens Radix with different processing methods [J]. China Pharm, 2020, 23(8): 1659-1662.
[17]	刘吉成, 梁爽, 陆国寿. HPLC法同时测定消瘤藤中茵芋苷、7-羟基香豆素、7-羟基-8-甲氧基香豆素的含量 [J]. 中国药师, 2021, 24(11): 2095-2098.
	Liu JC, Liang S, Lu GS. Simultaneous determination of skimmin, 7-hydroxycoumarin and 7-hydroxy-8-methoxycoumarin in Pileostegia tomentella hand. mazz.by HPLC [J]. China Pharm, 2021, 24(11): 2095-2098.
[18]	Walasek M, Grzegorczyk A, Malm A, et al. Bioactivity-guided isolation of antimicrobial coumarins from Heracleum mantegazzianum Sommier & Levier (Apiaceae) fruits by high-performance counter-current chromatography [J]. Food Chem, 2015, 186: 133-138.
[19]	Zhang YC, Bai PG, Zhuang YB, et al. Two O-methyltransferases mediate multiple methylation steps in the biosynthesis of coumarins in Cnidium monnieri [J]. J Nat Prod, 2022, 85(8): 2116-2121.
[20]	郭帅, 张孝军, 王泽, 等. 基于SSR分子标记的资丘独活种质资源遗传多样性分析 [J/OL]. 分子植物育种, 2022. .
	Guo S, Zhang XJ, Wang Z, et al. Genetic diversity analysis of angelicae pubescentis Radix from ziqiu germplasm resources based on SSR molecular markers [J/OL]. Mol Plant Breed, 2022. .
[21]	蒋小刚, 王华, 张美德. 白术种质资源遗传多样性的SRAP分析[J]. 北方园艺, 2022(1): 109-115.
	Jiang XG, Wang H, Zhang MD. Genetic diversity of Atractylodes macrocephala by SRAP markers [J]. North Hortic, 2022(1): 109-115.
[22]	Liu JY, Zhang YW, Han X, et al. An evolutionary population structure model reveals pleiotropic effects of GmPDAT for traits related to seed size and oil content in soybean [J]. J Exp Bot, 2020, 71(22): 6988-7002.
[23]	刘曼. 资丘独活的核心种质库构建 [D]. 武汉: 湖北中医药大学, 2021.
	Liu M. Constructing a core collection of Hubei genuine Angelica biserrata [D]. Wuhan: Hubei University of Chinese Medicine, 2021.
[24]	Frankel OH, Brown AHD. Plant genetic resources today: A critical appraisal[M]//Holden JHW, Williams JT, eds. Crop Genetic Resources: Conservation and Evaluation. London: George Allen and Unwin, 1984: 249-257.
[25]	马小军, 莫长明. 药用植物分子育种展望 [J]. 中国中药杂志, 2017, 42(11): 2021-2031.
	Ma XJ, Mo CM. Prospects of molecular breeding in medical plants [J]. China J Chin Mater Med, 2017, 42(11): 2021-2031.

性状 Traits	极小值 Minimal value	极大值 Maximum value	平均值 Mean value	中位数 Median value	标准差 Standard deviation	变异系数 Coefficient of variation （%）
蛇床子素OST	0.002	0.503	0.147	0.140	0.104	70.74
二氢欧山芹醇当归酸酯 CBD	0.021	0.814	0.278	0.242	0.166	59.71
异欧前胡素 IPT	0.000	0.139	0.012	0.010	0.014	116.67
二氢欧山芹醇 CBT	0.002	0.026	0.011	0.011	0.004	36.36
欧前胡素酚 OHN	0.000	0.436	0.138	0.133	0.068	49.28
伞形酮UMB	0.000	0.209	0.063	0.048	0.045	71.43
茎显色程度 JXS	1	4	3.228	3	0.782	24.22
花药显色程度 HYXS	2	3	2.179	3	0.384	17.62
茎生叶叶鞘茸毛丰富程度 YQRM	1	3	2.582	3	0.594	23.00
茎生叶叶鞘茸毛颜色 RMYS	1	3	1.363	1	0.620	45.49
叶鞘颜色YQYS	1	2	1.145	1	0.352	30.74

性状 Traits	极小值 Minimal value	极大值 Maximum value	平均值 Mean value	中位数 Median value	标准差 Standard deviation	变异系数 Coefficient of variation （%）
蛇床子素OST	0.002	0.503	0.147	0.140	0.104	70.74
二氢欧山芹醇当归酸酯 CBD	0.021	0.814	0.278	0.242	0.166	59.71
异欧前胡素 IPT	0.000	0.139	0.012	0.010	0.014	116.67
二氢欧山芹醇 CBT	0.002	0.026	0.011	0.011	0.004	36.36
欧前胡素酚 OHN	0.000	0.436	0.138	0.133	0.068	49.28
伞形酮UMB	0.000	0.209	0.063	0.048	0.045	71.43
茎显色程度 JXS	1	4	3.228	3	0.782	24.22
花药显色程度 HYXS	2	3	2.179	3	0.384	17.62
茎生叶叶鞘茸毛丰富程度 YQRM	1	3	2.582	3	0.594	23.00
茎生叶叶鞘茸毛颜色 RMYS	1	3	1.363	1	0.620	45.49
叶鞘颜色YQYS	1	2	1.145	1	0.352	30.74

性状 Traits	二氢欧山芹醇CBT	蛇床子素OST	异欧前胡素IPT	二氢欧山芹醇当归酸酯CBD	欧前胡素酚OHN	伞形酮UMB	茎显色程度JXS	花药显色程度HYXS	茎生叶叶鞘茸毛丰富程度YQRM	茎生叶叶鞘茸毛颜色RMYS	叶鞘颜色YQYS
二氢欧山芹醇CBT		0.074	-0.213	-0.279	0.369**	0.255*	0.051	-0.003	0.250*	-0.093	-0.070
蛇床子素OST	0.074		0.007	-0.185	0.023	0.156	0.089	0.013	0.126	-0.236	0.084
异欧前胡素IPT	-0.213	0.007		-0.118	-0.086	-0.056	-0.127	0.130	-0.065	-0.080	-0.070
二氢欧山芹醇当归酸酯CBD	-0.279	-0.185	-0.118		-0.047	-0.198	-0.176	0.123	-0.214	0.104	0.054
欧前胡素酚OHN	0.369**	0.023	-0.086	-0.047		0.390**	-0.076	0.110	0.246*	-0.129	-0.159
伞形酮UMB	0.255*	0.156	-0.056	-0.198	0.390**		-0.059	0.035	0.179	-0.174	-0.038
茎显色程度JXS	0.051	0.089	-0.127	-0.176	-0.076	-0.059		-0.050	0.150	0.435**	0.202
花药显色程度HYXS	-0.003	0.013	0.130	0.123	0.110	0.035	-0.050		0.108	-0.072	0.152
茎生叶叶鞘茸毛丰富程度YQRM	0.250*	0.126	-0.065	-0.214	0.246*	0.179	0.150	0.108		-0.213	-0.321
茎生叶叶鞘茸毛颜色RMYS	-0.093	-0.236	-0.080	0.104	-0.129	-0.174	0.435**	-0.072	-0.213		0.609**
叶鞘颜色YQYS	-0.070	0.084	-0.070	0.054	-0.159	-0.038	0.202	0.152	-0.321	0.609**

性状 Traits	二氢欧山芹醇CBT	蛇床子素OST	异欧前胡素IPT	二氢欧山芹醇当归酸酯CBD	欧前胡素酚OHN	伞形酮UMB	茎显色程度JXS	花药显色程度HYXS	茎生叶叶鞘茸毛丰富程度YQRM	茎生叶叶鞘茸毛颜色RMYS	叶鞘颜色YQYS
二氢欧山芹醇CBT		0.074	-0.213	-0.279	0.369**	0.255*	0.051	-0.003	0.250*	-0.093	-0.070
蛇床子素OST	0.074		0.007	-0.185	0.023	0.156	0.089	0.013	0.126	-0.236	0.084
异欧前胡素IPT	-0.213	0.007		-0.118	-0.086	-0.056	-0.127	0.130	-0.065	-0.080	-0.070
二氢欧山芹醇当归酸酯CBD	-0.279	-0.185	-0.118		-0.047	-0.198	-0.176	0.123	-0.214	0.104	0.054
欧前胡素酚OHN	0.369**	0.023	-0.086	-0.047		0.390**	-0.076	0.110	0.246*	-0.129	-0.159
伞形酮UMB	0.255*	0.156	-0.056	-0.198	0.390**		-0.059	0.035	0.179	-0.174	-0.038
茎显色程度JXS	0.051	0.089	-0.127	-0.176	-0.076	-0.059		-0.050	0.150	0.435**	0.202
花药显色程度HYXS	-0.003	0.013	0.130	0.123	0.110	0.035	-0.050		0.108	-0.072	0.152
茎生叶叶鞘茸毛丰富程度YQRM	0.250*	0.126	-0.065	-0.214	0.246*	0.179	0.150	0.108		-0.213	-0.321
茎生叶叶鞘茸毛颜色RMYS	-0.093	-0.236	-0.080	0.104	-0.129	-0.174	0.435**	-0.072	-0.213		0.609**
叶鞘颜色YQYS	-0.070	0.084	-0.070	0.054	-0.159	-0.038	0.202	0.152	-0.321	0.609**

种质来源 Germplasm source	观测等位基因数Na	有效等位基因数Ne	Nei’s基因多样性指数H	Shannon’s信息指数I	遗传分化系数Gst	基因流Nm
建始 Jianshi	1.883 3	1.490 0	0.293 8	0.445 2	0.112 6	3.939 4
巴东 Badong	1.983 3	1.494 1	0.298 5	0.457 2
五峰 Wufeng	1.883 3	1.520 9	0.304 3	0.456 4
均值 Mean value	1.916 6	1.500 7	0.298 9	0.452 9	-	-