Development of molecular markers for clubroot resistance and identification of germplasm resources for Brassica rapa

doi:10.6048/j.issn.1001-4330.2023.09.024

Xinjiang Agricultural Sciences ›› 2023, Vol. 60 ›› Issue (9): 2281-2290.DOI: 10.6048/j.issn.1001-4330.2023.09.024

• Plant Protection·Microbes • Previous Articles Next Articles

Development of molecular markers for clubroot resistance and identification of germplasm resources for Brassica rapa

ZHANG Liguo¹^,²(), GUO Chungui³, CHANG Lichun², GUO Xinlei², ZHANG Tao¹^,², WU Jian², LIANG Jianli², GAO Jie¹(), WANG Xiaowu²()

1. Laboratory of Germplasm Resources and High-efficient Production of Characteristic Horticultural Crops in Xinjiang / College of Horticulture, Xinjiang Agricultural University, Urumqi 830052, China
2. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
3. Ganzhou Agricultural College, Ganzhou Jiangxi 341000, China

Received:2022-12-11 Online:2023-09-20 Published:2023-09-19
Correspondence author: GAO Jie(1963-),male,native place:Tianjin,professor,research field:vegetable cultivation and physiology,(E-mail)ofc111@163.com;WANG Xiaowu(1966-),male,native place:Yudu,Jiangxi Province,researcher,research field:molecular genetics and genomics of vegetable crops,(E-mail)wangxiaowu@caas.cn
Supported by:
Key Discipline Fund of Horticulture Science of Xinjiang Uygur Autonomous Region(2016-10758-37);China Agriculture Research System of MOF and MARA(CARS-10-P19);Basic Scientific R & D Program Project of Chinese Academy of Agricultural Sciences(Y2022PT23)

白菜抗根肿病分子标记开发与种质资源鉴定

张立国¹^,²(), 郭春贵³, 常立春², 郭新磊², 张涛¹^,², 武剑², 梁建丽², 高杰¹(), 王晓武²()

1.新疆农业大学园艺学院/新疆特色园艺作物种质资源与高效生产实验室,乌鲁木齐 830052
2.中国农业科学院蔬菜花卉研究所,北京 100081
3.赣州农业学校,江西赣州 341000

通讯作者: 高杰(1963-),男,天津人,教授,硕士生/博士生导师,研究方向为蔬菜栽培与生理,(E-mail)ofc111@163.com; 王晓武(1966-),男,江西于都人,研究员,硕士生/博士生导师,研究方向为蔬菜作物分子遗传学与基因组学,(E-mail)wangxiaowu@caas.cn
作者简介:张立国(1994-),男,辽宁人,硕士研究生,研究方向为分子标记辅助育种,(E-mail)2516189164@qq.com
基金资助:
新疆维吾尔自治区园艺学重点学科建设项目(2016-10758-37);财政部和农业农村部:国家现代农业产业技术体系资助(CARS-10-P19);中国农业科学院基本科研业务费专项(Y2022PT23)

Abstract

Abstract:

【Objective】 To develop KASP (Kompetitive Allele Specific PCR) marker for efficiently identifying CR (clubroot resistance) genes in cabbage B.rapa germplasms by using KASP markers to explore the distribution of clubroot resistant genes in germplasm collected with a large number of B.rapa accessions.【Methods】 Based on the information of cloned clubroot resistance genes previously, KASP markers were developed using Primer 3 and DNAMAN and the highly reliable markers were screened using a representative population containing 62 B.rapa varieties.Then, these available markers were used to genotype the 862 B.rapa resource population for disease resistance.【Results】 Based on four cloned disease resistance genes (CRa, CRb, CRd, Crr1a), six gene-specific KASP markers related to clubroot disease resistance were developed and screened.Among the 862 B.rapa germplasm accessions, 105 accessions containing clubroot resistance genes were identified.Turnip and turnip rape accounted for 70.48%.CRa was found to be the clubroot resistance gene carried by most accessions among B.rapa germplasm resources, which accounted for 58.09% of all disease resistant materials.Additionally, we found 16 accessions that contained two or more resistance genes.【Conclusion】 In this study, six stable and reliable gene-specific KASP markers were developed.CRa was found to be the clubroot resistance genes carried by most accessions among the four CR genes.Importantly, a few germplasm resources were identified that carried multiple clubroot resistance genes.

Key words: Brassica rapa; clubroot resistance; KASP marker; germplasms

摘要：

【目的】开发可在白菜中高效鉴定根肿病抗病基因的KASP(Kompetitive Allele Specific PCR)标记,并利用这些标记研究大量白菜资源材料中抗病基因分布规律。【方法】根据已克隆白菜抗根肿病基因信息,使用Primer3与DNAMAN开发KASP标记。利用代表性的62份白菜材料组成测试群体筛选标记,获得高可靠性标记,对862份白菜资源进行抗病基因型鉴定。【结果】从4个已克隆的抗根肿病基因(CRa、CRb、CRd、Crr1a)内开发并筛选出6个基因特异性KASP标记。白菜资源中鉴定出105份含有抗病基因型材料,其中芜菁与油用白菜占70.48%;CRa分布最广,占所有抗病材料的58.09%;含有2个或2个以上抗病基因的材料16份。【结论】获得6个稳定可靠的抗根肿病基因特异性KASP标记。CRa分布最广,筛选出同时携带多个抗根肿病基因的资源材料。

关键词: 白菜, 根肿病, KASP分子标记, 种子资源

CLC Number:

S436.34

ZHANG Liguo, GUO Chungui, CHANG Lichun, GUO Xinlei, ZHANG Tao, WU Jian, LIANG Jianli, GAO Jie, WANG Xiaowu. Development of molecular markers for clubroot resistance and identification of germplasm resources for Brassica rapa[J]. Xinjiang Agricultural Sciences, 2023, 60(9): 2281-2290.

张立国, 郭春贵, 常立春, 郭新磊, 张涛, 武剑, 梁建丽, 高杰, 王晓武. 白菜抗根肿病分子标记开发与种质资源鉴定[J]. 新疆农业科学, 2023, 60(9): 2281-2290.

Figures/Tables 9

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种类 Type	数量 Number	材料占比 Percentage(%)
大白菜 Chinese cabbage	200	23.20
小白菜 Pak choi	177	20.53
芜菁 Turnip	146	16.94
菜心 CaiXin	125	14.50
油用白菜 Turnip rape	111	12.88
日本水菜 Mizuna	26	3.02
日本小菘菜 Japanese komatssuna	24	2.78
野生白菜 Others	23	2.67
意大利菜心 Broccoletto	14	1.62
薹菜 TaiCai	13	1.51
Grelos	3	0.35

种类 Type	数量 Number	材料占比 Percentage(%)
大白菜 Chinese cabbage	200	23.20
小白菜 Pak choi	177	20.53
芜菁 Turnip	146	16.94
菜心 CaiXin	125	14.50
油用白菜 Turnip rape	111	12.88
日本水菜 Mizuna	26	3.02
日本小菘菜 Japanese komatssuna	24	2.78
野生白菜 Others	23	2.67
意大利菜心 Broccoletto	14	1.62
薹菜 TaiCai	13	1.51
Grelos	3	0.35

引物名称 Primer name	染色体 Chr	物理位置 Pos	SNP来源 SNP Origin	标记序列 Primer sequences(5’→3’)
CRd-8	A03	15983074	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTCGGGTGAAGAAGTGCCTG
				A2:GAAGGTGACCAAGTTCATGCTCGGGTGAAGAAGTGCCTC
				C:AAGAGGCCTTGGAGTTAAACTGAT
CRd-7	A03	15983095	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTCTCTTCTTTACAACAAATATGCGGTATC
				A2:GAAGGTGACCAAGTTCATGCTCTCTTCTTTACAACAAATATGCGGTATT
				C:AAGAGGCCTTGGAGTTAAACTGAT
CRd-6	A03	15983125	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTGCATATTTGTTGTAAAGAAGAGGAGC
				A2:GAAGGTGACCAAGTTCATGCTGCATATTTGTTGTAAAGAAGAGGAGT
				C:ATGCAGAAGACTCGTAGCACTTCC
CRd-5	A03	15983231	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTCGCCGTCTCTCGGGACTAG
				A2:GAAGGTGACCAAGTTCATGCTCGCCGTCTCTCGGGACTAA
				C:TGTGCATCTAAAGATAGAAGGGAAGC
CRd-9	A03	15984613	克隆基因CRd	A1:GAAGGTGACCAAGTTCATGCTTGCAGAAAAAGAGTTCTTTCATTATCC
				A2:GAAGGTCGGAGTCAACGGATTTGCAGAAAAAGAGTTCTTTCATTATCT
				C:CAGGTCTACCCTTGACAGAGAAAT
CRd-1	A03	15985759	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTTTGCTCTCCCCTAACTACACTTCTC
				A2:GAAGGTGACCAAGTTCATGCTTTGCTCTCCCCTAACTACACTTCTT
				C:TGGATCCACTTCGTAGAAAATGGT
CRd-2G	A03	15985772	克隆基因CRd	A1:GAAGGTGACCAAGTTCATGCTCACCAGCTTGAAGAAGTGTAGTTA
				A2:GAAGGTCGGAGTCAACGGATTCACCAGCTTGAAGAAGTGTAGTTC
				C:CAGACAATCAAGGACAGCAATTGT
CRd-3	A03	15985811	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTAATTGCTGTCCTTGATTGTCTGATC
				A2:GAAGGTGACCAAGTTCATGCTAATTGCTGTCCTTGATTGTCTGATA
				C:AAGCGAGGTGAATCTATCGATCAA
CRd-4	A03	15985859	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTCACCTCGCTTGATCTCGTCA
				A2:GAAGGTGACCAAGTTCATGCTACCTCGCTTGATCTCGTCG
				C:TTTCTTAGCCACGTTCGAAAAGAG
CRa-1	A03	25545189	克隆基因CRa	A1:GAAGGTGACCAAGTTCATGCTGACAAAGCAGATGGAGTTGATGAA
				A2:GAAGGTCGGAGTCAACGGATTGACAAAGCAGATGGAGTTGATGAT
				C:TCCATCATCACCACCATCTTCTTT
CRa-2	A03	25545971	克隆基因CRa	A1:GAAGGTGACCAAGTTCATGCTGTGAATGGGAACTTCATTTAACTGAC
				A2:GAAGGTCGGAGTCAACGGATTGTGAATGGGAACTTCATTTAACTGAT
				C:AAAAATGTCTGCAATCTGTCCCAA
CRa-3.1	A03	25557155	克隆基因CRa	A1:GAAGGTGACCAAGTTCATGCTACGAGATTTGAGCAACCATTAAAAGT
				A2:GAAGGTCGGAGTCAACGGATTACGAGATTTGAGCAACCATTAAAAGA
				C:AAAGATTGCTCAAGTTTGGTGGAG
Crr1a-16	A08	12271667	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCATATATGAGAAAATAATAGTAAAACACCGA
				A2:GAAGGTCGGAGTCAACGGATTCATATATGAGAAAATAATAGTAAAACACCGG
				C:AAAGCTTGCATCTTGCTTGTTACT
Crr1a-20	A08	12272040	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTCGCTCCCACAGCTTCCAC
				A2:GAAGGTCGGAGTCAACGGATTTCGCTCCCACAGCTTCCAG
				C:CTCGCAGTTTTCTACATGCATGAA
Crr1a-19	A08	12272124	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGCGACACACGAATACAAGAAGTTG
				A2:GAAGGTCGGAGTCAACGGATTGCGACACACGAATACAAGAAGTTT
				C:TGTAGACGAGACATTTCCTTGACC
Crr1a-15	A08	12272160	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCATGCTTTTGACCTCATCACAG
				A2:GAAGGTCGGAGTCAACGGATTCATGCTTTTGACCTCATCACAA
				C:CCCATGGAGCAACTTCTTGTATTC
Crr1a-10	A08	12272182	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGTGATGAGGTCAAAAGCATGC
				A2:GAAGGTCGGAGTCAACGGATTTGTGATGAGGTCAAAAGCATGA
				C:TGGTCTCGTCTTGAAAGGTTACAT
Crr1a-7	A08	12272193	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCATATGTCATACAGTGAAAACCTCG
				A2:GAAGGTCGGAGTCAACGGATTCATATGTCATACAGTGAAAACCTCA
				C:CCCATGGAGCAACTTCTTGTATTC
Crr1a-11	A08	12272391	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGTTGATGTTCACTGGAAGGGA
				A2:GAAGGTCGGAGTCAACGGATTGTTGATGTTCACTGGAAGGGC
				C:GAATCTCGGAAATTGCTCAAAGCT
Crr1a-2.1	A08	12272412	克隆基因Crr1a	A1:GGAAGGTGACCAAGTTCATGCTTCATAATTTGAAGCAGTTGAATCTCG
				A2:GAAGGTCGGAGTCAACGGATTTCATAATTTGAAGCAGTTGAATCTCA
				C:TTCACTGGAAGGGACATTAGCTTT
Crr1a-3	A08	12272423	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTTGAGCAATTTCCGAGATTCAACT
				A2:GAAGGTCGGAGTCAACGGATTTTGAGCAATTTCCGAGATTCAACC
				C:CCCCTCTTCTATTGGGAATCTTCA
Crr1a-13	A08	12273132	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTTCCTTGAGATTGGTCATATTTCC
				A2:GAAGGTCGGAGTCAACGGATTTTCCTTGAGATTGGTCATATTTCT
				C:GAATCTTTCTGGATGCTCTAGCCT
Crr1a-12	A08	12273387	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTAAGCTCCTTTAGATATTTTGAATAAGACAA
				A2:GAAGGTCGGAGTCAACGGATTAAGCTCCTTTAGATATTTTGAATAAGACAG
				C:TATGCTTTAGCTGAGGATCGTCTT
Crr1a-14	A08	12274525	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTCATGGTTTGGTCCTGGAAGTA
				A2:GAAGGTCGGAGTCAACGGATTTCATGGTTTGGTCCTGGAAGTC
				C:ATATGATTGATCCGATGCGCATTC
Crr1a-4	A08	12274547	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCTACTTCCAGGACCAAACCATGAG
				A2:GAAGGTCGGAGTCAACGGATTCTACTTCCAGGACCAAACCATGAA
				C:TTAGTTGTTCTCGATGACGTGGAT
Crr1a-21	A08	12285308	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTGTTAATTGAAGCTGAAGAAACATTTT
				A2:GAAGGTCGGAGTCAACGGATTTGTTAATTGAAGCTGAAGAAACATTTC
				C:ATGATTCACCCAGATGGAGAAACA
Crr1a-1	A08	12285339	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTCTTCAGCTTCAATTAACATGGGG
				A2:GAAGGTCGGAGTCAACGGATTTCTTCAGCTTCAATTAACATGGGA
				C:AAATTGGGAGATTAAGGAATGCGG
Crr1a-18	A08	12285909	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGATGCACAAAGCTGGTTTCA
				A2:GAAGGTCGGAGTCAACGGATTGATGCACAAAGCTGGTTTCG
				C:TGTAATGAATCTGGAACTGTGGGA
Crr1a-8	A08	12285994	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGCGCAATTTCTTGAATTCCTT
				A2:GAAGGTCGGAGTCAACGGATTGCGCAATTTCTTGAATTCCTG
				C:TGGTCTCGTCTTGAAAGGTTACAT
Crr1a-22	A08	12286052	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTAAAGGTTACATATGTCATACAGCGAAG
				A2:GAAGGTCGGAGTCAACGGATTAAAGGTTACATATGTCATACAGCGAAA
				C:AATTCCTTTGTCGCTCAAGTGAAG
Crr1a-17	A08	12286186	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCTTCATCTCGGGGACTGCTG
				A2:GAAGGTCGGAGTCAACGGATTCTTCATCTCGGGGACTGCTC
				C:ACCATGACCTGATTGATTGAGGAA
Crr1a-9	A08	12286202	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCCAGCAGTCCCCGAGATG
				A2:GAAGGTCGGAGTCAACGGATTCCAGCAGTCCCCGAGATC
				C:TGAATGGTTGCTCAAAGCTAAAGG
Crr1a-6	A08	12286591	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTCTCGAGATTAGTCAGATTTCCAATAT
				A2:GAAGGTCGGAGTCAACGGATTTCTCGAGATTAGTCAGATTTCCAATAG
				C:CTTGTGGAGCTTCCCTGTTCTATT
Crr1a-5	A08	12286794	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTAGAGACAACATCGGGAGACTAGTA
				A2:GAAGGTCGGAGTCAACGGATTAGAGACAACATCGGGAGACTAGTC
				C:TGAAACTCCCTTCTTCTATTGGGG
CRBrG3-1	A03	24844527	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTATTGTGGATATAAAATCATTGCATGA
				A2:GAAGGTCGGAGTCAACGGATTATTGTGGATATAAAATCATTGCATGT
				C:AGTAGACTTTGGCAGGAACTATACT
CRBrG3-2	A03	24844558	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTACAATCTACTTGCAATCTTGCATAATAT
				A2:GAAGGTCGGAGTCAACGGATTACAATCTACTTGCAATCTTGCATAATAA
				C:AGTAGACTTTGGCAGGAACTATACT
CRBrG3-3	A03	24845823	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTGAGTATGAGCCACAATACTTTCTTGC
				A2:GAAGGTCGGAGTCAACGGATTGAGTATGAGCCACAATACTTTCTTGT
				C:AGGAACCAGTCTACTTTCGTCTTC
CRBrG3-4	A03	24846489	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTATTCAGGAACTCTTTTAACCGCT
				A2:GAAGGTCGGAGTCAACGGATTATTCAGGAACTCTTTTAACCGCC
				C:TCCTCAGTGGATGTTCAAAACTCA
CRBrG3-5	A03	24847312	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTTCTTCTTGGTTCCATAACCGC
				A2:GAAGGTCGGAGTCAACGGATTTCTTCTTGGTTCCATAACCGA
				C:AGGGAGGTTGGTTATGAATCATCC
CRBrG3-6	A03	24845563	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTCTTCAATGGAAACCCACCTG
				A2:GAAGGTCGGAGTCAACGGATTCTTCAATGGAAACCCACCTT
				C:TTGAATGAGACAACTACGCAGAGA
CRBrG3-7	A03	24849293	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTCTTCACGTGCTATCTCCTCGAT
				A2:GAAGGTCGGAGTCAACGGATTCTTCACGTGCTATCTCCTCGAC
				C:GATGAAAGCGTCGATCCCTTTC
CRBrG3-8	A03	24845969	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTCAACAGCTCGTTTCTCTTCCAG
				A2:GAAGGTCGGAGTCAACGGATTCAACAGCTCGTTTCTCTTCCAC
				C:ATGAGCATCCAGATACTGCAGATT

引物名称 Primer name	染色体 Chr	物理位置 Pos	SNP来源 SNP Origin	标记序列 Primer sequences(5’→3’)
CRd-8	A03	15983074	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTCGGGTGAAGAAGTGCCTG
				A2:GAAGGTGACCAAGTTCATGCTCGGGTGAAGAAGTGCCTC
				C:AAGAGGCCTTGGAGTTAAACTGAT
CRd-7	A03	15983095	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTCTCTTCTTTACAACAAATATGCGGTATC
				A2:GAAGGTGACCAAGTTCATGCTCTCTTCTTTACAACAAATATGCGGTATT
				C:AAGAGGCCTTGGAGTTAAACTGAT
CRd-6	A03	15983125	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTGCATATTTGTTGTAAAGAAGAGGAGC
				A2:GAAGGTGACCAAGTTCATGCTGCATATTTGTTGTAAAGAAGAGGAGT
				C:ATGCAGAAGACTCGTAGCACTTCC
CRd-5	A03	15983231	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTCGCCGTCTCTCGGGACTAG
				A2:GAAGGTGACCAAGTTCATGCTCGCCGTCTCTCGGGACTAA
				C:TGTGCATCTAAAGATAGAAGGGAAGC
CRd-9	A03	15984613	克隆基因CRd	A1:GAAGGTGACCAAGTTCATGCTTGCAGAAAAAGAGTTCTTTCATTATCC
				A2:GAAGGTCGGAGTCAACGGATTTGCAGAAAAAGAGTTCTTTCATTATCT
				C:CAGGTCTACCCTTGACAGAGAAAT
CRd-1	A03	15985759	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTTTGCTCTCCCCTAACTACACTTCTC
				A2:GAAGGTGACCAAGTTCATGCTTTGCTCTCCCCTAACTACACTTCTT
				C:TGGATCCACTTCGTAGAAAATGGT
CRd-2G	A03	15985772	克隆基因CRd	A1:GAAGGTGACCAAGTTCATGCTCACCAGCTTGAAGAAGTGTAGTTA
				A2:GAAGGTCGGAGTCAACGGATTCACCAGCTTGAAGAAGTGTAGTTC
				C:CAGACAATCAAGGACAGCAATTGT
CRd-3	A03	15985811	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTAATTGCTGTCCTTGATTGTCTGATC
				A2:GAAGGTGACCAAGTTCATGCTAATTGCTGTCCTTGATTGTCTGATA
				C:AAGCGAGGTGAATCTATCGATCAA
CRd-4	A03	15985859	克隆基因CRd	A1:GAAGGTCGGAGTCAACGGATTCACCTCGCTTGATCTCGTCA
				A2:GAAGGTGACCAAGTTCATGCTACCTCGCTTGATCTCGTCG
				C:TTTCTTAGCCACGTTCGAAAAGAG
CRa-1	A03	25545189	克隆基因CRa	A1:GAAGGTGACCAAGTTCATGCTGACAAAGCAGATGGAGTTGATGAA
				A2:GAAGGTCGGAGTCAACGGATTGACAAAGCAGATGGAGTTGATGAT
				C:TCCATCATCACCACCATCTTCTTT
CRa-2	A03	25545971	克隆基因CRa	A1:GAAGGTGACCAAGTTCATGCTGTGAATGGGAACTTCATTTAACTGAC
				A2:GAAGGTCGGAGTCAACGGATTGTGAATGGGAACTTCATTTAACTGAT
				C:AAAAATGTCTGCAATCTGTCCCAA
CRa-3.1	A03	25557155	克隆基因CRa	A1:GAAGGTGACCAAGTTCATGCTACGAGATTTGAGCAACCATTAAAAGT
				A2:GAAGGTCGGAGTCAACGGATTACGAGATTTGAGCAACCATTAAAAGA
				C:AAAGATTGCTCAAGTTTGGTGGAG
Crr1a-16	A08	12271667	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCATATATGAGAAAATAATAGTAAAACACCGA
				A2:GAAGGTCGGAGTCAACGGATTCATATATGAGAAAATAATAGTAAAACACCGG
				C:AAAGCTTGCATCTTGCTTGTTACT
Crr1a-20	A08	12272040	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTCGCTCCCACAGCTTCCAC
				A2:GAAGGTCGGAGTCAACGGATTTCGCTCCCACAGCTTCCAG
				C:CTCGCAGTTTTCTACATGCATGAA
Crr1a-19	A08	12272124	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGCGACACACGAATACAAGAAGTTG
				A2:GAAGGTCGGAGTCAACGGATTGCGACACACGAATACAAGAAGTTT
				C:TGTAGACGAGACATTTCCTTGACC
Crr1a-15	A08	12272160	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCATGCTTTTGACCTCATCACAG
				A2:GAAGGTCGGAGTCAACGGATTCATGCTTTTGACCTCATCACAA
				C:CCCATGGAGCAACTTCTTGTATTC
Crr1a-10	A08	12272182	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGTGATGAGGTCAAAAGCATGC
				A2:GAAGGTCGGAGTCAACGGATTTGTGATGAGGTCAAAAGCATGA
				C:TGGTCTCGTCTTGAAAGGTTACAT
Crr1a-7	A08	12272193	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCATATGTCATACAGTGAAAACCTCG
				A2:GAAGGTCGGAGTCAACGGATTCATATGTCATACAGTGAAAACCTCA
				C:CCCATGGAGCAACTTCTTGTATTC
Crr1a-11	A08	12272391	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGTTGATGTTCACTGGAAGGGA
				A2:GAAGGTCGGAGTCAACGGATTGTTGATGTTCACTGGAAGGGC
				C:GAATCTCGGAAATTGCTCAAAGCT
Crr1a-2.1	A08	12272412	克隆基因Crr1a	A1:GGAAGGTGACCAAGTTCATGCTTCATAATTTGAAGCAGTTGAATCTCG
				A2:GAAGGTCGGAGTCAACGGATTTCATAATTTGAAGCAGTTGAATCTCA
				C:TTCACTGGAAGGGACATTAGCTTT
Crr1a-3	A08	12272423	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTTGAGCAATTTCCGAGATTCAACT
				A2:GAAGGTCGGAGTCAACGGATTTTGAGCAATTTCCGAGATTCAACC
				C:CCCCTCTTCTATTGGGAATCTTCA
Crr1a-13	A08	12273132	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTTCCTTGAGATTGGTCATATTTCC
				A2:GAAGGTCGGAGTCAACGGATTTTCCTTGAGATTGGTCATATTTCT
				C:GAATCTTTCTGGATGCTCTAGCCT
Crr1a-12	A08	12273387	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTAAGCTCCTTTAGATATTTTGAATAAGACAA
				A2:GAAGGTCGGAGTCAACGGATTAAGCTCCTTTAGATATTTTGAATAAGACAG
				C:TATGCTTTAGCTGAGGATCGTCTT
Crr1a-14	A08	12274525	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTCATGGTTTGGTCCTGGAAGTA
				A2:GAAGGTCGGAGTCAACGGATTTCATGGTTTGGTCCTGGAAGTC
				C:ATATGATTGATCCGATGCGCATTC
Crr1a-4	A08	12274547	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCTACTTCCAGGACCAAACCATGAG
				A2:GAAGGTCGGAGTCAACGGATTCTACTTCCAGGACCAAACCATGAA
				C:TTAGTTGTTCTCGATGACGTGGAT
Crr1a-21	A08	12285308	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTGTTAATTGAAGCTGAAGAAACATTTT
				A2:GAAGGTCGGAGTCAACGGATTTGTTAATTGAAGCTGAAGAAACATTTC
				C:ATGATTCACCCAGATGGAGAAACA
Crr1a-1	A08	12285339	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTCTTCAGCTTCAATTAACATGGGG
				A2:GAAGGTCGGAGTCAACGGATTTCTTCAGCTTCAATTAACATGGGA
				C:AAATTGGGAGATTAAGGAATGCGG
Crr1a-18	A08	12285909	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGATGCACAAAGCTGGTTTCA
				A2:GAAGGTCGGAGTCAACGGATTGATGCACAAAGCTGGTTTCG
				C:TGTAATGAATCTGGAACTGTGGGA
Crr1a-8	A08	12285994	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTGCGCAATTTCTTGAATTCCTT
				A2:GAAGGTCGGAGTCAACGGATTGCGCAATTTCTTGAATTCCTG
				C:TGGTCTCGTCTTGAAAGGTTACAT
Crr1a-22	A08	12286052	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTAAAGGTTACATATGTCATACAGCGAAG
				A2:GAAGGTCGGAGTCAACGGATTAAAGGTTACATATGTCATACAGCGAAA
				C:AATTCCTTTGTCGCTCAAGTGAAG
Crr1a-17	A08	12286186	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCTTCATCTCGGGGACTGCTG
				A2:GAAGGTCGGAGTCAACGGATTCTTCATCTCGGGGACTGCTC
				C:ACCATGACCTGATTGATTGAGGAA
Crr1a-9	A08	12286202	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTCCAGCAGTCCCCGAGATG
				A2:GAAGGTCGGAGTCAACGGATTCCAGCAGTCCCCGAGATC
				C:TGAATGGTTGCTCAAAGCTAAAGG
Crr1a-6	A08	12286591	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTTCTCGAGATTAGTCAGATTTCCAATAT
				A2:GAAGGTCGGAGTCAACGGATTTCTCGAGATTAGTCAGATTTCCAATAG
				C:CTTGTGGAGCTTCCCTGTTCTATT
Crr1a-5	A08	12286794	克隆基因Crr1a	A1:GAAGGTGACCAAGTTCATGCTAGAGACAACATCGGGAGACTAGTA
				A2:GAAGGTCGGAGTCAACGGATTAGAGACAACATCGGGAGACTAGTC
				C:TGAAACTCCCTTCTTCTATTGGGG
CRBrG3-1	A03	24844527	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTATTGTGGATATAAAATCATTGCATGA
				A2:GAAGGTCGGAGTCAACGGATTATTGTGGATATAAAATCATTGCATGT
				C:AGTAGACTTTGGCAGGAACTATACT
CRBrG3-2	A03	24844558	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTACAATCTACTTGCAATCTTGCATAATAT
				A2:GAAGGTCGGAGTCAACGGATTACAATCTACTTGCAATCTTGCATAATAA
				C:AGTAGACTTTGGCAGGAACTATACT
CRBrG3-3	A03	24845823	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTGAGTATGAGCCACAATACTTTCTTGC
				A2:GAAGGTCGGAGTCAACGGATTGAGTATGAGCCACAATACTTTCTTGT
				C:AGGAACCAGTCTACTTTCGTCTTC
CRBrG3-4	A03	24846489	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTATTCAGGAACTCTTTTAACCGCT
				A2:GAAGGTCGGAGTCAACGGATTATTCAGGAACTCTTTTAACCGCC
				C:TCCTCAGTGGATGTTCAAAACTCA
CRBrG3-5	A03	24847312	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTTCTTCTTGGTTCCATAACCGC
				A2:GAAGGTCGGAGTCAACGGATTTCTTCTTGGTTCCATAACCGA
				C:AGGGAGGTTGGTTATGAATCATCC
CRBrG3-6	A03	24845563	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTCTTCAATGGAAACCCACCTG
				A2:GAAGGTCGGAGTCAACGGATTCTTCAATGGAAACCCACCTT
				C:TTGAATGAGACAACTACGCAGAGA
CRBrG3-7	A03	24849293	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTCTTCACGTGCTATCTCCTCGAT
				A2:GAAGGTCGGAGTCAACGGATTCTTCACGTGCTATCTCCTCGAC
				C:GATGAAAGCGTCGATCCCTTTC
CRBrG3-8	A03	24845969	克隆基因CRb	A1:GAAGGTGACCAAGTTCATGCTCAACAGCTCGTTTCTCTTCCAG
				A2:GAAGGTCGGAGTCAACGGATTCAACAGCTCGTTTCTCTTCCAC
				C:ATGAGCATCCAGATACTGCAGATT

引物名称 Primer name	物理位置 Physical position	抗病基因 Disease resistance genes	一致株数 Consistent number of plants	不一致株数 Number of inconsistent plants	总数 Total	一致率 Consistency rate(%)
CRa-3.1	A03:25557155	CRa	87	2	89	97.75
CRBrG3-8	A03:24845969	CRb	87	2	89	97.75
CRd2G	A03:15985772	CRd	86	3	89	96.63
CRd3	A03:15985811	CRd	89	0	89	100
CRd4	A03:15985859	CRd	86	3	89	96.63
Crr1-10	A08:12272182	Crr1a	89	0	89	100

Development of molecular markers for clubroot resistance and identification of germplasm resources for Brassica rapa

白菜抗根肿病分子标记开发与种质资源鉴定

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References 29

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	CRa-3.1	CRBrG3-8	CRd-2G	CRd-3	CRd-4	Crr1a-10
易感基因型(纯合) Susceptible genotype (homozygous)	800	817	805	828	845	809
抗病基因型(纯合) Resistance genotype (homozygous)	40	25	9	8	9	7
杂合基因型 Heterozygous genotype	21	19	44	18	6	41
未分型 Undetemined	1	1	4	8	2	5

编号 Number	品种类型 Type	抗病基因数量 Number of resistant genes	抗病基因 Resistant genes
SY18Q0717	芜菁	3	CRa/CRb/CRd
SY18Q0980	芜菁	2	CRd/Crr1a
SY18Q0994	芜菁	2	CRa/CRd
SY18Q0769	芜菁	2	CRa/CRd
SY18Q0699	芜菁	2	CRa/CRd
SY18Q0664	芜菁	2	CRa/CRd
SY18Q0985	芜菁	2	CRa/CRb
SY18Q0984	芜菁	2	CRa/CRb
SY18Q0972	芜菁	2	CRa/CRb
SY18Q0948	芜菁	2	CRa/CRb
SY18Q0913	Grelos	2	CRa/CRb
SY18Q0869	油用白菜	2	CRa/CRb
SY18Q0848	油用白菜	2	CRa/CRb
SY18Q0821	油用白菜	2	CRa/CRb
SY18Q0779	芜菁	2	CRa/CRb
SY18Q0668	芜菁	2	CRa/CRb

[1]	LI Junru, QIN Ning, LI Wenlong, DU Hui, LI Xihuan, ZHANG Caiying. Identification of Seed Oligosaccharide and Its Components and Screening for Elite Germplasms in Soybean [J]. Xinjiang Agricultural Sciences, 2022, 59(2): 353-360.
[2]	XU Geng, WANG Qing-hui, MA Yue-hong,GUO Hui, YANG Zhong-qiang, XIE Zhi-xuan. Experimental Research of the Hot Drying Process of Brassica rapa L. [J]. Xinjiang Agricultural Sciences, 2019, 56(10): 1869-1878.