Brassica Interspecies Hybridization and Polymerization Creates New Materials for High Resistance to Clubroot

doi:10.16420/j.issn.0513-353x.2024-0913

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (5): 1271-1284.doi: 10.16420/j.issn.0513-353x.2024-0913

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Brassica Interspecies Hybridization and Polymerization Creates New Materials for High Resistance to Clubroot

JIANG Shunshun¹^,², CHU Mengyu¹, YANG Jinxi¹, GAO Mengjie¹, FANG Zhiguo², WU Lei¹, REN Xuesong¹, LI Qinfei¹, SONG Hongyuan¹, SI Jun¹^,^*()

¹ Chongqing Key Laboratory of Vegetable Science，Key Laboratory of Agricultural Biosafety and Green Production of Upper Yangtze River，Ministry of Education，College of Horticulture and Landscape Architecture，Southwest University，Chongqing 400715，China
² Xiangyang Academy of Agricultural Sciences，Xiangyang，Hubei 441057，China

Received:2025-03-18 Revised:2025-04-11 Online:2025-05-23 Published:2025-05-21
Contact: SI Jun

Abstract

Abstract:

Cabbage 9LQ（CRb）was carried out with Chinese cabbage 8574（CRd）and turnip ECD4（PbBa8.1）by distantly hybridized combined embryo rescue technology，and the hybrid progeny was selected by molecular marker-assisted selection，and a total of 7 strains containing two resistance sites（two antibody materials）were screened，and the morphology and pollen vigor identification showed that the hybrid progeny were different from the parents，9LQ × ECD4 could not produce pollen，and the pollen fertility of 8574 × ECD4 was normal. A total of 27 hybrid progeny were obtained by hybridization with Z3（CRb^kato）with amphibiotic materials，and 4 strains containing 3 resistance sites（tri-antibody materials）were screened out by molecular marker-assisted selection，and the morphological and pollen vigor identification showed that the morphological characteristics of the hybrid progeny were different，the pollen fertility of 3R-9，3R-10 and 3R-19 was 100%，and the expression of 3R-26 was partially sterile. A total of 8 hybrid progeny were obtained by hybridizing the triplicate material with 9LQ，and four pairs of primers B0902，Yau389，A08-300 and OA were used for molecular marker-assisted selection，and one strain containing four resistance sites（4R-6）was successfully screened，and morphological identification showed that the quadruple antibody material grew robustly and the leaves began to have ball-setting traits. The identification of resistance to clubroot showed that the four resistant materials showed excellent resistance to the physiological races of Plasmodiophora brassicae from many regions.

Key words: Brassica, distant hybridization, clubroot disease, gene polymerization, molecular marker-assisted selection

JIANG Shunshun, CHU Mengyu, YANG Jinxi, GAO Mengjie, FANG Zhiguo, WU Lei, REN Xuesong, LI Qinfei, SONG Hongyuan, SI Jun. Brassica Interspecies Hybridization and Polymerization Creates New Materials for High Resistance to Clubroot[J]. Acta Horticulturae Sinica, 2025, 52(5): 1271-1284.

Figures/Tables 15

References 40

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培养基 Medium	蔗糖/g Sugar	琼脂/g Agar	MS/g	1/2MS/g	0.1 mg · mL^-1 NAA	1 mg · mL^-1 6-BA/mL	1 mol · L^-1 NaOH/μL
1/2MS	15	7.20		2.25			420
MS	30	7.20	4.47				420
MS分化培养基 MS differentiation medium	30	7.20	4.47		500 μL	3	420
生根培养基 Rooting medium	20	7.20		2.25	1.50 mL		420

培养基 Medium	蔗糖/g Sugar	琼脂/g Agar	MS/g	1/2MS/g	0.1 mg · mL^-1 NAA	1 mg · mL^-1 6-BA/mL	1 mol · L^-1 NaOH/μL
1/2MS	15	7.20		2.25			420
MS	30	7.20	4.47				420
MS分化培养基 MS differentiation medium	30	7.20	4.47		500 μL	3	420
生根培养基 Rooting medium	20	7.20		2.25	1.50 mL		420

引物名称 Primer	引物类型 Type of primer	片段大小/bp Fragment size	引物序列（5′-3′） Primer sequence	参考文献 Reference
OA	SCAR	401	F：TACACACGCTGATATACCAACA	何海艳,2018
OA	SCAR	401	R：TACACACGCTGCCCTGGAAA	何海艳,2018
Yau389	SSR	156	F：TCAAGTGGAGAGGGTCAGTC	付鹏宇,2019
Yau389	SSR	156	R：CAGTCACGAGGTCCCTAACA	付鹏宇,2019
A08-300	Indel	130	F：GTAGTGCGGGCCACAAAAT	战宗祥等,2016
A08-300	Indel	130	R：CACAATGGAGTGTTGAAATTCACT	战宗祥等,2016
B0902	SSR	241	F：AGCCTTGCGTAAAAGCAACTAC	沈舒,2019
B0902	SSR	241	R：GTTTGGAATCCGACAAATACATCCAT	沈舒,2019

引物名称 Primer	引物类型 Type of primer	片段大小/bp Fragment size	引物序列（5′-3′） Primer sequence	参考文献 Reference
OA	SCAR	401	F：TACACACGCTGATATACCAACA	何海艳,2018
OA	SCAR	401	R：TACACACGCTGCCCTGGAAA	何海艳,2018
Yau389	SSR	156	F：TCAAGTGGAGAGGGTCAGTC	付鹏宇,2019
Yau389	SSR	156	R：CAGTCACGAGGTCCCTAACA	付鹏宇,2019
A08-300	Indel	130	F：GTAGTGCGGGCCACAAAAT	战宗祥等,2016
A08-300	Indel	130	R：CACAATGGAGTGTTGAAATTCACT	战宗祥等,2016
B0902	SSR	241	F：AGCCTTGCGTAAAAGCAACTAC	沈舒,2019
B0902	SSR	241	R：GTTTGGAATCCGACAAATACATCCAT	沈舒,2019

处理 Treatment	杂交组合 Hybrid combination	授粉花蕾数 Number of pollinated buds	后代成活数 Number of survival offspring	成活率/% Survival rate
胚挽救 Embryo rescue	9LQ × ECD4	173	6	3.47
	ECD4 × 9LQ	185	0	0
	9LQ × 8574	86	0	0
	8574 × 9LQ	98	0	0
田间自然杂交 Natural hybridization in the field	8574 × ECD4	218	9	4.13
田间自然杂交 Natural hybridization in the field	ECD4 × 8574	206	0	0

Brassica Interspecies Hybridization and Polymerization Creates New Materials for High Resistance to Clubroot

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处理 Treatment	杂交组合 Hybrid combination	授粉花蕾数 Number of pollinated buds	收获角果数 Number of pods harvested	结荚率/% Pod setting rate	后代成活数 Number of survival offspring	成活率/% Survival rate
胚挽救Embryo rescue	（8574 × ECD4）× 9LQ	425	28	6.59	0	0
	9LQ ×（8574 × ECD4）	351	35	9.97	0	0
	（9LQ × ECD4）× 8574	441	0	0	0	0
	（9LQ × ECD4）× Z3	462	10	2.16	3	0.65
	（8574 × ECD4）× Z3	340	30	8.82	6	1.76
	Z3（8574 × ECD4）	390	226	57.95	9	2.31
田间自然杂交Natural hybridization in the field	（8574 × ECD4）× Z3	385	7	1.82	5	1.30
田间自然杂交Natural hybridization in the field	Z3 ×（8574 × ECD4）	393	6	1.53	4	1.02

编号 Number	叶片长/cm Blade length	叶片宽/cm Blade width	叶片形状 Blade shape	叶片颜色 Leaf color	叶尖 Leaf apex	花粉育性/% Pollen fertility
Z3（亲本Parents）	22.6	12.8	卵圆Oval	浅绿Light green	凹Concave	100
8574 × ECD4	18.6	13.7	卵圆Oval	浅黄绿Light yellowish green	凹Concave	100
3R-9	31.8	16.1	卵圆Oval	浅黄绿Light yellowish green	凹Concave	100
3R-10	33.0	14.7	卵圆Oval	浅绿Light green	凹Concave	100
3R-19	18.2	11.5	卵圆Oval	深绿Dark green	凹Concave	100
3R-26	20.8	12.4	卵圆Oval	深绿Dark green	平Flat	26.15

地区 Region	4R-6			感病对照（华良19号） Susceptible control（Hualiang 19）
地区 Region	发病率/% Disease incidence	病情指数 Disease index	抗性分级 Resistance grade	发病率/% Disease incidence	病情指数 Disease index	抗性分级 Resistance grade
重庆武隆Wulong，Chongqing	0	0	I	100	73.00	S
重庆涪陵Fuling，Chongqing	0	0	I	100	68.48	S
重庆万州Wanzhou，Chongqing	0	0	I	100	61.11	S
四川成都Chengdu，Sichuan	0	0	I	100	72.83	S
四川广元Guangyuan，Sichuan	0	0	I	100	56.94	S
四川攀枝花Panzhihua，Sichuan	70.00	22.5	T	100	48.81	S
湖北宜昌Yichang，Hubei	100	37.5	S	100	60.71	S
湖北恩施Enshi，Hubei	0	0	I	100	68.06	S

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