Creation and Identification of DH Lines for Chinese Cabbage Resistant to Clubroot Disease

doi:10.7668/hbnxb.20194742

Abstract

Abstract:

In order to quickly obtain homozygous and stable DH lines of Chinese cabbage resistant to clubroot disease,and to provide basic materials for breeding Chinese cabbage resistant varieties to clubroot disease,5 resistant genotypes of Chinese cabbage were used as materials for isolated microspore culture in this study,and phenotype and resistance of the obtained microspore regenerated plants were identified by combining molecular markers,morphological observation,and artificial inoculation of P.brassicae.The results showed that all 5 genotypes were induced to produce embryos,with a variation of 0.02 to 1.72 embryos per bud.Three genotypes,20aCR12,21aCR6,and 21aCR12,obtained regenerated plants,with regeneration rates of 27.09%,1.45%,and 26.07%,respectively.Resistance marker identification showed that molecular markers linked to CRa and CRb^kato all amplified the resistant bands in 50 obtained microspore plants.Phenotypic investigation showed that there were significant differences in the shape and color of basal leaves,bolting timing,and fertility among 50 microspore plants at reproductive growth stage,and 29 DH lines obtained through self-pollination also showed exhibited diversity in plant type,leaf traits,and heading traits at nutritional growth stage.Resistance inoculation showed that the disease index of 11 DH lines to races 4 and 1 of P.brassicae was less than 33.33,indicating resistance or tolerance to clubroot disease.Among them,there were 3 highly resistant DH lines with a disease index less than 5.0 for two races of P.brassicae,namely 20aCR12-23,20aCR12-29,and 21aCR12-39.The research results have shown that the homozygous DH lines resistant to clubroot can be obtained quickly by microspore culture combined with molecular marker assisted identification.

Key words: Chinese cabbage, Clubroot resistance, Microspore culture, Molecular marker, Double haploid

ZHANG Shuaiyu, CHANG Yucui, HAO Guanghua, WANG Yanhua, GU Aixia, LUO Shuangxia, MA Lisong, XUAN Shuxin, SHEN Shuxing. Creation and Identification of DH Lines for Chinese Cabbage Resistant to Clubroot Disease[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(3): 187-194. doi: 10.7668/hbnxb.20194742.

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References 29

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抗性基因 Resistant genes	分子标记 Molecular markers	正向引物序列(5'—3') Forward primer sequences(5'—3')	反向引物序列(5'—3') Reverse primer sequence(5'—3')
CRa	GC2360	AGTTTTGTAATTTTCACCCAAAGTATCA	CAGCTGGAGGAGCACTGCAACGGAGAGA
	GC3070	AGAAATAGCAAAGCTCAAACGG	TTTGGGTTTCCACAAAACAGATTACTTTA
CRb^kato	B1321	AGTGAAACAGACTATGATTAATGTTTT	GTTTCAACTTTGGTATAAGCGTTGAGA
	B1210	ATTGAAAAGTTGACTCCGTTGA	GTTTCTTCCTTGAAGTTGCTTCAGCTCTTC

抗性基因 Resistant genes	分子标记 Molecular markers	正向引物序列(5'—3') Forward primer sequences(5'—3')	反向引物序列(5'—3') Reverse primer sequence(5'—3')
CRa	GC2360	AGTTTTGTAATTTTCACCCAAAGTATCA	CAGCTGGAGGAGCACTGCAACGGAGAGA
	GC3070	AGAAATAGCAAAGCTCAAACGG	TTTGGGTTTCCACAAAACAGATTACTTTA
CRb^kato	B1321	AGTGAAACAGACTATGATTAATGTTTT	GTTTCAACTTTGGTATAAGCGTTGAGA
	B1210	ATTGAAAAGTTGACTCCGTTGA	GTTTCTTCCTTGAAGTTGCTTCAGCTCTTC

DH系编号 No. of DH lines	结球期性状 Traits at heading stage	根肿菌4号生理小种 Race 4 of P. brassicae			根肿菌1号生理小种 Race 1 of P. brassicae
DH系编号 No. of DH lines	结球期性状 Traits at heading stage	发病率/% Incidence	病情指数 Disease index	抗性水平 Resistance level	发病率/% Incidence	病情指数 Disease index	抗性水平 Resistance level
20aCR12-1	舒心,轻微干叶	16.67	11.11	T	43.48	23.19	T
20aCR12-4	不结球,干叶,腐烂	47.06	23.53	T	11.11	6.17	R
20aCR12-8	结球,合抱	28.57	19.05	T	20.00	6.67	R
20aCR12-9	结球,叠抱	79.17	31.94	T	41.67	13.89	T
20aCR12-23	轻微干叶,舒心不实	4.76	3.17	R	9.52	3.17	R
20aCR12-29	轻微干叶,不结球	0.00	0.00	I	5.00	1.67	R
20aCR12-34	结球,拧抱	16.67	9.26	R	16.67	11.11	T
20aCR12-39	结球,舒心	14.29	4.76	R	0.00	0.00	I
20aCR12-41	结球,舒心	16.67	9.26	R	12.50	4.17	R
21aCR12-3	结球,合抱	27.59	11.49	T	0.00	0.00	I
21aCR12-6	结球,叠抱	21.05	14.04	T	27.27	15.15	T
油绿3号 Youlü 3	结球,叠抱	100.00	97.44	HS	100.00	95.83	HS

DH系编号 No. of DH lines	结球期性状 Traits at heading stage	根肿菌4号生理小种 Race 4 of P. brassicae			根肿菌1号生理小种 Race 1 of P. brassicae
DH系编号 No. of DH lines	结球期性状 Traits at heading stage	发病率/% Incidence	病情指数 Disease index	抗性水平 Resistance level	发病率/% Incidence	病情指数 Disease index	抗性水平 Resistance level
20aCR12-1	舒心,轻微干叶	16.67	11.11	T	43.48	23.19	T
20aCR12-4	不结球,干叶,腐烂	47.06	23.53	T	11.11	6.17	R
20aCR12-8	结球,合抱	28.57	19.05	T	20.00	6.67	R
20aCR12-9	结球,叠抱	79.17	31.94	T	41.67	13.89	T
20aCR12-23	轻微干叶,舒心不实	4.76	3.17	R	9.52	3.17	R
20aCR12-29	轻微干叶,不结球	0.00	0.00	I	5.00	1.67	R
20aCR12-34	结球,拧抱	16.67	9.26	R	16.67	11.11	T
20aCR12-39	结球,舒心	14.29	4.76	R	0.00	0.00	I
20aCR12-41	结球,舒心	16.67	9.26	R	12.50	4.17	R
21aCR12-3	结球,合抱	27.59	11.49	T	0.00	0.00	I
21aCR12-6	结球,叠抱	21.05	14.04	T	27.27	15.15	T
油绿3号 Youlü 3	结球,叠抱	100.00	97.44	HS	100.00	95.83	HS

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