48份玉米自交系抗病性的精准鉴定

doi:10.3864/j.issn.0578-1752.2021.12.003

摘要/Abstract

摘要：

【目的】结合已克隆抗病基因的分子标记,对来自陕A群、陕B群的30份核心自交系和18份国内外优良种质进行抗病性鉴定,为玉米抗病育种奠定基础。【方法】2019年、2020年分别在陕西省不同地点开展茎腐病、穗腐病、大斑病和小斑病的田间接种鉴定,以及灰斑病的田间自然发病鉴定。试验采用随机区组设计,每个试验设置2—3个重复。采用苗期高粱粒接种法接种大斑病和小斑病,在乳熟后期,对大斑病、小斑病和灰斑病进行病情分级鉴定;在玉米抽雄期,采用土埋伤根法接种禾谷镰孢茎腐病,生理成熟后进行劈茎调查;采用花丝通道和针刺果穗接种法,分2次接种禾谷镰孢穗腐病,玉米生理成熟后进行调查。分别计算不同病害病情指数的最佳线性无偏预测（best linear unbiased predictions,BLUPs）,并分析两两病害之间的相关性。对已克隆抗病基因进行功能分子标记基因型鉴定。【结果】对48份自交系开展5种病害田间鉴定,筛选到9份高抗大斑病自交系、2份高抗小斑病自交系、10份抗灰斑病自交系、5份高抗禾谷镰孢茎腐病自交系和5份抗禾谷镰孢穗腐病自交系。1145、CML170、KA103等8份自交系兼抗3种叶斑病。对5种病害综合抗性表现优良的材料有7份,包括1145、CML170、KA105、KB020、X178、沈137和郑58。5种病害相关性分析发现,禾谷镰孢茎腐病与3种叶斑病抗性呈极显著正相关,与穗腐病抗性无相关性。对已知抗病基因鉴定表明,1145、KA081和沈137携带抗禾谷镰孢茎腐病的qRfg1位点,KB109携带抗炭疽茎腐病的Rcg1位点,带有抗大斑病基因Htn1和多抗小斑病、灰斑病基因ZmCCoAOMT2的材料较多。【结论】1145、CML170、KA105、KB020、X178、沈137和郑58对5种病害综合抗性表现良好,可作为供体亲本进行自交系的综合抗性改良。沈137携带抗病的qRfg1、Htn1和ZmCCoAOMT2等位基因,可用于分子标记辅助改良。

关键词: 玉米, 抗病性, 大斑病, 小斑病, 灰斑病, 茎腐病, 穗腐病

Abstract:

【Objective】 In order to provide valuable resources to the maize community for enhancing disease resistance, we selected 30 core maize inbred lines from Shaan A and Shaan B heterotic groups, and 18 major maize germplasms worldwide for disease resistance evaluation and resistance gene identification.【Method】Gibberella stalk rot, Gibberella ear rot, northern leaf blight (NLB) and southern leaf blight (SLB) were artificially inoculated, while gray leaf spot (GLS) was naturally infected in replicated field trials at different locations in 2019 and 2020. A randomized complete block design was adopted with two to three replicates being evaluated at each location. Sorghum grain inoculum was artificially inoculated to the whorl of each plant at the seedling stage for NLB and SLB. Gibberella stalk rot was artificially inoculated using corn grain inoculum buried next to the root of each plant at heading time, while Gibberella ear rot was artificially inoculated by injecting conidial suspension through the silk channel and the husk separately. The best linear unbiased predictions (BLUPs) were estimated and correlation coefficients were calculated between each pair of the disease resistance. Disease resistance genes were genotyped using functional markers. 【Result】Nine highly resistant (HR) lines for NLB, two HR lines for SLB, ten resistant (R) lines for GLS, five HR lines for stalk rot, and five R lines for ear rot resistance were identified out of the 48 maize germplasms. Eight inbred lines showed multiple disease resistance to the three foliar diseases, such as 1145, CML170, and so on. We identified seven elite inbred lines showing good performance for all the five diseases, including 1145, CML170, KA105, KB020, X178, Shen137, and Zheng58. NLB, SLB, GLS, and Gibberella stalk rot resistance were significantly positively correlated, while Gibberella ear rot was not correlated with the other four diseases. 1145, KA081, and Shen137 contain the resistant qRfg1 allele. KB109 harbors the resistant Rcg1 allele. Many inbred lines carry the resistant Htn1 and ZmCCoAOMT2 alleles.【Conclusion】1145, CML170, KA105, KB020, X178, Shen137, and Zheng58 could be selected for multiple disease resistance improvement in breeding program. Shen137 is the best line for marker-assisted selection for qRfg1, Htn1, and ZmCCoAOMT2 loci.

Key words: maize, disease resistance, northern leaf blight, southern leaf blight, gray leaf spot, stalk rot, ear rot

赵子麒,赵雅琪,林昌朋,赵永泽,余宇潇,孟庆立,曾广莹,薛吉全,杨琴. 48份玉米自交系抗病性的精准鉴定[J]. 中国农业科学, 2021, 54(12): 2510-2522.

ZHAO ZiQi,ZHAO YaQi,LIN ChangPeng,ZHAO YongZe,YU YuXiao,MENG QingLi,ZENG GuangYing,XUE JiQuan,YANG Qin. Precise Evaluation of 48 Maize Inbred Lines to Major Diseases[J]. Scientia Agricultura Sinica, 2021, 54(12): 2510-2522.

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大斑病 Northern leaf blight		小斑病 Southern leaf blight		灰斑病 Gray leaf spot		禾谷镰孢穗腐病 Gibberella ear rot		禾谷镰孢茎腐病 Gibberella stalk rot
感病面积 Diseased area (%)	抗性评价 Disease level	病情分级 Disease scale	抗性评价 Disease level	病情分级 Disease scale	抗性评价 Disease level	感病面积 Diseased area (%)	抗性评价 Disease level	病情分级 Disease scale	抗性评价 Disease level
0—10	HR	1—2	HR	1—2	HR	0—1	HR	1—2	HR
10—25	R	2—3	R	2—3	R	1—10	R	2—3	R
25—40	MR	3—4	MR	3—4	MR	10—25	MR	3—5	MR
40—65	S	4—5	S	4—5	S	25—50	S	5—7	S
65—100	HS	5—9	HS	5—9	HS	50—100	HS	7—9	HS

基因名称 Gene ID	病害 Disease	引物名称 Primer name	引物序列 Primer sequence (5′-3′)
qRfg1	禾谷镰孢茎腐病 Gibberella stalk rot	TED-F	GCACAAGAGAGATGGAGCATT
		TED-R	ATTCTCAATCCAAGGTGCAG
		TERB-F	CCTAAGAACCGTCGGAAACA
		TERB-R	CGAGCGTTTTCGACATAACA
		TELB-F	AAACGCTGACACTTCCGACT
		TELB-R	GTCGACACGTGTAGGAAGCA
Rcg1	炭疽茎腐病 Anthracnose stalk rot	FLP-111-F	TTCCTGTTCGTCTGTATCTGATCCG
		FLP-111-R	TTTGATTCCGGTCGAGTATAACCTG
		FLP-112-F	GAAACTGCCTTCCCAGAAAACAATG
		FLP-112-R	CAAGATCGGTGAAGTTGGTGCTTC
		FLP-113-F	ATCACAGATGGGTCTCAAGGATTGC
		FLP-A1-R	TTCCAAGCAATTCACAGCTC
Htn1	大斑病 Northern leaf blight	Htn1-F	TCTTCTCCCCGCCATGGC
Htn1	大斑病 Northern leaf blight	Htn1-R	CGGTGTAGCCGCAGGAGT
ZmCCoAOMT2	小斑病、灰斑病 Southern leaf blight, Gray leaf spot	CAMT-F	CCGTCCTGGACGACCTCGTG
ZmCCoAOMT2	小斑病、灰斑病 Southern leaf blight, Gray leaf spot	CAMT-R	TGCTGCGCGTCGTCTACGAT

病害 Disease	随机因子 Random factor	方差估计值（标准误） Estimate (standard error)	P值 P value	固定因子 Fixed factor	F测验 F test
大斑病 Northern leaf blight	重复Rep. (Env.)	0.00	NS	基因型Genotype	4.98***
	环境Env.	26.17(39.43)	NS
	基因型与环境互作 Genotype×Env.	51.55(17.12)	0.0038
	散粉期DTA	0.00	NS
	残差Residual	55.86(8.24)
小斑病 Southern leaf blight	重复Rep. (Env.)	0.01(0.01)	NS	基因型Genotype	11.89***
	环境Env.	0.00	NS
	基因型与环境互作 Genotype×Env.	0.11(0.04)	0.0047
	散粉期DTA	0.00	NS
	残差Residual	0.20(0.02)
灰斑病 Gray leaf spot	重复Rep. (Env.)	0.00	NS	基因型Genotype	3.42***
	环境Env.	0.00	NS
	基因型与环境互作 Genotype×Env.	0.18(0.08)	0.0093
	残差Residual	0.33(0.05)
禾谷镰孢菌茎腐病Gibberella stalk rot	重复Rep. (Env.)	0.33(0.25)	NS	基因型Genotype	4.44***
	环境Env.	0.93(1.14)	NS
	基因型与环境互作 Genotype×Env.	1.31(0.27)	<0.0001
	残差Residual	0.89(0.10)
禾谷镰孢穗腐病Gibberella ear rot	重复Rep. (Env.)	25.06(22.14)	NS	基因型Genotype	1.01
	环境Env.	20.41(41.66)	NS
	基因型与环境互作 Genotype×Env.	302.13(74.24)	<0.0001
	残差Residual	239.63(30.51)

自交系 Inbred line	类群划分 Sub-group	大斑病 Northern leaf blight	小斑病 Southern leaf blight	灰斑病 Gray leaf spot	禾谷镰孢茎腐病 Gibberella stalk rot	禾谷镰孢穗腐病 Gibberella ear rot
1145	P78599	HR	R	R	HR	MR
2082	NSSS	R	R	MR	R	MR
AMD43	NSSS	S	HS	HS	HS	S
B110	BSSS	S	HS	S	MR	MR
B73	BSSS	S	HS	HS	S	R
CML170	TST	HR	R	R	HR	MR
KA064	陕A群ShaanA	MR	HR	S	MR	MR
KA081	陕A群ShaanA	HR	R	MR	R	S
KA103	陕A群ShaanA	R	R	R	S	MR
KA105	陕A群ShaanA	R	R	MR	R	R
KA106	陕A群ShaanA	MR	MR	S	MR	MR
KA109	陕A群ShaanA	R	MR	MR	MR	S
KA115	陕A群ShaanA	HR	MR	MR	MR	MR
KA147	陕A群ShaanA	R	R	MR	R	S
KA203	陕A群ShaanA	R	R	MR	HR	HS
KA225	陕A群ShaanA	MR	S	MR	S	HS
KA327	陕A群ShaanA	R	R	R	MR	MR
KB019	陕B群ShaanB	MR	MR	MR	S	MR
KB020	陕B群ShaanB	HR	HR	MR	R	MR
KB024	陕B群ShaanB	R	HS	S	MR	MR
KB025	陕B群ShaanB	MR	HS	S	MR	MR
KB043	陕B群ShaanB	MR	MR	S	R	S
KB052	陕B群ShaanB	MR	S	S	S	MR
KB062	陕B群ShaanB	S	S	HS	R	S
KB081	陕B群ShaanB	S	MR	MR	MR	MR
KB089	陕B群ShaanB	MR	HS	S	HS	MR
KB102	陕B群ShaanB	R	R	R	R	S
KB106	陕B群ShaanB	MR	S	S	MR	S
KB107	陕B群ShaanB	MR	S	MR	S	R
KB109	陕B群ShaanB	R	S	MR	R	MR
KB128	陕B群ShaanB	MR	MR	MR	MR	MR
KB204	陕B群ShaanB	R	MR	MR	S	R
KB207	陕B群ShaanB	R	R	S	R	S
KB227	陕B群ShaanB	R	MR	MR	R	S
KB243	陕B群ShaanB	R	S	S	MR	MR
KB588	陕B群ShaanB	HR	MR	MR	MR	S
Mo17	Lan	R	MR	MR	MR	MR
NW-H537	Reid	MR	R	MR	R	MR
PH4CV	Lan	S	MR	MR	MR	HS
PH6WC	BSSS	R	MR	R	HR	HS
PHK42	NSSS	MR	R	MR	MR	MR
PHN11	NSSS	S	S	HS	MR	MR
PHT60	NSSS	R	S	MR	MR	S
X178	P78599	HR	R	R	HR	MR
昌7-2 Chang7-2	SPT	HR	MR	R	S	HS
黄早四HZS	SPT	MR	MR	MR	HS	R
沈137 Shen137	P78599	HR	R	R	R	MR
郑58 Zheng58	Reid	R	R	R	R	MR