Evaluation of Resistance to Spot Blotch in Core Germplasm Resources

doi:10.13304/j.nykjdb.2022.0183

Journal of Agricultural Science and Technology ›› 2022, Vol. 24 ›› Issue (11): 43-54.DOI: 10.13304/j.nykjdb.2022.0183

• BIOTECHNOLOGY & LIFE SCIENCE • Previous Articles Next Articles

Evaluation of Resistance to Spot Blotch in Core Germplasm Resources

Yifan SUN¹^,²(), Zhilei HUANG¹^,²(), Baochun LI¹^,³, Lirong YAO¹^,², Juncheng WANG¹^,², Erjing SI¹^,², Ke YANG¹^,², Yaxiong MENG¹^,², Xiaole MA¹^,², Huajun WANG¹^,²()

^1.State Key Laboratory of Aridland Crop Science，Gansu Provincial Key Laboratory of Crop Improvement & Germplasm Enhancement，Gansu Agricultural University，Lanzhou 730070，China
^2.College of Agronomy，Gansu Agricultural University，Lanzhou 730070，China
^3.College of Life Science and Technology，Gansu Agricultural University，Lanzhou 730070，China

Received:2022-03-11 Accepted:2022-05-23 Online:2022-11-15 Published:2022-11-29
Contact: Huajun WANG

大麦种质资源抗叶斑病评价

孙逸凡¹^,²(), 黄志磊¹^,²(), 李葆春¹^,³, 姚立蓉¹^,², 汪军成¹^,², 司二静¹^,², 杨轲¹^,², 孟亚雄¹^,², 马小乐¹^,², 王化俊¹^,²()

^1.甘肃农业大学省部共建干旱生境作物学国家重点实验室，甘肃省作物遗传改良与种质创新重点实验室，兰州 730070
^2.甘肃农业大学农学院，兰州 730070
^3.甘肃农业大学生命科学技术学院，兰州 730070

通讯作者: 王化俊
作者简介:孙逸凡 E-mail：790133094@qq.com
黄志磊 E-mail：752127656@qq.com第一联系人：孙逸凡和黄志磊为共同第一作者。
基金资助:
财政部和农业农村部国家现代农业产业技术体系建设项目(CARS-05-04B-2);甘肃省教育厅产业支撑计划项目(2021CYZC-12);国家自然科学基金项目(32001514);973计划前期研究专项(2014CB160313);甘肃省自然基金项目(20JR10RA507);甘肃农业大学伏羲青年英才项目(GANfx-03Y06)

Abstract

Abstract:

In order to clarify the resistance differences of different barley germplasms to leaf spot， the resistant germplasm materials were screened and related genetic loci were mined. In this study， 200 barley germplasms were inoculated with Z14484， a highly pathogenic strain of barley leaf spot， at three-leaf stage. After cultured for 9 d， the incidence of each material was counted， the disease resistance grade was clarified， and resistance identification and evaluation were carried out. Combined with Illumina 9K SNP chip， the population and significant loci were analyzed by Structure software and PLINK software to identify SNP loci significantly associated with traits. The results showed that there were significant differences in the resistance among the tested barley materials， and the population genetic structure could be divided into 3 subgroups at ΔK=3 level， including 100， 67 and 33 barley materials， respectively. 11_11436 and SCRI_RS_201075 located on chromosome 3H were most associated with resistance to barley leaf spot by genome-wide association study. This study provided a reference for barley leaf spot disease resistance gene mapping and disease resistance breeding.

Key words: spot blotch, resistance identification, SNP, genome-wide association study

摘要：

为明确不同大麦种质对叶斑病的抗性差异，筛选抗性种质材料和挖掘相关的遗传位点，在三叶期对200份大麦种质接种叶斑病强致病株Z14484，培养9 d后统计各材料发病情况，明确其抗病等级，进行抗性鉴定与评价。结合Illumina 9K SNP芯片，通过Structure和PLINK软件进行群体和显著位点分析，鉴定与性状显著关联的SNP位点。结果表明，供试大麦材料抗性差异显著，在ΔK=3水平上群体遗传结构可以分为3个亚群，分别包含100、67、33份大麦材料；全基因组关联分析（genome-wide association study，GWAS）共检测到12个与大麦叶斑病抗性相关的位点，其中位于3H染色体上的11_11436和SCRI_RS_201075与大麦叶斑病抗性关联最大。研究结果为大麦叶斑病抗病基因定位与抗病育种提供参考依据。

关键词: 大麦叶斑病, 抗性鉴定, SNP, 全基因组关联分析

CLC Number:

S512.3

Yifan SUN, Zhilei HUANG, Baochun LI, Lirong YAO, Juncheng WANG, Erjing SI, Ke YANG, Yaxiong MENG, Xiaole MA, Huajun WANG. Evaluation of Resistance to Spot Blotch in Core Germplasm Resources[J]. Journal of Agricultural Science and Technology, 2022, 24(11): 43-54.

孙逸凡, 黄志磊, 李葆春, 姚立蓉, 汪军成, 司二静, 杨轲, 孟亚雄, 马小乐, 王化俊. 大麦种质资源抗叶斑病评价[J]. 中国农业科技导报, 2022, 24(11): 43-54.

Figures/Tables 13

References 24

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等级 Level	病态特征 Symptoms of disease
0	完全没有发病 Completely without disease
1	圆形或椭圆形棕褐色坏死组织和斑点，面积小数量多 A small number of area which are round or oval brown necrotic tissue and spots
2	棕褐色斑点增大，病变周围叶片仍为绿色 Following brown spots increased， the lesion around the leaves are remain green
3	斑点增大，周围有不明显的萎黄区域 No obvious chlorosis area around the spots with tends to be more larger
5	斑点周围有明显萎黄区域并随时间增大 An increasing clearly chlorotic areas near leafspots flowing the elongation of time
7	斑点及萎黄区域变大 The spots and chlorosis area are varying larger
8	坏色组织和斑点数量少，面积大，直接影响叶片光合作用 A small number of spots with large areas of necrotic tissue on leaves， greatly affected by the leaf photosynthesis
9	病斑明显，严重萎黄，以致叶片死亡 The spots are obvious lesions and severe chlorosis， resulting in leaf death

等级 Level	病态特征 Symptoms of disease
0	完全没有发病 Completely without disease
1	圆形或椭圆形棕褐色坏死组织和斑点，面积小数量多 A small number of area which are round or oval brown necrotic tissue and spots
2	棕褐色斑点增大，病变周围叶片仍为绿色 Following brown spots increased， the lesion around the leaves are remain green
3	斑点增大，周围有不明显的萎黄区域 No obvious chlorosis area around the spots with tends to be more larger
5	斑点周围有明显萎黄区域并随时间增大 An increasing clearly chlorotic areas near leafspots flowing the elongation of time
7	斑点及萎黄区域变大 The spots and chlorosis area are varying larger
8	坏色组织和斑点数量少，面积大，直接影响叶片光合作用 A small number of spots with large areas of necrotic tissue on leaves， greatly affected by the leaf photosynthesis
9	病斑明显，严重萎黄，以致叶片死亡 The spots are obvious lesions and severe chlorosis， resulting in leaf death

抗病类型 Resistant type	病情指数DI
HR：高抗High resistant	0<DI≤5
R：抗性 Resistant	5<DI≤20
R-MR：抗-中抗 Resistant-moderate resistant	20<DI≤35
MR：中抗 Moderate resistant	35<DI≤50
MR-MS：中抗-中感 Moderate resistant- moderate susceptible	50<DI≤65
MS：中感 Moderate susceptible	65<DI≤80
MS-S：中感-高感Moderate susceptible-susceptible	80<DI≤95
S：高感 Susceptible	DI>95

抗病类型 Resistant type	病情指数DI
HR：高抗High resistant	0<DI≤5
R：抗性 Resistant	5<DI≤20
R-MR：抗-中抗 Resistant-moderate resistant	20<DI≤35
MR：中抗 Moderate resistant	35<DI≤50
MR-MS：中抗-中感 Moderate resistant- moderate susceptible	50<DI≤65
MS：中感 Moderate susceptible	65<DI≤80
MS-S：中感-高感Moderate susceptible-susceptible	80<DI≤95
S：高感 Susceptible	DI>95

编号Code	材料名称Material	抗性类型Resistance type	病情指数DI
1	六棱大麦Liulengdamai	R	6.44
2	兰坪大麦1号Lanpingdamai 1	MR	37.06
3	拉绥青稞Lasuiqingke	MR	39.06
4	浙秀12 Zhexiu 12	MR-MS	60.94
5	嘎香Gaxiang	MS-S	84.52
6	金钩大Jingouda	MR-MS	53.47
7	甘啤4Ganpi 4	MS-S	75.83
8	红芒大麦Hongmangdamai	MS-S	83.04
9	吉塘青稞Jitangqingke	MS-S	80.52
10	Zdm00190	MS-S	82.14
11	吉啤1号Jipi 1	MS	65.97
12	大麦2号Damai 2	MR	38.66
13	扬农啤8 Yangnongpi 8	MS-S	84.76
14	金华四角米大麦Jinhuasijiaomidamai	R	16.45
15	林口皮1号Linkoupi 1	R-M	32.34
16	南京374Nanjing 374	MS	70.56
17	谷城三月黄Guchengsabyuehuang	MR-MS	61.93
18	中品861428 Zhongpin 861428	R	19.39
19	六棱谷老麦Liulenggulaomai	MS-S	86.64
20	九龙白四棱Jiulongbaisileng	MR	45.69
21	黑大麦Heidamai	MS-S	84.27
22	江孜青稞Jiangziqingke	R-M	28.82
23	萨达青Sadaqing	MR	44.9
24	和田山青稞Hetianshanqingke	MS	70
25	贡乃Gongnai	R	17.67
26	六合四棱大麦Liuhesilengdamai	R-M	24.41

Evaluation of Resistance to Spot Blotch in Core Germplasm Resources

大麦种质资源抗叶斑病评价

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Figures/Tables 13

References 24

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编号Code	材料名称Material	抗性类型Resistance type	病情指数DI
27	垦啤麦5号Kenpimai 5	MS	69.44
28	Zdm01467	MS	69.27
29	嘎拉索珠Galasuozhu	MS-S	81.51
30	吉8703 Ji 8703	MS	65.7
31	达孜白青稞Dazibaiqingke	R	8.09
32	朗县钩芒青稞Langxiangoumangqingke	MR	37.98
33	普洱大麦Puerdamai	MR-MS	57.68
34	藏3002 Zang 3002	MR	48.15
35	德令哈青稞Delinghaqingke	MS	65.54
36	Zdm01678	MR-MS	59.38
37	肥城米大麦Feichengmidamai	R	10.9
38	乐都红六棱Leduhongliuleng	R	6.25
39	莱芜紫米大麦 Laiwuzimidamai	MR	49.62
40	洞四青稞 Dongsiqingke	MS	73.81
41	海南白六棱 Hainanbailiuleng	R-M	22.61
42	六轮大麦 Liulundamai	MS-S	82.43
43	吉青稞 Jiqingke	R-M	24.65
44	青农55 Qingnong 55	MR-MS	53.75
45	甘啤6号 Ganpi 6	MR-MS	59.18
46	中品861140 Zhongpin 861140	MS-S	87.5
47	酒大麦 Jiudamai	R-M	22.67
48	六十天成熟 Liushitianchengshu	R	8.65
49	老来唱 Laolaichang	MR-MS	51.02
50	甘啤3 Ganpi 3	MS-S	75.45
51	Zdm03379	MR	44.85
52	白青稞 Baiqingke	MR	42.86
53	尚志皮4号 Sahngzhipi 4	MS-S	76.07
54	Zdm03609	R-M	31.77
55	耐那 Naina	MR-MS	57
56	擦旺青稞 Cawangqingke	MS	70.45
57	钩芒紫白色青稞 Goumangzibaiqingke	MS	65.73
58	大麦 Damai	MR	47.92
59	春青稞 Chunqingke	MS-S	86.61
60	静宁青稞 Jingningqingke	MS	67.71
61	二月黄 Eryuehuang	MS-S	83.04
62	加久 Jiajiu	MR	49.58
63	洞嘎青稞 Donggaqingke	MS-S	75.71
64	红露仁大麦 Honglurengdamai	MS-S	76.44
65	红穗麦 Hongsuimai	MS	67.5
66	丹阳长穗晚元麦 Danyangchangsuiwan Yuanmai	MS-S	87.95
67	桔青稞 Juqingke	MS	69.61
68	Zdm04517	MS	67.59
69	Zdm04756	MS	68.9
70	新泰米大麦 Xintaimidamai	MS	65.06
71	中品861711 Zhongpin 861711	MR-MS	57.39
72	藏2994 Zang 2994	R-M	28.75
73	榕江老麦 Rongjianglaomai	R-M	30.43
74	桑早六 Sangzaoliu	R	18.75
75	六棱对芒大麦 Liulengduimangdamai	MS-S	81.47
76	吉862 Ji 862	R	12.98

编号Code	材料名称Material	抗性类型Resistance type	病情指数DI
77	丽水有芒裸麦 Lishuiyoumangluomai	MS-S	84.05
78	江宁1395 Jiangning 1395	MS	74.22
79	和林大麦 Helindamai	MS	71.63
80	鼓锤麦 Guchuimai	MR-MS	58.55
81	中品861132 Zhongpin 861132	MR-MS	55.5
82	Zdm05925	R-M	20.04
83	六棱麦 Liulengmai	MS-S	77.08
84	红原白青稞 Hongyuanbaiqingke	R-M	21.13
85	蒙啤麦1号 Mengpimai 1	S	95.16
86	蒙啤麦3号 Mengpimai 3	R	5.45
87	Zdm08028	MS	74.31
88	嘎拉索珠 Galasuozhu	MR-MS	60.44
89	Zdm08183	MS-S	80.41
90	合川大麦 Hechuandamai	MS	73.92
91	桦南裸1号 Huananluo 1	MR-MS	56.25
92	Zdm08385	MR	44.17
93	中品861596 Zhongpin 861596	MR-MS	63.78
94	青田红大麦 Qingtianhongdamai	R-M	26.45
95	甘啤2 Ganpi 2	MR-MS	50.82
96	黑山大麦 Heishandamai	MR	46.67
97	中品861616 Zhongpin 861616	MR-MS	63.94
98	奉化六棱 Fenghualiuleng	MR-MS	60.1
99	日喀则紫青稞 Rikazeziqingke	R-M	28.74
100	桑早六 Sangzaoliu	MR	41.85
101	沪麦8号Humai 8	MR-MS	54.61
102	贵德黑青稞Guideheiqingke	MS	73.61
103	Zdm08483	MR	47.58
104	洛甫青稞Luofuqingke	MR	42.32
105	达旗大麦Daqidamai	MR	47.97
106	Zdm08697	MR	46.23
107	壤塘黑颈白六棱Rangtangheijinbailiuleng	R-M	28.48
108	肚里黄Dulihuang	MR-MS	50.67
109	花22 Hua 22	MR-MS	55.3
110	Zdm08883	MS-S	81.25
111	青农83 Qingnong 83	R-M	25.88
112	Zdm08928	MS-S	78.19
113	互助藏青稞 Huzhuzangqingke	MS-S	77.39
114	曲松青稞 Qusongqingke	R-M	28.47
115	洞头有芒大麦Dongtouyoumangdamai	MR-MS	50.66
116	普陀二棱Putuoerleng	MR	38.24
117	化隆双槽棱Hualongshuangcaoleng	MR-MS	55.9
118	立新2号Lixin 2	R	11.92
119	中品861461 Zhongpin 861461	MR-MS	63.28
120	Zdm09952	MS-S	75.5
121	二长四短芒白青稞Erchangsiduanmang baiqingke	MS	67.72
122	大名三月黄Damingsanyuehuang	MS	66.35
123	莎车大麦Shachedamai	R-M	30.71
124	武城大麦Wuchengdamai	R-M	29.09
125	Zdm10006	R-M	33.46
126	中品861127 Zhongpin 861127	MR-MS	58.5

编号Code	材料名称Material	抗性类型Resistance type	病情指数DI
127	高原早1号 Gaoyuanzao 1	MR-MS	61.56
128	中品861587 Zhongpin 861587	MR-MS	63.36
129	庆元长芒大麦Qingyuanchangdamai	R-M	26.72
130	凉城洋草麦Liangchengyangcaomai	R	13.16
131	中品861106 Zhongpin 861106	MR-MS	58.24
132	莆田90天Putian 90 tian	MR	37.95
133	藏3001 Zang 3001	MR-MS	57.61
134	中品861437Zhongpin 861437	MR-MS	62.95
135	驻大麦4号Zhudamai 4	MR	40.54
136	中品861220 Zhongpin 861220	MR-MS	59.18
137	垦啤麦4号Kenpimai 4	MR	37.88
138	清水河草麦Qingshuihecaomai	R-M	26.67
139	迭部紫青稞 Diebuziqingke	MR	49.44
140	花30 Hua 30	MR-MS	55.47
141	密穗大麦 Misuidamai	R-M	34.87
142	中品861087 Zhongpin 861087	MR-MS	58.15
143	三月黄 Sanyuehuang	R-M	30
144	湟源白浪散青稞 Huangyuanbailangsan qingke	MR-MS	56.94
145	木选1号 Muxuan 1	MR	37.1
146	墨玉青稞 Moyuqingke	MR	35.85
147	光芒大麦 Guangmangdamai	MS	72.41
148	驻大麦3号 Zhudamai 3	MR	39.38
149	谷麦 Gumai	MS	71.43
150	嘎母 Gamu	MR-MS	60.87
151	格东有须老麦 Gedongyouxulaomai	MS	69.64
152	春大麦 Chundamai	MR	47.57
153	江达青稞 Jiangdaqingke	MS-S	83.15
154	青农94 Qingnong 94	R-M	25.93
155	驻大麦5 Zhudamai 5	MR	40.67
156	凤大麦6 Fengdamai 6	R-M	22.68
157	光头大麦 Guangtoudamai	MS	72.92
158	大通红六棱 Datonghongliuleng	MS	66.39
159	扬农啤6 Yangnongpi 6	R-M	24.53
160	果洛 Guoluo	MS	73.64
161	滇193 Dian 193	MR	49.43
162	乃那 Naina	R-M	31.4
163	中品86170 Zhongpin 86170	MR-MS	64.64
164	龙泉四棱 Longquansileng	MS	69.13
165	白大麦 Baidamai	MR	43.75
166	米林白青稞 Milinbaiqingke	R-M	32.86
167	滁县和尚头 Chuxianheshangtou	MR	47.18
168	老青稞 Laoqingke	R	10.67
169	驻大麦7 Zhudamai 7	MR	41.75
170	八宿青稞 Basuqingke	MR	43.27
171	六棱大麦 Liulengdamai	R	18.16
172	当地青稞 Dangdiqingke	MR	48.01
173	六棱米大麦 Liulengmidamai	R-M	20.14
174	长芒紫黑青稞 Changmangzuheiqingke	MR-MS	59.27
175	花11 Hua 11	MR-MS	55.29
176	Zym01448	R-M	20.83

编号Code	材料名称Material	抗性类型Resistance type	病情指数DI
177	Zym01472	R-M	23.71
178	列麦青稞 Liemaiqingke	R	13.39
179	中品861732 Zhongpin 861732	MS	65.28
180	曲水青稞 Qushuiqingke	R-M	27.61
181	黄光芒不规则型大麦 Huangguangmang Buguizexingdamai	MR-MS	56.52
182	华大麦8 Huadamai 8	R-M	23.54
183	贡嘎钩芒青稞 Gonggagoumangqingke	MS	69.71
184	中品861713 Zhongpin 861713	MS	65.18
185	黑连皮 Heilianpi	MR	46.43
186	谷大麦 Gudamai	MS	70.63
187	灯塔大麦 Dengtadamai	MR	49.26
188	米麦 Mimai	R-M	33.33
189	定日地方大麦 Dingridifangdamai	MR	49.55
190	Zym02413	MR	38.13
191	中品861375 Zhongpin 861375	MR-MS	61.76
192	Zym02507	MS-S	75.23
193	中品861592 Zhongpin 861592	MR-MS	63.75
194	黄麦 Huangmai	MR-MS	56.58
195	Zym02595	MS	69.64
196	红壳大麦 Hongkedamai	MR	46.79
197	Zym02611	MS	75
198	冬青6号 Dongqin 6	MR	49.57
199	Zym02830	MS	74.74
200	沪麦10 Humai 10	MR-MS	54.46

染色体 Chromosome	A/C	A/G	A/T	C/G	G/C	T/A	T/C	T/G	总计Total	碱基对比例 Base pair ratio/%
1H	54	236	11	33	20	10	226	52	642	9.95
2H	94	427	14	38	35	17	419	100	1 144	17.73
3H	99	355	14	43	32	15	346	83	987	15.29
4H	57	248	14	34	19	11	242	61	686	10.63
5H	124	494	26	45	36	21	420	122	1 288	19.96
6H	62	293	13	34	24	18	309	66	819	12.69
7H	88	333	18	37	29	12	307	64	888	13.76
合计Total	578	2 386	110	264	195	104	2 269	548	6 454	—
碱基对比例 Base pair ratio/%	8.96	36.97	1.70	4.09	3.02	1.61	35.16	8.49	—	—

SNP位点 SNP location	POS值 Pos value	SNP	染色体 Chromosome	-logP
SCRI_RS_168529	6.15	A/G	3H	5.70
11_20294	6.49	A/C	3H	3.80
SCRI_RS_159598	23.10	T/C	3H	3.50
11_10565	25.27	A/G	3H	3.90
SCRI_RS_159002	39.38	A/G	3H	1.97
SCRI_RS_171415	67.90	T/C	3H	4.42
SCRI_RS_202772	70.20	T/C	3H	1.65
BOPA1_ABC18746-1-2-109	90.51	A/G	3H	5.77
11_11436	145.65	C/G	3H	1.42
SCRI_RS_201075	145.69	A/G	3H	1.42
SCRI_RS_168096	57.20	A/G	6H	5.40
SCRI_RS_205103	58.24	T/G	6H	5.50

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