甘蔗抗黄锈病G1标记的分子检测及候选抗病基因WAK的分析

doi:10.3724/SP.J.1006.2021.04131

Abstract

Abstract:

Sugarcane orange rust is an important fungal disease caused by Puccinia kuehnii Butler, which could lead to a reduction in sugarcane production and sugar content and cause serious losses to the sugarcane industry in worldwide. In this study, the molecular marker G1 was used to detect orange rust resistance genes in cultivars, ancestral species and the related genus of Saccharum in the world. The representative amplified bands were cloned, sequenced, functionally annotated, and clustered, and the origin and evolution of resistance genes was then analyzed. The results showed that 83 and 34 were detected with G1 marker, accounting for 66.9% and 67.4% in 124 Chinese and 46 foreign sugarcane cultivars, respectively. Among 34 sugarcane ancestral species and the related genus of Saccharum, 17 were detected by G1 marker, accounting for 50%, of which the highest percentage (100%) was in Saccharum spontaneum. Functional annotation revealed that G1 target gene encoded a wall-associated receptor-like kinase (WAK), and three proteins with high similarity were identified from the haploid proteome database of sugarcane cultivars. These proteins all contain the extracellular domain, transmembrane domain and intracellular domain with kinase activity of typical cell wall receptor structures. Phylogenetic analysis of nucleotide sequences clearly showed the origin and evolution of the candidate resistance WAK genes. Specifically, the WAK genes amplified by G1 marker could be divided into three groups. The first group is from S. spontaneum and Saccharum robustum. The second group is from S. robustum, Saccharum officinarum and Narenga porphyrooma. The third group is from S. spontaneum, S. robustum, Saccharum sinense and cultivars. These results provided an important support for breeding of sugarcane cultivars resistant to orange rust, and lay a foundation for further analysis of molecular mechanism of resistance genes.

Key words: detection, molecular markers, wall-associated receptor-like kinase, orange rust, sugarcane

WANG Heng-Bo, CHEN Shu-Qi, GUO Jin-Long, QUE You-Xiong. Molecular detection of G1 marker for orange rust resistance and analysis of candidate resistance WAK gene in sugarcane[J].Acta Agronomica Sinica, 2021, 47(4): 577-586.

Figures/Tables 8

Table S1

Origins of 170 sugarcane cultivars and their PCR amplification results using G1 marker"

编号 No.	品种 Cultivar	来源 Origin	G1标记 Amplification result	备注 Note
1	赣南 99-591 Gannan 99-591	中国江西 Jiangxi, China	-	S. hybrid
2	四川 89-103 Sichuan 89-103	中国四川 Sichuan, China	+	S. hybrid
3	内江 00-118 Neijiang 00-118	中国四川 Sichuan, China	+	S. hybrid
4	内江 03-218 Neijiang 03-218	中国四川 Sichuan, China	-	S. hybrid
5	内江 92-44 Neijiang 92-44	中国四川 Sichuan, China	+	S. hybrid
6	内江 90-112 Neijiang 90-112	中国四川 Sichuan, China	+	S. hybrid
7	四川 73-219 Sichuan 73-219	中国四川 Sichuan, China	+	S. hybrid
8	新台糖 27号 ROC27	中国台湾 Taiwan, China	-	S. hybrid
9	新台糖 1号 ROC 1	中国台湾 Taiwan, China	+	S. hybrid
10	新台糖 16号 ROC 16	中国台湾 Taiwan, China	+	S. hybrid
11	新台糖 11号 ROC11	中国台湾 Taiwan, China	+	S. hybrid
12	新台糖 26号 ROC 26	中国台湾 Taiwan, China	+	S. hybrid
13	新台糖 95-0432 ROC 95-0432	中国台湾 Taiwan, China	+	S. hybrid
14	新台糖 95-8899 ROC 95-8899	中国台湾 Taiwan, China	+	S. hybrid
15	新台糖 20号 ROC 20	中国台湾 Taiwan, China	+	S. hybrid
16	新台糖 21号 ROC 21	中国台湾 Taiwan, China	+	S. hybrid
17	新台糖 23号 ROC 23	中国台湾 Taiwan, China	-	S. hybrid
18	新台糖 24号 ROC 24	中国台湾 Taiwan, China	-	S. hybrid
19	新台糖 10号 ROC10	中国台湾 Taiwan, China	-	S. hybrid
20	PT 43-52	中国台湾 Taiwan, China	+	S. hybrid
21	F108	中国台湾 Taiwan, China	-	S. hybrid
22	新台糖 22号 ROC 22	中国台湾Taiwan, China	+	S. hybrid
23	崖城 05-179 Yacheng 05-179	中国海南Hainan, China	+	S. hybrid
24	崖城 71-374 Yacheng 71-374	中国海南Hainan, China	+	S. hybrid
25	崖城 93-26 Yacheng 93-26	中国海南Hainan, China	-	S. hybrid
26	崖城 94-30 Yacheng 94-30	中国海南Hainan, China	+	S. hybrid
27	崖城 94-46 Yacheng 94-46	中国海南Hainan, China	+	S. hybrid
28	崖城 97-46 Yacheng 97-46	中国海南Hainan, China	-	S. hybrid
29	崖城 98-11 Yacheng 98-11	中国海南Hainan, China	-	S. hybrid
30	崖城 98-2 Yacheng 98-2	中国海南Hainan, China	+	S. hybrid
31	崖城 96-66 Yacheng 96-66	中国海南Hainan, China	-	S. hybrid
32	崖城 01-76 Yacheng 01-76	中国海南Hainan, China	+	S. hybrid
33	崖城 96-4 Yacheng 96-4	中国海南Hainan, China	-	S. hybrid
34	崖城 99-6 Yacheng 99-6	中国海南Hainan, China	+	S. hybrid
35	崖城 96-24 Yacheng 96-24	中国海南Hainan, China	+	S. hybrid
36	福农 02-3924 Funong 02-3924	中国福建Fujian, China	-	S. hybrid
37	福农 04-3504 Funong 04-3504	中国福建Fujian, China	+	S. hybrid
38	福农 99-20169 Funong 99-20169	中国福建Fujian, China	-	S. hybrid
39	福农 38号 Funong 38	中国福建Fujian, China	+	S. hybrid
40	福农 39号 Funong 39	中国福建Fujian, China	-	S. hybrid
41	福农 41号 Funong 41	中国福建Fujian, China	+	S. hybrid
42	闽糖 96-261 Mintang 96-261	中国福建Fujian, China	+	S. hybrid
43	农林 8 Nonglin 8	中国福建Fujian, China	+	S. hybrid
44	华南65-21 Huanan 65-21	中国广东Guangdong, China	-	S. hybrid
45	桂糖 94-119 Guitang 94-119	中国广西Guangxi, China	-	S. hybrid
46	桂糖 94-38 Guitang 94-38	中国广西Guangxi, China	-	S. hybrid
47	桂糖 96-211 Guitang 96-211	中国广西Guangxi, China	+	S. hybrid
48	柳城 01-1137 Liucheng 01-1137	中国广西Guangxi, China	+	S. hybrid
49	柳城 04-382 Liucheng 04-382	中国广西Guangxi, China	+	S. hybrid
50	柳城 05-129 Liucheng 05-129	中国广西Guangxi, China	+	S. hybrid
51	桂糖 02-761 Guitang 02-761	中国广西Guangxi, China	+	S. hybrid
52	桂糖 73-167 Guitang 73-167	中国广西Guangxi, China	+	S. hybrid
53	柳城 03-182 Liucheng 03-182	中国广西Guangxi, China	+	S. hybrid
54	桂糖 03-591 Guitang 03-591	中国广西Guangxi, China	+	S. hybrid
55	桂糖 03-1156 Guitang 03-1156	中国广西Guangxi, China	-	S. hybrid
56	桂糖 03-1403 Guitang 03-1403	中国广西Guangxi, China	+	S. hybrid
57	桂糖 03-2357 Guitang 03-2357	中国广西Guangxi, China	-	S. hybrid
58	桂糖 04-1007 Guitang 04-1007	中国广西Guangxi, China	+	S. hybrid
59	桂糖 08-278 Guitang 08-278	中国广西Guangxi, China	-	S. hybrid
60	桂糖 02-390 Guitang 02-390	中国广西Guangxi, China	+	S. hybrid
61	桂糖 05-2605 Guitang 05-2605	中国广西Guangxi, China	-	S. hybrid
62	桂糖 05-2743 Guitang 05-2743	中国广西Guangxi, China	+	S. hybrid
63	桂糖 05-322 Guitang 05-322	中国广西Guangxi, China	+	S. hybrid
64	桂糖 05-3661 Guitang 05-3661	中国广西Guangxi, China	+	S. hybrid
65	桂糖 05-375 Guitang 05-375	中国广西Guangxi, China	+	S. hybrid
66	桂糖 05-378 Guitang 05-378	中国广西Guangxi, China	+	S. hybrid
67	桂糖 05-3846 Guitang 05-3846	中国广西Guangxi, China	-	S. hybrid
68	桂糖 08-297 Guitang 08-297	中国广西Guangxi, China	+	S. hybrid
69	桂糖 35号 Guitang 35	中国广西Guangxi, China	-	S. hybrid
70	桂糖 05-827 Guitang 05-827	中国广西Guangxi, China	+	S. hybrid
71	桂糖 07-713 Guitang 07-713	中国广西Guangxi, China	-	S. hybrid
72	桂糖 02-1247 Guitang 02-1247	中国广西Guangxi, China	+	S. hybrid
73	桂糖 02-208 Guitang 02-208	中国广西Guangxi, China	-	S. hybrid
74	桂糖 02-619 Guitang 02-619	中国广西Guangxi, China	+	S. hybrid
75	桂糖 00-245 Guitang 00-245	中国广西Guangxi, China	+	S. hybrid
76	桂糖 96-211 Guitang 96-211	中国广西Guangxi, China	+	S. hybrid
77	桂糖 00-122 Guitang 00-122	中国广西Guangxi, China	-	S. hybrid
78	桂糖 02-351 Guitang 02-351	中国广西Guangxi, China	-	S. hybrid
79	桂糖 09-03 Guitang 09-03	中国广西Guangxi, China	+	S. hybrid
80	桂糖 96-167 Guitang 96-167	中国广西Guangxi, China	+	S. hybrid
81	桂糖 97-69 Guitang 97-69	中国广西Guangxi, China	-	S. hybrid
82	桂糖 00-173 Guitang 00-173	中国广西Guangxi, China	-	S. hybrid
83	桂糖 02-467 Guitang 02-467	中国广西Guangxi, China	+	S. hybrid
84	桂糖 29号 Guitang 29	中国广西Guangxi, China	+	S. hybrid
85	桂糖 30号 Guitang 30	中国广西Guangxi, China	-	S. hybrid
86	桂糖 31号 Guitang 31	中国广西Guangxi, China	-	S. hybrid
87	桂糖 92-66 Guitang 92-66	中国广西Guangxi, China	+	S. hybrid
88	桂糖 00-257 Guitang 00-257	中国广西Guangxi, China	+	S. hybrid
89	桂糖 94-119 Guitang 94-119	中国广西Guangxi, China	-	S. hybrid
90	桂糖35号 Guitang 35	中国广西Guangxi, China	+	S. hybrid
91	柳城 03-1137 Liucheng 03-1137	中国广西Guangxi, China	+	S. hybrid
92	德蔗 93-88 Dezhe 93-88	中国云南Yunnan, China	+	S. hybrid
93	云蔗 02-2332 Yunzhe 02-2332	中国云南Yunnan, China	-	S. hybrid
94	云蔗 02-2540 Yunzhe 02-2540	中国云南Yunnan, China	+	S. hybrid
95	云蔗 02-588 Yunzhe 02-588	中国云南Yunnan, China	+	S. hybrid
96	云蔗 03-194 Yunzhe 03-194	中国云南Yunnan, China	+	S. hybrid
97	云蔗 04-214 Yunzhe 04-214	中国云南Yunnan, China	+	S. hybrid
98	云蔗 05-51 Yunzhe 05-51	中国云南Yunnan, China	+	S. hybrid
99	云蔗 06-407 Yunzhe 06-407	中国云南Yunnan, China	+	S. hybrid
100	云蔗 94-343 Yunzhe 94-343	中国云南Yunnan, China	-	S. hybrid
101	云南 65-225 Yunzhe 65-225	中国云南Yunnan, China	+	S. hybrid
102	粤农 73-204 Yuenong 73-204	中国广东Guangdong, China	+	S. hybrid
103	粤甘 34号 Yuegan 34	中国广东Guangdong, China	-	S. hybrid
104	粤甘 16号 Yuegan 16	中国广东Guangdong, China	+	S. hybrid
105	粤农 73-204 Yuenong 73-204	中国广东Guangdong, China	-	S. hybrid
106	粤农 86-295 Yuenong 86-295	中国广东Guangdong, China	-	S. hybrid
107	粤糖 00-236 Yuetang 00-236	中国广东Guangdong, China	+	S. hybrid
108	粤糖 55 Yuetang 55	中国广东Guangdong, China	+	S. hybrid
109	粤糖 83-271 Yuetang 83-271	中国广东Guangdong, China	+	S. hybrid
110	粤糖 93-159 Yuetang 93-159	中国广东Guangdong, China	+	S. hybrid
111	粤糖 94-128 Yuetang 94-128	中国广东Guangdong, China	+	S. hybrid
112	粤糖 96-86 Yuetang 96-86	中国广东Guangdong, China	+	S. hybrid
113	湛蔗 19号 Zhanzhe 19	中国广东Guangdong, China	+	S. hybrid
114	湛蔗 22号 Zhanzhe 22	中国广东Guangdong, China	+	S. hybrid
115	湛蔗 24号 Zhanzhe 24	中国广东Guangdong, China	-	S. hybrid
116	湛蔗 74-141 Zhanzhe 74-141	中国广东Guangdong, China	+	S. hybrid
117	粤甘 24号 Yuegan 24	中国广东Guangdong, China	+	S. hybrid
118	粤糖 89-240 Yuetang 89-240	中国广东Guangdong, China	+	S. hybrid
119	湛蔗 92-126 Zhanzhe 92-126	中国广东Guangdong, China	-	S. hybrid
120	湛蔗 26号 Zhanzhe 26	中国广东Guangdong, China	+	S. hybrid
121	湛蔗 80-101 Zhanzhe 80-101	中国广东Guangdong, China	+	S. hybrid
122	粤糖 00-319 Guitang00-319	中国广东Guangdong, China	+	S. hybrid
123	粤糖 60号 Yuetang 60	中国广东Guangdong, China	-	S. hybrid
124	科5 Ke 5	中国广东Guangdong, China	+	S. hybrid
125	FR94-280	法国 France	+	S. hybrid
126	FR99-49	法国 France	-	S. hybrid
127	RB83-5089	巴西 Brazil	+	S. hybrid
128	Q208	澳大利亚 Australia	-	S. hybrid
129	Q117	澳大利亚 Australia	-	S. hybrid
130	Q200	澳大利亚 Australia	+	S. hybrid
131	Q209	澳大利亚 Australia	+	S. hybrid
132	Co1001	印度 India	+	S. hybrid
133	Co281	印度 India	+	S. hybrid
134	Co290	印度 India	+	S. hybrid
135	PINDAR	印度 India	-	S. hybrid
136	Co419	印度 India	+	S. hybrid
137	POJ2878	印度尼西亚 Indonesia	-	S. hybrid
138	NCO376	南非 South Africa	+	S. hybrid
139	NCO310	南非 South Africa	+	S. hybrid
140	CP67-412	美国 USA	+	S. hybrid
141	CP72-1210	美国 USA	+	S. hybrid
142	CP72-2086	美国 USA	-	S. hybrid
143	CP72-330	美国 USA	+	S. hybrid
144	CP72-3591	美国 USA	+	S. hybrid
145	CP82-1592	美国 USA	+	S. hybrid
146	CP84-1198	美国 USA	-	S. hybrid
147	CP85-1308	美国 USA	+	S. hybrid
148	CP86-1633	美国 USA	+	S. hybrid
149	CP88-1762	美国 USA	+	S. hybrid
150	CP89-1509	美国 USA	+	S. hybrid
151	CP89-2143	美国 USA	-	S. hybrid
152	CP92-1213	美国 USA	+	S. hybrid
153	CP94-1100	美国 USA	+	S. hybrid
154	CP94-1340	美国 USA	+	S. hybrid
155	CP96-1602	美国 USA	+	S. hybrid
156	H32-8560	美国 USA	+	S. hybrid
157	HoCP91-555	美国 USA	+	S. hybrid
158	HoCP92-624	美国 USA	-	S. hybrid
159	HoCP93-746	美国 USA	+	S. hybrid
160	HoCP95-988	美国 USA	-	S. hybrid
161	HoCP94-806	美国 USA	+	S. hybrid
162	HoCP95-950	美国 USA	-	S. hybrid
163	CP80-1277	美国 USA	+	S. hybrid
164	CP57-641	美国 USA	+	S. hybrid
165	CP92-1666	美国 USA	-	S. hybrid
166	CP93-182	美国 USA	-	S. hybrid
167	CP49-50	美国 USA	+	S. hybrid
168	CP67-412	美国 USA	+	S. hybrid
169	CP28-11	美国 USA	-	S. hybrid
170	R570	法国 France	-	S. hybrid

Table S1

Table S2

Origins of 34 germplasms from Saccharum and the related genus and their PCR amplification results using G1 marker"

编号 No.	品种 Name	来源 Origin	G1标记 Amplification result	备注 Note
1	贵州 78-2-12 Guizhou 78-2-12	中国贵州 Guizhou, China	+	Erianthus arundinaceus
2	云南 95-19 Yunnan 95-19	中国云南 Yunnan, China	-	Erianthus rockii
3	广西 79-8 Guangxi 79-8	中国广西 Guangxi, China	-	Miscanthus floridulus
4	云南 95-35 Yunnan 95-35	中国云南 Yunnan, China	-	Miscanthus sinensis
5	广西 89-13 Guangxi 89-13	中国广西 Guangxi, China	+	Narenga porphyrooma
6	广东 64号 Guangdong 64	中国广东 Guangdong, China	-	Narenga porphyrooma
7	HATUNI	印度 India	+	S. barberi
8	Katha	印度 India	-	S. barberi
9	Nangas	印度 India	-	S. barberi
10	LA-purple	印度尼西亚 Indonesia	-	S. officinarum
11	Badila	印度尼西亚 Indonesia	-	S. officinarum
12	路达仕 Loethers	美国 USA	-	S. officinarum
13	黑车里本 Black cheribon	美国 USA	-	S. officinarum
14	克里斯塔林娜 Crystalina	美国 USA	-	S. officinarum
15	云南大野Yunnandaye	中国云南 Yunnan, China	-	S. robustum
16	福建大野Fujiandaye	中国福建 Fujian, China	+	S. robustum
17	51NG3	印度尼西亚 Indonesia	-	S. robustum
18	57NG208	印度尼西亚 Indonesia	-	S. robustum
19	NG77004	印度尼西亚 Indonesia	-	S. robustum
20	MOL-6081	美国 USA	+	S. robustum
21	桂林竹蔗 Guilinzhuzhe	中国广西 Guangxi, China	+	S. sinense
22	Youba	中国云南 Yunnan, China	-	S. sinense
23	松溪竹蔗Songxizhuzhe	中国福建 Fujian, China	-	S. sinense
24	贵州 78-1-11 Guizhou 78-1-11	中国贵州 Guizhou, China	+	S. spontaneum
25	贵州 78-2-28 Guizhou 78-2-28	中国贵州 Guizhou, China	+	S. spontaneum
26	四川 79-1-26 Sichuan79-1-26	中国四川 Sichuan, China	+	S. spontaneum
27	四川 79-2-11 Sichuan 79-2-11	中国四川 Sichuan, China	+	S. spontaneum
28	云南 75-2-11 Yunnan 75-2-11	中国云南 Yunnan, China	+	S. spontaneum
29	广东 21号 Guangdong 21	中国广东 Guangdong, China	+	S. spontaneum
30	广东 30号 Guangdong 30	中国广东 Guangdong, China	+	S. spontaneum
31	福建 89-1-1 Fujian 89-1-1	中国福建 Fujian, China	+	S. spontaneum
32	福建 89-1-16 Fujian 89-1-16	中国福建 Fujian, China	+	S. spontaneum
33	印度1号 Yindu 1	印度 India	+	S. spontaneum
34	印度2号 Yindu 2	印度 India	+	S. spontaneum

Table S2

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

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Molecular detection of G1 marker for orange rust resistance and analysis of candidate resistance WAK gene in sugarcane

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