番茄颈腐根腐病抗性鉴定技术的建立及抗性种质资源筛选

doi:10.3864/j.issn.0578-1752.2022.04.007

Abstract

Abstract:

【Objective】The objective of this study is to explore the resistance identification technology of Fusarium crown and root rot (FCRR), carry out the resistance identification and analysis of tomato germplasm resources and varieties, enrich the available tomato resources resistant to FCRR, and to lay the foundation for the cultivation of tomato varieties that resistant to FCRR. 【Method】In the present study, four parameters that influence the inoculation effect including inoculum concentration, seedlings stage, environmental temperature and inoculation methods were studied using susceptible line Heinz 1706 infected with Fusarium oxysporum f. sp. radicis-lycopersici (Forl). Subsequently, 100 tomato materials were identified through artificial inoculation at seedling stage as well as the molecular markers linked to the resistance gene. 【Result】The disease incidence and disease index of FCRR increased in a certain range along with the increase of inoculum concentration, and the actual levels of plant resistance could be revealed with the inoculum of 10⁷spores/mL, for which the disease incidence and disease index were 100% and 89.2, respectively. The disease index was not significantly different among different seedling ages when the host was inoculated at 2 to 5 leaf stage. The effects of different environmental temperatures after inoculation on the disease index of FCRR were significantly different and the lower temperature (20℃) condition was more favorable to the occurrence of FCRR. The incidence and disease index were higher and the effect was stable using root dipping and root irrigation methods, which were significantly better than stem injection. The result of inoculation identification suggested that 38 materials showed resistance to FCRR among 100 tomato materials, which could be used for breeding or production in tomato for FCRR resistance. In addition, among the reported markers linked to Frl, SCAR_Frl has the lowest accuracy of 51%, while C2-25 has the accuracy of 59% and the accuracy of PNU-D4 is 83%, which is expected to be used in marker-assisted selection (MAS) for FCRR resistance. 【Conclusion】The established assessment system can identify levels of the resistance to FCRR in tomato seedlings, which can be used for identification and screening of tomato materials.

Key words: tomato, Fusarium crown and root rot (FCRR), resistance identification, molecular marker

WANG MengRui, LIU ShuMei, HOU LiXia, WANG ShiHui, LÜ HongJun, SU XiaoMei. Development of Artificial Inoculation Methodology for Evaluation of Resistance to Fusarium Crown and Root Rot and Screening of Resistance Sources in Tomato[J].Scientia Agricultura Sinica, 2022, 55(4): 707-718.

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Figures/Tables 5

Fig. 1

Table 1

Table 2

Table 3

The results of artificial inoculation and molecular detection of tomato germplasms"

编号 Number	材料 Material	世代 Generation	材料来源 Source	病情分级 DSS					病情指数 DI	抗级 RT	分子标记Molecular marker
编号 Number	材料 Material	世代 Generation	材料来源 Source	0级	1级	2级	3级	4级	病情指数 DI	抗级 RT	C2-25	PNU-D4	Tm-2	SCAR_Frl
1	VF-36	自交系IL	TGRC	0/0	0/0	0/0	4/0	4/8	93.8±8.8	HS	S	S	S	S
2	Moneymaker	自交系IL	TGRC	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	S	S	S	S
3	M82	自交系IL	TGRC	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	S	S	S	S
4	Hawaii 7998	自交系IL	TGRC	0/0	0/0	0/0	2/0	6/8	96.9±4.4	HS	S	S	S	S
5	LA2711	自交系IL	TGRC	0/0	0/0	1/0	4/1	3/7	89.1±11.0	HS	S	S	S	S
6	LA3008	自交系IL	TGRC	0/0	0/0	0/0	4/2	4/6	90.6±4.4	HS	S	S	S	S
7	Micro-Tom	自交系IL	TGRC	0/0	0/0	1/0	2/2	5/6	90.6±4.4	HS	S	S	S	S
8	E-6203	自交系IL	TGRC	0/0	1/0	1/0	1/1	5/7	89.1±11.0	HS	S	S	S	S
9	Ailsa Craig	自交系IL	TGRC	0/0	0/0	1/0	1/1	6/7	93.8±4.4	HS	S	S	S	S
10	KR2/9706	自交系IL	中国农业科学院 CAAS	0/0	0/0	0/0	1/1	7/7	96.9±0	HS	S	S	R	R
11	Momotaro yoku	自交系IL	TGRC	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	R	R
12	LA1589	自交系IL	TGRC	0/0	0/0	0/0	1/2	7/6	95.3±2.2	HS	S	S	S	S
13	TS-40	自交系IL	TGRC	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	S	S	R	R
14	TS-52	自交系IL	TGRC	0/0	0/0	0/0	4/0	4/8	93.8±8.8	HS	R	S	R	R
15	TS-151	自交系IL	TGRC	0/0	0/0	0/0	3/1	5/7	93.8±4.4	HS	S	S	S	S
16	TS-175	自交系IL	TGRC	0/0	0/0	0/0	3/0	5/8	95.3±6.6	HS	S	S	R	R
17	TS-211	自交系IL	TGRC	0/0	0/0	0/0	4/1	4/7	92.2±6.6	HS	R	R	R	R
18	Heinz1706	自交系IL	TGRC	0/0	0/0	0/0	1/2	7/6	95.3±2.2	HS	S	S	S	S
19	TS-272	自交系IL	TGRC	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	R	R	R	R
20	ZM-4	自交系IL	山东省农业科学院 SAAS	7/6	1/2	0/0	0/0	0/0	4.7±2.2	HR	S	R	R	R
21	ZM-6	自交系IL	山东省农业科学院 SAAS	8/8	0/0	0/0	0/0	0/0	0	I	S	R	S	S
22	ZM-7	自交系IL	山东省农业科学院 SAAS	8/8	0/0	0/0	0/0	0/0	0	I	S	R	S	S
23	12g-60	自交系IL	中国农业科学院 CAAS	8/8	0/0	0/0	0/0	0/0	0	I	R	R	R	R
24	LA1791^a	自交系IL	TGRC	8/8	0/0	0/0	0/0	0/0	0	I	R	R	R	R
25	LA2829^a	自交系IL	TGRC	8/8	0/0	0/0	0/0	0/0	0	I	R	R	R	R
26	LA3273^a	自交系IL	TGRC	8/8	0/0	0/0	0/0	0/0	0	I	R	R	R	R
27	LA3292^a	自交系IL	TGRC	8/8	0/0	0/0	0/0	0/0	0	I	S	R	R	R
28	LA3471^a	自交系IL	TGRC	8/8	0/0	0/0	0/0	0/0	0	I	R	R	R	R
29	强丰 Qiangfeng	自交系IL	中国农业科学院CAAS	0/0	0/0	0/0	2/0	6/8	96.9±4.4	HS	S	S	S	S
30	SD-1	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/0	3/1	5/7	93.8±4.4	HS	S	S	R	R
31	SD-2	自交系IL	山东省农业科学院 SAAS	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	S	R	S	S
32	SD-3	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/0	2/0	6/8	96.9±4.4	HS	S	S	S	S
编号 Number	材料 Material	世代 Generation	材料来源 Source	病情分级 DSS					病情指数 DI	抗级 RT	分子标记Molecular marker
编号 Number	材料 Material	世代 Generation	材料来源 Source	0级	1级	2级	3级	4级	病情指数 DI	抗级 RT	C2-25	PNU-D4	Tm-2	SCAR_Frl
33	SD-4	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/1	0/3	8/4	92.2±11.0	HS	S	S	R	R
34	SD-5	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	H	H
35	SD-6	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	R	S
36	SD-7	自交系IL	山东省农业科学院 SAAS	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	S	R	R	R
37	SD-8	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	H	H
38	SD-9	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	R	R
39	SD-10	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	S	S	S	S
40	SD-11	自交系IL	山东省农业科学院SAAS	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	H	H
41	SD-12	自交系IL	山东省农业科学院SAAS	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	S	S	R	R
42	SD-13	自交系IL	山东省农业科学院SAAS	0/0	0/0	1/0	2/0	5/8	93.8±8.8	HS	S	S	S	S
43	SD-14	自交系IL	山东省农业科学院SAAS	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	S	S	R	R
44	SD-15	自交系IL	山东省农业科学院SAAS	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	R	R
45	SD-16	自交系IL	山东省农业科学院SAAS	0/0	0/0	0/0	1/1	7/7	96.9±0	HS	R	R	R	R
46	SD-17	自交系IL	山东省农业科学院SAAS	5/3	3/5	0/0	0/0	0/0	12.5±4.4	R	S	R	S	S
47	SD-18	自交系IL	山东省农业科学院SAAS	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	R	R
48	SD-19	自交系IL	山东省农业科学院 SAAS	0/0	0/0	0/0	4/1	4/7	92.2±6.6	HS	R	S	S	S
49	SD-20	自交系IL	山东省农业科学院SAAS	0/0	0/0	0/0	3/0	5/8	95.3±6.6	HS	S	S	R	R
50	Ah2001	杂交种 Hybrid	宛东番茄（公司） WDT	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	H	H	H	H
51	Ah2002	杂交种 Hybrid	宛东番茄（公司） WDT	0/0	0/0	1/0	2/1	5/7	92.2±6.6	HS	S	S	H	H
52	Ah2003	杂交种 Hybrid	宛东番茄（公司） WDT	0/0	0/0	0/0	2/1	6/7	95.3±2.2	HS	H	H	R	R
53	Ah2004	杂交种 Hybrid	宛东番茄（公司） WDT	8/8	0/0	0/0	0/0	0/0	0	I	R	R	R	R
54	Ah2005	杂交种 Hybrid	宛东番茄（公司） WDT	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	S	H	H	H
55	Ah2006	杂交种 Hybrid	宛东番茄（公司） WDT	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	S	H	H	H
56	Ah2007	杂交种 Hybrid	宛东番茄（公司） WDT	8/8	0/0	0/0	0/0	0/0	0	I	H	H	H	H
57	Ah2008	杂交种 Hybrid	宛东番茄（公司） WDT	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	S	H	H	H
编号 Number	材料 Material	世代 Generation	材料来源 Source	病情分级 DSS					病情指数 DI	抗级 RT	分子标记Molecular marker
编号 Number	材料 Material	世代 Generation	材料来源 Source	0级	1级	2级	3级	4级	病情指数 DI	抗级 RT	C2-25	PNU-D4	Tm-2	SCAR_Frl
58	Ah2009	杂交种 Hybrid	宛东番茄（公司） WDT	5/3	1/2	2/1	0/0	0/2	26.6±15.5	MR	S	H	H	H
59	Ah2010	杂交种 Hybrid	宛东番茄（公司） WDT	8/8	0/0	0/0	0/0	0/0	0	I	S	H	H	S
60	Ah2011	杂交种 Hybrid	宛东番茄（公司） WDT	7/6	0/1	0/1	1/0	0/0	9.4±0	HR	S	H	H	H
61	Ah2012	杂交种 Hybrid	宛东番茄（公司） WDT	8/8	0/0	0/0	0/0	0/0	0	I	S	H	H	S
62	Ah2013	杂交种 Hybrid	宛东番茄（公司） WDT	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	R	R	R	R
63	Ah2014	杂交种 Hybrid	宛东番茄（公司） WDT	8/8	0/0	0/0	0/0	0/0	0	I	S	R	S	S
64	Ah2015	杂交种 Hybrid	宛东番茄（公司） WDT	8/4	0/4	0/0	0/0	0/0	6.3±8.8	HR	S	R	S	S
65	Ah2016	杂交种 Hybrid	宛东番茄（公司） WDT	8/8	0/0	0/0	0/0	0/0	0	I	S	R	S	S
66	Ah2017	杂交种 Hybrid	宛东番茄（公司） WDT	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	R	R
67	Ah2018	杂交种 Hybrid	宛东番茄（公司） WDT	8/6	0/2	0/0	0/0	0/0	3.1±4.4	HR	S	H	H	S
68	金棚砧木一号 Jinpengzhenmu-1	杂交种 Hybrid	伟丽种苗 WLS	8/6	0/2	0/0	0/0	0/0	3.1±4.4	HR	H	H	R	R
69	浙砧5号Zhezhen-5	杂交种 Hybrid	伟丽种苗 WLS	0/0	0/0	0/0	2/2	6/6	93.8±0	HS	H	S	S	S
70	砧木一号 Zhenmu-1	杂交种 Hybrid	伟丽种苗 WLS	8/8	0/0	0/0	0/0	0/0	0	I	S	H	R	R
71	萨瓦Sawa	杂交种 Hybrid	伟丽种苗 WLS	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	S	S	S	S
72	千叶Qianye	杂交种 Hybrid	伟丽种苗 WLS	8/8	0/0	0/0	0/0	0/0	0	I	S	H	R	R
73	久绿Jiulv	杂交种 Hybrid	伟丽种苗 WLS	8/8	0/0	0/0	0/0	0/0	0	I	H	H	H	H
74	育正泰Yuzhengtai	杂交种 Hybrid	伟丽种苗 WLS	8/8	0/0	0/0	0/0	0/0	0	I	H	R	R	R
75	天正1567 Tianzheng1567	杂交种 Hybrid	山东鲁蔬 Shandong Lushu	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	H	H	H
76	鲁蔬1907 Lushu1907	杂交种 Hybrid	山东鲁蔬 Shandong Lushu	0/0	0/0	0/0	2/2	6/6	93.8±0	HS	S	H	H	H
77	金棚8号 Jinpeng-8	杂交种 Hybrid	西安金棚 Xi’an Jinpeng	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	H	H	H	H
78	飞天Feitian	杂交种 Hybrid	海泽拉 Hazera	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	H	H
79	飞跃Feiyue	杂交种 Hybrid	海泽拉 Hazera	0/0	0/0	0/0	4/0	4/8	93.8±8.8	HS	H	H	R	R
80	罗拉Luola^b	杂交种 Hybrid	海泽拉 Hazera	8/8	0/0	0/0	0/0	0/0	0	I	S	H	H	H
81	圣罗兰3690 Shengluolan3690^b	杂交种 Hybrid	海泽拉 Hazera	8/6	0/2	0/0	0/0	0/0	3.1±4.4	HR	S	H	H	H
82	安纳西Annaxi^b	杂交种 Hybrid	海泽拉 Hazera	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	S	S	H	H
编号 Number	材料 Material	世代 Generation	材料来源 Source	病情分级 DSS					病情指数 DI	抗级 RT	分子标记Molecular marker
编号 Number	材料 Material	世代 Generation	材料来源 Source	0级	1级	2级	3级	4级	病情指数 DI	抗级 RT	C2-25	PNU-D4	Tm-2	SCAR_Frl
83	桃乐丝Taolesi^b	杂交种 Hybrid	海泽拉 Hazera	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	R	R	H	H
84	瓯秀806 Ouxiu806	杂交种 Hybrid	温州市农业科学研究院WAAS	0/0	0/0	0/0	2/0	6/8	96.9±4.4	HS	H	H	H	H
85	瑞星大宝 Ruixingdabao	杂交种 Hybrid	上海菲图 Shanghai Feitu	0/0	0/0	0/0	3/2	5/6	92.2±2.2	HS	S	S	H	H
86	瑞星5号 Ruixing-5	杂交种 Hybrid	上海菲图 Shanghai Feitu	0/0	0/0	2/0	4/1	2/7	85.9±15.5	HS	S	S	H	H
87	法拉利 Falali	杂交种 Hybrid	纽内姆 Nunhems	0/0	0/0	0/0	2/0	6/8	96.9±4.4	HS	S	S	H	S
88	粉宴1号 Fenyan-1	杂交种 Hybrid	纽内姆 Nunhems	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	H	S
89	齐达利 Qidali	杂交种 Hybrid	先正达 Syngenta	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	H	S	H	H
90	戴安娜 Daianna	杂交种 Hybrid	寿光旺林 Shouguang Wanglin	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	S	S	H	H
91	保罗塔 Baoluota	杂交种 Hybrid	先正达 Syngenta	0/0	0/0	0/0	2/0	6/8	96.9±4.4	HS	S	S	H	H
92	百利 Baili	杂交种 Hybrid	瑞克斯旺 Rijk Zwaan	0/0	0/0	0/0	3/1	5/7	93.8±4.4	HS	H	H	H	H
93	SV4224TH	杂交种 Hybrid	圣尼斯 Seminis	8/8	0/0	0/0	0/0	0/0	0	I	S	H	H	H
94	欧盾 Oudun	杂交种 Hybrid	圣尼斯 Seminis	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	H	H
95	103	杂交种 Hybrid	/	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	S	H	H	H
96	天禄1号 Tianlu-1	杂交种 Hybrid	沈阳谷雨 Shenyang Guyu	0/0	0/0	0/0	2/1	6/7	95.3±2.2	HS	H	H	R	R
97	安特莱斯 Antelaisi	杂交种 Hybrid	/	8/6	0/2	0/0	0/0	0/0	3.1±4.4	HR	H	H	R	R
98	普罗旺斯 Puluowangsi	杂交种 Hybrid	荷兰德奥特 De Ruiter	8/7	0/1	0/0	0/0	0/0	1.6±2.2	HR	H	H	R	R
99	粉太郎 Fentailang	杂交种 Hybrid	日本坂田 Sakata	0/0	0/0	0/0	0/0	8/8	100.0±0	HS	S	S	H	S
100	传奇 Chuanqi	杂交种 Hybrid	日本坂田 Sakata	0/0	0/0	0/0	1/0	7/8	98.4±2.2	HS	H	H	H	R

Table 3

Fig. 2

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引物名称 Primer name	引物序列 Primer sequence (5′-3′)	物理位置 Physical position (Mb)	参考文献 Reference
C2-25 (Apo I)	F: GTCATCCACATTCGTACTTC	5.5	STANIASZEK et al.^[18]
C2-25 (Apo I)	R: AATTTAGAGCACGTTTCATA	5.5	STANIASZEK et al.^[18]
PNU-D4 (Mbo I)	F: CAGCTGAAAGATGTCACCCA	6.1	KIM et al.^[20]
PNU-D4 (Mbo I)	R: TGATCATTTACAAGGCGGCA	6.1	KIM et al.^[20]
Tm-2 (Dde I)	F: TATGTCACTATCCCAAGCAA	13.6	/
Tm-2 (Dde I)	R: TAACGGTAGAATTGGACACTC	13.6	/
SCAR_Frl	F: GTAACAAGTGAAGTTAAAAATGCT	61.8	MUTLU et al.^[19]
SCAR_Frl	R: GTGTGGATTTGGGTTCAATTCC	61.8	MUTLU et al.^[19]

接种方法 Inoculation method	菌液浓度 Inoculum concentration (个/mL)	苗龄 Seedling stage	环境温度 Environmental temperature (℃)	发病率 Disease incidence (%)	病情指数 Disease index
浸根法 Root dipping	10⁶	2—3叶期2-3 leaf	20	90±0.1aA	66.7±1.4cC
	10⁷			100±0aA	89.2±1.4bB
	10⁸			100±0aA	96.7±1.4aA
	10⁷	2—3叶期2-3 leaf	20	100±0aA	91.7±3.8aA
		3—4叶期3-4 leaf		100±0aA	90.8±2.9aA
		4—5叶期4-5 leaf		100±0aA	89.2±1.4aA
	10⁷	2—3叶期2-3 leaf	20	100±0aA	91.7±2.9aA
			25	100±0aA	80.8±2.9bB
			30	100±0aA	71.7±1.4cC
	10⁷	2—3叶期2-3 leaf	20	100±0aA	97.5±2.5aA
灌根法Root irrigation				100±0aA	96.7±1.4aA
茎注射法Stem injection				100±0aA	81.7±11.5bA

Development of Artificial Inoculation Methodology for Evaluation of Resistance to Fusarium Crown and Root Rot and Screening of Resistance Sources in Tomato

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