一种基于叶片注射高效评价番茄青枯病抗性方法的建立与应用

doi:10.16420/j.issn.0513-353x.2024-1061

摘要/Abstract

摘要：

由青枯雷尔氏菌（Ralstonia solanacearum）侵染引起的青枯病是世界性的重大病害，在中国南方地区危害尤为严重。培育和种植抗病品种是防治该土传病害最为经济有效的措施，而种质资源的抗性筛选与鉴定是作物抗病育种的基础。本研究中选取福建地区5个具有地域代表性且毒力差异明显的R. solanacearum菌株组成鉴别菌株系，建立了基于叶片注射法的番茄青枯病抗性鉴定技术体系，筛选到砧6、砧8和砧10等3个抗病品种。比较了叶片注射法与传统伤根接种法、浸根法的优势，叶片注射法具有操作简便、接种量小、试验周期短、病症明显及结果稳定性好的优点。此外，验证了该方法适用于不同茄科作物抗病种质资源的筛选。

关键词: 番茄, 青枯病, 叶片注射法, 抗性鉴定, 青枯雷尔氏菌

Abstract:

Bacterial wilt，caused by Ralstonia solanacearum，is a significant crop disease worldwide，particularly in southern China. The adoption of resistant varieties remains the most economical and effective strategy for managing this soil-borne disease. Identification and screening of bacterial wilt-resistant germplasms are fundamental to crop disease resistance breeding. In this study，a leaf-injection inoculation method was established for the identification of bacterial wilt-resistant germplasms of tomato，while five highly virulent R. solanacearum strains with varied pathogenicity，collected from Fujian province were combined and utilized as an identification strain mix. Three germplasms，rootstock 6，8 and 10，were identified as resistant in the screening. Further，the leaf injection method was compared for its effectiveness with the traditional root injuring and root soaking inoculation methods. While the leaf injection method could be simply operated，results showed that it required less inocula and shorter experimental time to provide more clear and consistent symptoms than the traditional methods. In addition，this method was further evaluated and validated for its effectiveness on other crops in Solanaceae.

Key words: tomato, bacterial wilt of leaf injection, identification of resistance, Ralstonia solanacearum

黄欣冉, 王荣波, 张前荣, 李本金, 翁启勇, 刘裴清. 一种基于叶片注射高效评价番茄青枯病抗性方法的建立与应用[J]. 园艺学报, 2026, 53(4): 1215-1225.

HUANG Xinran, WANG Rongbo, ZHANG Qianrong, LI Benjin, WENG Qiyong, LIU Peiqing. Development and Implementation of an Efficient Method for Evaluating Tomato Resistance to Bacterial Wilt Based on Leaf Injection[J]. Acta Horticulturae Sinica, 2026, 53(4): 1215-1225.

图/表 8

表1 用于青枯菌检测的qPCR引物序列

Table 1 qPCR primer sequences for detection of Ralstonia solanacearum

基因名称Gene name	上游引物序列（5′-3′）Forward primer	下游引物序列（5′-3′）Reverse primer
16S rRNA	GTCGCCGTCAACTCACTTTCC	GTCGCCGTCAGCAATGCGGAATCG
SlEF1a	ACCTTTGCTGAATACCCTCCATTG	CACAGTTCACTTCCCCTTCTTCTG

图1 番茄AC和桂砧1号叶片注射接种青枯菌后的症状与病原菌积累 * P < 0.05

Fig. 1 Symptoms and pathogen accumulation of AC and Guizhen 1 tomatoes leaves after inoculation with Ralstonia solanacearum

图2 8份番茄材料叶片注射青枯菌后的表型

Fig. 2 Phenotypes of 8 tomato materials after injection of Ralstonia solanacearum

图3 采用叶片注射法和伤根接种法接种青枯菌的8份番茄材料植株死亡率 ns：不显著

Fig. 3 The plant mortality of 8 tomato materials inoculated with Ralstonia solanacearum by leaf injection method and root injury inoculation method ns：No significant

图4 基于叶片注射法鉴定番茄青枯病抗性技术体系示意图

Fig. 4 The schematic diagram of technical system for resistance evaluation of tomato bacterial wilt using the leaf injection inoculation method

图5 3种接种方法接种AC和桂砧1号后的症状

Fig. 5 Symptoms after inoculation of AC and Guizhen 1 by three inoculation methods

表2 供试番茄砧木种质叶片注射和伤根接种的病情指数及抗性评价

Table 2 Disease index and resistance evaluation of tomato rootstock germplasm resources by leaf injection and root injury inoculation

名称Name	材料Material	叶片注射法Leaf injection inoculation		伤根接种法Root injury inoculation
名称Name	材料Material	病情指数DI	表型Phenotype	病情指数DI	表型Phenotype
感病对照品种 Susceptible control varieties	AC	70.37	S	60.42	S
感病对照品种 Susceptible control varieties	千禧Cherry tomato	68.52	S	56.25	S
抗病对照品种 Disease-resistant control varieties	桂砧1号Guizhen 1	12.90	R	12.50	R
待测砧木材料 Rootstock material to be tested	番茄砧木6号Tomato rootstock 6	61.11	S	54.16	S
	番茄砧木405 Tomato rootstock 405	69.44	S	56.25	S
	浙砧1号Zhezhen 1	70.37	S	60.42	S
	砧1 Zhen 1	37.04	MR	22.92	R
	砧2 Zhen 2	70.37	S	58.33	S
	砧3 Zhen 3	41.66	MR	20.83	R
	砧4 Zhen 4	37.04	MR	31.25	MR
	砧5 Zhen 5	44.44	MR	35.41	MR
	砧6 Zhen 6	18.52	R	12.50	R
	砧7 Zhen 7	33.33	MR	22.92	R
	砧8 Zhen 8	20.37	R	16.67	R
	砧9 Zhen 9	25.92	MR	20.83	R
	砧10 Zhen 10	22.22	R	18.75	R

图6 6种茄科作物叶片注射接种青枯菌7 d后的症状

Fig.6 Symptoms of in leaves of six solanaceous crops 7 days after inoculation with Ralstonia solanacearum

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