大白菜苗期对Pantoea agglomerans抗性鉴定方法的建立与应用

doi:10.3969/j.issn.1004-1524.20221568

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (10): 2408-2414.DOI: 10.3969/j.issn.1004-1524.20221568

大白菜苗期对Pantoea agglomerans抗性鉴定方法的建立与应用

李家玺(), 顾冉, 刘焱, 张耀伟()

东北农业大学园艺园林学院,黑龙江哈尔滨 150030

收稿日期:2022-11-04 出版日期:2023-10-25 发布日期:2023-10-31
作者简介:李家玺(1999—),男,辽宁锦州人,硕士研究生,主要从事蔬菜遗传育种研究。E-mail:lijiaxi2019@126.com
通讯作者: ^*张耀伟,E-mail:zhyw1271@126.com
基金资助:
黑龙江省“百千万”工程科技重大专项课题(2019ZX16B02);黑龙江省自然科学基金(LH2019C016);黑龙江省创新人才项目(UNPYSCT-2020112);东北农业大学“学术骨干”项目

Establishment and application of the resistance identification method of Pantoea agglomerans during the seedling stage of Chinese cabbage

LI Jiaxi(), GU Ran, LIU Yan, ZHANG Yaowei()

College of Horticulture & Landscape Architecture, Northeast Agricultural University, Harbin 150030, China

Received:2022-11-04 Online:2023-10-25 Published:2023-10-31

摘要/Abstract

摘要：

2015年以来在哈尔滨市大白菜生产中发现一种新型软腐病,此病害逐年加重,经鉴定病原为成团泛菌(Pantoea agglomerans)。本研究以成团泛菌为接种病原,设置接种温度单因素试验,设置接种苗龄、接种湿度和接种方法L₉(3⁴)正交试验;以田间表现抗性有差异的3个品种为接种材料进行接种菌液浓度单因素试验,以期建立大白菜苗期对P. agglomerans抗性鉴定方法,并据此进行种质抗性评价。结果表明,最佳接种条件为温度25~30 ℃,苗龄7~8叶期,相对湿度90%~100%;接种方法为在中肋基部用刀片轻划“品”字形小伤口,并滴30 μL 1×10¹² CFU·mL^-1的菌液。按此方法在20份种质中鉴定出2份高抗和4份抗病大白菜种质,研究结果可以为大白菜抗病育种提供参考。

关键词: 大白菜, 软腐病, Pantoea agglomerans, 抗性

Abstract:

Since 2015, a new type of soft rot disease has been discovered in Chinese cabbage production in Harbin City. This disease has been increasing year by year, and the pathogen was identified as Pantoea agglomerans. In this study, P. agglomerans was used as the inoculant. The single-factor experiment was conducted to determine the optimal inoculation temperature, and the L₉(3⁴) orthogonal experiment was designed to investigate the effects of seedling age, humidity, and inoculation method. The single-factor experiment of inoculation concentration was carried out using 3 different varieties with varying resistance levels in the field. The aim was to establish a resistance identification method for Chinese cabbage against P. agglomerans during the seedling stage and evaluate the germplasm resistance based on this method. The results showed that the optimal conditions for inoculation were a temperature of 25-30 ℃, a seedling age of 7-8 leaves, and a relative humidity of 90%-100%. The recommended inoculation method was using a sterilized knife to scratch 3 small wounds and then dropping 30 μL of bacteria liquid with a concentration of 1×10¹² CFU·mL^-1. Using this method, 2 highly resistant and 4 resistant Chinese cabbage germplasm resources were identified from a total of 20 accessions. These research findings could provide valuable references for disease-resistant breeding of Chinese cabbage.

Key words: Chinese cabbage, soft rot, Pantoea agglomerans, resistance

中图分类号:

S436.341

李家玺, 顾冉, 刘焱, 张耀伟. 大白菜苗期对Pantoea agglomerans抗性鉴定方法的建立与应用[J]. 浙江农业学报, 2023, 35(10): 2408-2414.

LI Jiaxi, GU Ran, LIU Yan, ZHANG Yaowei. Establishment and application of the resistance identification method of Pantoea agglomerans during the seedling stage of Chinese cabbage[J]. Acta Agriculturae Zhejiangensis, 2023, 35(10): 2408-2414.

图/表 6

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水平 Level	因素Factor
水平 Level	A接种苗龄 Inoculation seeding stage	B接种湿度 Inoculation humidity/%	C接种方法 Inoculation method
1	3~4叶期Seedling age of 3-4 leaves	50~60	用刀片纵切5 mm Using knife to cut 5 mm longitudinally
2	5~6叶期Seedling age of 5-6 leaves	70~80	用针轻刺“品”字形小伤口
			Using needle to stab three small wounds like‘品’shape
3	7~8叶期Seedling age of 7-8 leaves	90~100	用刀片轻划“品”字形小伤口
			Using knife to scratch three small wounds like‘品’shape

水平 Level	因素Factor
水平 Level	A接种苗龄 Inoculation seeding stage	B接种湿度 Inoculation humidity/%	C接种方法 Inoculation method
1	3~4叶期Seedling age of 3-4 leaves	50~60	用刀片纵切5 mm Using knife to cut 5 mm longitudinally
2	5~6叶期Seedling age of 5-6 leaves	70~80	用针轻刺“品”字形小伤口
			Using needle to stab three small wounds like‘品’shape
3	7~8叶期Seedling age of 7-8 leaves	90~100	用刀片轻划“品”字形小伤口
			Using knife to scratch three small wounds like‘品’shape

试验号 Test No.	试验设计 Experimental design			发病率 Incidence rate/%	病情指数 Disease index
试验号 Test No.	A	B	C	发病率 Incidence rate/%	病情指数 Disease index
1	1	1	1	9.33	8.56
2	2	2	2	63.00	38.63
3	3	3	3	91.00	50.67
4	1	2	3	19.67	18.78
5	2	3	1	23.33	17.30
6	3	1	2	77.00	32.82
7	1	3	2	27.33	15.07
8	2	1	3	49.67	23.70
9	3	2	1	50.67	20.81

试验号 Test No.	试验设计 Experimental design			发病率 Incidence rate/%	病情指数 Disease index
试验号 Test No.	A	B	C	发病率 Incidence rate/%	病情指数 Disease index
1	1	1	1	9.33	8.56
2	2	2	2	63.00	38.63
3	3	3	3	91.00	50.67
4	1	2	3	19.67	18.78
5	2	3	1	23.33	17.30
6	3	1	2	77.00	32.82
7	1	3	2	27.33	15.07
8	2	1	3	49.67	23.70
9	3	2	1	50.67	20.81

因素 Factor	发病率Incidence rate/%
因素 Factor	K₁	K₂	K₃	R	优水平 Optimal level
A	42.41	79.63	104.3	61.89	3
B	65.08	78.22	83.04	17.96	3
C	46.67	86.52	93.15	46.48	3

大白菜苗期对Pantoea agglomerans抗性鉴定方法的建立与应用

Establishment and application of the resistance identification method of Pantoea agglomerans during the seedling stage of Chinese cabbage

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参考文献 21

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变异来源 Variation source	自由度 Degree of freedom	偏差平方和 Sum of squares of deviation from mean	方差 Variance	F	F_0.05	F_0.01
重复Repeat	2	0.13	0.07	0.21	3.55	6.01
A	2	17.46	8.73	27.21^**
B	2	2.43	1.22	3.79^*
C	2	10.87	5.44	16.94^**
误差Error	18	5.77	0.32
总变异Total variation	26	36.67

菌液浓度 Concentration of bacterial suspension/ (CFU·mL^-1)	不同时间的病情指数与(抗病等级) Disease index (resistance level) at different time
	5 d			7 d			9 d
	黄心冠军 Yellow heart champion	凯丰2号 Kaifeng No.2	珍亮 Zhenliang	黄心冠军 Yellow heart champion	凯丰2号 Kaifeng No.2	珍亮 Zhenliang	黄心冠军 Yellow heart champion	凯丰2号 Kaifeng No.2	珍亮 Zhenliang
10⁸	11.78 (R)	13.36 (R)	9.16 (HR)	23.93 (R)	23.22 (R)	19.90 (R)	35.21 (MR)	27.13 (R)	28.63 (R)
10¹⁰	20.26 (R)	22.87 (R)	20.05 (R)	48.67 (MR)	35.26 (MR)	23.33 (R)	53.07 (MR)	45.56 (MR)	27.33 (R)
10¹²	37.89 (MR)	30.04 (R)	23.11 (R)	62.78 (S)	47.21 (MR)	29.14 (R)	70.02 (S)	53.22 (MR)	32.05 (R)

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