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

doi:10.3969/j.issn.1004-1524.20221568

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (10): 2408-2414.DOI: 10.3969/j.issn.1004-1524.20221568

• Plant Protection • Previous Articles Next Articles

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

Abstract

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

CLC Number:

S436.341

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.

Figures/Tables 6

References 21

[1]	李晓颖, 田宇, 赵亮, 等. 京郊快菜细菌性软腐病病原鉴定[J]. 华北农学报, 2018, 33(3): 63-70.
	LI X Y, TIAN Y, ZHAO L, et al. Identification of the bacterial soft rot pathogens on Chinese cabbage in Beijing[J]. Acta Agriculturae Boreali-Sinica, 2018, 33(3): 63-70. (in Chinese with English abstract)
[2]	李晓颖, 田宇, 张瑾, 等. 大白菜软腐病新病原菌Pectobacterium aroidearum的鉴定及其生物学特性[J]. 植物病理学报, 2018, 48(4): 455-465.
	LI X Y, TIAN Y, ZHANG J, et al. Identification and characterization of a Pectobacterium aroidearum strain causing bacterial soft rot on Chinese cabbage (Brassica rapa L.ssp.pekinensis)[J]. Acta Phytopathologica Sinica, 2018, 48(4): 455-465. (in Chinese with English abstract)
[3]	OSKIERA M, KAŁU ŻNA M, KOWALSKA B, et al. Pectobacterium carotovorum subsp. odoriferum on cabbage and Chinese cabbage: identification, characterization and taxonomic relatedness of bacterial soft rot causal agents[J]. Journal of Plant Pathology, 2017, 99(1): 149-160.
[4]	CHEN C L, LI X Y, BO Z J, et al. Occurrence, characteristics, and PCR-based detection of Pectobacterium polaris causing soft rot of Chinese cabbage in China[J]. Plant Disease, 2021, 105(10): 2880-2887.
[5]	LI J X, ZHANG Y W. Biological characteristics and isolation of Pythium ultimum causing rot of Chinese cabbage[J]. Australasian Plant Pathology, 2020, 49(2): 201-207.
[6]	GUO M, LIU Y, LIU S N, et al. First report of bacterial soft rot caused by Pantoea agglomerans on Chinese cabbage in China[J]. Plant Disease, 2020, 104(1): 277.
[7]	保善存, 夏吾拉太, 殷光晶, 等. 杨树枝干病害病原物分离与鉴定[J]. 分子植物育种, 2021, 19(23): 7900-7904.
	BAO S C, XIAWULATAI, YIN G J, et al. Isolation and identification of pathogens of poplar stem diseases[J]. Molecular Plant Breeding, 2021, 19(23): 7900-7904. (in Chinese with English abstract)
[8]	佘小漫, 于琳, 蓝国兵, 等. 广东省冬枣叶枯病病原菌的分离与鉴定[J]. 植物病理学报, 2021, 51(3): 317-324.
	SHE X M, YU L, LAN G B, et al. Identification of the pathogen causing Ziziphus jujuba Mill. cv. Dongzao leaf necrotic disease in Guangdong Province[J]. Acta Phytopathologica Sinica, 2021, 51(3): 317-324. (in Chinese with English abstract)
[9]	童亚萍, 杨丙烨, 田茜, 等. 桃李细菌性穿孔病新病原的分离鉴定[J]. 福建农林大学学报(自然科学版), 2020, 49(3): 300-306.
	TONG Y P, YANG B Y, TIAN Q, et al. Isolation and identification of a new pathogen causing bacterial leaf shot hole disease on peach and plum[J]. Journal of Fujian Agriculture and Forestry University(Natural Science Edition), 2020, 49(3): 300-306. (in Chinese with English abstract)
[10]	严婉荣, 肖敏, 肖彤斌, 等. 海南火龙果细菌性腐烂病的病原鉴定及田间药剂筛选[J]. 基因组学与应用生物学, 2020, 39(3): 1185-1190.
	YAN W R, XIAO M, XIAO T B, et al. Pathogen identification and screening of field agents on dragon fruit bacterial rot disease in Hainan[J]. Genomics and Applied Biology, 2020, 39(3): 1185-1190. (in Chinese with English abstract)
[11]	肖波, 卢世栋, 杨斌, 等. 云南核桃细菌性黑斑病病原菌的分离与鉴定[J]. 贵州农业科学, 2017, 45(12): 55-58.
	XIAO B, LU S D, YANG B, et al. Isolation and identification of walnut bacterial black spot pathogen in Yunnan[J]. Guizhou Agricultural Sciences, 2017, 45(12): 55-58. (in Chinese with English abstract)
[12]	闫仁洁. 北京蔬菜学会推广大白菜优良品种[J]. 学会, 1997(6): 32.
	YAN R J. Beijing Vegetable Society promotes fine varieties of Chinese cabbage[J]. Society, 1997(6): 32. (in Chinese)
[13]	丁海凤. 大白菜优良品种北京新三号[J]. 农业知识, 2007(20): 7.
	DING H F. Beijing Xin No.3, an excellent Chinese cabbage variety[J]. Agriculture Knowlege, 2007(20): 7. (in Chinese)
[14]	王效睦, 白静, 谢坤, 等. 我国大白菜种质资源的研究与利用[J]. 山东农业科学, 2016, 48(3): 143-149.
	WANG X M, BAI J, XIE K, et al. Research and utilization of Chinese cabbage germplasm resources in China[J]. Shandong Agricultural Sciences, 2016, 48(3): 143-149. (in Chinese with English abstract)
[15]	朱琳, 孙素丽, 孙菲菲, 等. 绿豆尖镰孢枯萎病抗性鉴定方法[J]. 植物遗传资源学报, 2017, 18(4): 696-703.
	ZHU L, SUN S L, SUN F F, et al. The method for evaluation of mung bean resistance to Fusarium wilt[J]. Journal of Plant Genetic Resources, 2017, 18(4): 696-703. (in Chinese with English abstract)
[16]	孙叶烁, 郝玲玉, 张杰, 等. 大白菜菌核病抗性鉴定方法研究[J]. 西北农林科技大学学报(自然科学版), 2019, 47(12): 123-129.
	SUN Y S, HAO L Y, ZHANG J, et al. Identification method of resistance to Sclerotinia in Chinese cabbage[J]. Journal of Northwest A & F University(Natural Science Edition), 2019, 47(12): 123-129. (in Chinese with English abstract)
[17]	张鑫鑫, 张恩慧, 吴云锋, 等. 甘蓝黑腐病苗期抗性鉴定方法研究[J]. 中国瓜菜, 2021, 34(6): 39-46.
	ZHANG X X, ZHANG E H, WU Y F, et al. Study on the method of identification of resistance to black rot in cabbage at seedling stage[J]. China Cucurbits and Vegetables, 2021, 34(6): 39-46. (in Chinese with English abstract)
[18]	史国立, 崔崇士, 张耀伟. 大白菜对软腐病抗性的快速鉴定方法研究[J]. 植物保护, 2006, 32(6): 135-138.
	SHI G L, CUI C S, ZHANG Y W. Study on rapid identification method of Chinese cabbage resistance to soft rot[J]. Plant Protection, 2006, 32(6): 135-138. (in Chinese)
[19]	田宇, 李晓颖, 石妙涵, 等. 生菜细菌性软腐病抗性鉴定方法及其资源评价[J]. 华北农学报, 2019, 34(S1): 309-317.
	TIAN Y, LI X Y, SHI M H, et al. Assessment of identification method of lettuce bacterial soft rot resistance and evaluation of cultivar resistance[J]. Acta Agriculturae Boreali-Sinica, 2019, 34(S1): 309-317. (in Chinese with English abstract)
[20]	张耀伟. 大白菜抗软腐病生理生化指标研究[D]. 哈尔滨: 东北农业大学, 2002.
	ZHANG Y W. Study on physiological and biochemical indexes of Chinese cabbage resistance to soft rot[D]. Harbin: Northeast Agricultural University, 2002. (in Chinese with English abstract)
[21]	马燕勤, 李勤菲, 司军, 等. 甘蓝抗软腐病离体鉴定方法探究[J]. 植物保护, 2018, 44(6): 136-140.
	MA Y Q, LI Q F, SI J, et al. Assessment of inoculation method for identifying resistance to soft rot in cabbage[J]. Plant Protection, 2018, 44(6): 136-140. (in Chinese with English abstract)

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

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

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 21

Related Articles 15

Recommended Articles

Metrics

Comments

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)

[1]	LIU Guomin, ZHENG Xu, LIAO Yujiao, QIN Yexin, QIN Weizhi. Effect of grafting on cold resistance of potato seedlings under low temperature stress [J]. Acta Agriculturae Zhejiangensis, 2025, 37(8): 1615-1623.
[2]	WANG Yidi, WANG Jinglei, HU Tianhua, XU Yunmin, BAO Chonglai. Development of molecular markers for clubroot resistance and their application in Brassicaceae breeding [J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1272-1284.
[3]	YI Ming, SUN Hong, SHEN Qi, TANG Jiangwu. Research progress on ectopic fermentation system in treatment of fecal residue and waste water of livestock and poultry [J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1390-1396.
[4]	WANG Chao, LI Yanjie, NIU Yun, WEN Lianhao, CHEN Jingjing, WU Hongzhi, YANG Yuyong, WU Yandi. Biological characterization of black spot pathogens in Rosa chinensis and evaluation of varietal resistance [J]. Acta Agriculturae Zhejiangensis, 2025, 37(5): 1087-1096.
[5]	HU Xinrou, WANG Mei, ZHANG Yafen, CAI Weiming, JIN Qunli. Effect of abiotic stress on growth development and response mechanism of Ganoderma [J]. Acta Agriculturae Zhejiangensis, 2025, 37(5): 1182-1190.
[6]	ZHAO Jiahao, XU Xing, ZHOU Weidong, YANG Hua, ZHAO Xihong, WANG Wen. Characteristics of bacteria community and antibiotic resistance genes in piggery wastewater and surrounding water environment [J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 621-632.
[7]	XU Wenwu, WANG Zhenzhen, LU Lizhi. Analysis on egg-laying performance and stress resistance of four egg duck breeds [J]. Acta Agriculturae Zhejiangensis, 2024, 36(8): 1773-1778.
[8]	HAN Qingyu, CHENG Linrun, LI Yuehong, QIU Zhiling, HOU Meng, LOU Binggan. Evaluation of resistance to foot rot and analysis of disease resistance related indexes of 60 sweet potato germplasm resources [J]. Acta Agriculturae Zhejiangensis, 2024, 36(7): 1616-1625.
[9]	LI Jingjing, LI Chuang, LU Yanan, ZHENG Wenming. Identification and expression analysis of Thionin-like gene family in wheat [J]. Acta Agriculturae Zhejiangensis, 2024, 36(4): 729-737.
[10]	SONG Peng, LI Lixiang, JIANG Houlong, WANG Ru, LI Hui, ZHAO Pengyu, ZHANG Jun, QIN Pingwei, REN Jiangbo, CHEN Qingming. Effect of application of Brevibacillus laterosporus on potassium content of cured tobacco leaves and physiological characteristics of tobacco plants [J]. Acta Agriculturae Zhejiangensis, 2024, 36(3): 494-502.
[11]	CHAI Rongyao, YOU Yuxin, QIU Haiping, GUO Junning, ZHANG Zhen, LI Bin, SHEN Shengfa, WANG Yanli. Establishment of sweet potato stem rot resistance identification technology and analysis of germplasm resource resistance [J]. Acta Agriculturae Zhejiangensis, 2024, 36(3): 569-578.
[12]	GUO Nana, LI Wei, HUANG Lijuan, ZHANG Tao, WEI Bingqiang. Research progress on resistance of pepper to Tomato spotted wilt virus (TSWV) [J]. Acta Agriculturae Zhejiangensis, 2024, 36(10): 2416-2425.
[13]	ZOU Wenteng, LIU Zhenhong, YANG Jingxuan, LU Yangyang, PENG Kaisong. Genome-wide analysis of pathogenicity and drug resistance of Micropterus salmoides-derived Aeromonas veronii strain AV040 [J]. Acta Agriculturae Zhejiangensis, 2023, 35(9): 2068-2078.
[14]	LI Biyuan, YUE Zhichen, ZHAO Yanting, LEI Juanli, HU Qizan, TAO Peng. Identification and functional analysis of the BrLCYB gene of lycopene β-cyclase from Chinese cabbage [J]. Acta Agriculturae Zhejiangensis, 2023, 35(9): 2090-2096.
[15]	ZHANG Siyu, LIN Chaoyang, YE Yuxuan, SHEN Zhicheng. Characterization of transgenic insect resistance and glyphosate tolerance rice expressing cry1Ab-vip3Af2 and cp4-epsps [J]. Acta Agriculturae Zhejiangensis, 2023, 35(8): 1823-1833.