柠檬叶片溃疡病菌的分离鉴定及室内防治药剂筛选

doi:10.11924/j.issn.1000-6850.casb2020-0628

摘要/Abstract

摘要：

为了明确湛江地区柑橘溃疡病的病原类型并筛选出高效的防治药剂。以柠檬病叶为材料,用组织分离法对病样进行病原菌分离鉴定后,通过抑菌圈法测定了5种常用药剂对该病原菌的室内毒力。典型菌株zlm1908在NA培养基上菌落呈蜡黄色,圆形,全缘,有光泽,微隆起,黏稠,回接健康柠檬叶片后能产生明显的溃疡病斑,结合生理生化反应特点和特异性引物扩增序列分析确定菌株zlm1908为柑橘黄单胞柑橘亚种(Xanthomonas citri subsp.citri,Xcc)。5种药剂对菌株zlm1908的EC₅₀值分别为18.45 g/L、14.99 g/L、5.28 g/L、14.60 g/L和32.81 g/L。从EC₅₀的大小可以看出5种杀菌剂中枯草芽孢杆菌的抑制作用最强,其次为中生菌素,碱式硫酸铜和氢氧化铜,氯溴异氰尿酸抑制作用最弱。

关键词: 柑橘溃疡病菌, 病原鉴定, 杀菌剂, 毒力测定

Abstract:

The purpose of this study is to identify the pathogen types of citrus bacterial canker disease in Zhanjiang area and screen effective bactericides. Lemon leaves were used as materials, and the strains were isolated and identified by tissue separation. The indoor toxicity of five bactericides to this pathogen was determined by inhibition zone test. The colony morphology of classic strain zlm1908 on nutrient agar medium (NA) was wax yellow, circular, entire, glistening, convex and mucoid. Citrus canker disease symptoms appeared obviously after reinoculating healthy lemon leaves. The strain zlm1908 was identified as Xanthomonas citri subsp. citri (Xcc) by physiological and biochemical reaction, specific primer amplification and sequence analysis. The EC₅₀ of five bactericides was 18.45 g/L, 14.99 g/L, 5.28 g/L, 14.60 g/L and 32.81 g/L, respectively. According to the analysis of EC₅₀ value, the inhibiting effect of Bacillus subtilis is the strongest among the five bactericides, followed by zhongshengmycin, copper dihydroxosulphate, copper hydroxide, while chlorobromoisocyanurate has the least inhibiting effect.

Key words: Xanthomonas citri subsp. citri (Xcc), pathogen identification, bactericide, toxicity test

中图分类号:

李亚, 蔡永军, 余沁涵, 孔少连, 周晓晴, 张颖荷. 柠檬叶片溃疡病菌的分离鉴定及室内防治药剂筛选[J]. 中国农学通报, 2021, 37(24): 146-153.

Li Ya, Cai Yongjun, Yu Qinhan, Kong Shaolian, Zhou Xiaoqing, Zhang Yinghe. Citrus Bacterial Canker Disease: Pathogen Identification and Bactericides Screening[J]. Chinese Agricultural Science Bulletin, 2021, 37(24): 146-153.

图/表 12

参考文献 32

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引物名称	引物序列(5’-3’)
P8	GTTGCCCAGTTATCTCGCCCTGATC
P9	CCAATCCCTGCCGACGCTCAGGTAGT
JYF5	TTCGGCGTCAACAACCTG
JFR5	AACTCCAGCACATACGGGTC
Xac01	CGCCATCCCCACCACCACCACGAC
Xac02	AACCGCTCAATGCCATCCACTTCA
GGS1F	CCCTGCTGCTGACCTTCTTCT
GGS2R	GGTTGACCGTGGTTTCCCATA
27F	AGAGTTTGATCCTGGCTCAG
1492R	TACGGCTACCTTGTTACGACTT

引物名称	引物序列(5’-3’)
P8	GTTGCCCAGTTATCTCGCCCTGATC
P9	CCAATCCCTGCCGACGCTCAGGTAGT
JYF5	TTCGGCGTCAACAACCTG
JFR5	AACTCCAGCACATACGGGTC
Xac01	CGCCATCCCCACCACCACCACGAC
Xac02	AACCGCTCAATGCCATCCACTTCA
GGS1F	CCCTGCTGCTGACCTTCTTCT
GGS2R	GGTTGACCGTGGTTTCCCATA
27F	AGAGTTTGATCCTGGCTCAG
1492R	TACGGCTACCTTGTTACGACTT

名称	产酸	产气
D-果糖	+	—
D-甘露糖	—	—
D-半乳糖	—	—
蔗糖	—	—
蜜二糖	+	—
海藻糖	—	—
D-甘露醇	—	—
丙三醇	—	—
肌醇	—	—
核糖醇	—	—
山梨醇	—	—

名称	产酸	产气
D-果糖	+	—
D-甘露糖	—	—
D-半乳糖	—	—
蔗糖	—	—
蜜二糖	+	—
海藻糖	—	—
D-甘露醇	—	—
丙三醇	—	—
肌醇	—	—
核糖醇	—	—
山梨醇	—	—

试验	结果
淀粉水解	+
过氧化氢酶	+
明胶液化	+
硝酸盐还原	+
产氨	+
吲哚	—
V-P	—
甲基红	—
氧化酶	—