甘蓝抗软腐病离体鉴定方法探究
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摘要
通过分析不同因素对甘蓝软腐病离体接种后发病效率的影响,建立了一套甘蓝软腐病抗性的快速鉴定方法。结果发现,用浓度为6.4×10~8 cfu/mL的软腐病液体培养基悬浮液,采用针刺法接种苗龄为5～7叶期植株的离体叶片,接种后空气湿度为90%以上是最有利于软腐病发病的方法。利用该方法鉴定27份栽培甘蓝,发现高抗材料3份,抗病材料8份,耐病材料8份,感病材料8份。
A multifactor experiment with three duplicates was established to investigate three different inoculation techniques(transaction inoculation, injection and pin prick) by using pathogenic isolates of Pectobacterium carotovorum subsp. carotovorum in China against resistant and susceptible cabbage cultivars under controlled greenhouse conditions. It was found that the method with the pathogen suspended in the liquid NA medium at the density of 6.4×10~8cfu/mL, inoculating plants at the seedling age of 5-7-leaf old by pin prick in the growth chamber with a high moisture of over 90%, was suitable for the pathogen to infect the cabbage leaves. This method was used to identify resistance to soft rot in 27 cabbage cultivars, three of which showed high resistance, eight moderate resistance, eight tolerance and eight susceptibility to soft rot.
A multifactor experiment with three duplicates was established to investigate three different inoculation techniques(transaction inoculation, injection and pin prick) by using pathogenic isolates of Pectobacterium carotovorum subsp. carotovorum in China against resistant and susceptible cabbage cultivars under controlled greenhouse conditions. It was found that the method with the pathogen suspended in the liquid NA medium at the density of 6.4×10~8cfu/mL, inoculating plants at the seedling age of 5-7-leaf old by pin prick in the growth chamber with a high moisture of over 90%, was suitable for the pathogen to infect the cabbage leaves. This method was used to identify resistance to soft rot in 27 cabbage cultivars, three of which showed high resistance, eight moderate resistance, eight tolerance and eight susceptibility to soft rot.
引文
[1] 董金皋.农业植物病理学[M].第2版.北京:中国农业出版社,2007.
[2] 李志.十字花科蔬菜软腐病发生特点及防治技术[J].河北农业,2008(9):25.
[3] MA Bing, HIBBING M E, KIM H S, et al. Host range and molecular phylogenies of the soft rot enterobacterial genera Pectobacterium and Dickeya [J].Phytopathology,2007,97(9):1150-1163.
[4] GARDAN L, GOUY C, CHRISTEN R, et al. Elevation of three subspecies of Pectobacterium carotovorum to species level: Pectobacterium atrosepticum sp. nov., Pectobacterium betavasculorum sp. nov. and Pectobacterium wasabiae sp. nov[J]. International Journal of Systematic and Evolutionary Microbiology,2003,53(2):381-391.
[5] AVROVA A O, HYMAN L J, TOTH R L, et al. Application of amplified fragment length polymorphism fingerprinting for taxonomy and identification of the soft rot bacteria Erwinia carotovora and Erwinia chrysanthemi[J].Applied and Environmental Microbiology,2002,68(4):1499-1508.
[6] 田宇,马亚丽,何付新,等.北京地区芹菜细菌性软腐病菌鉴定及其致病力分析[J].植物病理学报,2016,46(4):433-442.
[7] CHARKOWSKI A O. The soft rot Erwinia [M]// Plant-Associated Bacteria. Springer Netherlands, 2007:423-505.
[8] 王茜,张宝海,赵亮,等.一株芹菜(Apium graveolens L.)软腐病菌Pectobacterium carotovorum subsp. odoriferum的特征[J].植物病理学报,2015,45(4):418-424.
[9] WANG Jing, WANG Yihong, ZHAO Tingchang, et al. Characterization of the pathogen causing a new bacterial vein rot disease in tobacco in China [J]. Crop Protection, 2017, 92: 93-98.
[10] NAZERIAN E, SIJAM K, ZAINAL A, et al. First report of lettuce soft rot caused by Pectobacterium carotovorum subsp. carotovorum in Malaysia[J]. Journal of Plant Pathology. 2011, 93(4):491-491.
[11] TSUDA K, TSUJI G, HIGASHIYAMA M, et al. Biological control of bacterial soft rot in Chinese cabbage by Lactobacillus plantarum strain BY under field conditions [J]. Biological Control, 2016, 100: 63-69.
[12] JEONG R D, CHU E H, PARK D H, et al. Control of postharvest bacterial soft rot by gamma irradiation and its potential modes of action [J].The Plant Pathology Journal,2016,32(2): 157-161.
[13] 张凤兰. 白菜对软腐病的室内抗性鉴定方法及抗源筛选[J]. 蔬菜, 1992(4):20.
[14] 张光明, 王翠花. 大白菜抗软腐病接种鉴定方法的初步研究[J]. 山东农业科学, 1995(5):39-40.
[15] 臧威, 崔崇士, 张耀伟. 大白菜软腐病苗期抗性鉴定方法的研究[J]. 北方园艺, 2003(3):57-58.
[16] 方中达. 植病研究方法[M]. 北京: 农业出版社, 1979.
[17] 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.
[18] 杨晓红, 徐文玲, 张晓明,等. 大白菜抗软腐病接种鉴定及抗源筛选研究[J]. 山东农业科学, 2009(5):67-69.
[19] 叶雪凌, 曹鸣庆, 马荣才. 拟南芥对细菌性软腐病抗性变异分析[J]. 中国生物工程杂志, 2008, 28(11):36-42.
[20] BURR T J, SCHROTH M N. Occurrence of soft rot Erwinia spp. In soil and plant material [J]. Phytopathology,1977,77(11):1382-1387.
[21] DYE D W. A taxonomic study of the genus Erwinia. I. The “amylovora” group [J]. New Zealand Journal of Agricultural Research, 1968, 11:590-607.
[2] 李志.十字花科蔬菜软腐病发生特点及防治技术[J].河北农业,2008(9):25.
[3] MA Bing, HIBBING M E, KIM H S, et al. Host range and molecular phylogenies of the soft rot enterobacterial genera Pectobacterium and Dickeya [J].Phytopathology,2007,97(9):1150-1163.
[4] GARDAN L, GOUY C, CHRISTEN R, et al. Elevation of three subspecies of Pectobacterium carotovorum to species level: Pectobacterium atrosepticum sp. nov., Pectobacterium betavasculorum sp. nov. and Pectobacterium wasabiae sp. nov[J]. International Journal of Systematic and Evolutionary Microbiology,2003,53(2):381-391.
[5] AVROVA A O, HYMAN L J, TOTH R L, et al. Application of amplified fragment length polymorphism fingerprinting for taxonomy and identification of the soft rot bacteria Erwinia carotovora and Erwinia chrysanthemi[J].Applied and Environmental Microbiology,2002,68(4):1499-1508.
[6] 田宇,马亚丽,何付新,等.北京地区芹菜细菌性软腐病菌鉴定及其致病力分析[J].植物病理学报,2016,46(4):433-442.
[7] CHARKOWSKI A O. The soft rot Erwinia [M]// Plant-Associated Bacteria. Springer Netherlands, 2007:423-505.
[8] 王茜,张宝海,赵亮,等.一株芹菜(Apium graveolens L.)软腐病菌Pectobacterium carotovorum subsp. odoriferum的特征[J].植物病理学报,2015,45(4):418-424.
[9] WANG Jing, WANG Yihong, ZHAO Tingchang, et al. Characterization of the pathogen causing a new bacterial vein rot disease in tobacco in China [J]. Crop Protection, 2017, 92: 93-98.
[10] NAZERIAN E, SIJAM K, ZAINAL A, et al. First report of lettuce soft rot caused by Pectobacterium carotovorum subsp. carotovorum in Malaysia[J]. Journal of Plant Pathology. 2011, 93(4):491-491.
[11] TSUDA K, TSUJI G, HIGASHIYAMA M, et al. Biological control of bacterial soft rot in Chinese cabbage by Lactobacillus plantarum strain BY under field conditions [J]. Biological Control, 2016, 100: 63-69.
[12] JEONG R D, CHU E H, PARK D H, et al. Control of postharvest bacterial soft rot by gamma irradiation and its potential modes of action [J].The Plant Pathology Journal,2016,32(2): 157-161.
[13] 张凤兰. 白菜对软腐病的室内抗性鉴定方法及抗源筛选[J]. 蔬菜, 1992(4):20.
[14] 张光明, 王翠花. 大白菜抗软腐病接种鉴定方法的初步研究[J]. 山东农业科学, 1995(5):39-40.
[15] 臧威, 崔崇士, 张耀伟. 大白菜软腐病苗期抗性鉴定方法的研究[J]. 北方园艺, 2003(3):57-58.
[16] 方中达. 植病研究方法[M]. 北京: 农业出版社, 1979.
[17] 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.
[18] 杨晓红, 徐文玲, 张晓明,等. 大白菜抗软腐病接种鉴定及抗源筛选研究[J]. 山东农业科学, 2009(5):67-69.
[19] 叶雪凌, 曹鸣庆, 马荣才. 拟南芥对细菌性软腐病抗性变异分析[J]. 中国生物工程杂志, 2008, 28(11):36-42.
[20] BURR T J, SCHROTH M N. Occurrence of soft rot Erwinia spp. In soil and plant material [J]. Phytopathology,1977,77(11):1382-1387.
[21] DYE D W. A taxonomic study of the genus Erwinia. I. The “amylovora” group [J]. New Zealand Journal of Agricultural Research, 1968, 11:590-607.