枣缩果病互隔链格孢菌的潜在寄主研究
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摘要
[目的]发掘枣缩果病菌可能存在的潜在寄主,为探究枣缩果病的初侵染来源寻找突破口。[方法]用gfp标记的枣缩果病初侵染互隔链格孢菌CN193菌株(CN193::gfp)的孢子悬浮液,以刺伤和无伤两种方式,接种选自枣园周围栽植的番茄、烟草、苹果、梨、毛白杨和加杨,然后,将发病部位病健交界处的组织置于加有10μg·mL~(-1)潮霉素B的PDA培养基上培养,荧光显微镜观察发病组织和从各寄主上分离获得的病菌菌丝。[结果]刺伤接种7 d后,除烟草外,其他刺伤的材料均发病,发病率依次为番茄果实45.8%、苹果果实79.2%、苹果叶片75.0%、梨果实56.3%、梨叶片52.1%、毛白杨叶片66.7%和加杨叶片54.2%;无伤处理的仅番茄发病,其他材料和对照组均未发病。番茄病部组织的荧光清晰可辨;在发病的番茄果实、苹果果实和叶片、梨果实和叶片、毛白杨和加杨离体叶片病部组织分离后长出的菌丝中,均可以观察到明显的荧光。[结论]枣缩果病菌能成功侵染番茄、苹果、梨、毛白杨和加杨,并可使各寄主植物产生与番茄黑斑病、苹果褐纹病、梨黑斑病和杨树叶枯病相同的症状,推断番茄、苹果、梨、毛白杨和加杨为枣缩果病菌的潜在寄主,扩大了枣缩果病菌初侵染来源的范围,为进一步开展枣缩果病的防治拓宽了渠道。
[Objective] To explore the potential hosts of Alternaria alternata, the pathogen of jujube shrunken-fruit disease(JSFD) and the sources of primary infection for JSFD. [Method] The spore suspension of a gfp-tagged strain of A. alternata CN193, designated as CN193::gfp, were inoculated with and without stab wound to the fruits and/or leaves of Lycopersivon esculentum, Nicotiana tabacum, Malus domestica cv. Red Fuji, Pyrus bretschneideri, Populus tomentosa and Populus canadensis from the surrounding area of jujube plantation. The marginal tissues with healthy and diseased parts were transferred onto potato dextrose agar media with supplement of 10 μg·mL~(-1) hygromycin B, and the infected tissues and the hyphae isolated from each of the inoculated material were observed under fluorescence microscope. [Result] All the inoculated materials with stabbing wound, except for N. tabacum plant, developed diseased symptoms after 7 days of inoculation. The disease rates of all the above materials inoculated with spore suspension were 45.8% of L. esculentum fruits, 79.2% of apple fruits, 75.0% of M. domestica cv. Red Fuji leaves, 56.3% of Pyrus bretschneideri fruits, 52.1% of Pyrus bretschneideri leaves, 66.7% of Populus tomentosa leaves and 54.2% of Populus canadensis leaves. Through non-wounded inoculation only L. esculentum fruits showed the symptoms, and all the other treated materials and the controls did not present any visible disease development. Strong fluorescence could be observed in the infected tissues of L. esculentum fruits, and in the hyphae of isolates from all infected materials, including the fruits of L. esculentum, the fruits and leaves of M. domestica cv. Red Fuji, the fruits and leaves of Pyrus bretschneideri, and the leaves of Populus tomentosa and Populus canadensis. [Conclusion] The pathogen of JSFD is able to successfully infect the L. esculentum, M. domestica cv. Red Fuji, Pyrus bretschneideri, Populus tomentosa and Populus canadensis, and it could cause the same symptoms as tomato melasma, apple brown spot, pear black spot, and poplar leaf blight, inferring that L. esculentum, M. domestica cv. Red Fuji, Pyrus bretschneideri, Populus tomentosa and Populus canadensis are the potential hosts of JSFD.
[Objective] To explore the potential hosts of Alternaria alternata, the pathogen of jujube shrunken-fruit disease(JSFD) and the sources of primary infection for JSFD. [Method] The spore suspension of a gfp-tagged strain of A. alternata CN193, designated as CN193::gfp, were inoculated with and without stab wound to the fruits and/or leaves of Lycopersivon esculentum, Nicotiana tabacum, Malus domestica cv. Red Fuji, Pyrus bretschneideri, Populus tomentosa and Populus canadensis from the surrounding area of jujube plantation. The marginal tissues with healthy and diseased parts were transferred onto potato dextrose agar media with supplement of 10 μg·mL~(-1) hygromycin B, and the infected tissues and the hyphae isolated from each of the inoculated material were observed under fluorescence microscope. [Result] All the inoculated materials with stabbing wound, except for N. tabacum plant, developed diseased symptoms after 7 days of inoculation. The disease rates of all the above materials inoculated with spore suspension were 45.8% of L. esculentum fruits, 79.2% of apple fruits, 75.0% of M. domestica cv. Red Fuji leaves, 56.3% of Pyrus bretschneideri fruits, 52.1% of Pyrus bretschneideri leaves, 66.7% of Populus tomentosa leaves and 54.2% of Populus canadensis leaves. Through non-wounded inoculation only L. esculentum fruits showed the symptoms, and all the other treated materials and the controls did not present any visible disease development. Strong fluorescence could be observed in the infected tissues of L. esculentum fruits, and in the hyphae of isolates from all infected materials, including the fruits of L. esculentum, the fruits and leaves of M. domestica cv. Red Fuji, the fruits and leaves of Pyrus bretschneideri, and the leaves of Populus tomentosa and Populus canadensis. [Conclusion] The pathogen of JSFD is able to successfully infect the L. esculentum, M. domestica cv. Red Fuji, Pyrus bretschneideri, Populus tomentosa and Populus canadensis, and it could cause the same symptoms as tomato melasma, apple brown spot, pear black spot, and poplar leaf blight, inferring that L. esculentum, M. domestica cv. Red Fuji, Pyrus bretschneideri, Populus tomentosa and Populus canadensis are the potential hosts of JSFD.
引文
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[12] 乔镜澄,刘宇,马敬昊,等.番茄黑斑病病原菌的鉴定及生物学特性研究[J].江苏农业科学,2017,45(10):94-97.
[13] 祖艳青,蒋士君,王海涛,等.河南省烟草赤星病病原鉴定[J].中国烟草学报,2013,19(4):73-77.
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[16] 任璐,史晓晶,姚众,等.苹果斑点落叶病菌对戊唑醇敏感基线建立及抗性突变体适合度[J].植物病理学报,2017,47(3):380-388.
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[18] 王宏.梨黑斑病菌(Alternaria alternata Japanese pear pathotype)生物学特性及其生物—化学协同控制技术的研究[D].南京:南京农业大学,2006.
[19] 苏晓华,张绮纹,沈瑞祥,等.美洲黑杨×青杨F_2代抗杨叶枯病遗传变异研究[J].林业科学研究,1998,11(6):4-7.
[20] 潘凤英,蓝霞,黄羽,等.广西地区葡萄黑痘病病原菌的分离与鉴定[J].植物病理学报,2017,47(1):9-14.
[21] Mukesh M,Andleeb Z,Prashant S,et al.Effect on lycopene,β-carotene,ascorbic acid and phenolic content in tomato fruits infected by Alternaria alternata and its toxins[J].Archives of Phytopathology and Plant Protection,2017,50(7-8):317-329.
[22] Hou Y J,Ma X,Wan W T,et al.Comparative genomics of pathogens causing brown spot disease of tobacco:Alternaria longipes and Alternaria alternata[J].Plos One,2016,11(5):1-21.
[23] Jung K H.Growth inhibition effect of pyroligneous acid on pathogenic fungus,Alternaria mali,the agent of Alternaria blotch of apple[J].Biotechnology and Bioprocess Engineering,2007,12(3):318-322.
[24] Ting T P,Da W S,Hong B P,et al.Detection of A.alternata from pear juice using surface-enhanced Raman spectroscopy based silver nanodots array[J].Journal of Food Engineering,2017:147-155.
[25] Zhi C J,Ji Q G,Yi M S,et al.Enhanced resistance to fungal pathogens in transgenic Populus tomentosa Carr.by overexpression of an nsLTP-like antimicrobial protein gene from motherwort (Leonurus japonicus)[J].Tree Physiology,2010,30(12):1599-1605.
[26] 张彩霞,陈莹,李壮,等.苹果斑点落叶病致病菌的鉴定及生物学特性研究[J].生物技术,2011,(4):58-61.
[27] 黄毅.杨树叶枯病生物防治初步研究[D].哈尔滨:东北林业大学,2010.
[28] 卢东升,杨霁虹.冷藏苹果褐斑病病原菌鉴定及致病性研究[J].河南农业科学,2006,(12):76-78.
[29] 王宏,常有宏,陈志谊.梨黑斑病病原菌生物学特性研究[J].果树学报,2006,(2):247-251.
[30] 赵乐,邢艺林,李正楠,等.枣缩果病初侵染链格孢菌的抗硫酸铜标记及稳定性[J].林业科学,2016,52(7):53-58.
[2] 郑晓莲,赵光耀,齐秋锁,等.枣干腰缩果病症状类型及侵染规律研究[J].植物保护,1998,24(4):17-19.
[3] 杨自民,姚忍让,王月丽,等.枣缩果病的发生原因及防治方法探讨[J].北方果树,2002,(4):33-34.
[4] 徐祥彬,赖童飞,景云飞,等.山西壶瓶枣缩果病病原菌分离和鉴定[J].植物病理学报,2009,39(3):225-230.
[5] 赵乐.枣缩果病菌的初侵染时期及防治研究[D].保定:河北农业大学,2015.
[6] 许瑛,姚兆群,王兰,等.阿拉尔地区枣园不同时期枣花枣果黑斑病菌带菌量检测[J].新疆农业科学,2017,54(10):1-8.
[7] 侯晓杰.枣缩果病病原和防治研究[D].保定:河北农业大学,2010.
[8] 韩党悦.枣缩果病初侵染病原及防治研究[D].保定:河北农业大学,2012.
[9] 许阳.枣缩果病初侵染病原的分子检测及防治研究[D].保定:河北农业大学,2014.
[10] 翟凤艳,张柯,李娟,等.新乡地区链格孢菌病害病原鉴定[J].河南科技学院学报,2015,43(5):30-35.
[11] Yang J,Sun C,Zhang Y,et al.Induced resistance in tomato fruit by γ-aminobutyric acid for the control of alternaria rot caused by Alternaria alternata[J].Food Chemistry,2017,221:1014-1020.
[12] 乔镜澄,刘宇,马敬昊,等.番茄黑斑病病原菌的鉴定及生物学特性研究[J].江苏农业科学,2017,45(10):94-97.
[13] 祖艳青,蒋士君,王海涛,等.河南省烟草赤星病病原鉴定[J].中国烟草学报,2013,19(4):73-77.
[14] 杨涛,黎妍妍,郑露,等.湖北烟区烤烟赤星病病原鉴定[J].中国烟草科学,2017,(5):32-38.
[15] Zhu L,Ni W,Liu S,et al.Transcriptomics analysis of apple leaves in response to Alternaria alternata apple pathotype infection[J].Frontiers in Plant Science,2017,8:22.
[16] 任璐,史晓晶,姚众,等.苹果斑点落叶病菌对戊唑醇敏感基线建立及抗性突变体适合度[J].植物病理学报,2017,47(3):380-388.
[17] Kan J,Liu T,Ma N,et al.Transcriptome analysis of Callery pear (Pyrus calleryana) reveals a comprehensive signalling network in response to Alternaria alternata.[J].Plos One,2017,12(9):e0184988.
[18] 王宏.梨黑斑病菌(Alternaria alternata Japanese pear pathotype)生物学特性及其生物—化学协同控制技术的研究[D].南京:南京农业大学,2006.
[19] 苏晓华,张绮纹,沈瑞祥,等.美洲黑杨×青杨F_2代抗杨叶枯病遗传变异研究[J].林业科学研究,1998,11(6):4-7.
[20] 潘凤英,蓝霞,黄羽,等.广西地区葡萄黑痘病病原菌的分离与鉴定[J].植物病理学报,2017,47(1):9-14.
[21] Mukesh M,Andleeb Z,Prashant S,et al.Effect on lycopene,β-carotene,ascorbic acid and phenolic content in tomato fruits infected by Alternaria alternata and its toxins[J].Archives of Phytopathology and Plant Protection,2017,50(7-8):317-329.
[22] Hou Y J,Ma X,Wan W T,et al.Comparative genomics of pathogens causing brown spot disease of tobacco:Alternaria longipes and Alternaria alternata[J].Plos One,2016,11(5):1-21.
[23] Jung K H.Growth inhibition effect of pyroligneous acid on pathogenic fungus,Alternaria mali,the agent of Alternaria blotch of apple[J].Biotechnology and Bioprocess Engineering,2007,12(3):318-322.
[24] Ting T P,Da W S,Hong B P,et al.Detection of A.alternata from pear juice using surface-enhanced Raman spectroscopy based silver nanodots array[J].Journal of Food Engineering,2017:147-155.
[25] Zhi C J,Ji Q G,Yi M S,et al.Enhanced resistance to fungal pathogens in transgenic Populus tomentosa Carr.by overexpression of an nsLTP-like antimicrobial protein gene from motherwort (Leonurus japonicus)[J].Tree Physiology,2010,30(12):1599-1605.
[26] 张彩霞,陈莹,李壮,等.苹果斑点落叶病致病菌的鉴定及生物学特性研究[J].生物技术,2011,(4):58-61.
[27] 黄毅.杨树叶枯病生物防治初步研究[D].哈尔滨:东北林业大学,2010.
[28] 卢东升,杨霁虹.冷藏苹果褐斑病病原菌鉴定及致病性研究[J].河南农业科学,2006,(12):76-78.
[29] 王宏,常有宏,陈志谊.梨黑斑病病原菌生物学特性研究[J].果树学报,2006,(2):247-251.
[30] 赵乐,邢艺林,李正楠,等.枣缩果病初侵染链格孢菌的抗硫酸铜标记及稳定性[J].林业科学,2016,52(7):53-58.