[1]陈慧杰,赵爽,张凯凯,等.菊花枯萎病病原菌的分离和鉴定及其粗毒素对切花菊‘神马’幼苗生长的影响[J].南京农业大学学报,2018,41(4):662-669.[doi:10.7685/jnau.201708009]
 CHEN Huijie,ZHAO Shuang,ZHANG Kaikai,et al.Isolation and identification of Fusarium oxysporum from chrysanthemum and the effects of crude toxin on growth of cut-chrysanthemum ‘Jimba’ seedlings[J].Journal of Nanjing Agricultural University,2018,41(4):662-669.[doi:10.7685/jnau.201708009]
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菊花枯萎病病原菌的分离和鉴定及其粗毒素对切花菊‘神马’幼苗生长的影响()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
41卷
期数:
2018年4期
页码:
662-669
栏目:
出版日期:
2018-07-09

文章信息/Info

Title:
Isolation and identification of Fusarium oxysporum from chrysanthemum and the effects of crude toxin on growth of cut-chrysanthemum ‘Jimba’ seedlings
作者:
陈慧杰 赵爽 张凯凯 邹忠幸 倪嘉琪 姜晓帆 陈发棣 房伟民
南京农业大学园艺学院/农业农村部景观设计重点实验室, 江苏 南京 210095
Author(s):
CHEN Huijie ZHAO Shuang ZHANG Kaikai ZOU Zhongxing NI Jiaqi JIANG Xiaofan CHEN Fadi FANG Weimin
College of Horticulture/Key Laboratory of Landscaping, Ministry of Agriculture and Rural Affairs, Nanjing Agricultural University, Nanjing 210095, China
关键词:
菊花尖孢镰刀菌分离和鉴定粗毒素生理响应
Keywords:
chrysanthemumFusarium oxysporumisolation and identificationcrude toxinphysiological response
DOI:
10.7685/jnau.201708009
摘要:
[目的]本文旨在通过对菊花枯萎致病菌的分离和鉴定,研究其粗毒素对切花菊‘神马’幼苗的化感作用。[方法]观察切花菊‘神马’枯萎病的田间发病症状,采用组织分离培养法从菊花病株上分离到1株致病真菌CFD-B2,提取CFD-B2粗毒素,设5个不同稀释倍数的粗毒素处理:对照、粗毒素原液(168.12 μg·mL-1)、粗毒素稀释5倍、粗毒素稀释10倍和粗毒素稀释25倍,测定不同处理中切花菊‘神马’幼苗株高、根长、茎粗,抗性物质含量,细胞膜相对透性以及根系过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)和多酚氧化酶(PPO)活性的变化。[结果]通过ITS基因核酸序列分析,鉴定CFD-B2为镰刀菌属尖孢镰刀菌(Fusarium oxporum)。尖孢镰刀菌CFD-B2不同稀释倍数的粗毒素处理对切花菊‘神马’幼苗的株高、茎粗、根长均有抑制作用,抑制程度由强到弱依次为:粗毒素原液处理、粗毒素稀释5倍处理、粗毒素稀释10倍处理、粗毒素稀释25倍处理。处理7 d时,粗毒素原液对菊花幼苗株高、茎粗、根长抑制效果最显著,比对照分别减少了28.87%、7.31%和42.48%。不同稀释倍数的粗毒素处理能增加切花菊‘神马’幼苗根系组织细胞膜透性及丙二醛(MDA)、可溶性糖和脯氨酸含量。不同稀释倍数的粗毒素处理能够增加根系保护酶活性,且效果从高到低表现为:粗毒素原液处理、粗毒素稀释5倍处理、粗毒素稀释10倍处理、粗毒素稀释25倍处理。同一稀释倍数的粗毒素处理短期内能够增加根系保护酶活性。粗毒素原液处理12 d时,PAL、POD和PPO活性达到最高,比对照分别提高了44.39%、36.96%和35.38%。[结论]尖孢镰刀菌CFD-B2粗毒素对切花菊‘神马’幼苗的化感作用主要表现在可抑制切花菊幼苗根、茎的正常生长,增加切花菊幼苗根系的细胞膜透性以及根系组织中MDA、可溶性糖和脯氨酸的含量,在短期内提高根系保护酶PAL、POD和PPO活性。
Abstract:
[Objectives]The paper aims to study the allelopathic effects of Fusarium oxysporum crude toxin on cut-chrysanthemum seedlings through the isolation and identification of cut-chrysanthemum Fusarium wilt pathogenic fungal.[Methods]Fusarium wilt symptoms in cut-chrysanthemum ‘Jimba’ were observed,then the tissue culture method was used to isolate the pathogenic fungi. The pathogenic fungus CFD-B2 was isolated. The crude toxin of CFD-B2 was extracted and diluted as below:CK,the crude toxin,5 times dilution,10 times dilution,15 times dilution,and 25 times dilution. The steam height,stem diameter,root length,resistant material content,membrane relative permeability,and activities of polyphenol oxidase(PPO),peroxidase(POD)and phenylalanine ammonia-lyase(PAL)in different treatments were investigated at different times.[Results]The results indicated that the strain CFD-B2 was isolated and identified as Fusarium oxysporum by ITS sequencing. The F. oxysporum crude toxin had significant effect on inhibiting the steam height,stem diameter and root length of cut-chrysanthemum seedlings. The inhibiting effect from strong to weak was:crude toxin,diluted 5 times,diluted 10 times,and diluted 25 times. Seven days after crude toxin spraying,the plant height,stem diameter and root length decreased by 28.87%,7.31% and 42.48% respectively when compared to the CK. The CFD-B2 crude toxin could increase the root cell membrane permeability,the content of MDA,soluble sugar and proline content in the root tissue. The CFD-B2 crude toxin had significant promotion effects on increasing the activities of the root cell protective enzyme PAL,POD and PPO. And the promoting effect from strong to weak was:crude toxin,diluted 5 times,diluted 10 times,and diluted 25 times. Compared with the control,12 days after crude toxin spraying,PAL,POD and PPO increased by 44.39%,36.96% and 35.38%,respectively.[Conclusions]Therefore,the allelopathic effects of F. oxysporum CFD-B2 crude toxin on cut-chrysanthemum seedlings were mainly in inhibiting root and stem growth in cut-chrysanthemum seedlings,destructing the root cell membrane permeability and increasing the contents of MDA,normal soluble sugar and proline,and promoting the activities of PAL,POD and PPO in root tissue of cut-chrysanthemum seedlings.

参考文献/References:

[1] 陈希,赵爽,姚建军,等. 微生物有机肥及杀菌剂对切花菊连作障碍的影响[J]. 应用生态学报,2015,26(4):1231-1236. Chen X,Zhao S,Yao J J,et al. Effects of bio-organic fertilizer and fungicide application on continuous cropping obstacles of cut chrysanthemum[J]. Chinese Journal of Applied Ecology,2015,26(4):1231-1236(in Chinese with English abstract).
[2] 刘新月,李凡,陈海如,等. 致病性尖孢镰刀菌生物防治研究进展[J]. 云南大学学报,2008,30(S1):89-93. Liu X Y,Li F,Chen H R,et al. Research progress in biological control of Fusarium oxysporum[J]. Journal of Yunnan University,2008,30(S1):89-93(in Chinese with English abstract).
[3] 陈希,赵爽,史亚东,等. 生物有机肥对‘滁菊’连作障碍的影响[J]. 南京农业大学学报,2015,38(1):50-56. DOI:10.7685/j.issn.1000-2030.2015.01.008. Chen X,Zhao S,Shi Y D,et al. Effects of bio-organic fertilizer application on the continuous cropping obstacles of Chrysanthemum morifolium Tzvel ‘Chuju’[J]. Journal of Nanjing Agricultural University,2015,38(1):50-56(in Chinese with English abstract).
[4] 郝晓娟,刘波,谢关林,等. 植物枯萎病生物防治研究进展[J]. 中国农学通报,2005,21(7):319-323. Hao X J,Liu B,Xie G L,et al. Research progress in biological control of Fusarium wilt disease[J]. Chinese Agricultural Science Bulletin,2005,21(7):319-323(in Chinese with English abstract).
[5] 高晓敏,王琚钢,马立国,等. 尖孢镰刀菌致病机理和化感作用研究进展[J]. 微生物学通报,2014,41(10):2143-2148. Gao X M,Wang J G,Ma L G,et al. Research advances on the mechanism of pathogenesis and allelopathy of Fusarium oxysporium[J]. Microbiology China,2014,41(10):2143-2148(in Chinese with English abstract).
[6] Chakrabarti D K,Chaudhary K C B. Correlation between virulence and fusaric acid production in Fusarium oxysporum f. sp. carthami[J]. Journal of Phytopathology,1980,99(1):43-46.
[7] 田雪亮,刘鸣韬,杨家荣,等. 黄瓜枯萎菌粗毒素对不同抗性黄瓜种子萌发及幼苗胁迫作用的研究[J]. 中国生态农业学报,2008,16(6):1495-1498. Tian X L,Liu M T,Yang J R,et al. Seed germination and growth of various resistant cucumber seedlings under Fusarium oxysporum crude toxin stress[J]. Chinese Journal of Eco-Agriculture,2008,16(6):1495-1498(in Chinese with English abstract).
[8] 韩鲁明,王丽君,曹莎,等. 西瓜砧木种传尖孢镰刀菌粗毒素研究[J]. 中国植保导刊,2013,33(2):1672-1680. Han L M,Wang L J,Cao S,et al. Study on the crude toxin produced by seed borne Fusarium oxysporum of rootstock for watermelon graftin[J]. China Plant Protection,2013,33(2):1672-1680(in Chinese with English abstract).
[9] 石凤梅. 植物病原真菌毒素的研究进展[J]. 黑龙江农业科学,2006(2):70-73. Shi F M. Research advances on the toxin of the plant pathogenic fungus[J]. Heilongjiang Agricultural Sciences,2006(2):70-73(in Chinese with English abstract).
[10] 张淑珍. 植物病原菌毒素研究进展[J]. 黑龙江农业科学,2001(2):42-43. Zhang S Z. Advances of research on the plant pathogenic toxin[J]. Heilongjiang Agricultural Sciences,2001(2):42-43(in Chinese with English abstract).
[11] 胡忠亮,邹东霞,黄敏仁,等. 镰刀菌毒素滤液对转基因香石竹抗枯萎病性能的测定[J]. 科学技术与工程,2016,16(27):131-136. Hu Z L,Zou D X,Huang M R,et al. Determination of Fusarium wilt resistance dealed with Fusarium toxins filtrate on transgenic resistance of carnation[J]. Science Technology and Engineering,2016,16(27):131-136(in Chinese with English abstract).
[12] 胡颖慧,龚束芳,李彩华,等. 枯萎病菌毒素培养滤液对唐菖蒲幼苗毒性的初步研究[J]. 植物病理学报,2012,42(5):497-504. Hu Y H,Gong S F,Li C H,et al. Poisoning effects of toxin culture filtrate of Fusarium wilt pathogen on seedling of gladiolus[J]. Acta Phytopathologica Sinica,2012,42(5):497-504(in Chinese with English abstract).
[13] 刘梅,卢志军,王文君,等. 甘蓝种传尖孢镰刀菌粗毒素的研究[J]. 中国农业大学学报,2010,15(3):63-69. Liu M,Lu Z J,Wang W J,et al. Effect of the crude toxin from seedborne Fusarium oxysporum on seed germination and seedlings of cabbage[J]. Journal of China Agricultural University,2010,15(3):63-69(in Chinese with English abstract).
[14] Song A P,Zhao S,Chen S S,et al. The abundance and diversity of soil fungi in continuously monocropped chrysanthemum[J]. The Scientific World Journal,2013(1):632920.
[15] Curir P,Guglieri L,Capponi A,et al. Fusaric acid production by Fusarium oxysporum f. sp. lilii and its role in the lily basal rot disease[J]. European Journal of Plant Pathology,2000,106(9):849-856.
[16] 周宝利,高艳新,林桂荣,等. 嫁接茄子抗病性与电导率、脯氨酸含量及苯丙氨酸解氨酶活性的关系[J]. 园艺学报,1998,25(3):300-302. Zhou B L,Gao Y X,Lin G R,et al. Relationship between disease resistance and electrolytic leakage,proline content and PAL activity in grafted eggplant[J]. Acta Horticulturae Sinica,1998,25(3):300-302(in Chinese with English abstract).
[17] 马国斌,林德佩,王叶筠,等. 西瓜枯萎病菌镰刀菌酸对西瓜苗作用机制的初步探讨[J]. 植物病理学报,2000,30(4):373-374. Ma G B,Lin D P,Wang Y Y,et al. Studies on the wilting mechanism of fusaric acids of Fusarium oxysporum on watermelon[J]. Acta Phytopathologica Sinica,2000,30(4):373-374(in Chinese with English abstract).
[18] 赵进红,王玉山,冯殿齐,等. 药物处理对疯枣枝过氧化物酶和苯丙氨酸解氨酶活性的影响[J]. 山东农业大学学报(自然科学版),2010,41(3):360-364. Zhao J H,Wang Y S,Feng D Q,et al. Effects of medcament treatment on peroxidase and phenylalanine ammonia lyase of jujube branches infected with witches broom disease[J]. Journal of Shandong Agricultural University(Natural Science),2010,41(3):360-364(in Chinese with English abstract).
[19] 张志良,翟伟菁. 植物生理学实验指导[M]. 北京:高等教育出版社,2003:69-71. Zhang Z L,Zhai W J. Plant Physiology Experiment Guide[M]. Beijing:Higher Education Press,2003:69-71(in Chinese).
[20] 高子勤,张淑香. 连作障碍与根际微生态研究Ⅰ. 根系分泌物及其生态效应[J]. 应用生态学报,1998,9(5):549-554. Gao Z Q,Zhang S X. Continuous cropping obstacle and rhizospheric microecology I. Root exudates and their eco-logical effects[J]. Chinese Journal of Applied Ecology,1998,9(5):549-554(in Chinese with English abstract).
[21] Astrom B,Gustafsson A,Gerhardson B. Characteristics of a plant deleterious rhizosphere pseudomonad and its inhibitory metabolite(s)[J]. Journal of Applied Bacteriology,1993,74:20-28.
[22] 杨媚,黄永辉,舒灿伟,等. 香蕉枯萎病菌4号生理小种粗毒素特性的研究[J]. 园艺学报,2012,39(3):545-551. Yang M,Huang Y H,Shu C W,et al. Studies on the characteristics of crude toxin produced by Fusarium oxysporum f. sp. cubense race 4[J]. Acta Horticulturae Sinica,2012,39(3):545-551(in Chinese with English abstract).
[23] 董鲜.土传香蕉枯萎病发生的生理机制及营养防控效果的研究[D]. 南京:南京农业大学,2014. Dong X. Study on physiological mechanism of soil-borne banana Fusarium wilt occurrence and nutrients control effect of wilt disease[D]. Nanjing:Nanjing Agricultural University,2014(in Chinese with English abstract).
[24] 朱虹,祖元刚,王文杰,等. 逆境胁迫条件脯氨酸对植物生长的影响[J]. 东北林业大学学报,2009,37(4):86-89. Zhu H,Zu Y G,Wang W J,et al. Effect of proline on plant growth under different stress conditions[J]. Journal of Northeast Forestry University,2009,37(4):86-89(in Chinese with English abstract).
[25] 赵娟,薛泉宏,杜军志,等. 两株镰孢菌的鉴定及其粗毒素对甜瓜幼苗的化感作用[J]. 应用生态学报,2013,24(1):142-148. Zhao J,Xue Q H,Du J Z,et al. Identification of two Fusarium isolates and their crude toxin allelopathic effect on Cucumis melo seedlings[J]. Chinese Journal of Applied Ecology,2013,24(1):142-148(in Chinese with English abstract).
[26] 秦国政,田世平,刘海波,等. 拮抗菌与病原菌处理对采后桃果实多酚氧化酶、过氧化物酶及苯丙氨酸解氨酶的诱导[J]. 中国农业科学,2003,61(3):89-93. Qin G Z,Tian S P,Liu H B,et al. Polyphenol oxidase,peroxidase and phenylalanine ammonium lyase in postharvest peach fruits induced by inoculation with Pichia membranefaciens or Rhizopus stolonifer[J]. Scientia Agricultura Sinica,2003,61(3):89-93(in Chinese with English abstract).

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备注/Memo

备注/Memo:
收稿日期:2017-08-07。
基金项目:江苏省产学研前瞻性联合研究项目(BY2016077-06);江苏省农业三新工程项目[XGC(2017)280];中央高校基本科研业务费专项资金(KYCYL201501);江苏省农业科技自主创新资金[CX(16)1025]
作者简介:陈慧杰,博士研究生。
通信作者:房伟民,教授,主要从事花卉遗传育种与栽培应用研究,E-mail:fangwm@njau.edu.cn
更新日期/Last Update: 1900-01-01