内生真菌司氏角担子菌B6对尖孢镰刀菌西瓜专化型菌株的拮抗作用
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摘要
近年来,尽管西瓜产业不断发展壮大,但轮作土壤种植西瓜易产生枯萎病害导致世界范围内的西瓜严重减产。通过对峙培养实验和抑菌实验探讨了重阳木内生真菌司氏角担子菌(Ceratobasidum stevensii)B6菌株对西瓜枯萎病病原菌尖孢镰刀菌西瓜专化型(Fusarium oxysporum f. sp.Niveum,FON)的拮抗作用,并初步分析了其作用机制。平板拮抗试验的结果表明,内生真菌B6生长过程中不是通过产生抑菌带来抑制FON菌丝,而是利用自身的生长优势将FON完全覆盖。显微观察B6与FON菌丝的接触部位,发现FON菌丝外侧附着B6顶端菌丝形成的胞样结构,表明FON菌丝生长仅受到B6菌丝的抑制。抑菌试验结果显示,B6产生的挥发性物质可以抑制FON的生长和产孢,并使其菌丝分枝明显减少;B6的发酵液对FON的生长和产孢没有抑制作用。因此,推测B6主要通过释放某些挥发性物质产生拮抗作用而抑制FON的生长。
In recent years, although the watermelon industry has developed and grown steadily, yet it is easy to produce serious wilt disease caused by continuous cropping soil and resulted in falling in production of watermelon. In this study a confrontation experiment and inhibition experiment were adopted to investigate the antagonism of endophytic fungus Ceratobasidum stevensii(B6) of Bischofia polycarpa against Fusarium oxysporum f. sp. Niveum(FON), and initially analyzed the effective mechanism. The results of plate antagonistic experiment showed that the endophytic fungus B6 did not inhibited FON hyphae through producing inhibition zone during its growth, but totally covered the mycelium of FON by its own growing superiority. It was found under microscopic observation that at the contact location between B6 and FON hyphae an appressoria-like structures were formed at the end hyphae of B6, indicated that the growth of FON hyphae was only inhibited by B6 hyphae. The inhibition experiment results showed that volatile substances produced by B6 could inhibit the growth of FON and sporulation, and made the mycelium branches obviously reduced, however, the fermentation broth of B6 did not have inhibition effects against the growth and spore production of FON. Therefore, it is inferred that B6 mainly inhibits the growth of FON by releasing antagonistic volatile substances.
In recent years, although the watermelon industry has developed and grown steadily, yet it is easy to produce serious wilt disease caused by continuous cropping soil and resulted in falling in production of watermelon. In this study a confrontation experiment and inhibition experiment were adopted to investigate the antagonism of endophytic fungus Ceratobasidum stevensii(B6) of Bischofia polycarpa against Fusarium oxysporum f. sp. Niveum(FON), and initially analyzed the effective mechanism. The results of plate antagonistic experiment showed that the endophytic fungus B6 did not inhibited FON hyphae through producing inhibition zone during its growth, but totally covered the mycelium of FON by its own growing superiority. It was found under microscopic observation that at the contact location between B6 and FON hyphae an appressoria-like structures were formed at the end hyphae of B6, indicated that the growth of FON hyphae was only inhibited by B6 hyphae. The inhibition experiment results showed that volatile substances produced by B6 could inhibit the growth of FON and sporulation, and made the mycelium branches obviously reduced, however, the fermentation broth of B6 did not have inhibition effects against the growth and spore production of FON. Therefore, it is inferred that B6 mainly inhibits the growth of FON by releasing antagonistic volatile substances.
引文
[1] 黄春艳,卜元卿,单正军,等.西瓜连作病害机理及生物防治研究进展[J].生态学杂志,2016,35(6):1670-1676.
[2] 康金磊,韩旭,张仕琦,等.一株来源于茼蒿根际的西瓜枯萎病拮抗菌的鉴定及特性研究[J].华北农学报,2016,31(S1):447-452.
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[8] 史应武,娄恺,李春.植物内生菌在生物防治中的应用[J].微生物学杂志,2009,29(6):61-64.
[9] 周开胜.厌氧还原土壤灭菌法抑制西瓜专化型FON[J].江苏农业学报,2015,(5):1006-1011.
[10] Ade C,Satejnberg A,Sabuquillo P,et al.Management Fusarium wilt on melon and watermelon by Penicillium oxalicum[J].Biological Control,2009,51(3):480-486.
[11] Ling N,Huang Q,Guo S,et al.Paenibacillus polymyxa SQR-21 systemically affects root exudates of watermelon to decrease the conidial germination of Fusarium oxysporum f.sp.niveum[J].Plant and Soil,2011,341(1-2):485-493.
[12] 赵丽明,丁延芹,路晓萌,等.西瓜根际枯萎病拮抗放线菌的筛选及鉴定[J].生物技术通报,2010,(5):107-110.
[13] 张丽萍,黄亚丽,程辉彩,等.复合生物制剂防治西瓜连作病害的研究[J].中国土壤与肥料,2006,(5):59-61.
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[15] 戴传超,余伯阳,王新风,等.驱杀菜青虫的内生菌鉴定与化合物研究[J].安徽农业科学,2006,34(4):694-697.
[16] Dai CC,Tian LS,Zhao YT,et al.Degradation of phenanthrene by the endophytic fungus Ceratobasidum stevensii found in Bischofia polycarpa[J].Biodegradation,2010,21(2):245-255.
[17] 肖逸,王兴祥,王宏伟,等.施加角担子菌B6对连作西瓜土壤微环境和西瓜生长的影响[J].生态学报,2012,32(4):1185-1192.
[18] 曹荣花,刘晓光,高克祥,等.芦苇内生真菌Choiromyces aboriginum Mü1w1c6 的拮抗作用及其生物防治潜力[J].植物保护学报,2008,35(2):107-112.
[19] 李江,袁月祥,闫志英,等.毛栓菌原生质体制备和再生及单核菌株产漆酶特性[J].菌物学报,2012,31(1):102-109.
[20] 郝玉娥,牟贵平,何爱桃,等.挥发性物质诱吸线虫的土壤细菌系统多样性特征及其活性挥发物鉴定[J].微生物学报,2011,51(11):1454-1460.
[21] 谷祖敏,毕卉,张兵,等.不同木霉菌株对黄瓜枯萎病菌的拮抗作用[J].西北农业学报,2018,27(3):426-431.
[22] 纪明山,李博强,陈捷,等.绿色木霉TR-8菌株对尖镰孢的拮抗机制[J].中国生物防治学报,2005,21(2):104-108.
[23] Combès A,Ndoye I,Bance C,et al.Chemical Communication between the Endophytic Fungus Paraconiothyrium variabile and the Phytopathogen Fusarium oxysporum[J].Plos One,2012,7(10):e47313.
[24] 姜华年,丁婷.厚朴内生真菌HPFJ3对小麦全蚀病的生防作用[J].江苏农业学报,2016,32(2):285-292.
[25] Gunatilaka AA.Natural products from plant-associated microorganisms:distribution,structural diversity,bioactivity,and implications of their occurrence[J].J Nat Prod,2006,69(3):509-526.
[26] Mitchell AM,Strobel GA,Moore E,et al.Volatile antimicrobials from Muscodor crispans,a novel endophytic fungus[J].Microbiology,2001,147(11):2943-2950.
[27] 谢洁,任慧爽,唐翠明,等.一株桑树内生细菌的鉴定和对桑椹核地杖菌的拮抗作用[J].蚕业科学,2015,41(5):815-824.
[28] Zhou JY,Zhao XY,Dai CC.Antagonistic mechanisms of endophytic Pseudomonas fluorescens against Athelia rolfsii[J].Journal of Applied Microbiology,2014,117(4):1144-1158.
[2] 康金磊,韩旭,张仕琦,等.一株来源于茼蒿根际的西瓜枯萎病拮抗菌的鉴定及特性研究[J].华北农学报,2016,31(S1):447-452.
[3] 沈宗专,孙莉,王东升,等.石灰碳铵熏蒸与施用生物有机肥对连作黄瓜和西瓜枯萎病及生物量的影响[J].应用生态学报,2017,28(10):3351-3359.
[4] 刘贵友,邹瑶,顾杨霞,等.内生真菌对连作花生土壤FON的拮抗作用[J].江苏农业科学,2016,44(12):171-174.
[5] Tziros GT,Lagopodi AL,Tzavellakolnari K.Reduction of Fusarium wilt in watermelon by Pseudomonas chlororaphis PCL1391 and P.fluorescens WCS365[J].Phytopathol Mediterr,2007,46(3):320-323.
[6] Ding T,Jiang T,Zhou J,et al.Evaluation of antimicrobial activity of endophytic fungi from Camptotheca acuminata (Nyssaceae).Genetics and Molecular Research,2010,9(4):2104-2112.
[7] 敖武,曾庆桂,李希茜,等.1株拮抗真菌的生防细菌Xj11的分离与鉴定[J].微生物学杂志,2013,33(3):45-48.
[8] 史应武,娄恺,李春.植物内生菌在生物防治中的应用[J].微生物学杂志,2009,29(6):61-64.
[9] 周开胜.厌氧还原土壤灭菌法抑制西瓜专化型FON[J].江苏农业学报,2015,(5):1006-1011.
[10] Ade C,Satejnberg A,Sabuquillo P,et al.Management Fusarium wilt on melon and watermelon by Penicillium oxalicum[J].Biological Control,2009,51(3):480-486.
[11] Ling N,Huang Q,Guo S,et al.Paenibacillus polymyxa SQR-21 systemically affects root exudates of watermelon to decrease the conidial germination of Fusarium oxysporum f.sp.niveum[J].Plant and Soil,2011,341(1-2):485-493.
[12] 赵丽明,丁延芹,路晓萌,等.西瓜根际枯萎病拮抗放线菌的筛选及鉴定[J].生物技术通报,2010,(5):107-110.
[13] 张丽萍,黄亚丽,程辉彩,等.复合生物制剂防治西瓜连作病害的研究[J].中国土壤与肥料,2006,(5):59-61.
[14] 戴传超,余伯阳,赵玉婷,等.大戟科4种植物内生真菌分离与抑菌研究[J].南京林业大学学报(自然科学版),2006,30(1):79-83.
[15] 戴传超,余伯阳,王新风,等.驱杀菜青虫的内生菌鉴定与化合物研究[J].安徽农业科学,2006,34(4):694-697.
[16] Dai CC,Tian LS,Zhao YT,et al.Degradation of phenanthrene by the endophytic fungus Ceratobasidum stevensii found in Bischofia polycarpa[J].Biodegradation,2010,21(2):245-255.
[17] 肖逸,王兴祥,王宏伟,等.施加角担子菌B6对连作西瓜土壤微环境和西瓜生长的影响[J].生态学报,2012,32(4):1185-1192.
[18] 曹荣花,刘晓光,高克祥,等.芦苇内生真菌Choiromyces aboriginum Mü1w1c6 的拮抗作用及其生物防治潜力[J].植物保护学报,2008,35(2):107-112.
[19] 李江,袁月祥,闫志英,等.毛栓菌原生质体制备和再生及单核菌株产漆酶特性[J].菌物学报,2012,31(1):102-109.
[20] 郝玉娥,牟贵平,何爱桃,等.挥发性物质诱吸线虫的土壤细菌系统多样性特征及其活性挥发物鉴定[J].微生物学报,2011,51(11):1454-1460.
[21] 谷祖敏,毕卉,张兵,等.不同木霉菌株对黄瓜枯萎病菌的拮抗作用[J].西北农业学报,2018,27(3):426-431.
[22] 纪明山,李博强,陈捷,等.绿色木霉TR-8菌株对尖镰孢的拮抗机制[J].中国生物防治学报,2005,21(2):104-108.
[23] Combès A,Ndoye I,Bance C,et al.Chemical Communication between the Endophytic Fungus Paraconiothyrium variabile and the Phytopathogen Fusarium oxysporum[J].Plos One,2012,7(10):e47313.
[24] 姜华年,丁婷.厚朴内生真菌HPFJ3对小麦全蚀病的生防作用[J].江苏农业学报,2016,32(2):285-292.
[25] Gunatilaka AA.Natural products from plant-associated microorganisms:distribution,structural diversity,bioactivity,and implications of their occurrence[J].J Nat Prod,2006,69(3):509-526.
[26] Mitchell AM,Strobel GA,Moore E,et al.Volatile antimicrobials from Muscodor crispans,a novel endophytic fungus[J].Microbiology,2001,147(11):2943-2950.
[27] 谢洁,任慧爽,唐翠明,等.一株桑树内生细菌的鉴定和对桑椹核地杖菌的拮抗作用[J].蚕业科学,2015,41(5):815-824.
[28] Zhou JY,Zhao XY,Dai CC.Antagonistic mechanisms of endophytic Pseudomonas fluorescens against Athelia rolfsii[J].Journal of Applied Microbiology,2014,117(4):1144-1158.