HOU Yangwei,WANG Xinyuan,YANG Linlin,et al.Determination of Cell Wall-degrading Enzyme Activity and Pathogenicity Analysis of Rotunda Pathogens[J].Northern Horticulture,2022,(19):106-113.[doi:10.11937/bfyy. 20221124]
地黄轮纹病病原菌的细胞壁降解酶活性测定及致病性分析
- Title:
- Determination of Cell Wall-degrading Enzyme Activity and Pathogenicity Analysis of Rotunda Pathogens
- 文献标志码:
- A
- 摘要:
- 以地黄叶片和地黄轮纹病病原菌Phoma herbarum和Fusarium solani为试材，研究了地黄轮纹病病原菌Phoma herbarum和Fusarium solani在离体培养及活体接种后对细胞壁降解酶活性的影响，以期为明确细胞壁降解酶在病原菌侵染地黄植株过程中的致病作用研究提供参考依据。结果表明：离体培养时，果胶诱导培养基中多聚半乳糖醛酸酶(PG)与果胶甲基半乳糖醛酸酶(PMG)活性较高，羧甲基纤维素（CMC）诱导培养基中羧甲基纤维素酶(Cx)活性较高，且Phoma herbarum的PG、PMG和Cx活性均高于Fusarium solani。活体接种后，Phoma herbarum的PG、PMG、Cx和β-葡萄糖苷酶活性最大值均大于Fusarium solani，且均于第8天时达到峰值。此时，其PG活性为7.83 U·mg-1，PMG活性为16.00 U·mg-1，Cx活性为13.65 U·mg-1，β-葡萄糖苷酶活性为12.01 U·mg-1。细胞壁降解酶在病原菌侵染过程中起重要作用，Phoma herbarum细胞壁降解酶活性强于Fusarium solani。PMG在侵染初期分泌并起作用，Cx和β-葡萄糖苷酶致病作用发生在后期。
- Abstract:
- The leaves and the pathogens (Phoma herbarum and Fusarium solani) of Rehmannia glutinosa were used as test materials,and the methods of in vitro induction culture and pathogen inoculation were used.The effects of cell wall-degrading enzyme activities of Phoma herbarum and Fusarium solani,which were pathogenic bacteria of Rehmannia glutinosa,in vitro culture and in vivo inoculation were studied,in order to provide reference for the study of the pathogenic role of cell wall-degrading enzymes in the process of pathogenic bacteria infecting Rehmannia glutinosa plants.The results showed that the activities of PG and PMG were higher in the pectin-induced medium,and the Cx activity was higher in the CMC-induced medium when cultured in vitro,and the activities of PG,PMG and Cx in Phoma herbarum were higher than those in Fusarium solani.After inoculation in vivo,the maximum values of PG,PMG,Cx and β-glucosidase activities of Phoma herbarum were higher than those of Fusarium solani,and all reached the peak on the 8th day.At this time,the PG activity was 7.83 U·mg-1,the PMG activity was 16.00 U·mg-1,the Cx activity was 13.65 U·mg-1,and the β-glucosidase activity was 12.01 U·mg-1.Cell wall degrading enzymes play an important role in the process of pathogenic bacteria infection.Phoma herbarum′s cell wall-degrading enzyme activity was stronger than that of Fusarium solani.PMG was secreted and acted at the early stage of infection,and the pathogenic effects of Cx and β-glucosidase occurred at the later stage.
参考文献/References:
[1]国家药典委员会.中华人民共和国药典.一部[M].北京:中国医药科技出版社,2020.[2]王飞.地黄轮纹病病原学及防治研究[D].武汉:华中农业大学,2012.[3]许园园.怀地黄活性内生菌在预防植物病害和克服连作障碍方面的应用[D].新乡:河南师范大学,2014.[4]冯韬.油菜菌核病病理与防治研究进展[J].作物研究,2014,28(3):316-320.[5]HEMATY K,CHERK C,SOMERVILLE S.Host-pathogen warfare at the plant cell wall[J].Current Opinion in Plant Biology,2009,12(4):406-413.[6]CANTU D,VICENTE A R,LABAVITCH J M,et al.Strangers in the matrix：Plant cell walls and pathogen susceptibility[J].Trends in Plant Science,2008,13(11)：610-617.[7]YANG Y Q,YANG M,LI M H,et al.Cloning and functional analysis of an endo-PG-encoding gene Rrspgl of Rhizoctonia solani,the causal agent of rice sheath blight[J].Canadian Journal of Plant Pathology,2012,34(3):43-47.[8]李明海,杨迎青,杨媚,等.井冈霉素对水稻纹枯病菌细胞壁降解酶活性和可溶性蛋白的影响[J].华中农业大学学报,2010,29(3):272-276.[9]TADASHI S,HIDEO N,KINJIRO T.Occurrence and some properties of cellulase in the filtrates of conidiospores and mycelia of Pyricularia oryzae Cavara[J].Agricultural and Biological Chemisty,1973,37(7):1651-1659.[10]ESHEL D,MIYARA I,AILING T,et al.pH regulates endoglucanase expression and virulence of Alternaria alternata in persimmon fruit[J].Molecular Plant-microbe Interactions,2002,15(8):774-779.[11]KOGANEZAWA H,SAKUMA T.Possible role of breakdown products of phloridzin in symptom development by Valsa ceratosperma[J].Japanese Journal of Phytopathology,1982,48(4):521-528.[12]NATSUME H,SETO H,〖AKO-〗TAKE N.Studies on apple canker disease.The necrotic toxins produced by Valsa ceratosperma[J].Agricultural Biological Chemistry,1982,46(8):2101-2106.[13]IKOTUN T.Cell wall-degrading enzymes produced by Penicillium oxalicum Curie et Thom[J].Mycopathologia,1984,88(1):15-21.[14]SIPOS B,BENK Z,DIENES D,et al.Characterisation of specific activities and hydrolytic properties of cell-wall-degrading enzymes produced by Trichoderma reesei Rut C30 on different carbon sources[J].Applied Biochemistry and Biotechnology,2010,161(1/8):347-364.[15]李宝聚,周长力,赵奎华,等.黄瓜黑星病菌致病机理的研究 Ⅱ 细胞壁降解酶及其在致病中的作用[J].植物病理学报,2000(1):13-18.[16]薛彩云.花生纹枯病病原学及其致病机制研究[D].沈阳:沈阳农业大学,2018.[17]BRADFORD M M.A rapid and sensitive method for the quantitation ofmicrogram quantities protein vtilizing the principle of protein-dyebinding[J].Analytical Biochemitry,1976,72(1/2)：248-254.[18]杨媚,杨迎青,郑丽,等.水稻纹枯病菌细胞壁降解酶组分分析、活性测定及其致病作用[J].中国水稻科学,2012,26(5):600-606.[19]车喜庆,傅俊范,李自博,等.人参核盘菌侵染对参根不同部位细胞壁降解酶及防御酶活性的影响[J].河南农业科学,2017,46(10):66-70.[20]金勤,周国英,刘君昂,等.细胞壁降解酶在油茶炭疽病菌致病过程中的作用研究[J].植物保护,2017,43(3):97-102.[21]张大智,詹儒林,柳凤,等.杧果细菌性角斑病菌细胞壁降解酶的致病作用[J].果树学报,2016,33(5):585-593.[22]COMSTOCK J C,MARTINSON C A,GENGENBACK B G,et al.Host specificity of a toxin from Phyllosticat maydis for Taxas cytoplasmically male-sterile maize[J].Phytopathology,1973,63(11):1357-1361.[23]BAILISS K W.The relationship of gibberellin content to cucumber mosaic virus infection of cucumber[J].Physiological Plant Pathology,1974,4(1):73-79.
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备注/Memo
第一作者简介：侯杨威（1994-），男，硕士研究生，研究方向为中药资源与栽培技术。E-mail:1744174697@qq.com.责任作者：董诚明（1963-），男，硕士，教授，现主要从事药用植物学、中药资源学等研究工作。E-mail:dcm371@sohu.com.基金项目：国家重点研发计划资助项目（2017YFC1702800）。收稿日期：2022-03-24