嗜果刀孢菌在野杏叶片致病过程中主要酶的种类及活性分析  

The kinds and activities of main enzymes in the pathogenic process of shot hole disease caused byWilsonomyces carpophilus in Prunus armeniaca

作　　者：刘楚丽 鲁海龙 陈虹瑾 孙洪涛 马荣[1] LIU Chuli;LU Hailonga;CHEN Hongjin;SUN Hongtao;MARong(College of Forestry and Landscape Architecture,Xinjiang Agricultural University,Urumqi 830052,Xinjiang,China)

机构地区：[1]新疆农业大学林学与风景园林学院,乌鲁木齐830052

出　　处：《果树学报》2023年第4期747-756,共10页Journal of Fruit Science

基　　金：新疆维吾尔自治区杰出自然科学基金项目(2022D01E47);新疆农业大学研究生科研创新项目(XJAUGRI2022034);国家级大学生创新创业训练计划项目(202210758013)。

摘　　要：【目的】明确引起野杏穿孔病的嗜果刀孢菌(Wilsonomyces carpophilus)在致病过程中主要酶的种类及活性。【方法】通过3,5-二硝基水杨酸(DNS)法开展了细胞壁降解酶的种类及其活性的测定,并分析了野杏叶片受嗜果刀孢菌侵染后抗氧化酶活性的变化。【结果】首次发现嗜果刀孢菌在体外不同诱导物培养条件下均可产生木聚糖酶、多聚半乳糖醛酸酶(PG)、聚甲基半乳糖醛酸酶(PMG)等6种细胞壁降解酶,且酶活性呈显著差异。嗜果刀孢菌侵染野杏叶片后产生的细胞壁降解酶中PG和PMG酶活性最高,羧甲基纤维素酶活性最低;过氧化物酶(POD)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)3种抗氧化酶活性随嗜果刀孢菌侵染时间的延长呈先上升后下降趋势。【结论】首次明确果胶酶在嗜果刀孢菌致病过程中起重要作用,野杏叶片受嗜果刀孢菌危害后体内抗氧化酶活性升高。【Objective】Prunus armeniaca is the origin species of apricot widely cultivated in the world,and an important germplasm resource of characteristic fruit trees in Xinjiang,China.The occurrence of perforation disease of drupes has become an important factor threatening the healthy growth of the wild fruit forest.Wilsonomyces carpophilus is the most serious pathogen causing perforation disease,which can not only damage the leaves of stone fruit plants,but also damage the fruits and shoots.The study aimed to clarify the kinds and activities of main enzymes in the pathogenic process of shot hole disease caused by W.carpophilus in P.armeniaca.【Methods】W.carpophilus were cultured for 10 days on PDA medium,the hyphae were added into induction media containing sucrose,pectin,CMC,cellulose particle,filter paper powder,and cotton pectin or P.armeniaca leaves respectively as inducers.The media were shaken in a shaker incubator set at 25℃for 7 days,vacuum filtered to remove hyphae and spores,and centrifuged at 4℃for 30 minutes at 10000 r·min-1.The healthy leaves of P.armeniaca were inoculated with W.carpophilus,and the blank culture medium was used as control.Samples were taken on 2,4,6,8,10,12,14 and 16 d after inoculation.The samples tissues were cut into slices and put into mortar,grinded with sodium chloride at 4℃,centrifuged at 4℃and 5000 r·min-1 after filtration,and the supernatant was collected as the crude enzyme for later use.The 3,5-dinitrosalicylic acid(DNS)and the Coomassie blue staining method was used to detect the kinds and activies of cell wall degrading enzymes.The absorbance of the reaction mixture was measured by UV-Vis spectrophotometer,and the the enzyme activities were calculated according to the reducing sugar released.The degradationof Apricot leaves by cell wall degrading enzymes produced by W.carpophilus.The obtained enzyme solutionsinduced by 7 inducers(20 mL)were inoculated into the leaves of P.armeniaca by in vitro needlingmethod,and the inactivated enzyme solution was boiled as t

关 键 词：野杏 穿孔病 嗜果刀孢菌 细胞壁降解酶 抗氧化酶 

分 类 号：S662.2[农业科学—果树学]