GFP标记的4种链格孢菌对枣树的侵染研究  

Infection of four GFP-labelled Alternaria species in Ziziphus jujuba

作　　者：田红雨[1,2] 史晓梦 张敏 王亚聪[1,2] 张书蔚[1,2] 冉隆贤 TIAN Hongyu;SHI Xiaomeng;ZHANG Min;WANG Yacong;ZHANG Shuyu;RAN Longxian(College of Forestry,Hebei Agricultural University,Baoding 071000,Hebei,China;Hebei Key Lab of Forest Germplasm Resources and Protection,Hebei Agricultural University,Baoding 071000,Hebei,China)

机构地区：[1]河北农业大学林学院,河北保定071000 [2]河北农业大学河北省林木种质资源与森林保护重点实验室,河北保定071000

出　　处：《经济林研究》2020年第2期84-92,共9页Non-wood Forest Research

基　　金：河北省自然科学基金项目(C2016204202)。

摘　　要：【目的】进一步探寻引起枣缩果病的潜在侵染性病原,为枣缩果病的有效防治提供理论依据。【方法】选取苹果斑点落叶病菌、梨黑斑病菌、马铃薯早疫病菌和番茄黑斑病菌为供试菌株,测定其对潮霉素B的敏感性后再进行GFP标记,并以枣缩果病互隔链格孢菌作为对照,将通过稳定性测试的菌株进行幼果刺伤接种和枣树花期喷雾接种试验,探明它们在枣树上的侵染途径。【结果】苹果斑点落叶病菌、梨黑斑病菌、马铃薯早疫病菌和番茄黑斑病菌对潮霉素B均较敏感,其有效中浓度EC50值分别为1.489、2.241、3.299和2.873μg·m L-1,4种链格孢菌均被GFP成功标记且其稳定性均良好;林间刺伤接种苹果斑点落叶病菌、梨黑斑病菌、马铃薯早疫病菌、番茄黑斑病菌和互隔链格孢菌(对照)的枣果均发病且其症状相同,其发病率分别为75.0%、87.5%、81.3%、83.3%与93.8%;将各处理病果的病组织放在含有10μg·mL-1潮霉素B的PDA培养基上培养后,其均有菌丝生长,且均可观察到带有荧光的菌丝;花期喷雾接种4种链格孢菌和互隔链格孢菌后,枣果均发病,将其病组织分离培养后均有菌丝长出,且在荧光显微镜下均可观察到荧光。【结论】苹果斑点落叶病菌、梨黑斑病菌、马铃薯早疫病菌和番茄黑斑病菌均能侵染枣树,都有可能成为枣缩果病的潜在病原菌,因此,在枣树周围应避免种植上述易被链格孢菌危害的果树和作物,这样可以阻止链格孢菌的侵染。【Objective】To further research potential infectious pathogens of jujube shrunken-fruit disease(JSFD)and to provide a theoretical basis for effective control of JSFD.【Method】Four Alternaria species were using as test materials,including A.mali ZN01(apple leaf blotch pathogen),A.kikuchiana ZN02(pear black spot pathogen),A.solani ZN03(potato early blight pathogen),and A.alternata ZN-XR(tomato black spot pathogen).Their sensitivities to hygromycin B were tested,and then were labelled with GFP marker.Some bacterial strains passing stability test were inoculated through stabbing young fruits and spraying at florescence,and A.alternata of JSFD was used as a control.Path of infecting Ziziphus jujuba was researched.【Result】A.mali ZN01,A.kikuchiana ZN02,A.solani ZN03,and A.alternata ZN-XR are all sensitive to hygromycin B,and their EC50 are 1.489,2.241,3.299 and 2.873μg·mL-1,respectively.The four Alternaria species can be successfully labelled by GFP marker and all of them show good stability.Through stabbing young fruits,all jujube fruits inoculated with A.mali ZN01,A.kikuchiana ZN02,A.solani ZN03,A.alternata ZN-XR and CK are diseased and have the same symptoms.Incidence rates are 75.0%,87.5%,81.3%,83.3%and 93.8%,respectively.In each treatment,infected tissues cultured in PDA plates with 10μg·mL-1 hygromycin B produce hyphae,and fluorescence of hyphae can be observed under fluorescence microscope.Through spraying at florescence,all jujube fruits inoculated with four Alternaria species and CK are diseased,infected tissues produce hyphae,and fluorescence of hyphae can be observed under fluorescence microscope.【Conclusion】A.mali ZN01,A.kikuchiana ZN02,A.solani ZN03,A.alternata ZN-XR can infect Ziziphus jujuba,and might also be potential pathogens of JSFD.Therefore,fruit trees and crops which are susceptible to Alternaria species should be avoided to cultivate around jujube,in order to prevent infection of Alternaria species.

关 键 词：枣树 枣缩果病 链格孢菌 绿色荧光蛋白 花期侵染 

分 类 号：S665.1[农业科学—果树学]