进境新西兰猕猴桃中猕猴桃果腐病菌的检测与鉴定
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摘要
对来自新西兰的猕猴桃进行了病原菌的分离培养,结果在果实中分离到2株疑似猕猴桃果腐病菌的菌株Na-3419和Na-4756。对其进行了病原菌形态学观察、致病性测定并结合分子生物学方法检测,实验结果表明,菌株Na-3419和Na-4756在PDA培养基上生长良好,能产生大量的分生孢子;经真菌通用引物(ITS4/ITS5)扩增和测序,菌株Na-3419与Gen Bank中登录号EU482221.1和EU482220.1的菌株序列同源性达到100%,菌株Na-4756与Gen Bank中登录号AY359230.1、AY359226.1和KR859079.1的菌株序列同源性达到100%;病菌接种猕猴桃3 d后开始发病。根据上述试验结果,将分离获得的菌株Na-3419和Na-4756鉴定为猕猴桃果腐病菌(Neofabraea actinidiae)。这是上海口岸首次在进境的猕猴桃果实上截获该有害生物。
The isolates Na-3419 and Na-4756 were isolated on imported Actinidia chinensis from New Zealand by traditional PDA method. They grew well on PDA,producing lots of conidia. The isolate DNA was amplificated and sequenced by universal primers. Compared with the identity of DNA sequence of Neofabraea actinidiae(Accession:AY359230.1,AY359226.1,KR859079.1,EU482221.1,EU482220.1) from NCBI Gen Bank databases by blast method,the result showed that its homology was up to 100%. Pathogenicity tests showed that kiwifruit was infected by Na-3419 and Na-4756. Based on morphological characteristics,molecular detection and pathogenicity test results,the isolates Na-3419 and Na-4756 were identified as Neofabraea actinidiae. This pest was intercepted on imported Actinidia chinensis for the first time in Shanghai.
The isolates Na-3419 and Na-4756 were isolated on imported Actinidia chinensis from New Zealand by traditional PDA method. They grew well on PDA,producing lots of conidia. The isolate DNA was amplificated and sequenced by universal primers. Compared with the identity of DNA sequence of Neofabraea actinidiae(Accession:AY359230.1,AY359226.1,KR859079.1,EU482221.1,EU482220.1) from NCBI Gen Bank databases by blast method,the result showed that its homology was up to 100%. Pathogenicity tests showed that kiwifruit was infected by Na-3419 and Na-4756. Based on morphological characteristics,molecular detection and pathogenicity test results,the isolates Na-3419 and Na-4756 were identified as Neofabraea actinidiae. This pest was intercepted on imported Actinidia chinensis for the first time in Shanghai.
引文
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[7]Robert A Spotts,Keith A Seifert,Kelly M Wallis,et al.Description of Cryptosporiopsis kienholzii and species profiles of Neofabraea in major pome fruit growing districts in the Pacific Northwest USA.Mycological Research,2009,113:1301-1311.
[8]王卫芳,胡佳,温庆秋,等.进境美国苹果牛眼果腐病病原真菌(Neofabraea perennans)的鉴定.果树学报,2016,33(4):457-465.
[9]李鑫,曹际娟,王有福,等.PCR检测引起苹果牛眼果腐的明孢盘菌属3个致病种.植物检疫,2013,27(3):75-79.
[10]王卫芳,胡佳,张永瑜,等.广东口岸首次截获美国苹果牛眼果腐病.植物检疫,2010,24(1):35-37.
[2]Ross E Beever,Stephanie L.Vegetative compatibility groups in the fungus Cryptosporiopsis actinidiae.New Zealand Journal of Crop and Horticultural Science,2007,35(35):67-72.
[3]Fullerton R A,Manning M A,Casonato S G,et al.Infection and rot expression by Cryptosporiopsis actinidiae and Phomopsis sp.in kiwifruit,Actinidia chinensis‘Hort16A’.Acta Horticulturae,2007,753(753):653-661.
[4]Davison E M,Cunnington J H,Tay F C S.First record of Cryptosporiopsis actinidiae in Australia.Australasian Plant Disease Notes,2009,4:66-67.
[5]James H.Three Neofabraea species on pome fruit in Australia.Australia Plant Pathology.2004,33:453-454.
[6]Sharon N de Jong,Gerard J M Verkley,Edwin C A Abeln,et al.Phylogenetic relationships among Neofabraea species causing tree cankers and bull's-eye rot of apple based on DNA sequencing of ITS nuclear r DNA,mitochondrial r DNA,and theβ-tubulin gene.Mycological Research.2001,105(6):658-669.
[7]Robert A Spotts,Keith A Seifert,Kelly M Wallis,et al.Description of Cryptosporiopsis kienholzii and species profiles of Neofabraea in major pome fruit growing districts in the Pacific Northwest USA.Mycological Research,2009,113:1301-1311.
[8]王卫芳,胡佳,温庆秋,等.进境美国苹果牛眼果腐病病原真菌(Neofabraea perennans)的鉴定.果树学报,2016,33(4):457-465.
[9]李鑫,曹际娟,王有福,等.PCR检测引起苹果牛眼果腐的明孢盘菌属3个致病种.植物检疫,2013,27(3):75-79.
[10]王卫芳,胡佳,张永瑜,等.广东口岸首次截获美国苹果牛眼果腐病.植物检疫,2010,24(1):35-37.