日本菜豆种子中菜豆晕疫病菌的检疫鉴定  

Detection and identification of Pseudomonas savastanoi pv. phaseolicola on kidney bean seed imported from Japan

作　　者：周密密 刘晓宇 杨静 钱茱希 许剑涛 王振华 吴翠萍 李彬 Zhou Mimi;Liu Xiaoyu;Yang Jing;Qian Zhuxi;Xu Jiantao;Wang Zhenhua;Wu Cuiping;Li Bin(Animal,Plant and Food Inspection Center,Nanjing Customs District,Nanjing 210000,China;Wuhan Customs District)

机构地区：[1]南京海关动植物与食品检测中心,江苏南京210000 [2]武汉海关

出　　处：《植物检疫》2022年第2期45-50,共6页Plant Quarantine

摘　　要：利用MT选择性培养基从来自日本的菜豆种子样品上分离到1株细菌分离物(编号1160),对该分离物进行培养性状观察、特异引物PCR检测、多位点序列分析、烟草过敏性反应和致病性测试。结果表明:该分离物菌落在MT上呈白色、略扁平、无水解圈,经菜豆晕疫病菌(Pseudomonas savastanoi pv. phaseolicola,Psp)特异引物PHA19/PHA95及P5.1/P3.1扩增,获到437 bp及500 bp目的条带,并与Psp菌株LMG2245及GenBank中Psp(基因登录号:AB237164、CP000058)的序列相似性均为100%。选择gap1、gltA、gyrB、rpoD 4个看家基因进行多位点序列分析(MLSA),系统发育树显示分离物1160与Psp菌株LMG2245、1448A聚在同一分支。该分离物人工接种菜豆叶片能引起典型的黄色晕疫病害症状,接种菜豆荚果引起水渍及褐变病害症状;接种烟草叶片可产生过敏性反应。根据上述试验结果,将分离物1160鉴定为菜豆晕疫病菌。这也是全国首次从进境商品中截获菜豆晕疫病菌。A bacterial isolate 1160 was obtained from Japan kidney bean seed with semi-selective MT(milk tween agar) medium,and identified by morphological character of pathogen,PCR using specific primers,multilocus sequence analysis,pathogenicity and hypersensitivity reaction on tobacco. The isolate1160 without hydrolysis ring was white and oblate on MT medium. With the specific primer PHA19/PHA95 and P5.1/P3.1 of Pseudomonas savastanoi pv. phaseolicola(Psp),437 bp and 500 bp products(100% sequence identity) were yielded respectively with the isolate 1160 and the Psp strain LMG2245,and the sequences were 100% identical to the Psp strains(AB237164、CP000058) in GenBank. Multilocus sequence analysis(MLSA) and phylogenetic analysis were carried out with the sequences from 4 housekeeping genes including gap1,gyrB,gltA,and rpoD. The isolate 1160 was clustered with strains Psp LMG2245 and 1448 A in the phylogenetic tree. The isolate 1160 could trigger hypersensitivity reaction on tobacco,and could cause halo blight on bean leaf and water-soaked symptom and brown symptom on bean pod by artificial inoculation. Based on the results above,the isolate 1160 was identified as Pseudomonas savastanoi pv. phaseolicola. This is the first interception of Psp from imported product in China.

关 键 词：菜豆种子 菜豆晕疫病菌 检测 细菌性病害 

分 类 号：S435.14[农业科学—农业昆虫与害虫防治]