进口小麦中链格孢菌的快速鉴定方法研究  

Study on the Rapid Identification Method of Alternaria alternata in Imported Wheat

作　　者：李毅然 薛佳欣 梁靓 兰茜 秦晋一 郭家琪 孙浩 LI Yiran;XUE Jiaxin;LIANG Liang;LAN Xi;QIN Jinyi;GUO Jiaqi;SUN Hao(Xi'an Customs Technical Center,Xi'an 710068,China;School of Civil Engineering,Chang5 an University,Xi*an 710061,China;College of Urban and Environmental Sciences,Northwest University,Xi5 an 710127,China)

机构地区：[1]西安海关技术中心,陕西西安710068 [2]长安大学建工学院,陕西西安710061 [3]西北大学城市与环境学院,陕西西安710127

出　　处：《河南农业科学》2021年第7期117-124,共8页Journal of Henan Agricultural Sciences

基　　金：海关总署科技计划项目(2018IK058)

摘　　要：口岸检疫中往往发现小麦携带真菌病原,为更快、更简便地对进口粮食携带的病原微生物进行检测,采用微生物细胞膜中具有遗传稳定性的磷脂脂肪酸(Phospholipid fatty acid,PLFA)作为重要标记进行鉴定。基于对微生物LB培养物的观察和DNA测序分析,将从进口小麦病变部位上分离纯化得到的病菌确定为链格孢菌,它是作物的主要致病菌之一。经过PLFA分析,比对响应值的显著性差异,明确检出该菌株的生物取样量介于0.070~0.090 g。经相关性分析和冗余分析,确定链格孢菌的特征脂肪酸为15:0 iso、15:0 anteiso、16:1 w7c和18:2 w6,9c。统计生物取样量与PLFA响应值之间的关系,建立Y1=0.047+6.183 × 10-5 × C15:0 anteiso+4.330 × 10-6 × C18:2w6,9c+1.386 × 10-7 × C16:1w7c 和 Y2=0.036+2.864 × 10-5 × C15:0 iso多元线性回归模型。Y1为15:0 anteiso、18:2 w6,9c、16:1 w7c回归计算出的生物量;Y2为15:0 iso回归计算出的生物量。该模型可用于估算小麦携带的病原微生物量,为建立进口粮食作物品质检疫检测标准提供理论数据和参考。Fungal diseases are usually found in wheat during port quarantine.In order to detect pathogenic microorganisms carried by imported grain more quickly and simply,the genetically stable phospholipid fatty acid(PLFA)in the microbial cell membrane was used as an important identification marker.Based on the observation of microbial LB culture and DNA sequencing analysis,it was determined that the strain isolated and purified from the diseased part of imported wheat was Alternaria alternata,which was one of the main pathogens of crops.After PLFA analysis,the significant difference in response value was compared,and the biological sampling range for detecting this strain was 0.070-0.090 g.After correlation analysis and redundancy analysis,it was determined that the characteristic fatty acids of Alternaria alternata were 15:0 iso,15:0 anteiso,16:1 w7c and 18:2 w6,9c.By analyzing the correlation between the biological sampling amount and the PLFA response value,the models of Y1=0.047+6.183×10-5×C15:0 anteoso+4.330×10-6×C18:2 w6,9c+1.386×10-7×C16:1w7c and Y2=0.036+2.864×10-5×C15:0 iso were established.Y1 is the biomass calculated by regression of 15:0 anteiso,18:2 w6,9c and 16:1w7c;Y2 is the biomass calculated by 15:0 iso regression.This model can predict the amount of pathogenic microorganisms carried by wheat,and provides theoretical data and references for establishing quality quarantine and testing standards for imported grains.

关 键 词：进口粮食 小麦 链格孢菌 磷脂脂肪酸PLFA分析 多元线性回归 

分 类 号：S512[农业科学—作物学]