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用于区分层出镰刀菌分离株的简单序列重复(SSR)标记的开发

Development of simple sequence repeat (SSR) markers for discrimination among isolates of Fusarium proliferatum.

作者信息

Moncrief I, Garzon C, Marek S, Stack J, Gamliel A, Garrido P, Proaño F, Gard M, Dehne H, Fletcher J

机构信息

Oklahoma State University, Stillwater, OK, United States.

Kansas State University, Manhattan, KS, United States.

出版信息

J Microbiol Methods. 2016 Jul;126:12-7. doi: 10.1016/j.mimet.2016.03.013. Epub 2016 Mar 25.

Abstract

The plant pathogen Fusarium proliferatum has a wide host range and occurs worldwide. Many isolates of the fungus produce mycotoxins in plant tissues, which, if ingested, can cause harm to animals and humans. In 2008, an outbreak of salmon blotch of onions, caused by F. proliferatum, was detected in southern Israel. The source and distribution of the fungus in Israel were unknown. Inter-simple sequence repeats (ISSR) were used to identify repetitive motifs present in seven isolates of F. proliferatum from Israel, Germany and Austria. ISSR repeat motifs were, used to develop 17 simple sequence repeat (SSR) loci. Six of these SSR markers were polymorphic in and consistently amplified from ten isolates collected in Israel, Germany, Austria and North America, from cucumber, onion, garlic, maize, and asparagus. These six polymorphic SSR alleles included 5 to 12 copies of di-, tri, and pentanucleotide motifs and yielded six to 9 alleles each. Sixteen of the SSR loci were amplified at least one of the seven Fusarium species, F. verticillioides, F. thapsinum, F. subglutinans, F. andiyazi, F. globosum, F. fujikoroi and F. oxysporum. The data demonstrate that these SSRs can be used for characterization of F. proliferatum isolates from diverse hosts and geographic locations and that they are transferable to other species of Fusarium.

摘要

植物病原菌层出镰刀菌(Fusarium proliferatum)寄主范围广泛,在全球均有发生。该真菌的许多分离株在植物组织中产生霉菌毒素,若被摄入,会对动物和人类造成危害。2008年,在以色列南部检测到由层出镰刀菌引起的洋葱鲑鱼色斑病爆发。该真菌在以色列的来源和分布情况尚不清楚。利用简单序列重复区间(ISSR)来鉴定来自以色列、德国和奥地利的7个层出镰刀菌分离株中存在的重复基序。ISSR重复基序被用于开发17个简单序列重复(SSR)位点。其中6个SSR标记在从以色列、德国、奥地利和北美收集的10个分离株(来自黄瓜、洋葱、大蒜、玉米和芦笋)中具有多态性且能持续扩增。这6个多态性SSR等位基因包含5至12个二核苷酸、三核苷酸和五核苷酸基序拷贝,每个等位基因产生6至9个等位基因。16个SSR位点在7种镰刀菌中的至少一种中得到扩增,这7种镰刀菌分别是轮枝镰孢(F. verticillioides)、沙栖镰孢(F. thapsinum)、胶孢镰孢(F. subglutinans)、安迪镰孢(F. andiyazi)、球形镰孢(F. globosum)、藤仓镰孢(F. fujikoroi)和尖孢镰孢(F. oxysporum)。数据表明,这些SSR可用于鉴定来自不同寄主和地理位置的层出镰刀菌分离株,并且它们可转移至其他镰刀菌物种。

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