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利用ITS1测序和简单重复序列区间法对从番茄中分离出的物种进行分子鉴定和遗传变异分析

Molecular identification and genetic variation of species isolated from tomatoes using ITS1 sequencing and inter simple sequence repeat methods.

作者信息

Mohammadi Abdelnasser, Bahramikia Seifollah

机构信息

Department of Biology, Faculty of Science, Lorestan University, Khorramabad, Iran.

出版信息

Curr Med Mycol. 2019 Jun;5(2):1-8. doi: 10.18502/cmm.5.2.1154.

Abstract

BACKGROUND AND PURPOSE

is one of the most abundant fungi that exists in numerous places around the world. This saprophytic fungus causes diseases in plants and accounts for the spoilage of cereals in warehouses. The aim of this study was to identify isolates based on their morphological characteristics and internal transcribed spacer ribosomal RNA (ITS rRNA) sequencing method. To this end, genetic diversity in the isolates was also examined using inter simple sequence repeat (ISSR) markers.

MATERIALS AND METHODS

To conduct this research, a total of 60 tomato samples with black spots were collected from supermarkets in Khorramabad City, Iran, in the winter of 2017. The specimens were cultured on a potato dextrose agar medium. After the purification of the fungus by the single-spore method, the identification of the species was carried out using morphological characteristics and ITS rRNA sequencing by polymerase chain reaction. The genetic diversity of the identified species with four primers was evaluated using the ISSR marker.

RESULTS

Based on the sequencing of the ITS1 region, all the isolates were identified as . Cluster diagrams for the ISSR marker were classified into six distinct groups. The mean polymorphism information content was obtained as 0.35, indicating the effectiveness of the primers in the separation of the isolates.

CONCLUSION

The sequencing of ITS1 led to the identification of species that are morphologically similar. The production of various mycotoxins by species leads to the contamination of livestock and human food. Regarding this, the investigation of the genetic diversity of species using the ISSR marker would facilitate the identification of suitable and effective strategies for controlling the fungal and mycotoxin contamination of human nutrition.

摘要

背景与目的

是世界上众多地方存在的最丰富的真菌之一。这种腐生真菌会引发植物病害,并导致仓库中谷物变质。本研究的目的是基于其形态特征和内部转录间隔区核糖体RNA(ITS rRNA)测序方法来鉴定分离株。为此,还使用简单序列重复区间(ISSR)标记检测了分离株的遗传多样性。

材料与方法

为开展本研究,2017年冬季从伊朗霍拉马巴德市的超市共采集了60个有黑斑的番茄样本。将标本接种在马铃薯葡萄糖琼脂培养基上。通过单孢法纯化真菌后,利用形态特征和聚合酶链反应进行ITS rRNA测序来鉴定物种。使用ISSR标记,用四种引物评估已鉴定物种的遗传多样性。

结果

基于ITS1区域的测序,所有分离株均被鉴定为 。ISSR标记的聚类图被分为六个不同的组。平均多态性信息含量为0.35,表明引物在分离分离株方面有效。

结论

ITS1测序有助于鉴定形态相似的 物种。 物种产生的各种霉菌毒素会导致家畜和人类食物受到污染。鉴于此,利用ISSR标记研究 物种的遗传多样性将有助于确定合适且有效的策略,以控制人类营养中的真菌和霉菌毒素污染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1c26/6626712/0b0f11bf9662/cmm-5-001-g001.jpg

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