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利用形态特征和系统发育分析从高海拔一些土壤无脊椎动物中分离的物种的鉴定和生物多样性模式。

Identification and biodiversity patterns of species isolated from some soil invertebrates at high altitude using morphological characteristics and phylogenetic analyses.

机构信息

Department of Biology, College of Science, Taif University, Taif, Saudi Arabia.

High Altitude Research Centre, Taif University, Taif, Saudi Arabia.

出版信息

PeerJ. 2023 Mar 28;11:e15035. doi: 10.7717/peerj.15035. eCollection 2023.

Abstract

BACKGROUND

The carcinogenic, mutagenic, and teratogenic chemicals such as aflatoxin are a worldwide health problem. spp., responsible for most cases of aflatoxin contamination, are common in the environment and spread easily to many different types of food. The objectives of this study were to conduct a survey of fungi associated with three soil invertebrates in Taif, Saudi Arabia, identify these isolates and explore mycotoxins formation.

METHODS

In total, 114 fungal isolates were collected from various soil invertebrates (millipedes, and ) in Taif, Saudi Arabia, among them, 22 isolates were identified as spp. based on morphological and molecular characteristics followed by both and

RESULTS

The sequences of ITS 1 and ITS 4 were utilized. Using bootstrap analysis, phylogenetic tree was split into two distinct clusters. Five sub clusters were included inside the first major cluster, and their bootstrap value was 99%. While, there were two small clusters in the second major cluster. All the tested strains were able to have a single PCR fragment amplified using the primer AspTef. TEF-1 DNA sequence bootstrap analysis with 1,000 replicates revealed two distinct groups. Additionally, the isolates were grouped into two different clusters with about 65% genetic similarity using ISSR-PCR analysis. The standard polymerase chain reaction was used to effectively amplify the and -A genes in aflatoxigenic strains. Four strains used in this investigation were shown to generate aflatoxin B1. While, three stains showed ochratoxin genes.

CONCLUSIONS

In conclusion, the results indicate significant differences in the fungal community between ecoregions and soil invertebrates. Moreover, mycotoxin detection and identification among isolates were elucidated. This study could shed light on the risk of mycotoxin contamination along the supply chain.

摘要

背景

黄曲霉毒素等致癌、致突变和致畸化学物质是一个全球性的健康问题。 spp.是导致大多数黄曲霉毒素污染的原因,它们在环境中很常见,很容易传播到许多不同类型的食物中。本研究的目的是调查沙特阿拉伯塔伊夫三种土壤无脊椎动物相关的真菌,鉴定这些分离株并探索其霉菌毒素的形成。

方法

从沙特阿拉伯塔伊夫的各种土壤无脊椎动物(千足虫和)中采集了 114 株真菌分离株,其中 22 株分离株根据形态和分子特征被鉴定为 spp.,随后进行了 和 ITS4 序列分析。使用bootstrap 分析,将系统发育树分为两个不同的聚类。第一个主要聚类中包含 5 个子聚类,其bootstrap 值为 99%。而第二个主要聚类中则有两个小聚类。使用引物 AspTef 对所有测试的菌株都能扩增出一个单一的 PCR 片段。使用 1000 次重复的 TEF-1 DNA 序列 bootstrap 分析显示出两个不同的聚类。此外,使用 ISSR-PCR 分析,将 分离株分为两个不同的聚类,遗传相似度约为 65%。标准聚合酶链反应有效地扩增了产黄曲霉毒素菌株的 和 -A 基因。本研究中使用的 4 株 菌株被证明能产生黄曲霉毒素 B1。而 3 株 菌株则显示出赭曲霉毒素基因。

结论

总之,结果表明不同生态区和土壤无脊椎动物之间的真菌群落存在显著差异。此外,还阐明了 分离株中霉菌毒素的检测和鉴定。本研究可以揭示沿供应链存在霉菌毒素污染的风险。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31b1/10075209/a0254e73df11/peerj-11-15035-g001.jpg

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