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在肯尼亚两个不同环境中从玉米分离出的群体内的遗传和产毒变异性。

Genetic and Toxigenic Variability within Population Isolated from Maize in Two Diverse Environments in Kenya.

作者信息

Okoth Sheila, De Boevre Marthe, Vidal Arnau, Diana Di Mavungu José, Landschoot Sofie, Kyallo Martina, Njuguna Joyce, Harvey Jagger, De Saeger Sarah

机构信息

School of Biological Sciences, University of Nairobi, Nairobi, Kenya.

Department of Bioanalysis, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.

出版信息

Front Microbiol. 2018 Jan 26;9:57. doi: 10.3389/fmicb.2018.00057. eCollection 2018.

Abstract

is the main producer of carcinogenic aflatoxins in agricultural commodities such as maize. This fungus occurs naturally on crops, and produces aflatoxins when environmental conditions are favorable. The aim of this study is to analyse the genetic variability among 109 isolates previously recovered from maize sampled from a known aflatoxin-hotspot (Eastern region, Kenya) and the major maize-growing area in the Rift Valley (Kenya), and to determine their toxigenic potential. DNA analyses of internal transcribed spacer (ITS) regions of ribosomal DNA, partial β-tubulin gene (benA) and calmodulin gene (CaM) sequences were used. The strains were further analyzed for the presence of four aflatoxin-biosynthesis genes in relation to their capability to produce aflatoxins and other metabolites, targeting the regulatory gene aflR and the structural genes aflP, aflD, and aflQ. In addition, the metabolic profile of the fungal strains was unraveled using state-of-the-art LC-MS/MS instrumentation. The three gene-sequence data grouped the isolates into two major clades, and . was most prevalent in Eastern Kenya, while was common in both regions. was represented by a single isolate collected from Rift Valley. Diversity existed within the population, which formed several subclades. An inconsistency in identification of some isolates using the three markers was observed. The calmodulin gene sequences showed wider variation of polymorphisms. The aflatoxin production pattern was not consistent with the presence of aflatoxigenic genes, suggesting an inability of the primers to always detect the genes or presence of genetic mutations. Significant variation was observed in toxin profiles of the isolates. This is the first time that a profound metabolic profiling of isolates was done in Kenya. Positive associations were evident for some metabolites, while for others no associations were found and for a few metabolite-pairs negative associations were seen. Additionally, the growth medium influenced the mycotoxin metabolite production. These results confirm the wide variation that exists among the group and the need for more insight in clustering the group.

摘要

是玉米等农产品中致癌黄曲霉毒素的主要产生者。这种真菌自然存在于作物上,在环境条件适宜时产生黄曲霉毒素。本研究的目的是分析先前从已知黄曲霉毒素热点地区(肯尼亚东部地区)和裂谷(肯尼亚)主要玉米种植区采集的玉米中分离出的109株菌株的遗传变异性,并确定它们的产毒潜力。使用核糖体DNA的内部转录间隔区(ITS)区域、部分β-微管蛋白基因(benA)和钙调蛋白基因(CaM)序列进行DNA分析。针对调控基因aflR和结构基因aflP、aflD和aflQ,进一步分析菌株中四个黄曲霉毒素生物合成基因的存在情况及其产生黄曲霉毒素和其他代谢产物的能力。此外,使用最先进的LC-MS/MS仪器解析真菌菌株的代谢谱。三个基因序列数据将分离株分为两个主要进化枝, 和 。 在肯尼亚东部最为普遍,而 在两个地区都很常见。 由从裂谷采集的单个分离株代表。 群体中存在多样性,形成了几个亚进化枝。观察到使用这三个标记对一些分离株进行鉴定时存在不一致。钙调蛋白基因序列显示出更广泛的多态性变异。黄曲霉毒素产生模式与产黄曲霉毒素基因的存在不一致,这表明引物无法始终检测到这些基因或存在基因突变。在分离株的毒素谱中观察到显著差异。这是首次在肯尼亚对 株分离株进行深入的代谢谱分析。一些代谢物之间存在明显的正相关,而其他一些则未发现相关性,少数代谢物对之间存在负相关。此外,生长培养基影响霉菌毒素代谢物的产生。这些结果证实了 组中存在广泛的变异,以及在对该组进行聚类时需要更多的深入了解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4980/5790802/515a5f0460c4/fmicb-09-00057-g0001.jpg

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