Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan.
J Sci Food Agric. 2011 Oct;91(13):2500-4. doi: 10.1002/jsfa.4507. Epub 2011 Jul 1.
Members of the genus Fusarium are well known as one of the most important plant pathogens causing food spoilage and loss worldwide. Moreover, they are associated with human and animal diseases through contaminated foods because they produce mycotoxins. To control fungal hazards of plants, animals and humans, there is a need for a rapid, easy and accurate identification system of Fusarium isolates with molecular methods.
To specify genes appropriate for identifying isolates of various Fusarium species, we sequenced the 18S rRNA gene (rDNA), internal transcribed spacer region 1, 5.8S rDNA, 28S rDNA, β-tubulin gene (β-tub), and aminoadipate reductase gene (lys2), and subsequently calculated the nucleotide sequence homology with pair-wise comparison of all tested strains and inferred the ratio of the nucleotide substitution rates of each gene. Inter-species nucleotide sequence homology of β-tub and lys2 ranged from 83.5 to 99.4% and 56.5 to 99.0%, respectively. The result indicated that sequence homologies of these genes against reference sequences in a database have a high possibility of identifying unknown Fusarium isolates when it is more than 99.0%, because these genes had no inter-species pair-wise combinations that had 100% homologies. Other markers often showed 100% homology in inter-species pair-wise combinations. The nucleotide substitution rate of lys2 was the highest among the six genes.
The lys2 is the most appropriate genetic marker with high resolution for identifying isolates of the genus Fusarium among the six genes we examined in this study.
镰刀菌属的成员是众所周知的最重要的植物病原体之一,在全球范围内导致食物变质和损失。此外,由于它们产生霉菌毒素,因此与受污染食物有关的人类和动物疾病。为了控制植物,动物和人类的真菌危害,需要使用快速,简便和准确的分子方法鉴定镰刀菌分离物。
为了确定适合鉴定各种镰刀菌种分离物的基因,我们对 18S rRNA 基因(rDNA),内部转录间隔区 1,5.8S rDNA,28S rDNA,β-微管蛋白基因(β-微管)和氨基己二酸还原酶基因(lys2)进行了测序,随后计算了与所有测试菌株进行两两比较的核苷酸序列同源性,并推断了每个基因的核苷酸替换率的比率。β-微管和 lys2 的种间核苷酸序列同源性分别为 83.5%至 99.4%和 56.5%至 99.0%。结果表明,当这些基因的序列同源性大于 99.0%时,对数据库中参考序列具有很高的可能性识别未知的镰刀菌分离物,因为这些基因没有种间具有 100%同源性的两两组合。其他标记物在种间的两两组合中通常表现出 100%的同源性。六个基因中 lys2 的核苷酸替换率最高。
在我们研究的六个基因中,lys2 是用于鉴定镰刀菌属分离物的最适合的遗传标记,具有高分辨率。