Abreu Lucas M, Moreira Gláucia M, Ferreira Douglas, Rodrigues-Filho Edson, Pfenning Ludwig H
Departamento de Fitopatologia, Universidade Federal de Lavras, PO Box 3037, 37200-000 Lavras, MG, Brazil.
Departamento de Fitopatologia, Universidade Federal de Lavras, PO Box 3037, 37200-000 Lavras, MG, Brazil.
Fungal Biol. 2014 Dec;118(12):1004-12. doi: 10.1016/j.funbio.2014.10.001. Epub 2014 Oct 14.
We assessed the species diversity among 45 strains of Clonostachys from different substrates and localities in Brazil using molecular phylogenetics, and compared the results with the phenotypic classification of strains obtained from matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Phylogenetic analyses were based on beta tubulin (Tub), ITS-LSU rDNA, and a combined Tub-ITS DNA dataset. MALDI-TOF MS analyses were performed using intact conidia and conidiophores of strains cultivated on oatmeal agar and 4% malt extract agar. Six known species were identified: Clonostachys byssicola, Clonostachys candelabrum, Clonostachys pseudochroleuca, Clonostachys rhizophaga, Clonostachys rogersoniana, and Clonostachys rosea. Two clades and two singleton lineages did not correspond to known species represented in the reference DNA dataset and were identified as Clonostachys sp. 1-4. Multivariate cluster analyses of MALDI-TOF MS data classified the strains into eight clusters and three singletons, corresponding to the ten identified species plus one additional cluster containing two strains of C. rogersoniana that split from the other co-specific strains. The consistent results of MALDI-TOF MS supported the identification of strains assigned to C. byssicola and C. pseudochroleuca, which did not form well supported clades in all phylogenetic analyses, but formed distinct clusters in the MALDI-TOF dendrograms.
我们利用分子系统发育学评估了来自巴西不同基质和地点的45株枝顶孢属菌株的物种多样性，并将结果与基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）获得的菌株表型分类结果进行了比较。系统发育分析基于β微管蛋白（Tub）、ITS-LSU rDNA以及Tub-ITS DNA组合数据集。MALDI-TOF MS分析使用了在燕麦片琼脂和4%麦芽提取物琼脂上培养的菌株的完整分生孢子和分生孢子梗。鉴定出六个已知物种：浅黄枝顶孢、烛台枝顶孢、假绿枝顶孢、根际枝顶孢、罗杰枝顶孢和粉红枝顶孢。两个进化枝和两个单系谱系与参考DNA数据集中代表的已知物种不对应，被鉴定为枝顶孢属1-4种。MALDI-TOF MS数据的多变量聚类分析将菌株分为八个簇和三个单株，对应于十个已鉴定的物种，另外还有一个簇包含两株与其他同物种菌株分离的罗杰枝顶孢。MALDI-TOF MS的一致结果支持了对归为浅黄枝顶孢和假绿枝顶孢的菌株的鉴定，这两种菌株在所有系统发育分析中均未形成得到充分支持的进化枝，但在MALDI-TOF树形图中形成了不同的簇。
