Genome-assisted development of nuclear intergenic sequence markers for entomopathogenic fungi of the Metarhizium anisopliae species complex
Genome-assisted development of nuclear intergenic sequence markers for entomopathogenic fungi of the Metarhizium anisopliae species complex
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DOI：
10.1111/1755-0998.12058
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发表时间：
2013-03-01
影响因子：
7.7
通讯作者：
Rehner, S. A.
中科院分区：
文献类型：
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作者：
Kepler, R. M.;Rehner, S. A.
绿僵菌属的昆虫病原真菌对世界各地重要的节肢动物害虫的生物防治方案是有用的。然而，物种之间的形态趋同阻碍了对这些生物真正的多样性和生态学的理解。分子技术的应用使得物种的分辨率比仅靠形态所允许的更高。特别是，常用的生防剂金龟子绿僵菌是由9个种组成的物种复合体。这项先前的工作是利用真菌系统发育中常用的标记(BTUB、RPB1、RPB2和TEF)进行的，这些标记可能不能充分代表绿僵菌复合体的多样性。利用麦长管藻和罗伯茨分枝杆菌核基因组的序列数据，我们确定了保守的基因共性区域，并开发了扩增7个基因座的基因间隔区的引物。使用前类型和鉴定的金龟子绿僵菌复合体物种的组织标本，我们证明了来自基因间基因座的序列数据比以前可用的标记更具变异性和系统发育信息。这些新的标记将促进对金龟子绿僵菌复合体的物种水平或物种水平以下的调查。这里使用的标记开发方法应该可以扩展到任何具有足够分散的基因组数据的群体。
Entomopathogenic fungi in the genus Metarhizium are useful for biological control programmes against economically important arthropod pests worldwide. However, understanding the true diversity and ecology of these organisms is hampered by convergent morphologies between species. The application of molecular techniques has enabled greater resolution of species than allowed by morphology alone. In particular, the commonly used biocontrol agent M. anisopliae was found to be a species complex composed of nine species. This prior work was conducted with commonly used markers in fungal phylogenetics (BTUB, RPB1, RPB2 and TEF), which likely under-represent diversity in the M. anisopliae complex. Using sequence data from nuclear genomes of M. acridum and M. robertsii we identified regions of conserved gene synteny and developed primers to amplify intergenic regions of seven loci. Using ex-type and authenticated tissue specimens for species in the M. anisopliae complex, we demonstrate that sequence data derived from intergenic loci is more variable and phylogenetically informative than previously available markers. These new markers will facilitate investigations at or below the species level for the M. anisopliae complex. The method of marker development employed here should be extendable to any group with sufficiently divergent genome data available.