Reclassification of Paenibacillus larvae subsp pulvifaciens and Paenibacillus larvae subsp larvae as Paenibacillus larvae without subspecies differentiation
Reclassification of Paenibacillus larvae subsp pulvifaciens and Paenibacillus larvae subsp larvae as Paenibacillus larvae without subspecies differentiation
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DOI：
10.1099/ijs.0.63928-0
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发表时间：
2006-03-01
影响因子：
2.8
通讯作者：
Fries, I
中科院分区：
文献类型：
--
作者：
Genersch, E;Forsgren, E;Fries, I
幼虫类芽孢杆菌两个亚种的多相分类研究。幼虫和类芽孢杆菌幼虫亚种pulvifaciens，支持将亚种重新分类为一个种，幼虫类芽孢杆菌，而不分离亚种。我们的结论是基于对6个参考菌株的分析。pulvifaciens和三个参考菌株和44个田间分离物。亚种幼虫后者来源于瑞典、芬兰和德国的临床患病蜂群或临床患病和无症状蜂群的育雏或蜂蜜。菌落和孢子形态，以及代谢的甘露醇和水杨苷，不允许一个明确的识别两个亚种和SDS-PAGE全细胞蛋白不支持亚种分化。对于基因组指纹图谱，使用ERIC引物和细菌DNA的PFGE进行重复元件PCR指纹图谱。后一种方法是一种高分辨率的DNA指纹法，被证明是上级的生化和分子分型的大多数其他方法，以前没有被用来表征幼虫。ERIC-PCR鉴定了4种不同的基因型，而PFGE揭示了两个主要的集群。一个集群包括大部分的P.幼虫subsp.幼虫田间分离物，以及所有的P. pulvifaciens参考菌株。另一个簇包括P. larvae subsp.幼虫测定了部分菌株的16 S rRNA基因序列。最后，暴露生物测定表明，参考菌株的P.幼虫亚种。粉化芽孢杆菌对蜜蜂幼虫是致病性的，产生与幼虫粉化芽孢杆菌亚种的参考菌株相似的症状。幼虫与幼虫疫霉亚亚种的模式菌株比较，幼虫，ATCC 9545（T），幼虫毕赤酵母亚种（P. larvae subsp.测试的粉化菌菌株毒性更强，因为它们显示出更短的LTA（100）。并对该种进行了订正描述。
A polyphasic taxonomic study of the two subspecies of Paenibacillus larvae, Paenibacillus larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens, supported the reclassification of the subspecies into one species, Paenibacillus larvae, without subspecies separation. Our conclusions are based on the analysis of six reference strains of P. larvae subsp. pulvifaciens and three reference strains and 44 field isolates of P. larvae. subsp. larvae. The latter originated from brood or honey of clinically diseased honey bee colonies or from honey of both clinically diseased and asymptomatic colonies from Sweden, Finland and Germany. Colony and spore morphology, as well as the metabolism of mannitol and salicin, did not allow a clear identification of the two subspecies and SDS-PAGE of whole-cell proteins did not support the subspecies differentiation. For genomic fingerprinting, repetitive element-PCR fingerprinting using ERIC primers and PFGE of bacterial DNA were performed. The latter method is a high-resolution DNA fingerprinting method proven to be superior to most other methods for biochemical and molecular typing and has not previously been used to characterize P. larvae. ERIC-PCR identified four different genotypes, while PFGE revealed two main clusters. One cluster included most of the P. larvae subsp. larvae field isolates, as well as all P. larvae subsp. pulvifaciens reference strains. The other cluster comprised the pigmented variants of P. larvae subsp. larvae. 16S rRNA gene sequences were determined for some strains. Finally, exposure bioassays demonstrated that reference strains of P. larvae subsp. pulvifaciens were pathogenic for honey bee larvae, producing symptoms similar to reference strains of P. larvae subsp. larvae. In comparison with the type strain for P. larvae subsp. larvae, ATCC 9545(T), the P. larvae subsp. pulvifaciens strains tested were even more virulent, since they showed a shorter LTA(100). An emended description of the species is given.