Valles Steven M, Strong Charles A, Hunter Wayne B, Dang Phat M, Pereira Roberto M, Oi David H, Williams David F
Center for Medical, Agricultural and Veterinary Entomology, USDA-ARS, Gainesville, FL 32608, USA.
J Invertebr Pathol. 2008 Sep;99(1):74-81. doi: 10.1016/j.jip.2008.01.004. Epub 2008 Feb 1.
An expression library was created and 2304 clones sequenced from a monogyne colony of Solenopsis invicta. The primary intention of the project was to utilize homologous gene identification to facilitate discovery of viruses infecting this ant pest that could potentially be used in pest management. Additional genes were identified from the ant host and associated pathogens that serve as an important resource for studying these organisms. After assembly and removal of mitochondrial and poor quality sequences, 1054 unique sequences were yielded and deposited into the GenBank database under Accession Nos. EH412746 through EH413799. At least nine expressed sequence tags (ESTs) were identified as possessing microsatellite motifs and 15 ESTs exhibited significant homology with microsporidian genes. These sequences most likely originated from Thelohania solenopsae, a well-characterized microsporidian that infects S. invicta. Six ESTs exhibited significant homology with single-stranded RNA viruses (3B4, 3F6, 11F1, 12G12, 14D5, and 24C10). Subsequent analysis of these putative viral ESTs revealed that 3B4 was most likely a ribosomal gene of S. invicta, 11F1 was a single-stranded RNA (ssRNA) virus contaminant introduced into the colony from the cricket food source, 12G12 appeared to be a plant-infecting tenuivirus also introduced into the colony as a field contaminant, and 3F6, 14D5, and 24C10 were all from a unique ssRNA virus found to infect S. invicta. The sequencing project illustrates the utility of this method for discovery of viruses and pathogens that may otherwise go undiscovered.
构建了一个表达文库，并从红火蚁单蚁后群体中对2304个克隆进行了测序。该项目的主要目的是利用同源基因鉴定来促进对感染这种害虫蚂蚁的病毒的发现，这些病毒有可能用于害虫治理。还从蚂蚁宿主和相关病原体中鉴定出了其他基因，这些基因是研究这些生物的重要资源。经过组装并去除线粒体序列和质量较差的序列后，得到了1054个独特序列，并以登录号EH412746至EH413799存入GenBank数据库。至少有9个表达序列标签（EST）被鉴定为具有微卫星基序，15个EST与微孢子虫基因表现出显著同源性。这些序列很可能源自索氏泰勒微孢子虫，这是一种已得到充分表征的感染红火蚁的微孢子虫。6个EST与单链RNA病毒（3B4、3F6、11F1、12G12、14D5和24C10）表现出显著同源性。对这些假定的病毒EST的后续分析表明，3B4很可能是红火蚁的核糖体基因，11F1是从蟋蟀食物来源引入群体的单链RNA（ssRNA）病毒污染物，12G12似乎是作为田间污染物也引入群体的一种感染植物的纤细病毒，而3F6、14D5和24C10均来自一种发现可感染红火蚁的独特ssRNA病毒。该测序项目说明了这种方法在发现否则可能未被发现的病毒和病原体方面的实用性。
