Fonseca Fernando Campos de Assis, Firmino Alexandre Augusto Pereira, de Macedo Leonardo Lima Pepino, Coelho Roberta Ramos, de Souza Júnior José Dijair Antonino, Silva-Junior Orzenil Bonfim, Togawa Roberto Coiti, Pappas Georgios Joannis, de Góis Luiz Avelar Brandão, da Silva Maria Cristina Mattar, Grossi-de-Sá Maria Fátima
Embrapa Recursos Genéticos e Biotecnologia, Brasília, Distrito Federal, Brazil; Universidade de Brasília, Brasília, Distrito Federal, Brazil.
Embrapa Recursos Genéticos e Biotecnologia, Brasília, Distrito Federal, Brazil; Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil.
PLoS One. 2015 Feb 23;10(2):e0118231. doi: 10.1371/journal.pone.0118231. eCollection 2015.
Sugarcane is a widely cultivated plant that serves primarily as a source of sugar and ethanol. Its annual yield can be significantly reduced by the action of several insect pests including the sugarcane giant borer (Telchin licus licus), a lepidopteran that presents a long life cycle and which efforts to control it using pesticides have been inefficient. Although its economical relevance, only a few DNA sequences are available for this species in the GenBank. Pyrosequencing technology was used to investigate the transcriptome of several developmental stages of the insect. To maximize transcript diversity, a pool of total RNA was extracted from whole body insects and used to construct a normalized cDNA database. Sequencing produced over 650,000 reads, which were de novo assembled to generate a reference library of 23,824 contigs. After quality score and annotation, 43% of the contigs had at least one BLAST hit against the NCBI non-redundant database, and 40% showed similarities with the lepidopteran Bombyx mori. In a further analysis, we conducted a comparison with Manduca sexta midgut sequences to identify transcripts of genes involved in digestion. Of these transcripts, many presented an expansion or depletion in gene number, compared to B. mori genome. From the sugarcane giant borer (SGB) transcriptome, a number of aminopeptidase N (APN) cDNAs were characterized based on homology to those reported as Cry toxin receptors. This is the first report that provides a large-scale EST database for the species. Transcriptome analysis will certainly be useful to identify novel developmental genes, to better understand the insect's biology and to guide the development of new strategies for insect-pest control.
甘蔗是一种广泛种植的植物,主要作为糖和乙醇的来源。其年产量会因多种害虫的侵害而显著降低,其中包括甘蔗大螟(Telchin licus licus),这是一种鳞翅目昆虫,生命周期长,使用杀虫剂控制其数量的效果不佳。尽管该物种具有经济重要性,但在GenBank中可获得的该物种DNA序列却很少。焦磷酸测序技术被用于研究该昆虫几个发育阶段的转录组。为了最大化转录本多样性,从昆虫全身提取了总RNA池,并用于构建标准化的cDNA数据库。测序产生了超过650,000条读数,这些读数被从头组装以生成一个包含23,824个重叠群的参考文库。经过质量评分和注释后,43%的重叠群在NCBI非冗余数据库中至少有一个BLAST匹配,40%与鳞翅目家蚕显示出相似性。在进一步的分析中,我们与烟草天蛾中肠序列进行了比较,以鉴定参与消化的基因转录本。与家蚕基因组相比,这些转录本中有许多在基因数量上出现了增加或减少。从甘蔗大螟(SGB)转录组中,基于与报道的作为Cry毒素受体的同源性,鉴定了一些氨肽酶N(APN)cDNA。这是第一份为该物种提供大规模EST数据库的报告。转录组分析对于鉴定新的发育基因、更好地理解昆虫生物学以及指导开发新的害虫控制策略肯定会很有用。