Key Laboratory of Molecular Biology of Crop Pathogens and Insects of Ministry of Agriculture, Institute of Insect Science, Zhejiang University, Hangzhou, China.
PLoS One. 2010 Dec 6;5(12):e14233. doi: 10.1371/journal.pone.0014233.
The brown planthopper (BPH) Nilaparvata lugens (Stål) is one of the most serious insect pests of rice in Asia. However, little is known about the mechanisms responsible for the development, wing dimorphism and sex difference in this species. Genomic information for BPH is currently unavailable, and, therefore, transcriptome and expression profiling data for this species are needed as an important resource to better understand the biological mechanisms of BPH.
METHODOLOGY/PRINCIPAL FINDINGS: In this study, we performed de novo transcriptome assembly and gene expression analysis using short-read sequencing technology (Illumina) combined with a tag-based digital gene expression (DGE) system. The transcriptome analysis assembles the gene information for different developmental stages, sexes and wing forms of BPH. In addition, we constructed six DGE libraries: eggs, second instar nymphs, fifth instar nymphs, brachypterous female adults, macropterous female adults and macropterous male adults. Illumina sequencing revealed 85,526 unigenes, including 13,102 clusters and 72,424 singletons. Transcriptome sequences larger than 350 bp were subjected to Gene Orthology (GO) and KEGG Orthology (KO) annotations. To analyze the DGE profiling, we mainly compared the gene expression variations between eggs and second instar nymphs; second and fifth instar nymphs; fifth instar nymphs and three types of adults; brachypterous and macropterous female adults as well as macropterous female and male adults. Thousands of genes showed significantly different expression levels based on the various comparisons. And we randomly selected some genes to confirm their altered expression levels by quantitative real-time PCR (qRT-PCR).
CONCLUSIONS/SIGNIFICANCE: The obtained BPH transcriptome and DGE profiling data provide comprehensive gene expression information at the transcriptional level that could facilitate our understanding of the molecular mechanisms from various physiological aspects including development, wing dimorphism and sex difference in BPH.
褐飞虱（BPH）Nilaparvata lugens（Stål）是亚洲水稻上最严重的害虫之一。然而，对于该物种的发育、翅二型性和性别差异的相关机制知之甚少。目前褐飞虱的基因组信息尚不可用，因此需要对该物种进行转录组和表达谱数据分析，作为更好地理解 BPH 生物学机制的重要资源。
方法/主要发现：在本研究中，我们使用短读测序技术（Illumina）与基于标签的数字基因表达（DGE）系统相结合，进行了从头转录组组装和基因表达分析。转录组分析组装了 BPH 不同发育阶段、性别和翅型的基因信息。此外，我们构建了六个 DGE 文库：卵、二龄若虫、五龄若虫、短翅雌成虫、长翅雌成虫和长翅雄成虫。Illumina 测序揭示了 85526 个 unigenes，包括 13102 个聚类和 72424 个单基因。长度大于 350bp 的转录组序列进行了基因同源（GO）和 KEGG 同源（KO）注释。为了分析 DGE 分析，我们主要比较了卵与二龄若虫、二龄与五龄若虫、五龄若虫与三种成虫、短翅雌成虫与长翅雌成虫以及长翅雌成虫与雄成虫之间的基因表达差异。根据各种比较，数千个基因表现出明显不同的表达水平。我们随机选择了一些基因，通过定量实时 PCR（qRT-PCR）验证其表达水平的改变。
结论/意义：获得的褐飞虱转录组和 DGE 分析数据提供了转录水平的全面基因表达信息，有助于我们从发育、翅二型性和性别差异等各个生理方面理解 BPH 的分子机制。
