Key Laboratory of Bio-resources and Eco-environment, Center for Functional Genomics and Bioinformatics, College of Life Sciences, Sichuan University, Chengdu, Sichuan, People's Republic of China.
PLoS One. 2012;7(4):e36234. doi: 10.1371/journal.pone.0036234. Epub 2012 Apr 27.
Sweet potato (Ipomoea batatas L. [Lam.]) ranks among the top six most important food crops in the world. It is widely grown throughout the world with high and stable yield, strong adaptability, rich nutrient content, and multiple uses. However, little is known about the molecular biology of this important non-model organism due to lack of genomic resources. Hence, studies based on high-throughput sequencing technologies are needed to get a comprehensive and integrated genomic resource and better understanding of gene expression patterns in different tissues and at various developmental stages.
METHODOLOGY/PRINCIPAL FINDINGS: Illumina paired-end (PE) RNA-Sequencing was performed, and generated 48.7 million of 75 bp PE reads. These reads were de novo assembled into 128,052 transcripts (≥ 100 bp), which correspond to 41.1 million base pairs, by using a combined assembly strategy. Transcripts were annotated by Blast2GO and 51,763 transcripts got BLASTX hits, in which 39,677 transcripts have GO terms and 14,117 have ECs that are associated with 147 KEGG pathways. Furthermore, transcriptome differences of seven tissues were analyzed by using Illumina digital gene expression (DGE) tag profiling and numerous differentially and specifically expressed transcripts were identified. Moreover, the expression characteristics of genes involved in viral genomes, starch metabolism and potential stress tolerance and insect resistance were also identified.
CONCLUSIONS/SIGNIFICANCE: The combined de novo transcriptome assembly strategy can be applied to other organisms whose reference genomes are not available. The data provided here represent the most comprehensive and integrated genomic resources for cloning and identifying genes of interest in sweet potato. Characterization of sweet potato transcriptome provides an effective tool for better understanding the molecular mechanisms of cellular processes including development of leaves and storage roots, tissue-specific gene expression, potential biotic and abiotic stress response in sweet potato.
番薯(Ipomoea batatas L. [Lam.]) 是世界上最重要的六大粮食作物之一。它在世界各地广泛种植,具有产量高、稳定性强、适应性强、营养丰富、用途广泛等特点。然而,由于缺乏基因组资源,人们对这种重要的非模式生物的分子生物学知之甚少。因此,需要基于高通量测序技术的研究来获得全面和综合的基因组资源,并更好地了解不同组织和不同发育阶段的基因表达模式。
方法/主要发现:进行了 Illumina 配对末端(PE)RNA 测序,生成了 4870 万条 75bp 的 PE 读段。通过组合组装策略,将这些读段从头组装成 128052 个转录本(≥100bp),对应 4110 万个碱基对。使用 Blast2GO 对转录本进行注释,51763 个转录本得到 BLASTX 命中,其中 39677 个转录本具有 GO 术语,14117 个具有与 147 个 KEGG 途径相关的 ECs。此外,还通过 Illumina 数字基因表达(DGE)标签分析了 7 种组织的转录组差异,鉴定了许多差异和特异性表达的转录本。此外,还鉴定了参与病毒基因组、淀粉代谢以及潜在的抗逆性和抗虫性的基因的表达特征。
结论/意义:组合从头转录组组装策略可应用于其他参考基因组不可用的生物体。这里提供的数据代表了番薯最全面和综合的基因组资源,可用于克隆和鉴定番薯中感兴趣的基因。番薯转录组的特征分析为更好地理解包括叶片和块根发育、组织特异性基因表达、潜在的生物和非生物胁迫反应在内的细胞过程的分子机制提供了有效工具。