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从头转录组测序和 SSR 标记开发用于 Cedrela balansae C.DC.,一种原产于阿根廷西北部的乡土树种。

De novo transcriptome sequencing and SSR markers development for Cedrela balansae C.DC., a native tree species of northwest Argentina.

机构信息

Instituto de Recursos Biológicos, Instituto Nacional de Tecnología Agropecuaria, Hurlingham, Buenos Aires Argentina.

Instituto de Biotecnología, Instituto Nacional de Tecnología Agropecuaria, Hurlingham, Buenos Aires, Argentina.

出版信息

PLoS One. 2018 Dec 7;13(12):e0203768. doi: 10.1371/journal.pone.0203768. eCollection 2018.

DOI:10.1371/journal.pone.0203768
PMID:30532149
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6285271/
Abstract

The endangered Cedrela balansae C.DC. (Meliaceae) is a high-value timber species with great potential for forest plantations that inhabits the tropical forests in Northwestern Argentina.Research on this species is scarce because of the limited genetic and genomic information available. Here, we explored the transcriptome of C. balansae using 454 GS FLX Titanium next-generation sequencing (NGS) technology. Following de novo assembling, we identified 27,111 non-redundant unigenes longer than 200 bp, and considered these transcripts for further downstream analysis. The functional annotation was performed searching the 27,111 unigenes against the NR-Protein and the Interproscan databases. This analysis revealed 26,977 genes with homology in at least one of the Database analyzed. Furthermore, 7,774 unigenes in 142 different active biological pathways in C. balansae were identified with the KEGG database. Moreover, after in silico analyses, we detected 2,663 simple sequence repeats (SSRs) markers. A subset of 70 SSRs related to important "stress tolerance" traits based on functional annotation evidence, were selected for wet PCR-validation in C. balansae and other Cedrela species inhabiting in northwest and northeast of Argentina (C. fissilis, C. saltensis and C. angustifolia). Successful transferability was between 77% and 93% and thanks to this study, 32 polymorphic functional SSRs for all analyzed Cedrela species are now available. The gene catalog and molecular markers obtained here represent a starting point for further research, which will assist genetic breeding programs in the Cedrela genus and will contribute to identifying key populations for its preservation.

摘要

受威胁的 Cedrela balansae C.DC.(楝科)是一种具有高价值的木材物种,具有很大的森林种植潜力,栖息在阿根廷西北部的热带森林中。由于可用的遗传和基因组信息有限,对该物种的研究很少。在这里,我们使用 454 GS FLX Titanium 下一代测序 (NGS) 技术探索了 C. balansae 的转录组。在从头组装之后,我们鉴定了 27,111 个长度超过 200bp 的非冗余非冗余基因,认为这些转录本可用于进一步的下游分析。通过对 27,111 个基因与 NR-Protein 和 Interproscan 数据库的同源性搜索,对这些转录本进行了功能注释。该分析显示,至少在分析的一个数据库中,有 26,977 个基因具有同源性。此外,使用 KEGG 数据库在 C. balansae 中鉴定了 142 个不同活性生物途径中的 7774 个基因。此外,经过计算机分析,我们检测到 2,663 个简单序列重复 (SSR) 标记。根据功能注释证据,选择了 70 个与重要“应激耐受”性状相关的 SSR 子集,用于在阿根廷西北部和东北部的 Cedrela 物种(Cedrela fissilis、Cedrela saltensis 和 Cedrela angustifolia)中进行湿 PCR 验证。可转移性在 77%到 93%之间,通过这项研究,现在为所有分析的 Cedrela 物种提供了 32 个多态功能 SSR。这里获得的基因目录和分子标记代表了进一步研究的起点,这将有助于 Cedrela 属的遗传育种计划,并有助于确定其保护的关键种群。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/72df4a343495/pone.0203768.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/0f2e230fe19b/pone.0203768.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/67ddbb81d2cd/pone.0203768.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/ec8b2dc7a291/pone.0203768.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/72df4a343495/pone.0203768.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/0f2e230fe19b/pone.0203768.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/67ddbb81d2cd/pone.0203768.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/ec8b2dc7a291/pone.0203768.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9c/6285271/72df4a343495/pone.0203768.g004.jpg

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