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基于单基因的RNA测序剖析芦竹对水分胁迫的早期转录反应

Dissection of early transcriptional responses to water stress in Arundo donax L. by unigene-based RNA-seq.

作者信息

Fu Yuan, Poli Michele, Sablok Gaurav, Wang Bo, Liang Yanchun, La Porta Nicola, Velikova Violeta, Loreto Francesco, Li Mingai, Varotto Claudio

机构信息

Department of Biodiversity and Molecular Ecology, Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, 38010 San Michele all'Adige, Trento Italy.

Dipartimento di Biotecnologie, Università degli Studi di Verona, Verona, Italy.

出版信息

Biotechnol Biofuels. 2016 Mar 8;9:54. doi: 10.1186/s13068-016-0471-8. eCollection 2016.

Abstract

BACKGROUND

Arundo donax L. (Poaceae) is considered one of the most promising energy crops in the Mediterranean region because of its high biomass yield and low input requirements, but to date no information on its transcriptional responses to water stress is available.

RESULTS

We obtained by Illumina-based RNA-seq the whole root and shoot transcriptomes of young A. donax plants subjected to osmotic/water stress with 10 and 20 % polyethylene glycol (PEG; 3 biological replicates/organ/condition corresponding to 18 RNA-Seq libraries), and identified a total of 3034 differentially expressed genes. Blast-based mining of stress-related genes indicated the higher responsivity of roots compared to shoots at the early stages of water stress especially under the milder PEG treatment, with a majority of genes responsive to salt, oxidative, and dehydration stress. Analysis of gene ontology terms underlined the qualitatively different responses between root and shoot tissues. Among the most significantly enriched metabolic pathways identified using a Fisher's exact test with FDR correction, a crucial role was played in both shoots and roots by genes involved in the signaling cascade of abscisic acid. We further identified relatively large organ-specific differences in the patterns of drought-related transcription factor AP2-EREBP, AUX/IAA, MYB, bZIP, C2H2, and GRAS families, which may underlie the transcriptional reprogramming differences between organs. Through comparative analyses with major Poaceae species based on Blast, we finally identified a set of 53 orthologs that can be considered as a core of evolutionary conserved genes important to mediate water stress responses in the family.

CONCLUSIONS

This study provides the first characterization of A. donax transcriptome in response to water stress, thus shedding novel light at the molecular level on the mechanisms of stress response and adaptation in this emerging bioenergy species. The inventory of early-responsive genes to water stress identified could constitute useful markers of the physiological status of A. donax and be a basis for the improvement of its productivity under water limitation. The full water-stressed A. donax transcriptome is available for Blast-based homology searches through a dedicated web server (http://ecogenomics.fmach.it/arundo/).

摘要

背景

芦竹(禾本科)因其高生物量产量和低投入需求,被认为是地中海地区最具潜力的能源作物之一,但迄今为止,尚无关于其对水分胁迫转录反应的信息。

结果

我们通过基于Illumina的RNA测序获得了遭受10%和20%聚乙二醇(PEG)渗透/水分胁迫的年轻芦竹植株的完整根和茎转录组(每个器官/条件3个生物学重复,对应18个RNA测序文库),共鉴定出3034个差异表达基因。基于Blast对胁迫相关基因的挖掘表明,在水分胁迫早期,尤其是在较温和的PEG处理下,根的反应性高于茎,大多数基因对盐、氧化和脱水胁迫有反应。基因本体术语分析强调了根和茎组织之间在质量上的不同反应。在使用经FDR校正的Fisher精确检验确定的最显著富集的代谢途径中,参与脱落酸信号级联的基因在茎和根中都发挥了关键作用。我们进一步在干旱相关转录因子AP2-EREBP、AUX/IAA、MYB、bZIP、C2H2和GRAS家族的模式中发现了相对较大的器官特异性差异,这可能是器官间转录重编程差异的基础。通过基于Blast与主要禾本科物种的比较分析,我们最终鉴定出一组53个直系同源基因,它们可被视为在该科中介导水分胁迫反应的进化保守基因核心。

结论

本研究首次对芦竹响应水分胁迫的转录组进行了表征,从而在分子水平上为这种新兴生物能源物种的胁迫反应和适应机制提供了新的见解。鉴定出的对水分胁迫早期响应的基因清单可构成芦竹生理状态的有用标记,并为提高其在水分限制下的生产力奠定基础。完整的水分胁迫下芦竹转录组可通过专用网络服务器(http://ecogenomics.fmach.it/arundo/)进行基于Blast的同源性搜索。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7bbf/4782572/698653ebb775/13068_2016_471_Fig1_HTML.jpg

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