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[通过抑制性消减杂交(SSH)与基因芯片分析相结合揭示岷江百合与尖孢镰刀菌百合专化型不亲和互作中的差异基因表达]

[Differential gene expression in incompatible interaction between Lilium regale Wilson and Fusarium oxysporum f. sp. lilii revealed by combined SSH and microarray analysis].

作者信息

Rao J, Liu D, Zhang N, He H, Ge F, Chen C

出版信息

Mol Biol (Mosk). 2014 Nov-Dec;48(6):915-26. doi: 10.7868/s0026898414060147.

Abstract

Fusarium wilt, caused by a soilborne pathogen Fusarium oxysporum f. sp. lilii, is the major disease of lily (Lilium L.). In order to isolate the genes differentially expressed in a resistant reaction to F. oxysporum in L. regale Wilson, a cDNA library was constructed with L. regale root during F. oxysporum infection using the suppression subtractive hybridization (SSH), and a total of 585 unique expressed sequence tags (ESTs) were obtained. Furthermore, the gene expression profiles in the incompatible interaction between L. regale and F. oxysporum were revealed by oligonucleotide microarray analysis of 585 unique ESTs comparison to the compatible interaction between a susceptible Lilium Oriental Hybrid 'Siberia' and F. oxysporum. The result of expression profile analysis indicated that the genes encoding pathogenesis-related proteins (PRs), antioxidative stress enzymes, secondary metabolism enzymes, transcription factors, signal transduction proteins as well as a large number of unknown genes were involved in early defense response of L. regale to F. oxysporum infection. Moreover, the following quantitative reverse transcription PCR (QRT-PCR) analysis confirmed reliability of the oligonucleotide microarray data. In the present study, isolation of differentially expressed genes in L. regale during response to F. oxysporum helped to uncover the molecular mechanism associated with the resistance of L. regale against F. oxysporum.

摘要

尖孢镰刀菌百合专化型引起的枯萎病是百合(Lilium L.)的主要病害。为了分离岷江百合(Lilium regale Wilson)对尖孢镰刀菌抗性反应中差异表达的基因,利用抑制性消减杂交技术(SSH)构建了尖孢镰刀菌侵染岷江百合根部时的cDNA文库,共获得585个独特的表达序列标签(EST)。此外,通过对585个独特EST进行寡核苷酸微阵列分析,揭示了岷江百合与尖孢镰刀菌不亲和互作中的基因表达谱,并与感病东方百合杂交种‘西伯利亚’与尖孢镰刀菌的亲和互作进行了比较。表达谱分析结果表明,编码病程相关蛋白(PR)、抗氧化应激酶、次生代谢酶、转录因子、信号转导蛋白以及大量未知基因的基因参与了岷江百合对尖孢镰刀菌侵染的早期防御反应。此外,随后的定量逆转录PCR(QRT-PCR)分析证实了寡核苷酸微阵列数据的可靠性。在本研究中,分离岷江百合对尖孢镰刀菌反应中差异表达的基因有助于揭示岷江百合对尖孢镰刀菌抗性相关的分子机制。

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