首页> 外文期刊>New Zealand Journal of Forestry Science >Heterologous hybridisation to a Pinus microarray: profiling of gene expression in Pinus radiata saplings exposed to ethephon
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Heterologous hybridisation to a Pinus microarray: profiling of gene expression in Pinus radiata saplings exposed to ethephon

机译:异种杂交到松树微阵列:暴露于乙烯利的辐射松树苗中的基因表达谱

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Background The design, construction and application of a Pinus microarray platform are described. The oligonucleotide microarray was developed using publicly available Pinus cDNA sequences mostly derived from Pinus taeda to test whether heterologous hybridisation of microarray probes will generate useful data when hybridised with cRNA constructed from the dominant New Zealand forestry species Pinus radiata. Methods A comprehensive consensus sequence collection of Pinus cDNA sequences was collated into a non-redundant database used for automated design of 60-mer oligonucleotide microarray probes. The microarray slides, manufactured by Agilent Technologies (Palo Alto, California), were used to monitor gene expression in an induction experiment using 2-chloroethylphosphonic acid, common name ethephon and the active ingredient of the plant growth regulator Ethrel? (Bayer Crop Science). The transcriptomes from tissues of 2-year old Pinus radiata saplings +/? ethephon treatment were compared by hybridisation onto the Pinus microarray slides. Results Statistically significant differentially expressed genes identified by heterologous hybridisation to the Pinus microarray following ethephon induction included the up-regulation of genes in the xylem that were related to the metabolism of phenylpropanoids and flavonoids, and also defence responses, specifically against fungal/insect attack and oxidative stress. Bark, mucilaginous xylem and xylem generated largely mutually exclusive cohorts of genes and Gene Ontology (GO) classes. The results are also interpreted in reference to gross and microscopic morphological changes. Samples of gene responses were validated by quantitative RT-PCR. Conclusion These results confirm the successful development of a Pinus microarray and demonstrate the utility of the microarray for transcriptomic research in Pinus radiata through heterologous hybridisation.
机译:背景介绍了Pinus微阵列平台的设计,构建和应用。寡核苷酸微阵列是使用主要来自taeda taeda的公共Pinus cDNA序列开发的,以测试微阵列探针的异源杂交在与从新西兰主要林业物种Pinus radiata构建的cRNA杂交时是否会产生有用的数据。方法将全面的Pinus cDNA序列共有序列收集到一个非冗余数据库中,该数据库用于自动设计60-mer寡核苷酸微阵列探针。由安捷伦科技公司(加利福尼亚州帕洛阿尔托)制造的微阵列玻片用于在诱导实验中监测基因表达,该实验使用了2-氯乙基膦酸,俗名乙烯利和植物生长调节剂Ethrel?的活性成分。 (拜耳作物科学)。 2岁辐射松幼树组织的转录组通过杂交到松属微阵列载玻片上比较乙烯利处理。结果在乙烯利诱导后通过与松属微阵列异源杂交鉴定出的统计学上显着差异表达的基因包括木质部中与苯基丙烷和类黄酮的代谢有关的基因上调,以及防御反应,特别是针对真菌/昆虫的侵袭和氧化应激。树皮,粘液木质部和木质部在很大程度上与基因和基因本体论(GO)类互斥。还参考总体和微观形态变化来解释结果。基因反应样品通过定量RT-PCR验证。结论这些结果证实了松属微阵列的成功开发,并证明了该微阵列通过异源杂交技术在辐射松中进行转录组学研究的实用性。

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