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    榛子正常发育与败育子房差异蛋白谱对比分析

    Comparative studies on differently expressed proteomes of developing and abortive ovary in hazelnut

    • 摘要:
      目的筛选参与调控榛子子房败育的候选蛋白,为榛子遗传改良研究提供科学依据。
      方法以平欧杂交榛‘达维’的正常发育与败育子房为材料,进行蛋白样品的同位素标记相对和绝对定量iTRAQ(isobaric tags for relative and absolute quantification)技术分析。对鉴定到的所有蛋白进行COG(Cluster of orthologous groups of proteins)功能分类,预测鉴定蛋白的功能。随后依据蛋白定量结果,筛选差异表达蛋白,进而开展GO(Gene ontology)功能富集与KEGG(Kyoto encyclopedia of genes and genomes)代谢路径富集分析以明确其分子功能和重要生物代谢路径。最后,主要从显著性富集路径中筛选可能参与子房败育调控的差异表达蛋白。
      结果蛋白鉴定共获得317068个二级谱图,特有多肽14267条,蛋白3538个。R、O、J、G和C类中的蛋白数量为最多,分别占有COG功能注释的蛋白总数的19.36%、9.97%、7.80%、7.67%和6.76%。共鉴定到249个差异表达蛋白,其中上调、下调表达蛋白分别为180和69个。GO富集分析结果表明,差异表达蛋白主要执行结合与催化分子功能。KEGG富集分析共找到11个显著性富集路径,最为显著的路径包括:苯丙素生物合成(ko00940),光合作用(ko00195),代谢路径(ko01100),光合作用-天线蛋白(ko00196),次生代谢产物生物合成(ko01110)。初步筛选获得可能参与调控榛子子房败育的候选蛋白37个。
      结论榛子败育子房的形成与光合作用、碳水化合物运输与代谢、能量合成与转换、花粉管生长与DNA甲基化等相关,本研究为深入解析榛子子房败育的分子机制提供了科学依据。

       

      Abstract:
      ObjectiveThe aim of this study is to screen candidate proteins which may be involved in the regulation of abortive ovary formation in hazelnut and provide scientific base for its genetic improvement.
      MethodHybrid hazelnut (Corylus heterophylla × C. avellana) cultivar 'Dawei' was used as study materials, and iTRAQ (Isobaric tags for relative and absolute quantification) technology was performed using protein extracted from abortive ovaries and developing ovaries. COG (Cluster of orthologous groups of proteins) functional classification was carried out using all the identified proteins, and their potential biological functions were predicted. Subsequently, significantly differently expressed proteins (DEPs) were identified according to protein quantification results, and GO (Gene ontology) and KEGG (Kyoto encyclopedia of genes and genomes) enrichment analysis of DEPs were carried out to explore their general molecular functions and important KEGG pathways. Finally, potential important proteins involved in abortive ovary formation were chosen mainly from significantly enriched KEGG pathways.
      ResultProtein identification generated 317068 total spectra, 14267 unique peptides and 3538 proteins. Functional class of R, O, J, G and C in COG analysis accounted for 19.36%, 9.97%, 7.80%, 7.67% and 6.76% of total proteins with COG annotations. In total, 249 DEPs were identified in the paired comparison of developing and abortive ovary, including 180 and 69 up- and down-regulated DEPs. Based on GO and KEGG enrichment analysis results, these DEPs mainly executed binding and catalysis molecular functions, and 11 significant enriched KEGG pathways were identified, including phenylpropanoid biosynthesis (ko00940), photosynthesis (ko00195), metabolic pathways (ko01100), photosynthesis-antenna proteins (ko00196) and biosynthesis of secondary metabolites (ko01110). Thirty-seven important candidate DEPs were identified and these DEPs may contribute to abortive ovary formation in hazelnut.
      ConclusionProteins related to photosynthesis, carbohydrate transport and metabolism, energy production and conversion, pollen tube growth and DNA methylation may regulate ovary abortion in hazel. Our findings provide insight into the molecular mechanisms of ovary abortion in hazelnut.

       

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