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在异源六倍体小黑麦中不表达的黑麦基因的鉴定与表征。

Identification and characterization of rye genes not expressed in allohexaploid triticale.

作者信息

Khalil Hala Badr, Ehdaeivand Mohammad-Reza, Xu Yong, Laroche André, Gulick Patrick J

机构信息

Department of Biology, Concordia University, 7141 Sherbrooke W., Montreal, Quebec, H4B 1R6, Canada.

Present addresses: Department of Genetics, Faculty of Agriculture, Ain-Shams University, Shoubra El-khema, Cairo, Egypt.

出版信息

BMC Genomics. 2015 Apr 10;16(1):281. doi: 10.1186/s12864-015-1480-x.

Abstract

BACKGROUND

One of the most important evolutionary processes in plants is polyploidization. The combination of two or more genomes in one organism often initially leads to changes in gene expression and extensive genomic reorganization, compared to the parental species. Hexaploid triticale (x Triticosecale) is a synthetic hybrid crop species generated by crosses between T. turgidum and Secale cereale. Because triticale is a recent synthetic polyploid it is an important model for studying genome evolution following polyploidization. Molecular studies have demonstrated that genomic sequence changes, consisting of sequence elimination or loss of expression of genes from the rye genome, are common in triticale. High-throughput DNA sequencing allows a large number of genes to be surveyed, and transcripts from the different homeologous copies of the genes that have high sequence similarity can be better distinguished than hybridization methods previously employed.

RESULTS

The expression levels of 23,503 rye cDNA reference contigs were analyzed in 454-cDNA libraries obtained from anther, root and stem from both triticale and rye, as well as in five 454-cDNA data sets created from triticale seedling shoot, ovary, stigma, pollen and seed tissues to identify the classes of rye genes silenced or absent in the recent synthetic hexaploid triticale. Comparisons between diploid rye and hexaploid triticale detected 112 rye cDNA contigs (~0.5%) that were totally undetected by expression analysis in all triticale tissues, although their expression was relatively high in rye tissues. Non-expressed rye genes were found to be strikingly less similar to their closest BLASTN matches in the wheat genome or in the other Triticum genomes than a test set of 200 random rye genes. Genes that were not detected in the RNA-seq data were further characterized by testing for their presence in the triticale genome by PCR using genomic DNA as a template.

CONCLUSION

Genes with low similarity between rye sequences and their closest matches in the Triticum genome have a higher probability to be repressed or absent in the allopolyploid genome.

摘要

背景

植物中最重要的进化过程之一是多倍体化。与亲本物种相比,一个生物体中两个或更多基因组的组合通常最初会导致基因表达的变化和广泛的基因组重组。六倍体小黑麦(×Triticosecale)是由硬粒小麦(T. turgidum)和黑麦(Secale cereale)杂交产生的人工杂交作物物种。由于小黑麦是一种新近合成的多倍体,它是研究多倍体化后基因组进化的重要模型。分子研究表明,由黑麦基因组中基因的序列消除或表达缺失组成的基因组序列变化在小黑麦中很常见。高通量DNA测序可以对大量基因进行检测,并且与先前使用的杂交方法相比,具有高度序列相似性的基因的不同同源拷贝的转录本能够得到更好的区分。

结果

分析了23,503个黑麦cDNA参考重叠群在来自小黑麦和黑麦的花药、根和茎的454 - cDNA文库中的表达水平,以及在从小黑麦幼苗地上部分、子房、柱头、花粉和种子组织创建的五个454 - cDNA数据集中的表达水平,以鉴定在新近合成的六倍体小黑麦中沉默或缺失的黑麦基因类别。二倍体黑麦和六倍体小黑麦之间的比较检测到112个黑麦cDNA重叠群(约0.5%),尽管它们在黑麦组织中的表达相对较高,但在所有小黑麦组织的表达分析中完全未检测到。与200个随机选择的黑麦基因测试集相比,发现未表达的黑麦基因与其在小麦基因组或其他小麦属基因组中最相似的BLASTN匹配序列的相似性明显更低。通过以基因组DNA为模板进行PCR检测其在小黑麦基因组中的存在情况,对RNA - seq数据中未检测到的基因进行了进一步表征。

结论

黑麦序列与其在小麦基因组中最相似匹配序列之间相似性较低的基因在异源多倍体基因组中被抑制或缺失的可能性更高。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfb6/4396786/0d395e679726/12864_2015_1480_Fig1_HTML.jpg

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