Department of Life Sciences, Ben-Gurion University, Beer-Sheva, 84105, Israel.
Plant J. 2013 Oct;76(2):201-10. doi: 10.1111/tpj.12285. Epub 2013 Aug 13.
Short interspersed nuclear elements (SINEs) are non-autonomous non-LTR retroelements that are present in most eukaryotic species. While SINEs have been intensively investigated in humans and other animal systems, they are poorly studied in plants, especially in wheat (Triticum aestivum). We used quantitative PCR of various wheat species to determine the copy number of a wheat SINE family, termed Au SINE, combined with computer-assisted analyses of the publicly available 454 pyrosequencing database of T. aestivum. In addition, we utilized site-specific PCR on 57 Au SINE insertions, transposon methylation display and transposon display on newly formed wheat polyploids to assess retrotranspositional activity, epigenetic status and genetic rearrangements in Au SINE, respectively. We retrieved 3706 different insertions of Au SINE from the 454 pyrosequencing database of T. aestivum, and found that most of the elements are inserted in A/T-rich regions, while approximately 38% of the insertions are associated with transcribed regions, including known wheat genes. We observed typical retrotransposition of Au SINE in the second generation of a newly formed wheat allohexaploid, and massive hypermethylation in CCGG sites surrounding Au SINE in the third generation. Finally, we observed huge differences in the copy numbers in diploid Triticum and Aegilops species, and a significant increase in the copy numbers in natural wheat polyploids, but no significant increase in the copy number of Au SINE in the first four generations for two of three newly formed allopolyploid species used in this study. Our data indicate that SINEs may play a prominent role in the genomic evolution of wheat through stress-induced activation.
短散在核元件(SINEs)是非自主的非长末端重复反转录元件,存在于大多数真核生物物种中。虽然 SINEs 在人类和其他动物系统中得到了深入研究,但在植物中,尤其是在小麦(Triticum aestivum)中研究甚少。我们使用各种小麦物种的定量 PCR 来确定一个称为 Au SINE 的小麦 SINE 家族的拷贝数,结合对 T. aestivum 的公开可用的 454 焦磷酸测序数据库的计算机辅助分析。此外,我们利用 57 个 Au SINE 插入的位点特异性 PCR、转座子甲基化显示和新形成的小麦多倍体的转座子显示,分别评估 Au SINE 的反转录活性、表观遗传状态和遗传重排。我们从 T. aestivum 的 454 焦磷酸测序数据库中检索到 3706 个不同的 Au SINE 插入,发现大多数元件插入在 A/T 丰富的区域,而大约 38%的插入与转录区域相关,包括已知的小麦基因。我们在新形成的小麦异源六倍体的第二代中观察到 Au SINE 的典型反转录,在第三代中观察到 Au SINE 周围的 CCGG 位点的大量超甲基化。最后,我们观察到二倍体小麦和拟南芥物种中拷贝数的巨大差异,以及在自然小麦多倍体中拷贝数的显著增加,但在所研究的三个新形成的异源多倍体物种中的两个中,Au SINE 的拷贝数在前四代中没有显著增加。我们的数据表明,SINEs 可能通过应激诱导的激活在小麦的基因组进化中发挥突出作用。