Ma Jin, Li Xiu-Qing
Potato Research Centre, Agriculture and Agri-Food Canada, 850 Lincoln Road, P.O. Box 20280, Fredericton, NB, E3B 4Z7, Canada.
Curr Genet. 2015 Nov;61(4):591-600. doi: 10.1007/s00294-015-0482-1. Epub 2015 Mar 18.
Little information is available about organellar genome copy numbers and integrity in plant roots, although it was reported recently that the plastid and mitochondrial genomes were damaged under light, resulting in non-functional fragments in green seedling leaves in a maize line. In the present study, we investigated organellar genome copy numbers and integrity, after assessing the cellular ploidy, in seedling leaves and roots of two elite maize (Zea mays) cultivars using both long-fragment polymerase chain reaction (long-PCR) and real-time quantitative polymerase chain reaction (qPCR, a type of short-PCR). Since maize leaf and root cells are mainly diploid according to chromosome number counting and the literature, the DNA amount ratio between the organellar genomes and the nuclear genome could be used to estimate average organellar genome copy numbers per cell. In the present study, both long-PCR and qPCR analyses found that green leaves had dramatically more plastid DNA and less mitochondrial DNA than roots had in both cultivars. The similarity in results from long-PCR and qPCR suggests that green leaves and roots during moderate maturation have largely intact plastid and mitochondrial genomes. The high resolution of qPCR led to the detection of an increase in copies in the plastid genome and a decrease in copies in the analyzed mitochondrial sub-genomes during the moderate maturation of seedling leaves and roots. These results suggest that green seedling leaves and roots of these two maize cultivars during moderate maturation had essentially intact organellar genomes, an increased copy number of the plastid genome, and decreased copy numbers of certain mitochondrial sub-genomes.
关于植物根中细胞器基因组的拷贝数和完整性,目前所知甚少,尽管最近有报道称,在光照下质体和线粒体基因组会受到损伤,导致一个玉米品系的绿色幼苗叶片中出现无功能的片段。在本研究中,我们使用长片段聚合酶链反应(long-PCR)和实时定量聚合酶链反应(qPCR,一种短PCR),在评估细胞倍性之后,研究了两个优良玉米(Zea mays)品种的幼苗叶片和根中细胞器基因组的拷贝数和完整性。由于根据染色体计数和文献报道,玉米叶片和根细胞主要是二倍体,因此细胞器基因组与核基因组之间的DNA量比可用于估计每个细胞中细胞器基因组的平均拷贝数。在本研究中,long-PCR和qPCR分析均发现,两个品种的绿色叶片中的质体DNA显著多于根,而线粒体DNA则少于根。long-PCR和qPCR结果的相似性表明,在适度成熟过程中,绿色叶片和根中的质体和线粒体基因组基本完整。qPCR的高分辨率使得在幼苗叶片和根的适度成熟过程中,检测到质体基因组的拷贝数增加,而分析的线粒体亚基因组的拷贝数减少。这些结果表明,这两个玉米品种的绿色幼苗叶片和根在适度成熟时,细胞器基因组基本完整,质体基因组拷贝数增加,某些线粒体亚基因组的拷贝数减少。