Lilly J W, Bartoszewski G, Malepszy S, Havey M J
Agricultural Research Service, US Department of Agriculture, University of Wisconsin, Madison 53706, USA.
Curr Genet. 2001 Sep;40(2):144-51. doi: 10.1007/s002940100238.
The cucumber mitochondrial genome is unique because of its large size, paternal transmission, and the existence of a paternally transmitted mosaic (MSC) phenotype spontaneously appearing after independent tissue culture experiments. Transmission studies eliminated paternal imprinting as the genetic basis for the MSC phenotype. We identified a 13 kb region (JLV5-DEL) in the wild-type mitochondrial genome that was absent from three MSC lines. This deleted region was paternally transmitted with the MSC phenotype through the F3 and test-cross generations. Southern hybridizations and PCR amplifications using primers within the JLV5 region revealed that rare wild-type sorters possessed the wild-type region. MSC plants possess the wild-type region at levels (approximately 0.002) below detection by standard PCR reactions and Southern hybridizations, using genomic DNA. Sequence analysis of the wild-type contig revealed no homologies to mitochondrial genes and no open reading frames within JLV5-DEL. PCR amplifications across JLV5-DEL using MSC mtDNA revealed that the loss of this region was not a simple deletion and may be associated with a rearrangement. Because no genic regions were identified within the wild-type JLV5 region, the specific lesion conditioning the MSC phenotype may be associated with the putative rearrangement or another mutation that occurred during tissue culture or exists substoichiometrically in the parental line and is transmitted together with JLV5 within the same mitochondrion.
黄瓜线粒体基因组很独特,这是由于其基因组规模大、父系遗传,以及在独立组织培养实验后自发出现的父系遗传嵌合(MSC)表型。遗传传递研究排除了父本印记作为MSC表型的遗传基础。我们在野生型线粒体基因组中鉴定出一个13 kb的区域(JLV5-DEL),该区域在三个MSC株系中不存在。这个缺失区域通过F3代和测交世代与MSC表型一起进行父系遗传。使用JLV5区域内引物的Southern杂交和PCR扩增显示,罕见的野生型分选体拥有野生型区域。MSC植株拥有野生型区域的水平(约为0.002)低于使用基因组DNA进行标准PCR反应和Southern杂交的检测水平。野生型重叠群的序列分析显示,JLV5-DEL内与线粒体基因无同源性,也没有开放阅读框。使用MSC线粒体DNA对JLV5-DEL进行PCR扩增表明,该区域的缺失不是简单的缺失,可能与重排有关。由于在野生型JLV5区域内未鉴定出基因区域,决定MSC表型的特定损伤可能与推定的重排或组织培养过程中发生的另一种突变有关,或者以亚化学计量存在于亲本系中,并与JLV5在同一线粒体内一起遗传。