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Characterization of maize B-chromosome-related transcripts isolated via cDNA-AFLP

机译:通过cDNA-AFLP分离的玉米B染色体相关转录本的表征

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The maize B-chromosome consists mainly of heterochromatin and is considered to be genetically inert. However, the B-chromosome contains euchromatin that carries control elements that direct its behaviors during cell division, such as nondisjunction during the second pollen mitosis. To determine the transcriptional activity of the B-chromosome, complementary DNA-amplified fragment length polymorphism analysis was applied to five inbred maize lines with and without B-chromosomes. Six putative B-chromosome-related transcripts were identified, four of which were cloned and characterized via Southern hybridization, fluorescence in situ hybridization, and sequence comparison to further confirm their B-chromosome origin. All the analyzed B-chromosome-related transcript sequences were repetitive and showed homology to A-chromosomes. Quantitative real-time reverse transcriptase-polymerase chain reaction revealed that the B-chromosome-specific transcribed sequences B3547-179 and B3849-212 were transcribed in a B-chromosome-dosage-dependent manner. Expression of B3849-189 and B3849-147 was not specific to the B-chromosome; however, the former showed a transcriptional pattern with B-chromosome dosage compensation, and the latter displayed down-regulation of transcription due to higher B-chromosome numbers. Using four B-10L translocations, B3849-212 was mapped to the B-chromosome region that contains the nondisjunction control elements of the B-chromosome. Taken together, our results suggested that the maize B-chromosome harbors few transcriptionally active sequences and might influence the transcription of A-chromosomes.
机译:玉米B染色体主要由异染色质组成,被认为具有遗传惰性。但是,B染色体含有常染色质,常染色质带有控制元件,这些控制元件指导其在细胞分裂过程中的行为,例如在第二个花粉有丝分裂期间不分离。为了确定B染色体的转录活性,将互补DNA扩增的片段长度多态性分析应用于具有和不具有B染色体的五个自交玉米品系。鉴定出六种与B染色体有关的转录本,其中四种通过Southern杂交,荧光原位杂交和序列比较进行克隆和表征,以进一步确认其B染色体来源。所有分析的B染色体相关的转录物序列都是重复的,并且显示出与A染色体的同源性。实时定量逆转录酶-聚合酶链反应显示,B染色体特异性转录序列B3547-179和B3849-212以B染色体剂量依赖性方式转录。 B3849-189和B3849-147的表达对B染色体不是特异的。但是,前者显示出具有B染色体剂量补偿的转录模式,而后者则由于较高的B染色体数目而显示出转录下调。使用四个B-10L易位,将B3849-212映射到B染色体区域,该区域包含B染色体的非分离控制元件。两者合计,我们的结果表明,玉米B染色体几乎没有转录活性序列,可能会影响A染色体的转录。

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