应用AFLP技术,筛选了40对引物,其中EcoR I AGC+M seI CGA,EcoR I AGC+M seI CAC,EcoR I AGG+M seI CAC,EcoR I AGG+M seI CCA,EcoR I ACG+M seI CAC,EcoR I ACG+M seI CCA等共12对引物经PCR扩增,效果理想,共扩增出4 227条带,带型清晰、...应用AFLP技术,筛选了40对引物,其中EcoR I AGC+M seI CGA,EcoR I AGC+M seI CAC,EcoR I AGG+M seI CAC,EcoR I AGG+M seI CCA,EcoR I ACG+M seI CAC,EcoR I ACG+M seI CCA等共12对引物经PCR扩增,效果理想,共扩增出4 227条带,带型清晰、丰富、可靠性强。在EcoR1-ACG+M seI-CAC这对引物下,果实性状优良的单株变异与普通对照(即普通冰糖橙)有明显的带型差异,多态性比例达10.2%。应用AFLP技术进行柑橘芽变的分析和鉴定获得成功。展开更多
As the largest transposable element in the plant genome, retrotransposons are thought to be involved in citrus genetic instability and genome evolution, especially in sweet orange, which is prone to bud mutation. In t...As the largest transposable element in the plant genome, retrotransposons are thought to be involved in citrus genetic instability and genome evolution, especially in sweet orange, which is prone to bud mutation. In the present study, the presence of copia-like retrotransposons, their heterogeneity, genomic distribution, and transcriptional activities in Citrus were investigated in 12 sweet orange （Citrus sinensis Osbeck） cultivars using a PCR assay designed to detect copia-like reverse transcriptase （RT） sequences. Twelve amplification products from each cultivar were cloned and sequenced. The cloned sequences showed great heterogeneity, except “Dream” navel and “Hamlin”, both of which shared the same sequence. Frame shifting, termination, deletion, and substitution accounted for the heterogeneity of RT sequences. Southern blot hybridization using the RT1 clone from the “Cara Cara” navel as a probe showed that multiple copies were integrated throughout the sweet orange genomes, which made the retrotransposon possible an effective molecular marker to detect citrus evolution events and to reveal its relationship with bud mutation. No transcriptional activities of the retrotransposon were detected by RT-PCR and Northern analysis in the fruits and leaves of either “Cara Cara” or “Seike” navels.展开更多
文摘应用AFLP技术,筛选了40对引物,其中EcoR I AGC+M seI CGA,EcoR I AGC+M seI CAC,EcoR I AGG+M seI CAC,EcoR I AGG+M seI CCA,EcoR I ACG+M seI CAC,EcoR I ACG+M seI CCA等共12对引物经PCR扩增,效果理想,共扩增出4 227条带,带型清晰、丰富、可靠性强。在EcoR1-ACG+M seI-CAC这对引物下,果实性状优良的单株变异与普通对照(即普通冰糖橙)有明显的带型差异,多态性比例达10.2%。应用AFLP技术进行柑橘芽变的分析和鉴定获得成功。
基金the National Natural Science Foundation of China，国家科技攻关项目
文摘As the largest transposable element in the plant genome, retrotransposons are thought to be involved in citrus genetic instability and genome evolution, especially in sweet orange, which is prone to bud mutation. In the present study, the presence of copia-like retrotransposons, their heterogeneity, genomic distribution, and transcriptional activities in Citrus were investigated in 12 sweet orange （Citrus sinensis Osbeck） cultivars using a PCR assay designed to detect copia-like reverse transcriptase （RT） sequences. Twelve amplification products from each cultivar were cloned and sequenced. The cloned sequences showed great heterogeneity, except “Dream” navel and “Hamlin”, both of which shared the same sequence. Frame shifting, termination, deletion, and substitution accounted for the heterogeneity of RT sequences. Southern blot hybridization using the RT1 clone from the “Cara Cara” navel as a probe showed that multiple copies were integrated throughout the sweet orange genomes, which made the retrotransposon possible an effective molecular marker to detect citrus evolution events and to reveal its relationship with bud mutation. No transcriptional activities of the retrotransposon were detected by RT-PCR and Northern analysis in the fruits and leaves of either “Cara Cara” or “Seike” navels.
