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首页> 美国卫生研究院文献>other >Karyotype Analysis of Diploid and Spontaneously Occurring Tetraploid Blood Orange Citrus sinensis (L.) Osbeck Using Multicolor FISH With Repetitive DNA Sequences as Probes
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Karyotype Analysis of Diploid and Spontaneously Occurring Tetraploid Blood Orange Citrus sinensis (L.) Osbeck Using Multicolor FISH With Repetitive DNA Sequences as Probes

机译：以重复DNA序列为探针的多色FISH对二倍体和自然发生的四倍体血橙Citrus sinensis（L.）Osbeck的染色体核型分析

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Blood orange [Citrus sinensis (L.) Osbeck] has been increasingly appreciated by consumers worldwide owing to its brilliant red color, abundant anthocyanin and other health-promoting compounds. However, there is still relatively little known about its cytogenetic characteristics, probably because of the small size and similar morphology of metaphase chromosomes and the paucity of chromosomal landmarks. In our previous study, a naturally occurring tetraploid blood orange plant was obtained via seedling screening. Before this tetraploid germplasm can be manipulated into a citrus triploid seedless breeding program, it is of great importance to determine its chromosome characterization and composition. In the present study, an integrated karyotype of blood orange was constructed using sequential multicolor fluorescence in situ hybridization (FISH) with four satellite repeats, two ribosomal DNAs (rDNAs), a centromere-like repeat and an oligonucleotide of telomere repeat (TTTAGGG)3 as probes. Satellite repeats were preferentially located at the terminal regions of the chromosomes of blood orange. Individual somatic chromosome pairs of blood orange were unambiguously identified by repetitive DNA-based multicolor FISH. These probes proved to be effective chromosomal landmarks. The karyotype was formulated as 2n = 2x = 18 = 16m+2sm (1sat) with the karyotype asymmetry degree belonging to 2B. The chromosomal distribution pattern of these repetitive DNAs in this spontaneously occurring tetraploid was identical to that of the diploid, but the tetraploid carried twice the number of hybridization sites as the diploid, indicating a possible pathway involving the spontaneous duplication of chromosome sets in nucellar cells. Our work may facilitate the molecular cytogenetic study of blood orange and provide chromosomal characterization for the future utilization of this tetraploid germplasm in the service of seedless breeding programs.
机译：血橙[Citrus sinensis（L.）Osbeck]由于其鲜艳的红色，丰富的花色苷和其他促进健康的化合物而受到全世界消费者的青睐。但是，关于其细胞遗传学特征的了解仍然相对较少，这可能是由于中期染色体的大小和相似形态以及染色体标志物的缺乏所致。在我们以前的研究中，通过幼苗筛选获得了天然四倍体血橙植物。在将该四倍体种质转化为柑橘三倍体无核育种程序之前，确定其染色体特征和组成非常重要。在本研究中，使用具有四个卫星重复序列，两个核糖体DNA（rDNA），着丝粒样重复序列和端粒重复序列寡核苷酸（TTTAGGG）的连续多色荧光原位杂交（FISH）构建了血橙的整合型核型。作为探针。卫星重复序列优选位于血橙染色体的末端区域。通过重复的基于DNA的多色FISH明确鉴定了血橙的个体体染色体对。这些探针被证明是有效的染色体标志。核型的不对称度为2B = 2n = 2x = 18 = 16m + 2sm（1sat）。这些自发出现的四倍体中这些重复DNA的染色体分布模式与二倍体相同，但四倍体所携带的杂交位点数量是二倍体的两倍，表明可能的途径涉及在细胞中自发复制染色体。我们的工作可能会促进血橙的分子细胞遗传学研究，并为该四倍体种质在无核育种计划中的未来利用提供染色体表征。

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