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番茄无色非成熟基因座的遗传分析与荧光原位杂交定位

Genetic analysis and FISH mapping of the Colourless non-ripening locus of tomato.

作者信息

Tör M., Manning K., King G. J., Thompson A. J., Jones G. H., Seymour G. B., Armstrong S. J.

机构信息

Horticulture Research International, Wellesbourne, Warwick CV35 9EF, UK.

出版信息

Theor Appl Genet. 2002 Feb;104(2-3):165-170. doi: 10.1007/s001220100760.

Abstract

Cnr ( Colourless non-ripening) is a dominant pleiotropic ripening mutation of tomato ( Lycopersicon esculentum) which has previously been mapped to the proximal region of tomato chromosome 2. We describe the fine mapping of the Cnr locus using both linkage analysis and fluorescence in situ hybridisation (FISH). Restriction fragment length polymorphism (RFLP)-, amplified restriction fragment polymorphism (AFLP)-, and cleaved amplified polymorphic sequence (CAPS)-based markers, linked to the Cnrlocus were mapped onto the long arm of chromosome 2. Detailed linkage analysis indicated that the Cnr locus was likely to lie further away from the top of the long arm than previously thought. This was confirmed by FISH, which was applied to tomato pachytene chromosomes in order to gain an insight into the organisation of hetero- and euchromatin and its relationship to the physical and genetic distances in the Cnr region. Three molecular markers linked to Cnr were unambiguously located by FISH to the long arm of chromosome 2 using individual BAC probes containing these single-copy sequences. The physical order of the markers coincided with that established by genetic analysis. The two AFLP markers most-closely linked to the Cnr locus were located in the euchromatic region 2.7-cM apart. The physical distance between these markers was measured on the pachytene spreads and estimated to be approximately 900 kb, suggesting a bp:cM relationship in this region of chromosome 2 of about 330 kb/cM. This is less than half the average value of 750 kb/cM for the tomato genome. The relationship between genetic and physical distances on chromosome 2 is discussed.

摘要

无色非成熟(Cnr)是番茄(Lycopersicon esculentum)的一种显性多效成熟突变体,此前已被定位到番茄第2号染色体的近端区域。我们利用连锁分析和荧光原位杂交(FISH)描述了Cnr基因座的精细定位。与Cnr基因座连锁的基于限制性片段长度多态性(RFLP)、扩增限制性片段多态性(AFLP)和酶切扩增多态性序列(CAPS)的标记被定位到第2号染色体的长臂上。详细的连锁分析表明,Cnr基因座可能比之前认为的更远离长臂顶端。这通过FISH得到了证实,FISH应用于番茄粗线期染色体,以便深入了解异染色质和常染色质的组织及其与Cnr区域物理距离和遗传距离的关系。使用含有这些单拷贝序列的单个BAC探针,通过FISH将与Cnr连锁的三个分子标记明确地定位到第2号染色体的长臂上。标记的物理顺序与遗传分析确定的顺序一致。与Cnr基因座最紧密连锁的两个AFLP标记位于常染色质区域,相距2.7厘摩。在粗线期铺展上测量了这些标记之间的物理距离,估计约为900 kb,这表明在第2号染色体的这个区域,碱基对与厘摩的关系约为330 kb/cM。这不到番茄基因组平均750 kb/cM值的一半。讨论了第2号染色体上遗传距离与物理距离的关系。

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