Alpert K B, Tanksley S D
Department of Plant Breeding and Biometry, Cornell University, Ithaca, NY 14853.
Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15503-7. doi: 10.1073/pnas.93.26.15503.
A high-resolution physical and genetic map of a major fruit weight quantitative trait locus (QTL), fw2.2, has been constructed for a region of tomato chromosome 2. Using an F(2) nearly isogenic line mapping population (3472 individuals) derived from Lycopersicon esculentum (domesticated tomato) x Lycopersicon pennellii (wild tomato), fw2.2 has been placed near TG91 and TG167, which have an interval distance of 0.13 +/- 0.03 centimorgan. The physical distance between TG91 and TG167 was estimated to be </= 150 kb by pulsed-field gel electrophoresis of tomato DNA. A physical contig composed of six yeast artificial chromosomes (YACs) and encompassing fw2.2 was isolated. No rearrangements or chimerisms were detected within the YAC contig based on restriction fragment length polymorphism analysis using YAC-end sequences and anchored molecular markers from the high-resolution map. Based on genetic recombination events, fw2.2 could be narrowed down to a region less than 150 kb between molecular markers TG91 and HSF24 and included within two YACs: YAC264 (210 kb) and YAC355 (300 kb). This marks the first time, to our knowledge, that a QTL has been mapped with such precision and delimited to a segment of cloned DNA. The fact that the phenotypic effect of the fw2.2 QTL can be mapped to a small interval suggests that the action of this QTL is likely due to a single gene. The development of the high-resolution genetic map, in combination with the physical YAC contig, suggests that the gene responsible for this QTL and other QTLs in plants can be isolated using a positional cloning strategy. The cloning of fw2.2 will likely lead to a better understanding of the molecular biology of fruit development and to the genetic engineering of fruit size characteristics.
已构建了番茄2号染色体一个区域的主要果实重量数量性状位点(QTL)fw2.2的高分辨率物理图谱和遗传图谱。利用从栽培番茄(Lycopersicon esculentum)×潘那利番茄(Lycopersicon pennellii)(野生番茄)衍生而来的F2近等基因系作图群体(3472个个体),fw2.2被定位在TG91和TG167附近,它们之间的间隔距离为0.13±0.03厘摩。通过番茄DNA的脉冲场凝胶电泳估计TG91和TG167之间的物理距离≤150 kb。分离出了一个由6个酵母人工染色体(YAC)组成并包含fw2.2的物理重叠群。基于使用YAC末端序列和来自高分辨率图谱的锚定分子标记的限制性片段长度多态性分析,在YAC重叠群内未检测到重排或嵌合现象。基于遗传重组事件,fw2.2可被缩小到分子标记TG91和HSF24之间小于150 kb的区域,并包含在两个YAC中:YAC264(210 kb)和YAC355(300 kb)。据我们所知,这是首次将一个QTL定位到如此精确的程度并限定在一段克隆DNA片段上。fw2.2 QTL的表型效应可定位到一个小间隔这一事实表明,该QTL的作用可能归因于单个基因。高分辨率遗传图谱与物理YAC重叠群的结合表明,可使用定位克隆策略分离负责该QTL以及植物中其他QTL的基因。fw2.2的克隆可能会使人们更好地理解果实发育的分子生物学,并实现果实大小特征的基因工程。