Frelichowski James E, Palmer Michael B, Main Dorrie, Tomkins Jeffrey P, Cantrell Roy G, Stelly David M, Yu John, Kohel Russell J, Ulloa Mauricio
USDA-ARS, W.I.C.S. Res. Unit, Cotton Enhancement Program, Shafter, CA, USA.
Mol Genet Genomics. 2006 May;275(5):479-91. doi: 10.1007/s00438-006-0106-z. Epub 2006 Feb 25.
Fine mapping and positional cloning will eventually improve with the anchoring of additional markers derived from genomic clones such as BACs. From 2,603 new BAC-end genomic sequences from Gossypium hirsutum Acala 'Maxxa', 1,316 PCR primer pairs (designated as MUSB) were designed to flank microsatellite or simple sequence repeat motif sequences. Most (1164 or 88%) MUSB primer pairs successfully amplified DNA from three species of cotton with an average of three amplicons per marker and 365 markers (21%) were polymorphic between G. hirsutum and G. barbadense. An interspecific RIL population developed from the above two entries was used to map 433 marker loci and 46 linkage groups with a genetic distance of 2,126.3 cM covering approximately 45% of the cotton genome and an average distance between two loci of 4.9 cM. Based on genome-specific chromosomes identified in G. hirsutum tetraploid (A and D), 56.9% of the coverage was located on the A subgenome while 39.7% was assigned to the D subgenome in the genetic map, suggesting that the A subgenome may be more polymorphic and recombinationally active than originally thought. The linkage groups were assigned to 23 of the 26 chromosomes. This is the first genetic map in which the linkage groups A01 and A02/D03 have been assigned to specific chromosomes. In addition the MUSB-derived markers from BAC-end sequences markers allows fine genetic and QTL mapping of important traits and for the first time provides reconciliation of the genetic and physical maps. Limited QTL analyses suggested that loci on chromosomes 2, 3, 12, 15 and 18 may affect variation in fiber quality traits. The original BAC clones containing the newly mapped MUSB that tag the QTLs provide critical DNA regions for the discovery of gene sequences involved in biological processes such as fiber development and pest resistance in cotton.
随着源自基因组克隆(如细菌人工染色体,BAC)的其他标记的定位,精细定位和位置克隆最终将会得到改进。从陆地棉阿卡拉‘马克扎’(Gossypium hirsutum Acala 'Maxxa')的2603个新的BAC末端基因组序列中,设计了1316对PCR引物(命名为MUSB),使其位于微卫星或简单序列重复基序序列两侧。大多数(1164对或88%)MUSB引物对成功扩增了三种棉花的DNA,每个标记平均有三个扩增子,365个标记(21%)在陆地棉和海岛棉(G. barbadense)之间具有多态性。利用由上述两个品种构建的种间重组自交系群体,对433个标记位点和46个连锁群进行了定位,遗传距离为2126.3厘摩,覆盖了约45%的棉花基因组,两个位点之间的平均距离为4.9厘摩。基于在陆地棉四倍体(A和D)中鉴定出的基因组特异性染色体,在遗传图谱中,56.9%的覆盖区域位于A亚基因组上,而39.7%被分配到D亚基因组上,这表明A亚基因组可能比原先认为的具有更多的多态性和重组活性。这些连锁群被分配到26条染色体中的23条上。这是第一张将连锁群A01和A02/D03分配到特定染色体的遗传图谱。此外,来自BAC末端序列标记的MUSB衍生标记允许对重要性状进行精细遗传和QTL定位,并首次实现了遗传图谱和物理图谱的整合。有限的QTL分析表明,染色体2、3、12、15和18上的位点可能影响纤维品质性状的变异。包含新定位的标记QTL的MUSB的原始BAC克隆为发现参与棉花纤维发育和抗虫性等生物学过程的基因序列提供了关键的DNA区域。