Perry D J, Isabel N, Bousquet J
Centre de recherche en biologie forestière, Université Laval, Sainte-Foy, Québec, Canada G1K 7P4.
Heredity (Edinb). 1999 Sep;83 ( Pt 3):239-48. doi: 10.1038/sj.hdy.6885740.
We examined the amount and nature of variation revealed by cDNA-based sequence-tagged-site (STS) markers in Norway spruce (Picea abies (L.) Karst.) using 39 pairs of heterologous primers that were based upon arbitrary genes in black spruce (Picea mariana (Mill.) B.S.P.). A panel of 22 diverse Norway spruce genotypes was screened for variation that could be observed directly using standard agarose gel electrophoresis, without additional manipulation of amplification products. Examination of marker segregation among haploid megagametophytes revealed that nine markers behaved in a codominant manner, two markers had codominant length polymorphisms and null alleles, and four others had dominant length polymorphisms. DNA sequencing of codominant alleles at seven loci indicated that most insertions/deletions (indels) were in noncoding regions and that alleles often differed by the presence or absence of direct repeats that ranged in size from three to 23 bp. The nine markers that showed exclusively codominant polymorphisms in Norway spruce had an average observed heterozygosity of 0.30 and an average of 2.9 alleles in the panel of 22 trees. These levels of variation are similar to those previously found for similar sets of markers in other spruces, and appear to be at least as high as those revealed by polymorphic allozyme markers in Norway spruce. Polymorphisms at one STS locus suggested a higher affinity between Norway spruce and white spruce (Picea glauca (Moench) Voss) than between either of these spruces and black spruce. The STS markers described in this report should be useful in a variety of applications in Norway spruce, including population studies and genome mapping.
我们使用基于黑云杉(Picea mariana (Mill.) B.S.P.)中任意基因的39对异源引物,检测了挪威云杉(Picea abies (L.) Karst.)中基于cDNA的序列标签位点(STS)标记所揭示的变异数量和性质。对一组22种不同的挪威云杉基因型进行筛选,以寻找可通过标准琼脂糖凝胶电泳直接观察到的变异,而无需对扩增产物进行额外操作。对单倍体大配子体间标记分离的检测表明,9个标记表现为共显性,2个标记具有共显性长度多态性和无效等位基因,另外4个具有显性长度多态性。对7个位点的共显性等位基因进行DNA测序表明,大多数插入/缺失(indel)位于非编码区,等位基因通常因存在或不存在大小从3到23 bp的直接重复序列而有所不同。在挪威云杉中仅表现出共显性多态性的9个标记,在22株树的样本中平均观察杂合度为0.30,平均有2.9个等位基因。这些变异水平与之前在其他云杉中类似标记组所发现的水平相似,并且似乎至少与挪威云杉中多态性等位酶标记所揭示的水平一样高。一个STS位点的多态性表明,挪威云杉与白云杉(Picea glauca (Moench) Voss)之间的亲和力高于这两种云杉与黑云杉之间的亲和力。本报告中描述的STS标记在挪威云杉的各种应用中应该是有用的,包括种群研究和基因组作图。