Ashley M V, Wilk J A, Styan S M N, Craft K J, Jones K L, Feldheim K A, Lewers K S, Ashman T L
Department of Biological Sciences, University of Illinois at Chicago, 60608, USA.
Theor Appl Genet. 2003 Nov;107(7):1201-7. doi: 10.1007/s00122-003-1370-5. Epub 2003 Aug 8.
The objectives of the present study were to develop microsatellite markers for the wild strawberry, Fragaria virginiana, to evaluate segregation patterns of microsatellite alleles in this octoploid species, and assess genetic variability at microsatellite loci in a wild population. A genomic library was screened for microsatellite repeats and several PCR primers were designed and tested. We also tested the use of heterologous primers and found that F. virginiana primers amplified products in cultivated strawberry, Fragaria x ananassa Duch. and Fragaria chiloensis. Similarly, microsatellite loci developed from cultivated strawberry also successfully amplified F. virginiana loci. We investigated four microsatellite loci in detail, three developed from F. virginiana and one from cultivated strawberry. A survey of 100 individuals from a population of F. virginiana in Pennsylvania demonstrated high heterozygosities (H(e) or gene diversity ranged from 0.80 to 0.88 per locus) and allelic diversity (12-17 alleles per locus), but individual plants had no more than two alleles per locus. Segregation patterns in parents and progeny of two controlled crosses at these four loci were consistent with disomic Mendelian inheritance. Together these findings suggest that the genome of F. virginiana is "highly diploidized" and at least a subset of microsatellite loci can be treated as codominant, diploid markers. Significant heterozygote deficiencies were found at three of the four loci for hermaphroditic individuals but for only one locus among females in this gynodioecious species.
本研究的目的是开发野生草莓弗州草莓(Fragaria virginiana)的微卫星标记,评估该八倍体物种中微卫星等位基因的分离模式,并评估野生种群中微卫星位点的遗传变异性。对一个基因组文库进行微卫星重复序列筛选,并设计和测试了几种PCR引物。我们还测试了异源引物的使用情况,发现弗州草莓引物能在栽培草莓凤梨草莓(Fragaria x ananassa Duch.)和智利草莓(Fragaria chiloensis)中扩增出产物。同样,从栽培草莓开发的微卫星位点也成功扩增了弗州草莓的位点。我们详细研究了四个微卫星位点,其中三个是从弗州草莓开发的,一个是从栽培草莓开发的。对宾夕法尼亚州一个弗州草莓种群的100个个体进行的调查显示出高杂合度(每个位点的H(e)或基因多样性范围为0.80至0.88)和等位基因多样性(每个位点12 - 17个等位基因),但单个植株每个位点的等位基因不超过两个。这四个位点在两个控制杂交的亲本和后代中的分离模式与二体孟德尔遗传一致。这些发现共同表明,弗州草莓的基因组“高度二倍体化”,并且至少一部分微卫星位点可被视为共显性的二倍体标记。在这个雌雄异株物种中,雌雄同体个体的四个位点中有三个发现了显著的杂合子不足,但雌性个体中只有一个位点存在这种情况。