基于SNP标记的桃矮化基因精细定位

doi:10.3864/j.issn.0578-1752.2017.18.013

Abstract

Abstract: 【Objective】 Dwarfing peach, due to the small trees, shorter internodes, is an important genetic resources in ornamental and rootstock breeding program. The fine mapping of peach dwarfing genes and explication of the genetic mechanism will provide a basis for the establishment of a molecular-assisted selection system and desired variety improvement, which could be used in breeding dwarf ornamental peach and rootstock varieties. 【Method】 Based on the results of genetic analysis in this study, 395 individuals generated from a self-pollinated population of 05-2-144 were selected for fine mapping the dwarfing gene in peach (Prunus persica (L.) Batsch). By referencing the peach genome sequence, SNP markers were developed in the parents and progenies to generate markers linked to the locus and used to map the gene based on Sanger sequencing. Subsequently, the parent was resequenced to generate SNP markers in the mapping region and acquire heterozyous SNPs for fine mapping. Within the fine mapping region, SNP primers were designed to verify the phenotype of 89 individuals generated from the F₁ segregation population of 10-7×96-5-1.【Result】The segregating population of 05-2-144 was generated to assess the genetic characteristics, resulting in observed 3:1 (300 standard type and 95 dwarfing type individuals) Mendelian ratio fitting with the expected ratio for a monogenic recessive genetic control (P=0.67, χ²=0.19). Moreover, the ratio of standard type to dwarf type corresponded to the expected 3:1 (66 standard type and 23 dwarfing type individuals) segregation (P=0.854, χ²=0.034) for molecular detection. Based on Sanger sequencing results, the linked SNP markers were identified in the position 25 230 425 bp and 27 191 090 bp on Pp06 of peach genome and the locus were on the right side of these two markers. For fine mapping, the parent of this segregation population was resequenced with 66.89X depth and identified the heterozygous SNP to develop SNP markers. Totally, fifteen SNP primers were designed and 12 SNPs (80.0%) were consistent with resequencing data. The dwarfing locus of this type was narrowed on Pp06 between the position 28 712 165 bp and 28 899 661 bp with genetic distances of 0.38 cM and 0.13 cM. The physical region of fine mapping was 277 kb containing 54 known transcripts. Within 28 108 436bp and 29 247 763 bp on Pp06, SNP markers were developed and detected the progeny phenotype of a segregation population of 10-7×96-5-1 with 100% accuracy, constituted of 89 individuals. 【Conclusion】 The dwarfing gene was fine mapped on Pp06 within a physical distance 277 kb. The results of this study will be helpful to clone dwarfing gene and select parents for breeding dwarfing ornamental peach and rootstock varieties.

Key words: peach, dwarfing gene, SNP, fine mapping

LU ZhenHua, NIU Liang, ZHANG NanNan, YAO JiaLong, CUI GuoChao, ZENG WenFang, PAN Lei, WANG ZhiQiang. Fine Mapping of Dwarfing Gene for Peach Based on SNP Markers[J].Scientia Agricultura Sinica, 2017, 50(18): 3572-3580.

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Fine Mapping of Dwarfing Gene for Peach Based on SNP Markers

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