本文關(guān)鍵詞： 桃 矮化基因 SNP 精細(xì)定位　出處：《中國農(nóng)業(yè)科學(xué)》2017年18期 　論文類型：期刊論文

【摘要】：【目的】矮化型桃樹體矮小、節(jié)間短,是盆栽觀賞和砧木育種的重要遺傳資源。明確矮化性狀形成的遺傳機(jī)制并對桃矮化基因進(jìn)行精細(xì)定位,是建立目標(biāo)性狀分子輔助選種體系和遺傳改良的前提,可為有目標(biāo)的選育矮化觀賞桃和砧木品種奠定基礎(chǔ)�！痉椒ā恳浴�05-2-144’(‘97矮’ב鴛鴦垂枝’桃)套袋自交獲得的395個后代單株構(gòu)建的分離群體為材料。參考桃基因組信息并基于Sanger技術(shù)開發(fā)的SNP標(biāo)記對親本和后代單株進(jìn)行分析,在擴(kuò)大群體單株中進(jìn)行連鎖關(guān)系分析,確定連鎖的SNP標(biāo)記,初步定位目標(biāo)基因。在定位區(qū)域內(nèi)基于二代測序技術(shù)開發(fā)更多的基因型和表型一致的SNP標(biāo)記,對后代單株進(jìn)行基因分型,完成目標(biāo)性狀的精細(xì)定位。然后在精細(xì)定位區(qū)域內(nèi)開發(fā)SNP標(biāo)記,即雜交群體雙親均為Aa雜合基因型,對‘10-7’ב96-5-1’雜交后代89個單株進(jìn)行分子鑒定,以驗證基因定位結(jié)果的準(zhǔn)確性。【結(jié)果】通過對桃單株‘05-2-144’自交后代實生苗表型鑒定表明,普通型和矮化型單株數(shù)分別為300株和95株,性狀分離比例接近3㑳1(P值為0.67;χ2為0.19),符合孟德爾遺傳規(guī)律,桃矮化性狀受隱性單基因控制。用于分子鑒定的單株來源于‘10-7’(普通型)ב96-5-1’(普通型)雜交組合,共獲得89個后代單株,其中普通型66株;矮化型23株(P值為0.854;χ2為0.034)。基于Sanger測序技術(shù)開發(fā)了SNP標(biāo)記,在桃基因組數(shù)據(jù)庫Pp06上25 230 425 bp和27 191 090 bp處獲得了連鎖的SNP標(biāo)記,且目標(biāo)基因位于這兩個標(biāo)記的右側(cè),初步獲得了連鎖的SNP分子標(biāo)記。在此基礎(chǔ)上,對親本進(jìn)行66.89X深度測序,繼續(xù)開發(fā)符合Aa雜合基因型的SNP標(biāo)記位點。根據(jù)參考基因組和物理距離區(qū)間共設(shè)計了15對SNP引物,其中12對引物與重測序結(jié)果中SNP的類型一致,3對引物與重測序結(jié)果中SNP的類型不一致,連鎖標(biāo)記的基因分型成功率為80.0%。通過基于SNP基因分型分析,最終完成了目標(biāo)性狀的精細(xì)定位,位點位于Pp06的28 712165 bp(引物為JXSNP-5)和28 899 661 bp(引物為JXHRM-SNP-3)之間,遺傳距離分別為0.38 c M和0.13 c M,物理距離約為277 kb,精細(xì)定位區(qū)域內(nèi)有54個已知轉(zhuǎn)錄本。在定位區(qū)域內(nèi)桃基因組Pp06的28 108 436 bp處和29 247 763 bp處開發(fā)SNP標(biāo)記用于雜交后代表型的鑒定,結(jié)果表明所有后代單株基因型和表型鑒定結(jié)果完全一致,鑒定準(zhǔn)確率為100%�！窘Y(jié)論】本研究精細(xì)定位了桃矮化基因,物理距離約為277 kb,為基因克隆、親本早期篩選以選育矮化觀賞桃和砧木品種等奠定了基礎(chǔ)。
[Abstract]:[objective] Dwarf peach is an important genetic resource for potted ornamental and rootstock breeding because of its short body and short internodes. It is the premise of establishing molecular aided seed selection system and genetic improvement for target traits. It can lay a foundation for the selection of dwarf ornamental peach and rootstock varieties. The isolated populations of 395 progenies constructed by bagging self-crossing were used as materials. Referring to the genomic information of peach and based on the SNP markers developed by Sanger technology, the parents and progenies were analyzed. The linkage relationship analysis was carried out in the expanded population, and the linkage SNP markers were determined. Based on the second generation sequencing technology, more genotypic and phenotypic SNP markers were developed for genotyping of single progeny. Then SNP markers were developed in the fine mapping area, that is, both parents of the hybrid population were AA heterozygote genotypes. The molecular identification of 89 individual plants in the progenies of 10 ~ (-7'脳 10 ~ (-7)'脳 10 ~ (-7)'脳 10 ~ (-7)'脳. In order to verify the accuracy of gene mapping results. [results] the phenotypic identification of self-bred seedlings of Peach per plant 05-2-144'showed that the number of common type and dwarf type per plant were 300 and 95, respectively. The segregation ratio of traits is close to 3? 1P value was 0.67; 蠂 2 was 0.19, which was consistent with Mendelian inheritance. The dwarf traits of peach were controlled by recessive single gene. The single plant for molecular identification was derived from the cross combination of 10-7 (common type) 脳 96-5-1 (common type), and 89 progenies were obtained. Among them, 66 were common type; P value of 23 dwarf strains was 0.854; 蠂 2 was 0.034. SNP markers were developed based on Sanger sequencing technique. Linkage SNP markers were obtained at 25 230,425 BP and 27 191 090 BP in peach genome database Pp06, and the target gene was located to the right of the two markers. The linkage SNP markers were obtained, and the parents were further sequenced by 66.89X. Further development of SNP marker sites consistent with AA heterozygote genotype was carried out. A total of 15 pairs of SNP primers were designed according to reference genomes and physical distances. The type of SNP in 12 pairs of primers was the same as that in the result of resequencing. 3 pairs of primers were not consistent with the type of SNP in the result of resequencing. The success rate of genotyping of linkage markers was 80.0%. Based on SNP genotyping analysis, the precise mapping of target traits was finally completed. The loci were between 28 712165 BP (primer JXSNP-5) and 28 899,661bp (primer JXHRM-SNP-3) of Pp06. The genetic distance was 0.38 cm and 0.13 cm, and the physical distance was about 277 kb. There are 54 known transcripts in the fine location region. In the localized region of peach genome Pp06, 28 108 436 BP and 29 247 763. SNP markers were developed to identify the representative type after hybridization. The results showed that the results of genotypic and phenotypic identification of all progenies were identical, and the accuracy of identification was 100. [conclusion] in this study, the dwarf gene of peach was located with a physical distance of 277kb. For gene cloning, early screening of parents for breeding dwarf ornamental peach and rootstock varieties laid the foundation.
【作者單位】： 中國農(nóng)業(yè)科學(xué)院鄭州果樹研究所/國家桃葡萄品種改良中心/農(nóng)業(yè)部果樹育種技術(shù)重點實驗室;新西蘭植物與食品研究所;
【基金】：國家自然科學(xué)基金(31500558) 中國農(nóng)業(yè)科學(xué)院創(chuàng)新工程(CAAS-ASTIP-2017-ZFRI) 中央級科研院所基本科研業(yè)務(wù)費專項(1610192017702)
【分類號】：S662.1
【正文快照】： 0引言【研究意義】桃[Prunus persica(L.)Batsch]是中國栽培面積較大的落葉果樹之一。2014年中國桃栽培面積約799 500 hm2,居世界桃栽培第一位(FAO)。其中,矮化型桃因其節(jié)間短、成花好是觀賞桃和砧木育種的發(fā)展方向之一。通過對桃矮化基因進(jìn)行精細(xì)定位是建立目標(biāo)性狀分子輔助

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