本文選題：甘藍(lán)型油菜 + 耐鹽性狀�。� 參考：《中國農(nóng)業(yè)科學(xué)》2017年07期

【摘要】：【目的】通過對甘藍(lán)型油菜耐鹽相關(guān)性狀進(jìn)行全基因組關(guān)聯(lián)分析,尋找可能與油菜耐鹽性相關(guān)的SNP位點(diǎn),發(fā)掘與油菜耐鹽性有關(guān)的候選基因。【方法】以1.2%NaCl溶液作為培養(yǎng)液,去離子水為對照,對307個(gè)不同品系的甘藍(lán)型油菜進(jìn)行發(fā)芽試驗(yàn)。播種后7 d測定幼苗根長、鮮重及發(fā)芽率,計(jì)算鹽脅迫下各性狀相對值,并作為評價(jià)耐鹽的指標(biāo)。結(jié)合油菜60K SNP芯片,利用SPAGeDi v1.4軟件對該群體307份甘藍(lán)型油菜進(jìn)行親緣關(guān)系分析,并計(jì)算親緣關(guān)系值的矩陣。利用軟件STRUCTURE v2.3.4對關(guān)聯(lián)群體進(jìn)行了群體結(jié)構(gòu)分析。為有效排除假關(guān)聯(lián)的影響,將群體結(jié)構(gòu)和材料間的親緣關(guān)系考慮進(jìn)關(guān)聯(lián)分析中,同時(shí)進(jìn)行了PCA+K模型、Q+K模型以及K模型3種混合線性模型分析和比較,根據(jù)所有SNP的 lg(P)觀察值和期望值,確定每個(gè)性狀GWAS分析的最優(yōu)模型。采用TASSEL 5.0軟件,在最優(yōu)模型下對307份材料耐鹽各性狀的相對值分別與SNP標(biāo)記進(jìn)行全基因組關(guān)聯(lián)分析。利用油菜基因組數(shù)據(jù)庫,在顯著SNP位點(diǎn)側(cè)翼序列200 kb范圍內(nèi)提取基因。根據(jù)擬南芥中已經(jīng)明確功能的耐鹽相關(guān)基因,篩選出目標(biāo)基因組區(qū)段內(nèi)與耐鹽相關(guān)的油菜同源基因。【結(jié)果】全基因組關(guān)聯(lián)分析共檢測到164個(gè)與根長顯著關(guān)聯(lián)的SNP位點(diǎn),23個(gè)與鮮重顯著關(guān)聯(lián)的SNP位點(diǎn),38個(gè)與發(fā)芽率顯著關(guān)聯(lián)的SNP位點(diǎn)。其中與根長、鮮重、發(fā)芽率最顯著關(guān)聯(lián)的SNP位點(diǎn)分別位于染色體A08、A02和A06,貢獻(xiàn)率分別為23.84%、18.59%和31.81%。在這些顯著SNP位點(diǎn)側(cè)翼序列200 kb范圍內(nèi)發(fā)掘出可能與油菜耐鹽性有關(guān)的50個(gè)候選基因。這些候選基因主要包括轉(zhuǎn)錄因子MYB、WRKY、ABI1、b ZIP、ERF1、CZF1、XERICO等以及一些下游受轉(zhuǎn)錄因子調(diào)控的不同功能基因NHX1、PTR3、CAT1、HKT、CAX1、ACER、STH、STO等。在根長和發(fā)芽率2個(gè)不同耐鹽性狀的分析結(jié)果中均篩選出位于A03染色體上的耐鹽基因BnaA03g14410D。另外,這些耐鹽候選基因中包含兩組串聯(lián)重復(fù)基因,分別是位于A03染色體上的BnaA03g18900D和BnaA03g18910D,位于C09染色體上的BnaC09g19080D、BnaC09g19090D和BnaC09g19100D。除此之外,耐鹽候選基因中還包含2個(gè)距離非常近(中間只間隔2個(gè)基因)的重復(fù)基因BnaC02g39600D和BnaC02g39630D�！窘Y(jié)論】共檢測到225個(gè)與耐鹽性狀顯著關(guān)聯(lián)的SNP位點(diǎn),篩選出50個(gè)可能與油菜耐鹽性有關(guān)的候選基因。
[Abstract]:[objective] to identify the SNP sites associated with salt tolerance in Brassica napus by genome association analysis, and to identify candidate genes related to salt tolerance in Brassica napus. [methods] 1.2%NaCl solution was used as culture medium. The germination test of 307 different lines of Brassica napus was carried out in deionized water as control. The root length, fresh weight and germination rate of seedlings were measured 7 days after sowing, and the relative values of the characters under salt stress were calculated and used as an index to evaluate salt tolerance. Combined with 60K SNP microarray, SPAGeDi v1.4 software was used to analyze the phylogenetic relationship of 307 Brassica napus in this population, and the matrix of relationship value was calculated. The group structure of associated population was analyzed by software STRUCTURE v2.3.4. In order to eliminate the influence of false association, the relationship between population structure and material was taken into account in the association analysis, and the PCA K model, QK model and K model were analyzed and compared. According to the observed values and expected values of all SNP, the optimal model of GWAS analysis for each trait was determined. Under the optimal model, TASSEL 5.0 software was used to analyze the whole genome association between the relative values of each trait of salt tolerance and SNP markers. Using rapeseed genome database, the gene was extracted from the flanking sequence of significant SNP locus within 200kb. According to the salt-tolerant genes in Arabidopsis thaliana, The homologous genes related to salt tolerance in the target genome region were screened. [results] A total of 164 SNP loci were detected in the whole genome association analysis, 23 SNP loci were significantly associated with fresh weight, 38 loci were associated with hair and root length. Bud rate was significantly associated with the SNP locus. The SNP loci, which were most significantly related to root length, fresh weight and germination rate, were located on chromosome A08A02 and A06respectively, and the contribution rates were 23.848.59% and 31.81%, respectively. Fifty candidate genes which may be related to brassica napus salt tolerance were identified within the range of 200kb flanking sequence of these significant SNP loci. These candidate genes mainly include the transcription factor MYBX WRKYN ABI1nb ZIPERF1 CZF1 + XERICO, and some downstream genes, NHX1PTR3, CAT1, HKT1, CAX1, ACERCER, STHSTO, and so on. Two salt-tolerant genes, BnaA03g14410D, which were located on chromosome A03, were screened from two different salt-tolerant traits of root length and germination rate. In addition, these salt tolerance candidate genes contain two sets of tandem repeat genes, BnaA03g18900D and BnaA03g18910D located on chromosome A03, BnaC09g19080D on chromosome C09 and BnaC09g19090D and BnaC09g19100D. In addition, two repeat genes, BnaC02g39600D and BnaC02g39630D. [conclusion] 225 SNP sites were found to be significantly associated with salt tolerance traits. Fifty candidate genes that may be related to salt tolerance in rape were screened.
【作者單位】： 西南大學(xué)農(nóng)學(xué)與生物科技學(xué)院;
【基金】：國家重點(diǎn)研發(fā)計(jì)劃“七大農(nóng)作物育種”重點(diǎn)專項(xiàng)(2016YFD0100202) 國家自然科學(xué)基金(31671729、31301351) 中央高�；究蒲袠I(yè)務(wù)費(fèi)(XDJK2016B031)
【分類號】：S565.4

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