本文選題：甘藍型油菜 + 耐鹽性狀��； 參考：《中國農業(yè)科學》2017年07期

【摘要】：【目的】通過對甘藍型油菜耐鹽相關性狀進行全基因組關聯(lián)分析,尋找可能與油菜耐鹽性相關的SNP位點,發(fā)掘與油菜耐鹽性有關的候選基因�！痉椒ā恳�1.2%NaCl溶液作為培養(yǎng)液,去離子水為對照,對307個不同品系的甘藍型油菜進行發(fā)芽試驗。播種后7 d測定幼苗根長、鮮重及發(fā)芽率,計算鹽脅迫下各性狀相對值,并作為評價耐鹽的指標。結合油菜60K SNP芯片,利用SPAGeDi v1.4軟件對該群體307份甘藍型油菜進行親緣關系分析,并計算親緣關系值的矩陣。利用軟件STRUCTURE v2.3.4對關聯(lián)群體進行了群體結構分析。為有效排除假關聯(lián)的影響,將群體結構和材料間的親緣關系考慮進關聯(lián)分析中,同時進行了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標記進行全基因組關聯(lián)分析。利用油菜基因組數(shù)據(jù)庫,在顯著SNP位點側翼序列200 kb范圍內提取基因。根據(jù)擬南芥中已經明確功能的耐鹽相關基因,篩選出目標基因組區(qū)段內與耐鹽相關的油菜同源基因�！窘Y果】全基因組關聯(lián)分析共檢測到164個與根長顯著關聯(lián)的SNP位點,23個與鮮重顯著關聯(lián)的SNP位點,38個與發(fā)芽率顯著關聯(lián)的SNP位點。其中與根長、鮮重、發(fā)芽率最顯著關聯(lián)的SNP位點分別位于染色體A08、A02和A06,貢獻率分別為23.84%、18.59%和31.81%。在這些顯著SNP位點側翼序列200 kb范圍內發(fā)掘出可能與油菜耐鹽性有關的50個候選基因。這些候選基因主要包括轉錄因子MYB、WRKY、ABI1、b ZIP、ERF1、CZF1、XERICO等以及一些下游受轉錄因子調控的不同功能基因NHX1、PTR3、CAT1、HKT、CAX1、ACER、STH、STO等。在根長和發(fā)芽率2個不同耐鹽性狀的分析結果中均篩選出位于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個距離非常近(中間只間隔2個基因)的重復基因BnaC02g39600D和BnaC02g39630D�！窘Y論】共檢測到225個與耐鹽性狀顯著關聯(lián)的SNP位點,篩選出50個可能與油菜耐鹽性有關的候選基因。
[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】： 西南大學農學與生物科技學院;
【基金】：國家重點研發(fā)計劃“七大農作物育種”重點專項(2016YFD0100202) 國家自然科學基金(31671729、31301351) 中央高校基本科研業(yè)務費(XDJK2016B031)
【分類號】：S565.4

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