人工合成甘蓝型油菜早期世代基因组变异的AFLP和MSAP标记

摘要

为了揭示人工甘蓝型油菜早期世代遗传和表观遗传变异规律,以A组合(大黄油菜×中花芥蓝)S0世代、B组合(大黄油菜×中迟芥蓝)S0和S1世代人工甘蓝型油菜为材料,分别利用AFLP和MSAP技术检测基因组变化及甲基化模式变化情况.结果表明,16对引物在A组合S0扩增到523条带,其中4对引物扩增出9条变异带,包括7条亲本缺失带和2条新增带,分别占S0总条带的1.33％和0.38％; 45对引物在B组合双亲植株扩增到1093条带,只有1对引物检测到1条父本带型在所有S0植株中缺失,约占S0总条带的0.09％;在B9子代F19-1～F19-16总共扩增得到1092条带,变异带有10条,占总条带的0.915％,其中包括9条缺失带和1条新增带,9条缺失带全部位于C基因组.MSAP检测发现,B组合S0植株中有3个位点发生了甲基化模式的改变,全部位于A基因组,甲基化模式改变位点占总检测位点的1.37％.研究还发现B组合S0世代一个植株出现可遗传的花色变异,推测该表型变异与B组合人工甘蓝型油菜中C基因组变异有关.%In order to reveal pattern of genetic and epigenetic changes following allopolyploidization of A and C genomes in Brassica,AFLP (amplified fragment length polymorphism) was utilized to detect level of genome changes for S0 plants of cross A (Qinghai Dahuang×Zhonghua Jielan) and S0,S1 plants of cross B (Qinghai Dahuang×Zhongchi Jielan).Moreover,MSAP (methylation sensitive amplification polymorphism) was utilized to detect level of methylation changes for S0 plants of cross B.Five hundred and twenty three bands were amplified in S0 plants of cross A with 16 pairs of primer,nine out of these bands amplified showed significant differences between S0 and parents plants,mainly including deletion of seven parental restriction fragments and gain of two new fragments,accounting for 1.33％ and 0.38％ of total fragments,respectively.One thousand and ninety three bands were amplified in parental plants of B cross with 45 pairs of primer,only one from C genome out of these fragments lost in all S0 plants,accounting for 0.09％ of total bands.In 16 S1 plants from the B9 plant,10 out of 1092 bands amplified showed variations,consisting of the deletion of nine fragments from C genome and the gain of one new fragment.By MSAP approach,three loci with cytosine methylation changes were founded on A genome of S0 plants of B cross,which accounted for 1.37％ of loci detected.In addition,a plant with flower color mutation in S0 generation of B cross was observed; we speculated that the flower color mutation was probably caused by genetic changes on C genome.This study provided theoretical informations for innovation of rapeseed germplasm by resynthesizing Brassica napus.