转基因甘蓝型油菜品系W-4高油酸性状遗传分析

Inheritance of high oleic acid trait in the transgenic line W-4 of Brassica napus

为研究转基因甘蓝型油菜(Brassica napus L.)高油酸品系W-4的高油酸性状的遗传,用W-4(P1)及其野生型Westar(P2)为亲本,配置正反杂交组合,衍生获得F1、RF1、F2、RF2,B1、B2和RB1、RB2等世代群体。应用气相色谱定量分析方法,检测亲本及衍生世代群体成熟种子中油酸含量;同时,在苗期检测各个世代群体对卡那霉素(Kan)抗性的反应。结果显示:W-4种子中油酸平均含量为(85.10±0.73)%,Westar为(62.16±3.68)%;F1、RF1群体油酸平均含量分别为(77.55±1.30)%和(77.34±2.46)%,均表现为高油酸含量(≥69.5%),无细胞质效应;B1、RB1群体全部表现为高油酸(≥69.5%)。B2、RB2群体中,高油酸个体分别为55个和52个,低油酸个体分别为45个和48个,均呈1∶1分离(χ2=1.00,χ2=0.16);F2、RF2分离群体中,高油酸与低油酸个体呈3∶1分离(χ2=0.96,χ2=0.11)。表明,P1的高油酸性状受1对显性基因控制,呈孟德尔遗传,无细胞质遗传效应。此外,苗期Kan抗性检测结果表明,P1、F1、RF1、B1、RB1群体植株对Kan反应均表现为抗;P2对Kan反应表现为感;B2、RB2群体中抗、感植株呈1∶1分离,F2、RF2群体中抗、感植株呈3∶1分离。表明P1的卡那霉素抗性受1对显性基因控制。

To investigate the inheritance of high oleic acid content of W-4, a transgenic line of Brassica napus L. transformed with a ihpRNAi vector targeting the delta-12 oleate desaturase （fad2） gene, the generations derived from W-4 （ PI ） Westar（ wild type, P2 ） including F1 （ RF1 ）, F2 （ RF2 ） , B1 （ RB1 ）, B2 （ RB2 ） were collected. Oleie acid con- tents of the self-pollinated seeds harvested from parents and derived generations were analyzed by gas chromatography, and the plant resistances of parents and derived generations to kanamycin （Kan） were investigated at seedling stage. Results showed that the oleic acid contents in the seeds of P1 and P2 were （85.10±0.73）% and （62. 16±3.68）%, respectively, and those of F1 and RF1 were （ 77.55 ± 1.30 ） % and （ 77. 34 ± 2.46 ） %, respectively, which were high（ ≥69. 5% ）. The seeds of Bl and RBl all had high contents of oleic acid （ 〉169. 5% ）. The seeds from the populations of B2and RB2 gave a segregation ratio of 1 ： 1 （ χ2 = 1.0, χ2 = 0. 16）for high oleic acid （ ≥69. 5% ） ： low oleic acid （ 〈69. 5 % ） content, and those of F2 and RF2 gave a segregation ratio of 3 ： 1 （χ2 = 0.96,X2 = 0. 11 ）. All these suggested that the oleic acid content of W-4 was controlled by one pair of dominant gene and inherited in the mode of Mendelian. There was no cytoplasmic effect on the oleic content in this transgenic high oleic cross in Brassica napus L. In addition, the plants of P1 ,F1, RF1 ,B1, and RB1 exhibited kanamycin-resistant whereas the plants of P2 were sensitive to kanamycin. And the ratios of kanamycin-resistant to kanamycin-sensitive for the plants of B2 and RB2 were 1 ： 1 and 3 ： 1 for F2 and RF2. These indicated that the resistance of P1 to kanamicin was controlled by one pair of dominant gene.