棉花转基因体细胞胚胎的诱导及植株再生

Abstract

以陆地棉种质系Coker 312为材料，探讨了三种氨基酸对棉花转基因胚性愈伤生长、胚状体诱导、萌发及NAA对转基因再生植株生根的影响，建立一个高效的转基因体细胞胚萌发系统。结果表明：单独使用Pro、Glu时能提高转基因胚性愈伤增长、胚状体诱导及萌发能力，但以综合使用效果更为显著，Ala的附加没有显著影响；当生根培养基NAA的浓度为0.3 mg/L，根的诱导数量及质量都优于对照。但NAA的浓度不宜过高，过高则加重了根的褐化程度。对得到的转基因再生植株进行Kan涂抹实验，Kan抗性苗获得率可达到11.53%；PCR检测约83.78%的Kan抗性苗呈阳性。 In this paper, the effect of three amino acids on the growth of embryonic calli, embryo germination after transformation and the effect of Naphthlcetic acid (NAA) on rooting in cotton (Gossypium hirsutum L.) were investigated. The results showed that: embryogenic callus growth and embryo germination ability were effectively improved on the medium with Glu and Pro, especially with 0.5 mg/LGlu and 0.5 mg/L Pro at the same time. However, the additional of Alahad no significant impact; Using 0.3 mg/L NAA in the rooting medium could produce more strong roots, but too high NAA concentration increased the level of root browning. The percentage of Kan-resistant regenerated plantlets reached 11.53% according to the Kanamycin smear experiments and about 83.78% of the Kan-resistant plantlets were proven to be PCR-positive. The successful transformation and regeneration protocol established in this study can be used to improve cotton cultivars by genetic engineering.