基因枪法转化籼稻有关因素的评价

 利用基因枪轰击水稻未成熟胚及未成熟胚诱导来源的愈伤组织，对国内外16个籼稻品种进行了转化。设置不同的基因枪转化参数，并在培养基中添加各种植物激素、甘露醇以及2-N-吗啉-乙基磺酸(简称MES)来改善继代过程中籼稻愈伤组织的生长状态。基因枪转化的操作参数（包括金粉浓度和DNA含量、基因枪氦气压力、轰击受体的不同状态）对转化频率有重要影响；在继代培养基中添加MES（500 mg/L）作为缓冲剂，可以减少籼稻愈伤组织褐化现象的发生；在诱导和继代培养基中补加NAA(0.5 mg/L），KT(0.2 mg/L)可改进愈伤组织质量，进而提高再生频率；在继代培养基中补加低浓度的 ABA(1 mg/L)和甘露醇（20 g/L），能普遍改善愈伤组织生长状况，减少褐化现象的产生，得到较多结构致密的颗粒状抗性愈伤。获得了再生的转化植株，经检测表现出对Basta的抗性。

Factors Involved in Microprojectile-Mediated Transformation of Indica Rice

Immature embryos and embryogenic calli of 16 indica rice cultivars have been transformed usingparticle bombardment. We set up a variety of parameters to optimize the important factors involved in biolistictransformation of indica rice, and supplement various hormones , mannitol and MES to NB medium in order to improve growth state of subcultured calli. Results indicate that operation parameters (include concentration of AuDNA complex, helium pressure, and receptor state) involved in microprojectile-mediated transformat ion are veryimportant to transformation rate; MES (500 mg/L) added to the subculture medium as a buffer can mitigatebrowning of indica rice calli; supplementing NAA(0. 5 mg/L),KT(0. 2 mg/L) to the induce medium and the subculture medium can improve the quality of calli and increase the regeneration rate ; ABA(1 mg/L) and mannitol (20g/L) can improve the growth state of calli, mitigate browning and obtain more granular Basta calli that are fineand close in texture. The transgenic plants resistant to Basta were obtained.