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以 70 个高代玉米自交系的幼胚为材料在 NB 培养基上进行离体培养,通过多次继代选择,从中筛选出了胚性愈伤组织诱导率高、克隆能力强的 10 个自交系,将这 10 个自交系的一部分胚性愈伤组织用作绿苗再分化,研究其再生能力;另一部分用农杆菌 C58 和 GV3301 转化,利用植物载体中携带的 gus 和 gfp 报告基因的瞬时表达为指标研究玉米基因型对农杆菌的敏感性。绿苗再分化结果表明,10 个自交系都具有绿苗再分化能力,其中自交系 6010、6038、6015、6051 和 6060的绿苗再分化力相对较高,分别为 61.11%、31.94%、45%、33.33%和 56.94%。gus 瞬时表达率方差分析结果表明:玉米基因型对农杆菌 C58 的敏感性存在极显著差异,在自交系 6034、6038 的胚性愈伤组织上没有检测到 gus 瞬时表达,即这两个基因型对农杆菌 C58 不敏感,不能被其转化;其他 7 个基因型的平均 gus 瞬时表达率均大于 70%,说明这 7 个基因型对农杆菌 C58 敏感。GFP 荧光检测结果表明,在自交系 6034、6038、6069 和 6010 的胚性愈伤组织上没有检测到 gfp 瞬时表达,在其他 6 个自交系上检测到绿色荧光蛋白的表达。因此,认为自交系 6015、6051 和 6060 是对农杆菌 GV3301 敏感且具有高再生能力的玉米转基因受体材料;自交系 6051、6010、6015、6060 和 6050 是再生能力强且对农杆菌 C58 敏感的玉米转基因受体材料。
The embryos of 70 elite inbred lines of Maize were cultured in vitro on NB medium. After several successive generations of selection, the embryogenic callus induction rate was high and the clonal ability was strong Inbred lines, a part of these 10 inbred lines of embryogenic callus were used as redifferentiation of green shoots to study their regeneration ability; the other part was transformed with Agrobacterium tumefaciens C58 and GV3301 and the gus and gfp carried in plant vectors Transient expression of genes as an indicator of corn genotype Agrobacterium sensitivity. The redifferentiation of green plantlets showed that all of the 10 inbred lines had the ability of redistribution of green plantlets, of which the redistillation ability of green plantlets in inbred lines 6010, 6038, 6015, 6051 and 6060 were relatively high, which were 61.11% and 31.94% , 45%, 33.33% and 56.94% respectively. The results of variance analysis of instantaneous expression rate of gus showed that the sensitivity of Agrobacterium to C58 was significantly different, and no transient expression of gus was detected on the embryogenic callus of inbred lines 6034 and 6038, that is, Were not sensitive to Agrobacterium C58 and could not be transformed by Agrobacterium tumefaciens C58. The average gus transient expression rates of the other seven genotypes were more than 70%, indicating that these seven genotypes were sensitive to Agrobacterium C58. GFP fluorescence detection results showed that gfp transient expression was not detected in the embryogenic callus of inbred lines 6034, 6038, 6069 and 6010, and the expression of green fluorescent protein was detected in the other six inbred lines. Therefore, it is considered that inbred lines 6015, 6051 and 6060 are maize transgenic materials that are sensitive to Agrobacterium tumefaciens GV3301 and have high regenerative ability. Inbred lines 6051, 6010, 6015, 6060 and 6050 are highly regenerative and resistant to Agrobacterium tumefaciens C58 Sensitive maize transgene receptor material.