作物学报 ›› 2014, Vol. 40 ›› Issue (02): 355-361.doi: 10.3724/SP.J.1006.2014.00355
廖芳蕾1,王鲁2,辛可行1,陈文荣1,郭卫东1,*
LIAO Fang-Lei1,WANG Lu2,XIN Ke-Xing1,CHEN Wen-Rong1,GUO Wei-Dong1,*
摘要:
为探讨反义寡聚核苷酸抑制(antisense oligodeoxynucleotides inhibition)技术在植物材料上的应用, 以已知功能的NtGNL1 (Nicotiana tabacum GNOM-Like1)为目标基因, 寻找反义寡聚核苷酸抑制最适作用体系, 并进一步分析NtGNL1的具体作用。根据目标基因mRNA序列设计反义寡聚核苷酸序列, 商业合成后并将其添加到烟草胚珠、种子和花粉管离体培养的培养基中, 以抑制NtGNL1的表达。结果表明, 将反义寡聚核苷酸引入离体培养系统, 短时间内抑制了目标基因mRNA的表达, 但对胚胎发育过程的影响和种子萌发的抑制都不明显。在花粉离体萌发系统中, 反义寡聚核苷酸抑制能够高效地引起目标基因mRNA的下调。显微缩时对FM4-64染色的花粉管观察发现, NtGNL1的下调能够引起囊泡分布和运输方向的改变。对花粉管膜流相关的5个基因的半定量分析也显示反义寡聚核苷酸进入花粉管后, 导致3个基因的表达下调, 暗示NtGNL1抑制表达会影响花粉管囊泡运输的多个节点。
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