Kunz C, Schöb H, Leubner-Metzger G, Glazov E, Meins F
Friedrich Miescher Institute, Basel, Switzerland.
Planta. 2001 Jan;212(2):243-9. doi: 10.1007/s004250000383.
Nicotiana sylvestris Speg. & Comes transformed with a tobacco class-I beta-1,3-glucanase (GLU I ) cDNA driven by CaMV 35S RNA expression signals exhibits posttranscriptional gene silencing (PTGS) which is triggered between the cotyledon and two-leaf stages of seedling development and is postmeiotically reset to the high-expressing state during seed development. The incidence of GLU I PTGS in sibling plants differed for the two different transformants tested and increased with the number of T-DNA loci. Comparison of host class-I and class-II beta-1,3-glucanase gene expression suggests that a similarity of 60-70% in the coding-region is required for PTGS of the homologous host genes. The GLU I transformants exhibited a spatial gradient in PTGS, in which expression of the silent phenotype gradually increased in successive leaves toward the bottom of the plant. In contrast, transformants carrying an unrelated tobacco class I chitinase (CHN I) cDNA in the same expression vector exhibited discontinuous patterns of PTGS with adjacent high-expressing and silent leaves. The GLU I- and CHN I-specific patterns were maintained in hybrids homozygous for both T-DNA's indicating that two different transgenes present in the same genome can exhibit independent and distinctive patterns of PTGS. This implies that the nature of the transgene rather than a general pre-pattern of competence for PTGS or propagation of the silent state are important for pattern determination.
用由花椰菜花叶病毒35S RNA表达信号驱动的烟草I类β-1,3-葡聚糖酶(GLU I)cDNA转化的野生烟草(Nicotiana sylvestris Speg. & Comes)表现出转录后基因沉默(PTGS),该沉默在幼苗发育的子叶期和两叶期之间触发,并在种子发育过程中减数分裂后重置为高表达状态。在所测试的两种不同转化体中,同胞植株中GLU I PTGS的发生率不同,并且随着T-DNA位点数量的增加而增加。宿主I类和II类β-1,3-葡聚糖酶基因表达的比较表明,同源宿主基因的PTGS需要编码区60-70%的相似性。GLU I转化体在PTGS中表现出空间梯度,其中沉默表型的表达在朝着植株底部的连续叶片中逐渐增加。相反,在同一表达载体中携带不相关的烟草I类几丁质酶(CHN I)cDNA的转化体表现出PTGS的不连续模式,相邻叶片有高表达和沉默的情况。对于两个T-DNA均为纯合的杂种,GLU I和CHN I特异性模式得以维持,这表明存在于同一基因组中的两个不同转基因可以表现出独立且独特的PTGS模式。这意味着转基因的性质而非PTGS能力的一般预模式或沉默状态的传播对于模式确定很重要。