Aukerman M J, Schmidt R J
Department of Biology, University of California, San Diego, La Jolla 92093.
Plant Mol Biol. 1993 Jan;21(2):355-62. doi: 10.1007/BF00019950.
From a directed transposon tagging of the maize Opaque-2 gene (O2), we have isolated a stable mutant o2 allele, o2-23. Cloning and molecular analysis of the allele revealed a 168 nucleotide insertion in the third exon of o2. The sequence of this small insertion indicated identity with the 5' and 3' ends of the 8.3 kb Suppressor-mutator/Enhancer (Spm/En) transposable element. This represents the smallest deletion derivative of Spm (dSpm) thus far characterized in maize. Genetic crosses of plants homozygous for o2-23 with plants homozygous for both an o2 null allele (o2-R) and an autonomous Spm produce stable opaque seed having no apparent sectors of vitreous endosperm. DNA fragments of the size expected if the dSpm were to excise were not detectable by Southern analysis, suggesting that this element is unable to transpose. Northern analysis detected an o2-23 mRNA that was much more abundant in o2-23 seeds lacking Spm than in o2-23 seeds containing Spm, consistent with the idea that Spm transacting functions can suppress the accumulation of the o2-23 transcript.
通过对玉米不透明-2基因(O2)进行定向转座子标签实验,我们分离出了一个稳定的突变体o2等位基因o2-23。对该等位基因的克隆和分子分析显示,o2的第三个外显子中有一个168个核苷酸的插入。这个小插入片段的序列与8.3 kb抑制子-突变体/增强子(Spm/En)转座元件的5'和3'末端相同。这代表了迄今为止在玉米中鉴定出的最小的Spm缺失衍生物(dSpm)。o2-23纯合植株与o2无效等位基因(o2-R)和自主Spm均纯合的植株进行遗传杂交,产生的稳定不透明种子没有明显的玻璃质胚乳区域。Southern分析未检测到如果dSpm切除预期大小的DNA片段,这表明该元件无法转座。Northern分析检测到一种o2-23 mRNA,在不含Spm的o2-23种子中比在含有Spm的o2-23种子中丰富得多,这与Spm反式作用功能可以抑制o2-23转录本积累的观点一致。