Wisman E, Koornneef M, Chase T, Lifshytz E, Ramanna M S, Zabel P
Department of Molecular Biology, Agricultural University, Wageningen, The Netherlands.
Mol Gen Genet. 1991 Apr;226(1-2):120-8. doi: 10.1007/BF00273595.
Treatment of tomato seeds with ethyl methanesulphonate (EMS) followed by allyl alcohol selection of M2 seeds has led to the identification of one plant (B15-1) heterozygous for an alcohol dehydrogenase (Adh) null mutation. Genetic analysis and expression studies indicated that the mutation corresponded to the structural gene of the Adh-1 locus on chromosome 4. Homozygous Adh-1 null mutants lacked ADH-1 activity in both pollen and seeds. Using an antiserum directed against ADH from Arabidopsis thaliana, which cross-reacts with ADH-1 and ADH-2 proteins from tomato, no ADH-1 protein was detected in seeds of the null mutant. Northern blot analysis showed that Adh-1 mRNA was synthesized at wild-type levels in immature seeds of the null mutant, but dropped to 25% in mature seeds. Expression of the Adh-2 gene on chromosome 6 was unaffected. The potential use of the Adh-1 null mutant in selecting rare transposon insertion mutations in a cross with "mutable" Adh-1+ tomato lines is discussed.
用甲磺酸乙酯(EMS)处理番茄种子,随后对M2种子进行烯丙醇筛选,已鉴定出一株对酒精脱氢酶(Adh)无效突变杂合的植株(B15 - 1)。遗传分析和表达研究表明,该突变对应于4号染色体上Adh - 1位点的结构基因。纯合的Adh - 1无效突变体在花粉和种子中均缺乏ADH - 1活性。使用针对拟南芥ADH的抗血清,其与番茄的ADH - 1和ADH - 2蛋白发生交叉反应,在无效突变体的种子中未检测到ADH - 1蛋白。Northern印迹分析表明,Adh - 1 mRNA在无效突变体未成熟种子中以野生型水平合成,但在成熟种子中降至25%。6号染色体上Adh - 2基因的表达未受影响。讨论了Adh - 1无效突变体在与“可变”Adh - 1 +番茄品系杂交中用于筛选罕见转座子插入突变的潜在用途。