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An Allele of the Ripening-Specific 1-Aminocyclopropane-1-Carboxylic Acid Synthase Gene (ACS1) in Apple Fruit with a Long Storage Life1

机译:特定于成熟的等位基因 1-氨基环丙烷-1-羧酸合酶基因(ACS1) 苹果果实中的保质期长1

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摘要

An allele of the 1-aminocyclopropane-1-carboxylic acid (ACC) synthase gene (Md-ACS1), the transcript and translated product of which have been identified in ripening apples (Malus domestica), was isolated from a genomic library of the apple cultivar, Golden Delicious. The predicted coding region of this allele (ACS1-2) showed that seven nucleotide substitutions in the corresponding region of ACS1-1 resulted in just one amino acid transition. A 162-bp sequence characterized as a short interspersed repetitive element retrotransposon was inserted in the 5′-flanking region of ACS1-2 corresponding to position −781 in ACS1-1. The XhoI site located near the 3′ end of the predicted coding region of ACS1-2 was absent from the reverse transcriptase-polymerase chain reaction product, revealing that exclusive transcription from ACS1-1 occurs during ripening of cv Golden Delicious fruit. DNA gel-blot and polymerase chain reaction analyses of genomic DNAs showed clearly that apple cultivars were either heterozygous for ACS1-1 and ACS1-2 or homozygous for each type. RNA gel-blot analysis of the ACS1-2 homozygous Fuji apple, which produces little ethylene and has a long storage life, demonstrated that the level of transcription from ACS1-2 during the ripening stage was very low.
机译:从苹果的基因组文库中分离出1-氨基环丙烷-1-羧酸(ACC)合酶基因(Md-ACS1)的等位基因,其转录本和翻译产物已在成熟的苹果(Malus domestica)中鉴定出。品种,金冠。该等位基因(ACS1-2)的预测编码区域显示,ACS1-1对应区域中的七个核苷酸取代仅导致一个氨基酸过渡。在与ACS1-1中的位置-781相对应的ACS1-2的5'侧翼区域中插入162bp的序列,其特征为短的散布的重复元件逆转录转座子。逆转录酶-聚合酶链反应产物中没有位于ACS1-2预测编码区3'末端附近的XhoI位点,表明ACS1-1独家转录发生在cv Golden Delicious果实成熟期间。基因组DNA的DNA凝胶印迹和聚合酶链反应分析清楚地表明,苹果品种对于ACS1-1和ACS1-2是杂合的,或者对于每种类型都是纯合的。 ACS1-2纯合的富士苹果的RNA凝胶印迹分析结果表明,乙烯苹果几乎不产生乙烯并且具有长的存储寿命,因此在成熟阶段从ACS1-2转录的水平非常低。

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