Positional cloning of rice semidwarfing gene, sd-1:: Rice "Green revolution gene" encodes a mutant enzyme involved in gibberellin synthesis
Positional cloning of rice semidwarfing gene, sd-1:: Rice "Green revolution gene" encodes a mutant enzyme involved in gibberellin synthesis
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DOI：
10.1093/dnares/9.1.11
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发表时间：
2002-02-28
期刊：
影响因子：
4.1
通讯作者：
Minobe, Y
中科院分区：
文献类型：
--
作者：
Monna, L;Kitazawa, N;Minobe, Y
通过定位克隆获得了水稻半矮秆基因SD-1，被称为“绿色革命基因”，编码赤霉素生物合成途径中的关键酶赤霉素20-氧化酶。利用几种基于PCR的标记技术，包括裂解扩增多态序列、衍生CAPS和单核苷酸多态，对3477个分离物进行了分析，发现在6kb的候选区间内有1个ORF。正常类型水稻品种在该区域具有相同的序列，由3个外显子(558、318和291个核苷酸)和2个内含子(105和1471个核苷酸)组成。Dee-Geo-Woo-Gen型SD-1突变体在基因组中有383个碱基的缺失(从表达序列中缺失278个碱基)，从外显子1的中部到外显子2的上游，包括一个105个碱基的内含子，导致帧移位，在缺失位点之后产生一个终止密码子。辐射诱导的SD-1突变体CalRose 76在外显子2有1个碱基的替换，导致氨基酸替换(Leu[CTC]到Phe[TTC])。表达分析表明，至少还有一个赤霉素20-氧化酶基因座存在，这可能阻止了sd-1突变体的严重矮化。
A rice semidwarfing gene, sd-1, known as the "green revolution gene," was isolated by positional cloning and revealed to encode gibberellin 20-oxidase, the key enzyme in the gibberellin biosynthesis pathway. Analysis of 3477 segregants using several PCR-based marker technologies, including cleaved amplified polymorphic sequence, derived-CAPS, and single nucleotide polymorphisms revealed 1 ORF in a 6-kb candidate interval. Normal-type rice cultivars have an identical sequence in this region, consisting of 3 exons (558, 318, and 291 bp) and 2 introns (105 and 1471 bp). Dee-Geo-Woo-Gen-type sd-1 mutants have a 383-bp deletion from the genome (278-bp deletion from the expressed sequence), from the middle of exon 1 to upstream of exon 2, including a 105-bp intron, resulting in a frame-shift that produces a termination codon after the deletion site. The radiation-induced sd-1 mutant Calrose 76 has a 1-bp substitution in exon 2, causing an amino acid substitution (Leu [CTC] to Phe [TTC]). Expression analysis suggests the existence of at least one more locus of gibberellin 20-oxidase which may prevent severe dwarfism from developing in sd-1 mutants.