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鉴定决定洋葱显性白色鳞茎颜色的 I 位点的候选基因。

Identification of a candidate gene for the I locus determining the dominant white bulb color in onion (Allium cepa L.).

机构信息

Department of Horticulture, Biotechnology Research Institute, Chonnam National University, Gwangju, 61186, Republic of Korea.

Genomics Division, National Institute of Agricultural Sciences, RDA, Jeonju, 54874, Republic of Korea.

出版信息

Theor Appl Genet. 2024 May 6;137(6):118. doi: 10.1007/s00122-024-04626-9.

Abstract

Through a map-based cloning approach, a gene coding for an R2R3-MYB transcription factor was identified as a causal gene for the I locus controlling the dominant white bulb color in onion. White bulb colors in onion (Allium cepa L.) are determined by either the C or I loci. The causal gene for the C locus was previously isolated, but the gene responsible for the I locus has not been identified yet. To identify candidate genes for the I locus, an approximately 7-Mb genomic DNA region harboring the I locus was obtained from onion and bunching onion (A. fistulosum) whole genome sequences using two tightly linked molecular markers. Within this interval, the AcMYB1 gene, known as a positive regulator of anthocyanin production, was identified. No polymorphic sequences were found between white and red AcMYB1 alleles in the 4,860-bp full-length genomic DNA sequences. However, a 4,838-bp LTR-retrotransposon was identified in the white allele, in the 79-bp upstream coding region from the stop codon. The insertion of this LTR-retrotransposon created a premature stop codon, resulting in the replacement of 26 amino acids with seven different residues. A molecular marker was developed based on the insertion of this LTR-retrotransposon to genotype the I locus. A perfect linkage between bulb color phenotypes and marker genotypes was observed among 5,303 individuals of segregating populations. The transcription of AcMYB1 appeared to be normal in both red and white onions, but the transcription of CHS-A, which encodes chalcone synthase and is involved in the first step of the anthocyanin biosynthesis pathway, was inactivated in the white onions. Taken together, an aberrant AcMYB1 protein produced from the mutant allele might be responsible for the dominant white bulb color in onions.

摘要

通过图谱克隆方法,鉴定出一个编码 R2R3-MYB 转录因子的基因是控制洋葱显性白色鳞茎颜色的 I 位点的一个因果基因。洋葱的白色鳞茎颜色由 C 或 I 位点决定。C 位点的因果基因已被分离出来,但负责 I 位点的基因尚未被鉴定出来。为了鉴定 I 位点的候选基因,从洋葱和分葱(A. fistulosum)全基因组序列中使用两个紧密连锁的分子标记获得了大约 7-Mb 的基因组 DNA 区域,该区域包含 I 位点。在这个区间内,鉴定出 AcMYB1 基因,它是花青素生物合成的正调控因子。在 4860-bp 全长基因组 DNA 序列中,白色和红色 AcMYB1 等位基因之间没有发现多态性序列。然而,在白色等位基因中,在终止密码子上游的 79-bp 编码区中发现了一个 4838-bp 的 LTR-逆转录转座子。这个 LTR-逆转录转座子的插入产生了一个提前终止密码子,导致 26 个氨基酸被 7 个不同的残基取代。基于这个 LTR-逆转录转座子的插入,开发了一个分子标记来对 I 位点进行基因分型。在分离群体的 5303 个个体中,观察到鳞茎颜色表型和标记基因型之间存在完美的连锁。在红色和白色洋葱中,AcMYB1 的转录似乎是正常的,但参与花青素生物合成途径第一步的 CHS-A 的转录在白色洋葱中失活。总的来说,突变等位基因产生的异常 AcMYB1 蛋白可能是洋葱显性白色鳞茎颜色的原因。

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