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基于图谱的黄瓜果实白色不成熟颜色控制基因 w 的克隆、鉴定和特征分析。

Map-based cloning, identification and characterization of the w gene controlling white immature fruit color in cucumber (Cucumis sativus L.).

机构信息

College of Horticulture, Northwest A&F University, Yangling, Shaanxi, 712100, China.

出版信息

Theor Appl Genet. 2016 Jul;129(7):1247-1256. doi: 10.1007/s00122-016-2700-8. Epub 2016 Mar 2.

Abstract

A single-nucleotide insertion resulted in a premature stop codon that is responsible for white immature fruit color in cucumber. Despite our previous progress in the mapping of the gene controlling white color in immature cucumber fruit and the identification of candidate genes, the specific gene that governs chlorophyll metabolism and its regulatory mechanism remains unknown. Here, we generated a mapping population consisting of 9497 F2 plants to delimit the controlling gene to an 8.2-kb physical interval that defines a sole candidate gene, APRR2. Sequencing the full-length DNA and cDNA of APRR2 allowed for identification of an allele, aprr2, encoding a truncated 101-amino acid protein due to a frameshift mutation and a premature stop codon. Gene structure prediction indicated that these 101 residues are located in a domain necessary for the function of the protein. The expression patterns of APRR2 were entirely consistent with the visual changes in green color intensity during fruit development. A microscopic observation of the fruit pericarp revealed fewer chloroplasts and a lower chloroplast chlorophyll storage capacity in Q24 (white) than in Q30 (green). A single-base insertion in the white color gene w, which leads to a premature stop codon, is hypothesized to have disabled the function of this gene in chlorophyll accumulation and chloroplast development. These findings contribute to basic research and the genetic improvement of fruit color.

摘要

一个单核苷酸的插入导致了一个提前的终止密码子,这是黄瓜白色未成熟果实颜色的原因。尽管我们之前在定位控制未成熟黄瓜果实白色的基因和鉴定候选基因方面取得了进展,但控制叶绿素代谢及其调控机制的特定基因仍然未知。在这里,我们生成了一个由 9497 个 F2 植物组成的作图群体,将控制基因限定在一个 8.2kb 的物理区间内,该区间定义了一个唯一的候选基因 APRR2。对 APRR2 的全长 DNA 和 cDNA 进行测序,发现一个等位基因 aprr2 ,由于移码突变和提前终止密码子,编码一个截断的 101 个氨基酸的蛋白质。基因结构预测表明,这 101 个残基位于该蛋白功能所必需的结构域中。APRR2 的表达模式与果实发育过程中绿色强度的视觉变化完全一致。果皮的微观观察显示,与 Q30(绿色)相比,Q24(白色)的叶绿体数量较少,叶绿体叶绿素储存能力较低。白色基因 w 中的一个单碱基插入导致了一个提前的终止密码子,据推测,这个基因在叶绿素积累和叶绿体发育中的功能已经丧失。这些发现为基础研究和果实颜色的遗传改良做出了贡献。

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