Department of Genetics and Plant Breeding, School of Agriculture, Aristotle University of Thessaloniki, Greece.
BMC Plant Biol. 2011 Mar 14;11:46. doi: 10.1186/1471-2229-11-46.
Grafting is a widely used technique contributing to sustainable and ecological production of many vegetables, but important fruit quality characters such as taste, aroma, texture and shape are known for years to be affected by grafting in important vegetables species including pepper. From all the characters affected, fruit shape is the most easily observed and measured. From research in tomato, fruit shape is known to be controlled by many QTLs but only few of them have larger effect on fruit shape variance. In this study we used pepper cultivars with different fruit shape to study the role of a pepper Ovate-like gene, CaOvate, which encodes a negative regulator protein that brings significant changes in tomato fruit shape.
We successfully cloned and characterized Ovate-like genes (designated as CaOvate) from two pepper cultivars of different fruit shape, cv. "Mytilini Round" and cv. "Piperaki Long", hereafter referred to as cv. "Round" and cv. "Long" after the shape of their mature fruits. The CaOvate consensus contains a 1008-bp ORF, encodes a 335 amino-acid polypeptide, shares 63% identity with the tomato OVATE protein and exhibits high similarity with OVATE sequences from other Solanaceae species, all placed in the same protein subfamily as outlined by expert sequence analysis. No significant structural differences were detected between the CaOvate genes obtained from the two cultivars. However, relative quantitative expression analysis showed that the expression of CaOvate followed a different developmental profile between the two cultivars, being higher in cv. "Round". Furthermore, down-regulation of CaOvate through VIGS in cv. "Round" changes its fruit to a more oblong form indicating that CaOvate is indeed involved in determining fruit shape in pepper, perhaps by negatively affecting the expression of its target gene, CaGA20ox1, also studied in this work.
Herein, we clone, characterize and study CaOvate and CaGA20ox1 genes, very likely involved in shaping pepper fruit. The oblong phenotype of the fruits in a plant of cv. "Round", where we observed a significant reduction in the expression levels of CaOvate, resembled the change in shape that takes place by grafting the round-fruited cultivar cv. "Round" onto the long-fruited pepper cultivar cv. "Long". Understanding the role of CaOvate and CaGA20ox1, as well as of other genes like Sun also involved in controlling fruit shape in Solanaceae plants like tomato, pave the way to better understand the molecular mechanisms involved in controlling fruit shape in Solanaceae plants in general, and pepper in particular, as well as the changes in fruit quality induced after grafting and perhaps the ways to mitigate them.
嫁接是一种广泛应用的技术,有助于许多蔬菜的可持续和生态生产,但多年来,人们已经知道,包括辣椒在内的重要蔬菜物种的重要果实品质特征,如味道、香气、质地和形状,都会受到嫁接的影响。在所有受影响的特征中,果实形状是最容易观察和测量的。从番茄的研究中可知,果实形状由许多 QTL 控制,但只有少数 QTL 对果实形状变异有较大影响。在这项研究中,我们使用了具有不同果实形状的辣椒品种来研究辣椒卵形基因 CaOvate 的作用,该基因编码一种负调控蛋白,可显著改变番茄果实的形状。
我们成功地从两种不同果实形状的辣椒品种 cv."Mytilini Round"和 cv."Piperaki Long"中克隆并表征了卵形基因(命名为 CaOvate),简称 cv."Round"和 cv."Long",因为这两个品种成熟果实的形状就是它们的名称。CaOvate 保守序列包含一个 1008bp 的 ORF,编码一个 335 个氨基酸的多肽,与番茄 OVATE 蛋白有 63%的同一性,并与其他茄科植物的 OVATE 序列表现出高度相似性,所有这些序列都在专家序列分析中归入同一蛋白亚家族。然而,从两个品种获得的 CaOvate 基因之间没有检测到明显的结构差异。然而,相对定量表达分析表明,CaOvate 的表达在两个品种之间遵循不同的发育模式,在 cv."Round"中表达水平更高。此外,通过 VIGS 下调 cv."Round"中的 CaOvate 会使其果实变成更椭圆形,表明 CaOvate 确实参与了辣椒果实形状的决定,可能通过负调控其靶基因 CaGA20ox1 的表达,这也是本研究中研究的基因。
本文中,我们克隆、表征并研究了 CaOvate 和 CaGA20ox1 基因,它们很可能参与了辣椒果实的塑造。在 cv."Round"的植株中,果实呈现出长椭圆形,我们观察到 CaOvate 的表达水平显著降低,其果实形状与将圆形果实品种 cv."Round"嫁接到长果实品种 cv."Long"上所发生的形状变化相似。了解 CaOvate 和 CaGA20ox1 以及其他基因(如 Sun)的作用,这些基因也参与了控制番茄等茄科植物果实形状,为我们理解茄科植物,特别是辣椒果实形状的分子机制铺平了道路,也为嫁接后果实品质的变化以及可能的缓解方法提供了线索。