Bretó M P, Ruiz C, Pina J A, Asíns M J
Instituto Valenciano de Investigaciones Agrarias, Ctra. Moncada-Náquera, Km 4.5, 46113 Moncada, Valencia, Spain.
Mol Phylogenet Evol. 2001 Nov;21(2):285-93. doi: 10.1006/mpev.2001.1008.
Clementines, due to their high quality, are one of the most important cultivated citrus mandarins. As in the case of sweet orange and satsuma mandarins, genetic variability within this species is minimal when analyzed by molecular markers, because the existing varieties have not been obtained through hybridization, but through the selection of spontaneous mutations affecting traits of agronomic interest. This would explain, at least in part, the greater diversity for agronomic traits when compared to the variability for molecular markers. Another possible (nonexclusive) reason is that the types of molecular marker used are not focused on the kind of molecular change mainly involved in the origination of new clementine cultivars; i.e., are all sources of variation equally involved in the diversification of these plants? To answer this question, different kinds of markers based on primers of random sequence, simple sequence repeats, and retrotransposon sequences that may reveal point mutations, and somatic recombination and transposon activity, respectively, were used to compare the level of variability among 24 clementine varieties. Their ISSR, RAPD, and AFLP analysis provided only two polymorphic bands, distinguishing just two varieties. No variability was found by SSRs, i.e., no new allele arising through somatic recombination was detected. Instead, the amplification of sequences adjacent to retrotransposons yielded a higher number of polymorphisms (14.6 vs 2.4% for the previous mentioned marker types). Two geographical distant groups, one from North Africa and the other from Spain, have evolved in agreement with polymorphisms based on IRAP markers anchored to, at least, two different Copia-like retrotransposon sequences. Therefore, this study suggests that the DNA of this type of mobile elements is evolving faster than the DNA of other markers in this clonal lineage.
由于其高品质,克莱门氏小柑橘是最重要的栽培柑橘类宽皮橘之一。与甜橙和温州蜜柑的情况一样,通过分子标记分析时,该物种内的遗传变异性极小,因为现有的品种并非通过杂交获得,而是通过选择影响农艺性状的自发突变获得。这至少可以部分解释,与分子标记的变异性相比,农艺性状的多样性为何更大。另一个可能(并非唯一)的原因是,所使用的分子标记类型并未聚焦于主要参与新克莱门氏小柑橘品种起源的分子变化类型;也就是说,所有变异来源在这些植物的多样化过程中所起的作用是否相同?为了回答这个问题,分别使用了基于随机序列引物、简单序列重复和反转录转座子序列的不同类型标记,这些标记可能分别揭示点突变、体细胞重组和转座子活性,以比较24个克莱门氏小柑橘品种之间的变异水平。它们的ISSR、RAPD和AFLP分析仅提供了两条多态性条带,仅区分出两个品种。通过SSR未发现变异性,即未检测到通过体细胞重组产生的新等位基因。相反,反转录转座子相邻序列的扩增产生了更多的多态性(上述标记类型分别为14.6%和2.4%)。两个地理上相距遥远的群体,一个来自北非,另一个来自西班牙,其进化与基于至少锚定在两种不同类Copia反转录转座子序列上的IRAP标记的多态性一致。因此,本研究表明,在这个克隆谱系中,这类可移动元件的DNA比其他标记的DNA进化得更快。