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Xoconostle or acidic prickly pear is an important fruit in Mexico; it is produced by a group of Opuntia plants known for their nutritional qualities and adapted to harsh environmental conditions. In this study, we report for the first time the estimation of genetic diversity within a set of 24 xoconostle accessions using inter simple sequence repeats (ISSR) markers. Five ISSR primers were selected to detect DNA polymorphisms in these plants. Broad genetic diversity was detected (0.41-0.82, Dice coefficient) and the unweighted pair group method with the arithmetic averaging (UPGMA) analysis allowed discriminating all genotypes and enabled their clustering into six groups; the 'Matizado' accession was significantly divergent from all tested genotypes. In addition, as shown by the clustering analysis and principal component analysis (PCA), the tested genotypes did not group according to their origin, to culture conditions, or to the current species classification. This work, on one hand, demonstrates the high genetic variation among genotypes of xoconostles, and on the other hand, it suggests a taxonomic revision and raises doubts about the number of species reported for these plants.

ABSTRACT. Molecular methods are powerful tools in characterizing and determining relationships between plants. The aim of this study was to study genetic divergence between 103 accessions of Mexican Opuntia. To accomplish this, polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis of three chloroplast intergenic spacers (atpB-rbcL, trnL-trnF, and psbA-trnH), one chloroplast gene (ycf1), two nuclear genes (ppc and PhyC), and one mitochondrial gene (cox3) was conducted. The amplified products from all the samples had very similar molecular sizes, and there were only very small differences between the undigested PCR amplicons for all regions, with the exception of ppc. We obtained 5850 bp from the seven regions, and 136 fragments were detected with eight enzymes, 37 of which (27.2%) were polymorphic. We found that 40% of the fragments from the chloroplast regions were polymorphic, 9.8% of the bands detected in the nuclear genes were polymorphic, and 20% of the bands in the mitochondrial locus were polymorphic. trnL-trnF and psbA-trnH were the most variable regions. The Nei and Li/Dice distance was very short, and ranged from 0 to 0.12; indeed, 77 of the 103 genotypes had the same genetic profile. All the xoconostle accessions (acidic fruits) were grouped together without being separated from three genotypes of prickly pear (sweet fruits). We assume that the genetic divergence between prickly pears and xoconostles is very low, and question the number of Opuntia species currently considered in Mexico.

Opuntia, an important horticultural crop in Mexico, is cultivated mainly for its two fruitsvariants: sweet fruits or prickly pears (tunas) and acidic fruits (xoconostles). The interretrotransposonamplified polymorphism (IRAP) technique was applied to evaluate geneticdiversity of Opuntia varieties and to differentiate xoconostle fruits from tunas. Five IRAPprimers previously described for other plant species and classified into three retrotransposonfamilies, namely Copia, Gypsy and TRIM, were analysed in 43 Opuntia varieties (eightxoconostles and 35 tunas). The five individual IRAP primers generated a total of 264fragments, where 64.8 % of them were polymorphics. The retrotransposon of the Gypsyfamily (60 fragments) was more represented than Copia (average of 52 fragments) or TRIM(48 fragments) families. Moreover, the percentage of polymorphic fragments was higher (61.9%) in xoconostles than in tunas (56.5 %). A larger number of total amplified fragments (262)was found among tunas, compared to those amplified from xoconostle varieties (257fragments). In contrast, a lower number of polymorphic bands were counted among tunas(148) than among xoconostle varieties (159). Unlike the UPGMA analysis, where three of thexoconostle-producing varieties were grouped with other tunas, the PCoA analysis allowed abetter separation of all xoconostle varieties. These results suggest a potential role of thetransposable elements in genetic divergence within the Opuntia genus.