N the species of Carthamus and Phonus, we discovered only HinfI

The HinfI sequences of V. lippii belonged to subfamilies I, II, III or V, mainly to sort III (nine sequences out of 14). In O different prime-boost vaccine regimens. Comparison addition, most HinfI sequences of V. crupinoides belonged to subfamilies II, III and IV. Nevertheless, many of the sequences of V. muricata and V. tubuliflora and 3 of V. crupinoides belonged to subfamily VI. Three species of Cheirolophus (C. intybaceus, C. sempervirens and C. sventenii) shared two forms of HinfI monomers in their genomes, these of subfamilies VII and VIII. Cheirolophus sventenii also had a type II repeat. Cheirolophus falcisectus had only sequences of subfamily VII. In contrast, amongst the sequences isolated from C. teydis, 12 belonged to subfamily III and the other 3 to subfamily II (Fig. two). We tested the reliability in the CenHinf1 and CenHinf2 primers to detect the eight subfamilies in these species. For this, we designed four additional primer pairs for the particular amplification of repeats of subfamilies I, III, V and VI, and probed them in a set of representative species. We obtained benefits similar to these obtained together with the basic primers. Nonetheless, some variations were identified (Fig. two; Supplementary Information Fig. S2). For example, subfamily I is present in Centaurea granatensis and Crupina vulgaris (the only species of Centaurea and Crupina for which we didn't get repeats of subfamily I just after sequencing of amplified product employing the primers CenHinf1 and CenHinf2). The subfamily III is located in species of Volutaria and Cheirolophus besides these previously detected using the `general' primers. Nonetheless, the primers employed had been not in a position to amplify repeats of subfamily III in Carduncellus (even though we detected a few repeats of this kind working with the CenHinf1 and CenHinf2 primers). Subfamily V is found not only in Klasea and Rhaponticum but additionally in Volutaria, as we previously discovered by sequencing the CenHinf1/CenHinf2 amplified item. Subfamily VI was discovered only in Volutaria, as detected by CenHinf1 and CenHinf2 primers. Phylogenetic reconstruction by ML and Bayesian inference resulted in related trees. A Bayesian majority-rule consensus tree with Bayesian PPs is shown in Supplementary Data Fig. S3. sempervirens and C. sventenii) shared two types of HinfI monomers in their genomes, those of subfamilies VII and VIII. Cheirolophus sventenii also had a kind II repeat. Cheirolophus falcisectus had only sequences of subfamily VII. In contrast, amongst the sequences isolated from C. teydis, 12 belonged to subfamily III plus the other 3 to subfamily II (Fig. two). We tested the reliability of the CenHinf1 and CenHinf2 primers to detect the eight subfamilies in these species. For this, we developed four added primer pairs for the precise amplification of repeats of subfamilies I, III, V and VI, and probed them within a set of representative species. We obtained benefits related to those obtained using the common primers. Nevertheless, some differences had been located (Fig. two; Supplementary Information Fig. S2). By way of example, subfamily I is present in Centaurea granatensis and Crupina vulgaris (the only species of Centaurea and Crupina for which we did not acquire repeats of subfamily I right after sequencing of amplified product working with the primers CenHinf1 and CenHinf2). The subfamily III is identified in species of Volutaria and Cheirolophus other than these previously detected together with the `general' primers.