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

S2). For RDEA594 biological activity example, subfamily I is present in Centaurea granatensis and Crupina vulgaris (the only species of Centaurea and Crupina for which we did not get repeats of subfamily I after sequencing of amplified solution using the primers CenHinf1 and CenHinf2). 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 created 4 further primer pairs for the distinct amplification of repeats of subfamilies I, III, V and VI, and probed them inside a set of representative species. We obtained benefits similar to those obtained with the general primers. Nevertheless, some differences had been located (Fig. 2; 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 didn't receive repeats of subfamily I following sequencing of amplified product working with the primers CenHinf1 and CenHinf2). The subfamily III is identified in species of Volutaria and Cheirolophus besides those previously detected together with the `general' primers. Even so, the primers made use of had been not in a position to amplify repeats of subfamily III in Carduncellus (despite the fact that we detected a few repeats of this form employing the CenHinf1 and CenHinf2 primers). Subfamily V is discovered not just in Klasea and Rhaponticum but additionally in Volutaria, as we previously identified by sequencing the CenHinf1/CenHinf2 amplified product. Subfamily VI was found only in Volutaria, as detected by CenHinf1 and CenHinf2 primers.N the species of Carthamus and Phonus, we located only HinfI type III sequences. Subfamily IV is characteristic of Carduncellus. Nevertheless, some species of this genus also have low-copy repeats of subfamilies III and VI in their genomes. Subfamily V was discovered in Rhaponticum and Klasea. These species had only this kind of HinfI repeat, except R. acaule, which also contained sequences of subfamilies II, IV and VI in its genome. We analysed 4 species of Volutaria (Table 1). The HinfI sequences of V. lippii belonged to subfamilies I, II, III or V, primarily to form III (nine sequences out of 14). In addition, most HinfI sequences of V. crupinoides belonged to subfamilies II, III and IV. However, a lot of the sequences of V. muricata and V. tubuliflora and three of V. crupinoides belonged to subfamily VI. Three species of Cheirolophus (C. intybaceus, C. sempervirens and C. sventenii) shared two sorts of HinfI monomers in their genomes, those of subfamilies VII and VIII. Cheirolophus sventenii also had a sort 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 three 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 4 additional primer pairs for the specific amplification of repeats of subfamilies I, III, V and VI, and probed them inside a set of representative species. We obtained results equivalent to those obtained using the basic primers. Nevertheless, some variations had been located (Fig. two; Supplementary Data Fig.