Application of ISSR markers for verification of F1 hybrids in mungbean (Vigna radiata)

Abstract

ABSTRACT. Mungbean improvement via hybridization requires the identification of true F 1 hybrids from controlled crosses before further generations of selfing/crossing and selection. We utilized inter-simple sequence repeat (ISSR) markers for identifying putative F 1 hybrids from six cross combinations whose morphological characteristics were very similar to those of their respective female parents and could not be visually discriminated from the self-pollinated progeny. Based on 10 ISSR primers, polymorphisms were found between female and male parents of all six cross combinations. The highest value of genetic differentiation (21.4%) was found between male and female parents of the SUT3 x M5-1 cross. These 10 ISSR primers gave 2.8-25.0% polymorphism between male and female parents, with a mean of 12.1%, and 0-13.0% polymorphism between F 1 hybrid and female parents, with a mean of 4.8%. F 1 hybrids of all six cross combinations could be differentiated from the self-pollinated progeny of their female parents by using only either ISSR 841 or 857 primers, together with the ISSR 835 primer. We conclude that ISSR markers are useful and efficient for identifying mungbean F 1 hybrids in controlled crosses

Genetic Diversity of Indigofera tinctoria L. in Java and Madura Islands as Natural Batik Dye based on Inter-simple Sequence Repeat Markers

“…The percentage range of band polymorphism was 50-90% (Table 2). Compared to previous studies on Sesbania, Vigna radiata and Vigna savi assessment [10][11][12], the observed primer produced less bands, but the percentage of polymorphic bands was higher (50-90%) than the previous one (0-25%). Note: Y = (C, T); R = (A, G).…”

“…Previous studies have shown that V. radiata hybrid, Phaseolus vulgaris, Pandanus, and Freycinetia also yielded abundant bands from (AG)8T primer amplification. Thus, the complementary sequences of (AG)8T primer are widely distributed in other plants and (AG)8T primer can be used as a universal ISSR primer [6,11,15].…”

“…Polymerase chain reaction assays were set up in a final volume of 25 μL containing 2.5 μL DNA template, 1 μL ISSR primer, 12.5 μL of 2X GoTaq® Reaction Buffer pH 8.5 (consisting of 400 μM dATP, 400 μM dGTP, 400 μM dCTP, 400 μM dTTP, 3 mM MgCl2) and 9 μL nuclease-free water (Promega, USA). The amplification process consisted of initial denaturation at 94 °C for 3 min, followed by 30 cycles of denaturation at 94 °C for 1 min, annealing at 45-51 °C for 45-60 s, extension at 72 °C for 2 min, and 10 min of final extension at 72 °C [10][11][12]. The amplification products were resolved in a 1% agarose gel with 1X TBE buffer at 80 V for 90 min.…”

Molecular Characterization of Green Gram [Vigna radiata (L.) Wilczek] for Future Breeding Programmes

“…Based on 10 ISSR primers, polymorphisms were found between female and male parents of all six cross combinations. F1 hybrids of all six cross combinations could be differentiated from the self-pollinated progeny of their female parents by using only either ISSR 841 or 857 primers, together with the ISSR 835 primer (Khajudparn et al, 2012). Five species of Pinellia endemic in China were analyzed using inter-simple sequence repeat (ISSR) marker examined genetic relationship among.…”

Analysis of genetic relationships and identification of lily cultivars based on inter-simple sequence repeat markers

“…In addition, the ISSR technique is easy and economical, and it has been used successfully in genetic diversity studies of many plants, including Jatropha curcas (Grativol et al, 2011), spring orchid (Wang et al, 2009a), Auricularia auricula (Tang et al, 2010), clematis (Gardner and Hokanson, 2005), and loquats . ISSR markers can also be used to identify cultivars of Curcuma (Taheri et al, 2012) and strawberry (Arnau et al, 2002); detect the authenticity of hybrid offspring in Coffea (Ruas et al, 2003), mungbean (Khajudparn et al, 2012), and Phyllostachys (Lin et al, 2010) including lily (Wang et al, 2009b;Wu et al, 2009); and test asexual reproduction strain stability in lily (Liu and Yang, 2012;Xi et al, 2012). To date, there have been very few reports using ISSRs to differentiate varieties of lily hybrids.…”