Development and application of SCAR markers for sex identification in the dioecious species Ginkgo biloba L.

Abstract

There is an urgent need for early sex identification to support field planting in Ginkgo biloba L., due to the different economic and medicinal values between male and female trees. An easy, rapid and reliable molecular method for sex type determination of G. biloba was reported in the paper. Random amplification of polymorphic DNA (RAPD) and sequence-characterized amplified region (SCAR) were used to search for specific molecular markers linked to the sex locus. A total of 48 primers were used for screening of specific RAPD markers in six male and three female samples. Only one primer, S10, showed different amplification band patterns associated with sex types. Then the sex-specific bands, S10-BandA and S10-BandB, were cloned and sequenced. Based on the sequences two pairs of SCAR primers, GBA and GBB, were designed. The GBA primers amplify a single 571 bp band in male samples but not in female samples, and DNA amplification using GBB primers could generate a 688 bp band only in the female individuals. Finally, the SCAR primers were used to test 16 sex-unknown samples. SCAR primers developed in this paper can be used as effective, convenient and reliable molecular markers for sex identification in G. biloba.

CHARACTERIZATION OF GENETIC MARKERS LINKED TO SEX DETERMINATION IN THE HAPLOID‐DIPLOID RED ALGA GRACILARIA CHILENSIS1

“…Indeed, some studies report a very low success rate in screening for sex markers, as in Populus, where none of the 1,219 markers tested were linked to sex (McLetchie and Tuskan 1994); in A. officinalis, with only one marker linked to sex of the 760 markers tested (0.13%, Jiang and Sink 1997); or in S. viminalis, where two markers linked to sex of the 953 markers tested (0.21%, Alstrom-Rapaport et al 1998 andGunter et al 2003), which correspond to one sex-linked band over 1,080 scored bands (0.09%, Alstrom-Rapaport et al 1998). On the other hand, a high rate of success was reported in white campion (four over 60 markers tested, 7%, Mulcahy et al 1992), A. deliciosa (eight over 34 markers tested, 23.5%; corresponding to eight over 430 bands scrored, 1.9%; Shirkot et al 2002), palmyrah (B. flabellifer, three over 180 markers tested, 1.7%; George et al 2007), P. fascicularis (one over 89 markers tested, 1.1%, corresponding to one over 813 bands scored, 0.1%; Vinod et al 2007), and G. biloba (one over 48 markers tested, 2.1%; Liao et al 2009). Interestingly, the three species with a low rate of success in screening sex markers show single dominant locus, two locus epistatic system, or incipient sex chromosome gender determination mechanisms, whereas heteromorphic X ⁄ Y sex chromosomes are encountered in white campion (Vyskot and Hobza 2004).…”

“…2007), P. fascicularis (one over 89 markers tested, 1.1%, corresponding to one over 813 bands scored, 0.1%; Vinod et al. 2007), and G. biloba (one over 48 markers tested, 2.1%; Liao et al. 2009).…”

“…Specific sex-linked markers have been used for sex determination in many commercially important plants (Asparagus, Reamon-Bü ttner et al 1998;papaya, Deputy et al 2002;kiwifruit, Shirkot et al 2002;palmyrah, George et al 2007;padang, Vinod et al 2007; Ginkgo, Liao et al 2009). However, very few are available for sex determination in cultivated seaweeds (G. gracilis, Martinez et al 1999;Saccharina japonica, Liu et al 2009; Undaria pinnatifida, Shan and Pang 2010).…”

Identification of EeSt-genome species in Pseudoroegneria and Elytrigia (Poaceae: Triticeae) by using SCAR markers from ITS sequences

“…Recently, reliable SCAR markers have been used in molecular analysis (Dai et al, 2005;Paxton et al, 2005;Rahman et al, 2007;Liao et al, 2009;Bandyopadhyay and Raychaudhuri, 2010;Duan et al, 2011;Lee et al, 2011;Yu et al, 2011a,b). In the current study, SCAR primers were designed based on ITS characterization to amplify ITS fragments of E e St-genome species.…”

Development and application of a SRAP marker for the identification of sex in Buchloe dactyloides

“…Therefore, cytological methods cannot be used to identify sex in most dioecious plants. With the development of molecular biology, molecular markers amplified using polymerase chain reaction (PCR) have been widely used to identify the sexes in many dioecious species, including Silene latifolia (Mulcahy et al 1992;Stilio et al 1998), Asaparagus officinalis (Jiang and Sink 1997;Reamon-Buttner and Jung 2000;Gao et al 2007), Salix viminalis (AlstromRapaport et al 1998;Gunter et al 2003), Actinidia chinensis (Gill et al 1998;Yao et al 2005), Atriplex garrerrii (Ruas et al 1998), Carica papaya (Parasnis et al 1999(Parasnis et al , 2000Urasaki et al 2002;Deputy et al 2003;Ren et al 2007;Chaves-Bedoya and Nunez 2007), Cannabis sativa (Mandolino et al 1998;Torjek et al 2002;Cristiana Moliterni et al 2004), Ginkgo biloba (Liao et al 2009), Podocarpus macrophyllus (Liang et al 2002), Pisacia vera (Hormaza et al 1994;Tang et al 2003;Yakubov et al 2005), Rumex acetosa (Korpelainen 2002), Mercurialis annua (Khadka et al 2002), Eucommia ulmoide (Xu et al 2004), Ficus fulva (Parrish et al 2004), and Aucuba japonica (Masayuki 2009). …”