Development of Microsatellite Markers in Melon (Cucumis melo L.) and Their Application to Major Cucurbit Crops.

Abstract

We developed 31 sets of melon (Cucumis melo L.) microsatellite markers in this study. Primers for these markers were designed from clones containing simple sequence repeats (SSRs) of (GA/CT)n or (GT/CA)n isolated from a melon genomic library. These microsatellite loci were examined for the detection of alleles among twelve lines and cultivars from six varieties of C. melo. A total of 28 marker loci showed polymorphism by 2.0 % agarose TBE gel separation. And these markers were also examined for cross-species amplification applicable to nine other species in Cucurbitaceae. Among them, 13 marker loci were commonly detected within the major cucurbit crops, cucumber, pumpkin and watermelon using the present primer sets. The species to which the melon microsatellite markers would be most applicable was Momordica charantia (24 markers), followed by Cucumis sativus (20 markers) and Cucurbita maxima (18 markers). These conserved microsatellite loci could be used as anchor markers in studies on synteny in Cucurbitaceae.

Molecular Analysis of the Genetic Diversity of Chinese Hami Melon and Its Relationship to the Melon Germplasm from Central and South Asia

“…Sixteen SSR markers showing distinct stable polymorphism were selected among 177 SSR markers developed by Danin-Poleg et al (2000), Akashi et al (2001), Chiba et al (2003), Ritschel et al (2004), andFukino et al (2007). Table 3 shows details of the SSR markers used.…”

Development of Simple Sequence Repeat Markers for Identification of Japanese Gentian Cultivars

“…Thus, we attempted to develop SSR markers containing di-nucleotide repeats applicable to Japanese gentian. To isolate SSR loci from plants lacking genomic and cDNA sequence information, several protocols, such as the screening of a genomic library by SSR probes (Chiba et al, 2003;Suwabe et al, 2002), utilization of SSR markers from closed species (Hayashi et al, 2008;Shiran et al, 2007), construction of an SSR-enriched library (Díaz et al, 2006;Nunome et al, 2006), and sequencing of amplified ISSR fragments (Hayden et al, 2004;Lian et al, 2001Lian et al, , 2006, have been developed. Our preliminary experiments using suppression PCR (Lian et al, 2001) to isolate SSRs in Japanese gentian were inefficient, although the reason was not clearly understood (data not shown).…”

“…SSRs are tandemly repeated sequence motifs 1-6 bp in length and are widely present in the eukaryotic genome. Therefore, SSR markers have been applied to discriminate cultivars in many crops, such as soybean (Hossain et al, 2000), fruit trees including apple (Goulão and Oliveira, 2001), olive (Díaz et al, 2006), peach (Dirlewanger et al, 2002;Yamamoto et al, 2003), sweet cherry (Dirlewanger et al, 2002), pear (Kimura et al, 2002), Japanese apricot (Hayashi et al, 2008), and almond (Shiran et al, 2007), in vegetable crops including melon (Chiba et al, 2003), strawberry (Shimomura and Hirashima, 2006), eggplant (Nunome et al, 2009), Brassica rapa (Suwabe et al, 2002, and cucumber (Fukino et al, 2008), and floricultural plants including pelargonium (Becher et al, 2000) and carnation (Smulders et al, 2003). In Gentiana, ten SSR markers were developed for the Chinese medicinal plant G. crassicaulis (Li et al, 2007); however, since G. crassicaulis belongs to a different taxonomic section from the Japanese cultivated gentians G. triflora and G. scabra (Ho and Liu, 2001), these SSR markers could not be successfully applied to Japanese gentians.…”

Genetic Mapping and Molecular Breeding in Cucurbits

“…However, once they are identified and tested, SSRs constitute good quality markers. Melon SSR markers have been largely developed by Katzir et al (1996), Danin-Poleg et al (2001), Chiba et al (2003), and Ritschel et al (2004), while SSRs in cucumber and watermelon have been studied by Fazio et al (2002) and Jarret et al (1997), respectively. SNP markers are also being developed for melon by Morales et al (2004) and have been used in cucumber mapping (Fazio et al 2003b).…”