Table 1. Primer sequence information.
Locus
name 		Primer sequence (5’ to 3’) Reference Label GenBank accession number Expected size range Amplified size range Concentration of primers in multiplex PCR (µM)
WGA 001 F: ATTGGAAGGGAAGGGAAATG
R: CGCGCACATACGTAAATCAC 		Dangel et al. 2005 6 FAM AY465952 180–192 186–192 0.3
WGA 004 F: TGTTGCATTGACCCACTTGT
R: TAAGCCAACATGGTATGCCA 		Dangel et al. 2005 NED AY465953 228–238 228–230 0.15
WGA 005 F: CAGTTTGTCCCACACCTCCT
R: AACCCATGGTGAGAGTGAGC 		Woeste et al. 2002 6 FAM No data 240–266 Multiallele pattern -
WGA 009 F: CATCAAAGCAAGCAATGGG
R: CCATTGCTCTGTGATTGGG 		Dangel et al. 2005 VIC AY465954 229–252 236–244 0.15
WGA 027 F: AACCCTACAACGCCTTGATG
R: TGCTCAGGCTCCACTTCC 		Woeste et al. 2002 6 FAM AY333951 192–210 202–208 -
WGA 032 F: CTCGGTAAGCCACACCAATT
R: GGACCCAGCTCCTCTTCTCT 		Woeste et al. 2002 VIC AY333952 120–196 Multiallele pattern -
WGA 071 F: ACCCGAGAGATTTCTGGGAT
R: GGACCCAGCTCCTCTTCTCT 		Woeste et al. 2002 6 FAM No data 136–210 205–207 -
WGA 069 F: TTAGATTGCAAACCCACCCG
R: AGATGCACAGACCAACCCTC 		Dangel et al. 2005 PET AY333953 158–180 164–184 0.15
WGA 089 F: ACCCATCTTTCACGTGTGTG
R: TGCCTAATTAGCAATTTCCA 		Dangel et al. 2005 6 FAM AY352440 212–220 212–220 0.25
WGA 118 F: TGTGCTCTGATCTGCCTCC
R: GGGTGGGTGAAAAGTAGCAA 		Dangel et al. 2005 NED AY479958 184–209 184–206 0.1
WGA 148 F: GGTGAACTCCCATAGGGGTA
R: CCAATGCTACTTGCAGAACC 		Woeste et al. 2006 VIC DQ307431 253–271 252–296 0.2
WGA 202 F: CCCCATCTACCGTTGCACTTT
R: GCTGGTGGTTCTATCATGGG 		Dangel et al. 2005 VIC AY479959 238–275 251–275 -
WGA 204 F: GGGTCTCGCCTTCTTTTCTT
R: CACAGAGAGAAGCACGGGTA 		Woeste et al. 2007 NED DQ307432 172–178 174–176 0.1
WGA 225 F: AATCCCTCTCCTGGGCAG
R: TGTTCCACTGACCACTTCCA 		Dangel et al. 2005 PET AY479960 190–202 192–204 -
WGA 256 F: TGAAGACAACAAAACTGCGC
R: CCGGCATTGTTTCTGAAAAT 		Woeste et al. 2009 PET DQ307432 227–253 249–291 0.2
WGA 276 F: CTCACTTTCTCGGCTCTTCC
R: GGTCTTATGTGGGCAGTCGT 		Dangel et al. 2005 6 FAM AY479961 143–192 168–194 -
WGA 321 F: TCCAATCGAAACTCCAAAGG
R: GTCCAAAGACGATGATGGA 		Dangel et al. 2005 NED AY479962 222–225 223–239 -
WGA 331 F: TCCCCCTGAAATCTTCTCCT
R: CGGTGGTGTAAGGCAAATG 		Dangel et al. 2005 VIC AY479963 273–277 271–275 -
WGA 332 F: ACGTCGTTCTGCACTCCTCT
R: GCCACAGGAACGAGTGCT 		Foroni et al. 2006 PET AY479964 214–225 215–225 0.1
WGA 349 F: GTGGCGAAAGTTTATTTTTTGC
R: ACAAATGCACAGCAGCAAAC 		Woeste et al. 2002 6 FAM AY479965 258–274 262–274 0.25
WGA 376 F: GCCCTCAAAGTGATGAACGT
R: TCATCCATATTTACCCCTTTCG 		Woeste et al. 2002 NED AY479966 218–254 230–252 -
1

Fig. 1. Genotype profile (electropherogram) of the eleven microsatellite loci (SSR markers) of common walnut generated by multiplex PCR. The x-axis represents DNA fragment size in base pairs, and the y-axis represents fluorescence units. The SSR profile is displayed in the blue (6-FAM labeled primers), green (VIC labeled primers), yellow (NED labeled primers) and red (PET labeled primers) channels of a five-color fluorescent system, with the orange (LIZ labeled fragments) channel being used for a size marker (not shown). View larger in new window/tab.

Table 2. Gene diversity parameters of eleven common walnut SSR markers, sample size (N), number of observed alleles (Na), effective number of alleles (Ne), observed heterozygosity (Ho), expected heterozygosity (He), fixation index (FIS), and Hardy-Weinberg equilibrium (HW).
Locus N Na Ne Ho He FIS HW
WGA 001 15 4 2,514 0,200 0,602 0,668 ***
WGA 004 15 2 1,923 0,533 0,480 –0,111 ns
WGA 009 15 3 2,761 0,533 0,638 0,164 ns
WGA 069 15 7 4,091 0,333 0,756 0,559 *
WGA 089 15 4 3,103 0,667 0,678 0,016 ns
WGA 118 15 4 2,571 0,667 0,611 –0,091 ns
WGA 148 15 6 2,586 0,400 0,613 0,348 ***
WGA 204 15 2 1,069 0,067 0,064 –0,034 ns
WGA 256 15 4 2,432 0,667 0,589 –0,132 ns
WGA 332 15 3 2,761 0,667 0,638 –0,045 ns
WGA 349 15 3 2,381 0,467 0,580 0,195 ns
Mean 15 3,818 2,563 0,473 0,568 0,140  
SE 0 0,464 0,222 0,062 0,054 0,084  
ns = not significant, * P<0.05, ** P<0.01, *** P<0.001