ISHS Acta Horticulturae 871: IV International Symposium on Cucurbits
POLYMORPHISM ANALYSES OF WATERMELON (CITRULLUS LANATUS) MAPPING PARENTS USING RAPD AND ISSR MOLECULAR MARKERS |
| Authors: | X.P. Yang, G. Liu, J.H. Xu, C.Z. Gao, X.L. Hou |
| Keywords: | watermelon, RAPD, ISSR, system optimization, polymorphism analyses |
| DOI: | 10.17660/ActaHortic.2010.871.92 |
| Abstract:
Genomic DNA of watermelon was analyzed by optimizing the concentration of Mg2+, dNTPs, and primer in RAPD and ISSR analysis systems. The screened optimization system of RAPD for watermelon genomic DNA was 20 μl, which contained primer 0.4 μΜ L-1, dNTPs 200 μΜ L-1, Mg2+ 2.0 mΜ L-1, DNA 20 ng, 1×buffer, Taq E 1.0 U. The optimization system of ISSR was 16 μl, which contained primer 0.25 μΜ L-1, dNTPs 160 μΜ L-1, Mg2+ 1.0 mΜ L-1, DNA 20 ng, 1×buffer, Taq E 0.64 U. RAPD and ISSR were applied in polymorphism analyses of watermelon mapping parents, 64-1Ø-3 and 6Ø-2, and 23Ø-1 and 4-1Ø-2, which were inbred lines of district variety in China. The results of mapping parents 64-1Ø-3 and 6Ø-2 showed that 170 out of 181 RAPD primers generated 525 amplified bands, 55 produced 77 polymorphic bands, and the polymorphism rate was 14.67%; 45 out of 62 ISSR primers generated 188 amplified bands with an average of 4.18 bands per primer. And 15 polymorphic primers produced 23 polymorphic loci. The polymorphism rate of ISSR analyses was 12.23%. In mapping parents 23Ø-1 and 4-1Ø-2, 456 out of 499 RAPD primers amplified 1431 bands and 142 polymorphic primers produced 240 polymorphic loci, so the polymorphism rate was 16.77%. These showed that the genetic background of watermelon mapping parents was narrow. | |
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