Whole Genome Amplification of Single Pollen Grains from a Sugarcane Cultivar and Analysis of the Genetic Relatedness based on SCoT Markers
Single pollen grains were isolated from an intact anther of a sugarcane cultivar and collected using a pair of special forceps.The single pollen grains were lysed in an alkali/detergent solution respectively.The resulting solution was used as the template for Whole Genome Amplification.Genomic DNA of individual pollen grains was amplified successfully and WGA products of single pollen grains were tested by 5S rRNA-ITS primers to select those with high fidelity to the donor plant.The WGA products selected out were analyzed with SCoT markers and produced a mount of DNA polymorphism.Genetic similarity calculation was performed using the software NTSYSpc 2.1.The similarity coefficient values ranged from 0.0888～0.3654 and 69 percent of total pollen grains were clustered into two groups at the value of 0.187.The results indicated a high scale of pollen grains shared same gene loci during the meiotic segregation stage.The 52 single pollen grain samples were clustered into four groups based on the homology tree.A probability distribution of the pollen grains was calculated based on the number of pollen grains in the four groups.