Hybrid testing of sweet corn and waxy corn using seed storage proteins by ultrathin-layer isoelectric focusing technique
2010
Manum, P., Kasetsart Univ., Nakhon Patohom 73140 (Thailand). Center for Agricultural Biotechnology | Thongket, T., Kasetsart Univ., Nakhon Pathom 73140 (Thailand). Dept. of Horticulture | Chanprame, S., Kasetsart Univ., Nakhon Patom 73140 (Thailand). Center for Agricultural Biotechnology
The hybrid test of F1 hybrid is very important in seed production industry especially for the cross pollination crop such as corn. To ensure farmer's satisfactory, seed companies have to intensively test their hybrid seeds. Ultrathin-layer isoelectric focusing (UTLIEF) technique is the one that was considered as reliable, convenient and low cost. Three different protein extraction solvents including water, phosphate buffer and 4M urea were tested with mature seeds of six sweet corn cultivars. The results revealed that these six sweet corn varieties could be distinguished by UTLIEF technique using any of these three solvents. However, protein bands from water extraction had clearer and sharper resolution than extract with phosphate buffer or 4M urea. Thus, water was selected as solvent for the hybrid tested in sweet corn and waxy corn. The inbred lines and their F-1 hybrids were subjected to hybridity tested using UTLIEF. Seed storage proteins were extracted with water and were separated on the pH 2-9 acrylamide gel according to their isoelectric when the electrical voltage of 2500V was applied. The results demonstrated that this technique could be successfully used for hybrid test in both sweet and waxy corns. For sweet corn, ten distinct protein bands could be used to indicate parental lines and their F-1 hybrid. For waxy corn, even only four protein bands could be observed but theywere clear cut for hybridity test. In the waxy corn seed, there was a distinct band which indicated that the seed was not the F-1 hybrid seed. Thus, the UTLIEF technique not only offers the means for hybrid test but the genetic purity test also.
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