Development of Biochemical and Molecular Markers for Polygamodioecious Character in Simarouba Glauca DC

Simarouba glauca, an oil seed tree, is polygamodioecious, a condition in which trees have both bisexual and purely male flowers on certain branches of same tree. It segregates into male, female and andromonoecious during flowering. Since controlling the sex ratio of the tree population is important, an attempt was made to develop markers for polygamodioecious character in Simarouba. Nine enzyme systems were screened with four different tissue, four extraction buffers and four electrode buffers. With use of Feret extraction buffer containing ascorbic acid, Tris-Borate- EDTA electrode buffer, and flower tissues, five enzyme systems gave good resolution and staining. These enzyme systems were studied using starch gel electrophoresis and no polymorphism was detected. To access variability, plants were analyzed using randomly amplified polymorphic DNA (RAPD). Out of one hundred and fifty decamer primers used, seventeen decamer primers were selected based on amplification polymorphism. The binary data obtained were subjected to cluster, 2D-PCA and 3D-PCA analyses, which showed five clusters. The t-test showed that two primers OPT-7 and OPW-3 could be used to differentiate between males and females. One primer, OPS-6 gave a characteristic band only in 100% males and not in others.