Genetic Analysis of Some Indonesian Black Rice Cultivars (Oryza sativa L.) Using RAPD (Random Amplified Polymorphic DNA) Engga Dwi Supriyanto1, Gatot Subroto1, Denna Eriani Munandar1, and Tri Handoyo1,2*
1Faculty of Agriculture, University of Jember, INDONESIA. 2Center for Development of Advanced Science and Technology, University of Jember, INDONESIA. (*E-mail: trihandoyo.faperta[at]unej.ac.id)
Abstract
Background: Rice (Oryza sativa. L) is the main commodity of Indonesian food crops and become a staple food for the community. Black rice has high anthocyanin content, so it is very beneficial for the community. The relatively long age of black rice and low productivity are weaknesses that must be overcome. The development of black rice varieties is needed to assemble new plants that have a short life and high productivity. Plant assembly is strongly influenced by the level of plant kinship. Plant assembly is often done incorrectly due to close kinship, so diversity analysis is needed so that the development of black rice varieties is appropriate. Phylogenetic analysis describes genetic distance or differences between individuals in an organism or species. Phylogenic analysis can be carried out using morphological, physiological and molecular characteristics. Morphological characteristics are based on differences in the morphological characteristics of each plant. Molecular identification use RAPD (Random Amplified Polymorphic DNA) based on DNA sequences that are homologous with a single short (10-12 base) oligonucleotide sequence. Molecular characters are based on genotypic characters, namely by looking at the different patterns of DNA bands that owned by plants. Aims: To determine the level of kinship and diversity in 10 varieties of black rice (Oryza sativa L.) based on morphological and molecular identification. Materials and Methods: In this study, we determinated the level of kindship of 10 varieties of black rice included Hitam Purbalingga, Hitam Purwekerto, Hitam Lumajang, Hitam Blitar, Hitam Bantul, Hitam Banjarnegara, Hitam Toraja, Hitam Melik, Pari Ireng, and Here Lahok/Timor Leste based on morphological and molecular identification. Morphological identification uses 7 variables i.e plant height, number of productive tillers, weight of 100 seeds, shape of grain, colour of grain, colour of rice age of flowering and age of harvest. The analysis results realized in the form of a dendogram. Molecular identification using RAPD marker with 9 primers included OPB-01, OPB-04, OPB-08, OPB-10, OPB-11, OPB12,OPB-13, OPB-17 and OPB-18. The DNA band profile translated into binary data with the provision of a value of 0 for no band and 1 for the presence of DNA bands in certain fragments. Both of data analysed into cluster using NTSYSpc software with DIST coefficient for morphological data and Jaccard-s Index for molecular data. The analysis results are realized in the form of dendogram and genetic similarity values. Results: Dendrogram of morphological data showed in 2 clusters, 2 groups and 2 subgroups. Cluster I was accopied by Hare Lahok and Hitam Purwekerto. Cluster II devided into 2 groups. First group include Hitam Blitar and second group include Hitam Melik, Pari Ireng, Hitam Bantul, Hitam Lumajang, Hitam Toraja, Hitam Purbalingga. Molecular kinship analysis used to determine the level of genetic similarity of 10 black rice vari
Keywords: Black Rice, Genetic Diversity, Morphological Characteristics, and RAPD
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