Penggunaan Marka Molekuler RAPD pada Hasil Persilangan Tanaman Karet (Hevea brasiliensis Muell. Arg.) untuk Analisis Genetik dan Identifikasi Kemurnian Genotipe

Abstract

The objective of research was to find genotype information data and phenotype characteristic of each Fl plant from crossing result between RRIM 600 x PN 1546 clone, identification genotypes purification based on moleculer, and to find DNA markers that will be used as first stage in developing the genetic linkage maps of rubber plant. The research was done in Sungei Putih Research Centre for observation latex yield, girth, bark thickness, number of latex vessel, diameter of latex vessel, content of sucrose, thiol, Pi (Phosphate inorganic), PI (Plugging Index), DRC (Dry Rubber Content), DNA quantity. While DNA extraction, DNA quality, primer screening, PCR was done in Laboratory ofMoleculer Biology, Marihat-Oil Palm Research Institute (IOPRI). Research was conducted on August 2005 to August 2006. Plant material used in this research consist of two parents were RRIM 600 (Wickham 1876 population) with PN 1546 (IRRDB 1981 germ plasm population) and 25 progenies of crossing result. Plant material were seven years old, planted with 2 meter x 2 meter planting distance in Seedling Evaluation Trial (SET) of Sungei Putih Research Center. Phenotype variation among progenies of two parents recombination was highest for some characteristics such as thiol, latex yield, Pi (Phosphat inorganic), and sucrose. This showed that rubber plant had heterozygous character. Girth, bark thickness, number of latex vessel and diameter of latex vessel characters were significantly correlated to production. While latex physiological characters (sucrose, thiol, phosphate inorganic, DRC) had indirect effect to production. Screening primers on this research was done to 24 primers, as a result are 14 primers had polymorphism character (Abi-117.7, OPC-05, OPC-14, OPC-16, OPC-20, OPD-05, OPJ-04, OPJ-11, OPJ-14, OPJ-20, OPH-04, OPH-18, OPN-18, OPN-05, OPM-20, two primers monomorphisme characters (OPJ-13, OPH-14), and eight primers can not be amplified (OPC-19, OPC-13, OPD-01, OPD-03, OPD-04, OPJ -03, OPJ-08, OPH-10). Result of grouping analysis based on moleculer data showed that,

grouping devided in two big groups (A and B). Group A consist of five sub- groups. Based on grouping male and female parents on one group but not in one

sub-group, therefore had far genetic distance. Group B consist of two individues and had far genetic distance to male and female parents as well as to another progenies. Among progenies that had highest similiraty was found on genotype No. 13 (26/689) and 14 (28/794) with similiraty coefficient of 78% or genetic distance of 22%. Genotype purification could be observed from deviation value, analysis result to showed that two genotypes can be categorized as off type, that were No. 18(53/666) and 27 (37/1078). Result of linkage maps constraction showed that one linkage map group formed from polymorphisme to female on OPC14-650 and OPC14-600 (LOD 3.0, 8= 0.25) with distant 4.2 cM. While polymorphisme to male parents formed two linkage map groups (LOD 1.50, 8=0.25) on OPC 16-650 and OPD 16-600 with distance 32.7 cM and OPN05-500 and OPH04-375 with distance 32.7 cM. Linkage map group formed by OPC14 primer on locus 600-650 bp in female parent could be promoted as marker of high production