Isolation of genomic difference of rubber trees with different Susceptibility to Common Disease in Field.

• Main Author: Liew Kuen Harng
• Format: Undergraduate Final Project Report
• Published: 2012
• Online Access: http://discol.umk.edu.my/id/eprint/5613/

in this study, representational difference analysis (RDA) was applied to isolate the genomic difference of rubber trees that is susceptible and resistant to common disease of Colletotrichurn gloeosporioides and Oidium heyecie infection. Samples were collected and categories into six groups based on the severity of disease infection. Samples which are susceptible to disease include group A, B and C. Samples which are tolerance to disease include group D, E and F. DNA extraction was conducted to extract the DNA from the leaves samples with two protocols, which were extraction with liquid nitrogen and extraction with ethanol. Extracted DNA from extraction with ethanol was choosing for further conducted of RDA. RDA was carried out by preparation of amplicons for driver and tester genome. Driver genome consists of the gene pooling of disease susceptible samples and tester genome consists of the gene pooling of disease tolerance samples. After that, kinetic enrichment of DNA through hybridization and amplification was conducted to obtain the genomic difference of resistance gene. For hybridization, tester and driver genome was mixed in the ratio of 1:50 and 1:100 (tester/driver). Distinct fragment of isolate gene was observed in 1.5% agarose gel after the first round of hybridization-amplification process. The result obtained from hybridization with ratio 1-.50 was better than hybridization with ratio 1:100 because hybridization with ratio 1:50 had shorter length of sequence smearing.