Development of C319-allele specific marker for EPSPS gene from Eleusine indica

Abstract

The herbicides resistant weeds become a major threat in Malaysia's plantation. Eleusine indica (L.) Gaertn appeared as resistance to glyphosate in 1997. Hence, the development of C319 allele specific marker will help in controlling the weed in the effective ways. Genomic DNA extracted from susceptible and different level of resistances of goosegrass was used in this study. EPSPS allele specific clones were generated by cloning of the PCR fragments. EPSPS-C319 marker was optimized and used in screening of different resistant level of goosegrass with the EMC-F/ES2-R primer combination. The PCR optimization showed that the optimum condition was at 35 cycles and annealing temperature of 61 °C. Susceptible biotypes showed the presence of C allele in all ten samples. The resistant of 6-fold and 4-fold also showed the existence of C allele at specific band of 628 bp while no band produced in resistant 8-fold. Only C319 allele specific marker was successfully developed in this study thus further research on other allele specific marker could be carried out in the future.