摘要
Glyphosate has been used worldwide for nearly 40 years,and 30 types of resistant weeds have been reported.Glyphosate is mass-produced and widely used in China,but few studies and reports on glyphosate-resistant weeds and resistance mechanisms exist.Previous studies found a goosegrass species with high glyphosate resistance from orchards in South China and its glyphosate resistant mechanism was described in this study.The cDNAof 5-enolpyruvylshikimate-3-phosphate synthase（EPSPS,EC 2.5.1.19）,the target enzyme of glyphosate,was cloned from the glyphosate-resistant and-susceptible goosegrass,respectively,and referred as EPSPS-R and EPSPS-S.The Pro106 residue was known to be involved in the glyphosate resistance in most goosegrass populations.However,sequence analysis did not find the mutation at the Pro106 residue in the R biotype EPSPS amino acid sequence.The residue 133 and 382 was mutated in the R biotype EPSPS amino acid sequence instead,but it did not affect the EPSPS-S and EPSPS-R genes sensitivities to glyphosate.RT-PCR and Western blot analyses suggested that EPSPS mRNA and protein are mainly present in the shoot tissues both in the R and S goosegrass biotypes.The EPSPS-R rapidly responds to the glyphosate in R-biotype goosegrass and the induced expression was detected at 12 h post glyphosate treatment.The mRNA and protein expression of EPSPS-R increased constantly as the increasing concentration of glyphosate.However,the expression of the EPSPS-S was not induced significantly by glyphosate in the S goosegrass biotype.Quantification of real-time PCR results showed that the copy number of the EPSPS in R-biotype goosegrass was 4.7 times higher than that in the S goosegrass biotype.All the results implied that EPSPS gene amplification might mainly caused the glyphosate resistance of a goosegrass population collected from orchards in South China.
Glyphosate has been used worldwide for nearly 40 years,and 30 types of resistant weeds have been reported.Glyphosate is mass-produced and widely used in China,but few studies and reports on glyphosate-resistant weeds and resistance mechanisms exist.Previous studies found a goosegrass species with high glyphosate resistance from orchards in South China and its glyphosate resistant mechanism was described in this study.The cDNAof 5-enolpyruvylshikimate-3-phosphate synthase（EPSPS,EC 2.5.1.19）,the target enzyme of glyphosate,was cloned from the glyphosate-resistant and-susceptible goosegrass,respectively,and referred as EPSPS-R and EPSPS-S.The Pro106 residue was known to be involved in the glyphosate resistance in most goosegrass populations.However,sequence analysis did not find the mutation at the Pro106 residue in the R biotype EPSPS amino acid sequence.The residue 133 and 382 was mutated in the R biotype EPSPS amino acid sequence instead,but it did not affect the EPSPS-S and EPSPS-R genes sensitivities to glyphosate.RT-PCR and Western blot analyses suggested that EPSPS mRNA and protein are mainly present in the shoot tissues both in the R and S goosegrass biotypes.The EPSPS-R rapidly responds to the glyphosate in R-biotype goosegrass and the induced expression was detected at 12 h post glyphosate treatment.The mRNA and protein expression of EPSPS-R increased constantly as the increasing concentration of glyphosate.However,the expression of the EPSPS-S was not induced significantly by glyphosate in the S goosegrass biotype.Quantification of real-time PCR results showed that the copy number of the EPSPS in R-biotype goosegrass was 4.7 times higher than that in the S goosegrass biotype.All the results implied that EPSPS gene amplification might mainly caused the glyphosate resistance of a goosegrass population collected from orchards in South China.
基金
supported by the National Natural Science Foundation of China(31301683)
the Science and Technology Planning Project of Guangdong Province,China(2012A020100009)
