TY - JOUR
T1 - Mutant selection from progeny of gamma-ray-irradiated rice by DNA heteroduplex cleavage using Brassica petiole extract
AU - Sato, Yutaka
AU - Shirasawa, Kenta
AU - Takahashi, Yoshinobu
AU - Nishimura, Minoru
AU - Nishio, Takeshi
PY - 2006
Y1 - 2006
N2 - Selection of mutants by the reverse genetic approach is a useful tool for gene function analysis and crop improvement. TILLING (Targeting Induced Local Lesions in Genomes), in which mutants are selected from the progeny of EMS-treated plants by the SNP analysis using heteroduplex cleavage enzyme from celery, has been applied to various plant species. Since gamma-ray-induced point mutations have not been identified by the reverse genetic approach, we attempted to select mutants from a population of the gamma-ray-irradiated rice. A heteroduplex between wild-type DNA and mutant DNA was cleaved by an endonuclease extracted from petioles of Brassica rapa, and cleaved DNA was detected using agarose gel electrophoresis. M2 plants derived from 2,130 M1 plants irradiated by gamma rays were used for the screening of mutants having nucleotide changes in 25 regions from 1.0 kb to 1.5 kb. Six mutations were identified and the rate of mutation induced by gamma rays was estimated to be one mutation per 6,190 kb. Four of the mutations were single nucleotide substitutions and two were 2-bp and 4-bp deletions. These results suggest that the point mutation rate with gamma rays was quite lower than that with EMS, but that the rate of knockout mutations among detected mutations generated by the gamma-ray irradiation was higher than that by the EMS treatment.
AB - Selection of mutants by the reverse genetic approach is a useful tool for gene function analysis and crop improvement. TILLING (Targeting Induced Local Lesions in Genomes), in which mutants are selected from the progeny of EMS-treated plants by the SNP analysis using heteroduplex cleavage enzyme from celery, has been applied to various plant species. Since gamma-ray-induced point mutations have not been identified by the reverse genetic approach, we attempted to select mutants from a population of the gamma-ray-irradiated rice. A heteroduplex between wild-type DNA and mutant DNA was cleaved by an endonuclease extracted from petioles of Brassica rapa, and cleaved DNA was detected using agarose gel electrophoresis. M2 plants derived from 2,130 M1 plants irradiated by gamma rays were used for the screening of mutants having nucleotide changes in 25 regions from 1.0 kb to 1.5 kb. Six mutations were identified and the rate of mutation induced by gamma rays was estimated to be one mutation per 6,190 kb. Four of the mutations were single nucleotide substitutions and two were 2-bp and 4-bp deletions. These results suggest that the point mutation rate with gamma rays was quite lower than that with EMS, but that the rate of knockout mutations among detected mutations generated by the gamma-ray irradiation was higher than that by the EMS treatment.
KW - Gamma rays
KW - Heteroduplex cleavage
KW - Mutation
KW - Reverse genetics
KW - TILLING
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U2 - 10.1270/jsbbs.56.179
DO - 10.1270/jsbbs.56.179
M3 - Article
AN - SCOPUS:33745147174
VL - 56
SP - 179
EP - 183
JO - Breeding Science
JF - Breeding Science
SN - 1344-7610
IS - 2
ER -