In vitro mutagenesis and identification of mutants via ISSR in lily (Lilium longiflorum).
An efficient in vitro mutagenesis protocol for Lilium longiflorum Thunb. cv. White fox has been established. The effect of 6-BA and NAA on adventitious bud formation from the bulblet-scale thin cell layers was tested. Results showed that the optimal medium for adventitious bud induction is MS basal medium supplemented with 2.0 mg/l 6-BA and 0.1 mg/l NAA. The differentiation frequency and the average number of adventitious buds reached 95.55% and 3.00, respectively. Various doses (0.0, 0.5, 1.0, 1.5, 2.0, and 2.5 Gy) of gamma rays were applied to investigate the effect of radiation on adventitious bud formation from bulblet-scale thin cell layers. The forming capacity of the adventitious buds significantly decreased with the increase of radiation dose. The results suggested that the optimal irradiation dose is 1.0 Gy. Dose of 1.0 Gy treatment resulted in 55.33% survival of irradiated bulblet-scale thin cell layers and 39.27% mutagenesis rate. The genetic variations among the morphological mutants were evaluated by DNA fingerprinting using ISSR molecular marker. The genetic variation frequency reached 36.06% using seven ISSR primers. Out of the 50 mutant lines transferred to the greenhouse, 9 were observed to have significantly different morphological characters than those of the controls.
Key Laboratory of Forest Genetics and Biotechnology of the Ministry of Education, Nanjing Forestry University, Nanjing 210037, People's Republic of China.
Mengli
M
Xi
Lina
L
Sun
Shuai
S
Qiu
Juanjuan
J
Liu
Jin
J
Xu
Jisen
J
Shi
D001593:Benzyl Compounds; D004307:Dose-Response Relationship, Radiation; D035264:Flowers / Q000187:drug effects / Q000254:growth & development / Q000528:radiation effects; D005720:Gamma Rays; D007701:Kinetin / Q000494:pharmacology; D027762:Lilium / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D018895:Microsatellite Repeats / Q000235:genetics*; D016296:Mutagenesis / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D009154:Mutation / Q000235:genetics*; D009280:Naphthaleneacetic Acids / Q000494:pharmacology; D010641:Phenotype; D011687:Purines; D013242:Sterilization
D001593:Benzyl Compounds; D009280:Naphthaleneacetic Acids; D011687:Purines; C034182:1-naphthaleneacetic acid; C480551:benzylaminopurine; D007701:Kinetin