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目的建立甜叶菊甲基磺酸乙酯(EMS)离体诱变体系,通过SRAP检测鉴定获得甜叶菊耐盐突变体。方法将“皖甜1号”-B1甜叶菊组培苗茎段接种在含有不同质量浓度Na Cl的MS培养基中,进行耐盐临界Na Cl质量浓度的筛选;用不同浓度的EMS处理甜叶菊组培苗茎段不同时间,筛选EMS诱变甜叶菊的适宜浓度和时间;将经过EMS诱变过的甜叶菊组培苗茎段接种在含有临界Na Cl质量浓度的MS培养基上进行耐盐变异体的筛选,对存活的变异株通过SRAP标记进行耐盐性鉴定。结果甜叶菊组培苗耐盐临界Na Cl质量浓度为1.0%;EMS诱变甜叶菊组培苗茎段的适宜浓度和时间范围为0.8%~1.0%EMS处理8~10 h;筛选出41株甜叶菊耐盐变异株,SRAP分子标记表明,4株在DNA水平上发生了变异,突变比率为9.76%。结论初步建立了甜叶菊EMS离体诱变体系,为甜叶菊高产耐盐新品种的选育提供了一种新的育种途径。
Aim To establish the in vitro mutagenesis system of stevia ethylmethanesulfonate (EMS) and obtain the salt-tolerant mutant of stevia rebaudiana by SRAP. Methods The shoots of tissue culture plantlets of “Sweet Sweet 1” - B1 were inoculated on MS medium containing different concentrations of NaCl, and then were screened for the Na salt concentration critical to salt tolerance. EMS treatment The appropriate concentration and time of EMS-mutagenized Stevia was screened at different time points in the tissue culture seedlings of Stevia rebaudianae. The stems of the tissue-cultured Stevia rebaudiana seedlings were inoculated on the MS medium containing the critical NaC1 concentration Salt-tolerant variants were screened, and surviving variants were identified for salt tolerance by SRAP markers. Results The concentration of NaCl in the tissue culture seedlings with salt tolerance was 1.0%. The optimum concentration and time of EMS in the treatment of Stevia rebaudianae tissue culture seedlings were 0.8% -1.0% EMS for 8-10 h. 41 strains The salt tolerance mutant of Stevia rebaudiana and SRAP molecular marker showed that 4 strains mutated at the DNA level with a mutation rate of 9.76%. Conclusion The in vitro mutagenesis system of Stevia rebaudiana was preliminarily established, which provided a new breeding method for the breeding of new varieties of high yield and salt tolerant stevia.