EMS离体诱变及抗草莓灰霉病愈伤组织的筛选

EMS MUTAGENESIS in vitro AND SCREENING OF CALLI FROM STRAWBERRY LEAVES OF RESISTANCE TO Botrytis chinerea Pers.

用甲基磺酸乙酯(EMS)处理草莓愈伤组织后通过草莓灰霉病病原毒素(Toxin)离体筛选获得抗病愈伤组织并诱导成苗。EMS诱变处理‘幸香’草莓离体叶片产生的愈伤组织,用经毒力检测的草莓灰霉菌毒素粗提液进行离体筛选,通过在愈伤组织继代培养中附加不同浓度毒素粗提液,以确定适宜的抗病选择压,获得抗灰霉病突变体材料。结果表明,将愈伤组织直接进行EMS处理的成活率高于将愈伤组织切割成小块继代培养后再经EMS处理的愈伤组织;且0.2%EMS处理1h为最佳选择。在含80%毒素粗提液的培养基中培养10 d为抗性筛选的适宜选择压。诱变处理后的867块愈伤组织,在经不连续二次筛选后有195块成活,成活率22.5%,其中有82块再生出了不定芽,再生率为42.1%。不定芽经生根后移栽至田间进一步进行抗性鉴定。

Calli from leaves of strawberry （Fragaria ananassa Duch. cv. Sachinoka）leaves were mutagenized in vitro by EMS （Ethyl methyl sulfonate） and screened by toxin extracts after the toxicity testing of Botrytis cinerea Pers. on strawberry. The results showed that the calli treated by EMS directly had higher survival rate than that of subcultured calli from small callus pieces and treated by EMS. The appropriate treatment was 0.2% EMS and for 1 hour, and the appropriate selection pressure was that the callus pieces were cultured for 10 days in medium supplemented with 80% of the toxic culture filtrate. There were 195 survival in 867 callus pieces after mutation and twice discontinuous screening the survival rate was 22.5 %, and of which 82 were regenerated adventitious shoots, regeneration rate was 42.1%. Some of resistant calli produced shoots were cultured, rooted and then transferred to field to test the resistance to Botrytis cinerea Pers.