Eutrema salsugineum is an Arabidopsis-Related Model Species naturally adapted to harsh environments andtolerant to cold and high soil salinity. Growth requirements are very similar to those of Arabidopsis thaliana;however key differences in life cycle require special care. To generate a mutagenized population for forwardgenetic studies, we optimized the EMS (Ethyl Methanesulfonate) mutagenesis procedure for this species.We used ver162 mutant, developed in Dr. Jian-Kang Zhu's laboratory, which does not require vernalizationand is therefore easier to handle in high throughput studies. Different EMS concentrations were tested anda refined protocol was developed based on the adverse effect on seed germination and the presence ofalbino sections on leaves. When bulk mutagenesis was performed using 0.6% EMS for 6 hours, we foundover 12 plants that required vernalization in the resulting M-1 population, suggesting a possible reversion ofthe phenotype compared to background ver162, a further indication of the success of the mutagenesis. M-2progeny was harvested in pools and screening in vitro for NaCl-hypersensitive phenotypes is now inprogress.
Using Arabidopsis-Related Model Species (ARMS): Growth, Mutagenesis and Screening of Eutrema Salsugineum
Ruggiero A;Costa A;Aurilia V;De Palma M;Batelli G
2016-01-01
Abstract
Eutrema salsugineum is an Arabidopsis-Related Model Species naturally adapted to harsh environments andtolerant to cold and high soil salinity. Growth requirements are very similar to those of Arabidopsis thaliana;however key differences in life cycle require special care. To generate a mutagenized population for forwardgenetic studies, we optimized the EMS (Ethyl Methanesulfonate) mutagenesis procedure for this species.We used ver162 mutant, developed in Dr. Jian-Kang Zhu's laboratory, which does not require vernalizationand is therefore easier to handle in high throughput studies. Different EMS concentrations were tested anda refined protocol was developed based on the adverse effect on seed germination and the presence ofalbino sections on leaves. When bulk mutagenesis was performed using 0.6% EMS for 6 hours, we foundover 12 plants that required vernalization in the resulting M-1 population, suggesting a possible reversion ofthe phenotype compared to background ver162, a further indication of the success of the mutagenesis. M-2progeny was harvested in pools and screening in vitro for NaCl-hypersensitive phenotypes is now inprogress.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
