Techniques for the regeneration and genetic transformation of Arabidopsis pumila: an ephemeral plant suitable for investigating the mechanisms for adaptation to desert environments

Key message Establishment of tissue culture in ephemeral plant Arabidopsis pumila and efficient Agrobacterium-mediated genetic transformation for investigating the mechanisms for adaptation to desert environments. Arabidopsis pumila is a type of cruciferous ephemeral plant, which in China mainly grows in the desert environments of northern Xinjiang. A. pumila not only has a short growth duration, but also has high photosynthetic efficiency, seed yield, salt tolerance, and drought resistance. It is an ideal species for the study of environmental adaptations in ephemeral plants. We induced callus tissue formation on the roots and hypocotyls of 8-day-old seedlings, and on the leaves and petioles of 4-week-old seedlings, and obtained multiple adventitious shoots on these tissues grown on Murashige and Skoog induction medium supplemented with 0.5 mg/L 6-Benzylaminopurine and 0.1 mg/L alpha-Naphthalene acetic acid. Young roots, hypocotyls, leaves, and petioles could all induce calluses, but the induction rate was highest on young roots. In addition, the leaves and petioles of 4-week-old seedlings were used as explants, the Delta 1-pyrroline-5-carboxylic acid synthase gene 1 of A. pumila controlled by 35S promoter of cauliflower mosaic virus was used as target gene, and hygromycin B was used as screening antibiotic to explore Agrobacterium tumefaciens GV3101 mediated transformation. The results showed that the callus induction rate of petiole explants was the highest when they were treated with Agrobacterium suspension (OD600 = 0.6) for 10 min and thenco-cultured in dark for 2 days. The qRT-PCR results showed that the ApP5CS1.1 gene was overexpressed in the transgenic plants. These protocols provide working research methods for exploring the cellular level adaptative mechanisms of this species to desert environments.

Automated summary

Translation: Establishment of tissue culture and efficient Agrobacterium-mediated genetic transformation in ephemeral plant Arabidopsis pumila for studying adaptation mechanisms to desert environments. Callus tissue formation was induced on different parts of the plant, and multiple adventitious shoots were obtained. The highest callus induction rate was observed in petiole explants. Using Agrobacterium tumefaciens GV3101-mediated transformation, the target gene ApP5CS1.1 was found to be overexpressed in the transgenic plants. These protocols provide a research method for studying the cellular-level adaptive mechanisms of A. pumila to desert environments.