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

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Abstract

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 α-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 Δ1-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.

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.

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Abbreviations

MS:

Murashige and Skoog

6-BA:

6-Benzylaminopurine

NAA:

α-Naphthalene acetic acid

Carb:

Carbenicillin

P5CS1 :

Delta-1-pyrroline-5-carboxylic acid synthase 1

qRT-PCR:

Quantitative real-time PCR

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Acknowledgements

This work is supported by the grants from the National Natural Science Foundation of China (Grant No. U1303302), the Shihezi University International Cooperation Program (Grant No. GJHZ201806), the Academic Leaders of Anhui University of Science and Technology Introduction Starting Fund (Grant No. NXYJ202001), and the Research and Innovation Project of Postgraduates in Xinjiang Uygur Autonomous Region (Grant No. XJ2019G083).

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Authors and Affiliations

  1. College of Life Sciences, Shihezi University, Shihezi, 832000, Xinjiang, China

    Yuhuan Jin, Li Guo, Danqing Liu & Yongguang Li

  2. Center for Crop Biotechnology, College of Agriculture, Anhui University of Science and Technology, Anhui Crop Biotechnology Center, Fengyang, 233100, China

    Yuhuan Jin, Hao Ai & Xianzhong Huang

  3. Special Plant Genomics Laboratory, College of Life Sciences, Shihezi University, Shihezi, 832000, Xinjiang, China

    Xianzhong Huang

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  1. Yuhuan Jin
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  2. Li Guo
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Contributions

XH and YJ conceived and designed research. YJ, LG, DL, and HA conducted the experiments. LZ and AC analyzed the data and revise the manuscript. All authors read and approved the manuscript.

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Correspondence to Xianzhong Huang.

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Jin, Y., Guo, L., Liu, D. et al. Techniques for the regeneration and genetic transformation of Arabidopsis pumila: an ephemeral plant suitable for investigating the mechanisms for adaptation to desert environments. Plant Cell Tiss Organ Cult 150, 237–246 (2022). https://doi.org/10.1007/s11240-022-02264-x

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