J Integr Plant Biol. ›› 2023, Vol. 65 ›› Issue (4): 907-917.DOI: 10.1111/jipb.13427

• New Technology • Previous Articles     Next Articles

Efficient genotype-independent cotton genetic transformation and genome editing

Xiaoyang Ge1,2,3†, Jieting Xu4†, Zhaoen Yang1,2†, Xiaofeng Yang4†, Ye Wang2, Yanli Chen2, Peng Wang2 and Fuguang Li1,2,3*   

  1. 1. Zhengzhou Research Base, State Key Laboratory of Cotton Biology, School of Agricultural Sciences, Zhengzhou University, Zhengzhou 450001, China;
    2. State Key Laboratory of Cotton Biology, Institute of Cotton Research, the Chinese Academy of Agricultural Sciences, Anyang 455000, China;
    3. Hainan Yazhou Bay Seed Laboratory, Sanya 572024, China;
    4. WIMI Biotechnology Co. Ltd, Changzhou 213000, China
    These authors contributed equally to this work.
    *Correspondence: Fuguang Li (aylifug@caas.cn)
  • Received:2022-11-08 Accepted:2022-12-02 Online:2022-12-07 Published:2023-04-01

Abstract: Cotton (Gossypium spp.) is one of the most important fiber crops worldwide. In the last two decades, transgenesis and genome editing have played important roles in cotton improvement. However, genotype dependence is one of the key bottlenecks in generating transgenic and gene‐edited cotton plants through either particle bombardment or Agrobacterium‐mediated transformation. Here, we developed a shoot apical meristem (SAM) cell‐ mediated transformation system (SAMT) that allowed the transformation of recalcitrant cotton genotypes including widely grown upland cotton (Gossypium hirsutum), Sea island cotton (Gossypium barbadense), and Asiatic cotton (Gossypium arboreum). Through SAMT, we successfully introduced two foreign genes, GFP and RUBY, into SAM cells of some recalcitrant cotton genotypes. Within 2–3 months, transgenic adventitious shoots generated from the axillary meristem zone could be recovered and grown into whole cotton plants. The GFP fluorescent signal and betalain accumulation could be observed in various tissues in GFP‐ and RUBY‐positive plants, as well as in their progenies, indicating that the transgenes were stably integrated into the genome and transmitted to the next generation. Furthermore, using SAMT, we successfully generated edited cotton plants with inheritable targeted mutagenesis in the GhPGF and GhRCD1 genes through CRISPR/Cas9‐mediated genome editing. In summary, the established SAMT transformation system here in this study bypasses the embryogenesis process during tissue culture in a conventional transformation procedure and significantly accelerates the generation of transgenic and gene‐edited plants for genetic improvement of recalcitrant cotton varieties.

Key words: cotton (Gossypium spp.), genotype dependence, shoot apical meristem, ultrasonic treatment

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