Skip to main content
Log in

A rapid and efficient Agrobacterium-mediated transient transformation system in grape berries

  • Original Article
  • Published:
Protoplasma Aims and scope Submit manuscript

Abstract

Transient transformation is extremely useful for rapid in vivo assessment of gene function, especially for fruit-related genes. Grape berry, while an important fruit crop, is recalcitrant to transient transformation, due to the high turgor pressure in its mesocarp cells that limits the ability of Agrobacterium to penetrate into the tissue. It is urgent to establish a simple transient transformation system for rapid analysis of gene function. In this study, different injection methods, grape genotypes, and developmental stages were tested in order to develop a rapid and efficient Agrobacterium-mediated transient transformation methodology for grape berries. Two injection methods, namely punch injection and direct injection, were evaluated using the β-glucuronidase (GUS) gene and by x-gluc tissue staining and 4-methylumbelliferyl-β-D-glucuronide fluorescence analysis. The results indicated that there were no significant differences on transformation effects between the two methods, but the latter was more suitable because of its simplicity and convenience. Six grape cultivars ('Hanxiangmi', 'Moldova', 'Zijixin', 'Jumeigui', 'Shine-Muscat', and 'A17') were tested for transient transformation. 'Hanxiangmi', 'Moldova', and 'Zijixin' grape berries were not suitable for agroinfiltration due to frequently fruit cracking, browning, and formation of scar skin. The fruit integrity rates of 'Jumeigui', 'Shine-Muscat', and 'A17' berries were all above 80%, and GUS activity was detected in the berries of the three cultivars 3–14 days after injection with the Agrobacterium culture, while higher GUS activities were observed in the 'Jumeigui' berries. The levels of GUS activity in injected berries at 7–8 weeks after full blooming (WAFB) were more than twice at 6 WAFB. In subsequent assays, the over-expression of MYB transcription factor VvMYB44 via transient transformation accelerated the anthocyanin accumulation and fruit coloring through raising the expression levels of VvLAR1, VvUFGT, VvLDOX, VvANS, and VvDFR, which verified the effectiveness of this transformation system. These experiments finally identified the reliable grape cultivars and suitable operational approach for transient transformation and further indicated that this Agrobacterium-mediated transient transformation system was efficient and suitable for the elucidation of gene function in grape berries.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Data availability

The datasets for all the figures and tables in this paper are available from the corresponding author on reasonable request.

References

Download references

Funding

This research was funded by the Natural Science Foundation of Henan Province (Grant No. 222300420457), the National Natural Science Foundation of China (Grant No. 32002017), and the Henan Province Outstanding Foreign Scholar Program (Grant No. GZS2020007).

Author information

Authors and Affiliations

Authors

Contributions

J.X. and C.H. performed the experiments and writing—original draft. M.L., S.H., and J.Q. completed the data analysis. Z.L. and B.T. provided experimental materials. X.Z and J.C. prepared the graphs. W.W. and J.L. helped in editing and revising the manuscript. X.Y. and J.F. conceived and designed the experiments. All the authors approved the finally version of the manuscript.

Corresponding authors

Correspondence to Jiancan Feng or Xia Ye.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Communicated by Handling Editor: Peter Nick

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xie, J., He, C., Li, Z. et al. A rapid and efficient Agrobacterium-mediated transient transformation system in grape berries. Protoplasma (2024). https://doi.org/10.1007/s00709-024-01938-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00709-024-01938-x

Keywords

Navigation