Abstract
The ability of Orobanchaceae to establish a parasitic relationship is based on the development of a specific organ called haustorium. Previous studies in Phelipanche aegyptiaca and P. ramosa have underlined the interest of producing calli from germinated seeds as an efficient genetic transformation system, but they also pointed the need to improve the attachment rate of infectious calli to the host plant as well as the seed yield of the regenerated plants following attachment. It was previously shown that haustorium formation in P. ramosa is triggered by cytokinins. The present work demonstrates that one month-old microcalli produced from germinated seeds displayed a response to c/tZ similar to germinated seeds, as shown by the temporal expression profile of markers genes of cytokinin-related haustorium initiation, including PrRR5, PrCKX2, PrCKX4, PrTRN2 and PrZFP6. In addition, a 48 h treatment of microcalli with c/tZ (10−7 M) before infestation triggered a 4-fold increase in the attachment rate of microcalli to tomato roots after 18 days in minirhizotrons when compared to untreated microcalli. Finally, it also outlines that increasing the aggressiveness of microcalli with a c/tZ treatment followed by the transfer of parasitized tomato plants into pots significantly improved seed yield of regenerated P. ramosa plants. Indeed, high amounts of viable seeds which germinated at more than 90% in response to GR24 were harvested after a 10 weeks-co-cultivation period. According to these results, cytokinin treated P. ramosa microcalli thus appear to be a good tool for further functional studies in holoparasitic plants, especially on haustorium formation.
Key message
Cytokinin treated microcalli show a high ability to attach to host roots and to regenerate seed-producing plants. They are of great interest for functional studies especially for studying haustorium formation.
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Change history
01 May 2020
The caption to Fig. 4 belonged to Fig. 5 and vice versa in the initial online publication. The original article has been corrected.
Abbreviations
- c/tZ:
-
cis–trans-Zeatin
- MS/MES:
-
Murashige & Skoog medium containing Nitsch vitamins and MES buffer
- PPM:
-
Plant preservative mixture
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Acknowledgements
EB and VG were financially supported by PhD grants from RFI Objectif Végétal program (Region Pays de Loire, France) and the French Ministry of Education and Research, respectively. The authors also thank Johannes Schmidt and Sabine Delgrange for their technical support.
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Funding was provided by Conseil Régional des Pays de la Loire (Grant No. Ministère de l'Enseignement supérieur, de la Recherche et de l'Innovation).
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This work was supervised by PS and GM. PS, GM, EB, VG conceived the design experiments. EB, VG performed the experiments and analysed the data. EB, PS, PD, GM wrote the manuscript.
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Communicated by Mohammad Faisal.
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The original version of this article has been revised: The captions to Figs. 4 and 5 have been switched to match the correct figures.
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Billard, E., Goyet, V., Delavault, P. et al. Cytokinin treated microcalli of Phelipanche ramosa: an efficient model for studying haustorium formation in holoparasitic plants. Plant Cell Tiss Organ Cult 141, 543–553 (2020). https://doi.org/10.1007/s11240-020-01813-6
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DOI: https://doi.org/10.1007/s11240-020-01813-6