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Cell Wall Epitopes and Endoploidy as Reporters of Embryogenic Potential in Brachypodium Distachyon Callus Culture

1
Department of Plant Anatomy and Cytology, Faculty of Biology and Environmental Protection, University of Silesia in Katowice, Katowice 40-007, Poland
2
Department of Genetics, Faculty of Biology and Environmental Protection, University of Silesia in Katowice, Katowice 40-007, Poland
3
Department of Cell Biology, Faculty of Biology and Environmental Protection, University of Silesia in Katowice, Katowice 40-007, Poland
4
National Plant Phenomics Centre, IBERS, Aberystwyth University, Aberystwyth SY23 3EE, UK
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2018, 19(12), 3811; https://doi.org/10.3390/ijms19123811
Received: 31 October 2018 / Revised: 26 November 2018 / Accepted: 27 November 2018 / Published: 29 November 2018
(This article belongs to the Section Molecular Plant Sciences)
Effective regeneration of callus tissue into embryos and then into whole plants is essential for plant biotechnology. The embryonic potential is often low and can further decrease with time in culture, which limits the utilisation of calli for transformation procedures and in vitro propagation. In this study, we show that the loss of embryogenic potential in callus cultures of Brachypodium distachyon is progressive over time. Flow cytometry analyses indicated endoploidy levels increased in 60- and 90-day-old calli with effective loss of the 2C DNA content peak in the latter. Analysis of indolic compounds content revealed a decrease in 60- and 90-day-old calli compared to either freshly isolated explants or 30-day-old calli. Immunohistochemical analysis revealed a decrease in arabinogalactan proteins (AGP) signal with the time of culture, but extensin (EXT) epitopes either increased (JIM12 epitopes) or decreased (JIM11 epitopes). The transcript accumulation levels of AGPs and EXTs confirmed these results, with most of AGP and EXT transcripts gradually decreasing. Some chimeric EXT transcripts significantly increased on the 30th day of culture, perhaps because of an increased embryogenic potential. Selected somatic embryogenesis-related genes and cyclins demonstrated a gradual decrease of transcript accumulation for YUCCA (YUC), AINTEGUMENTA-LIKE (AIL), BABY BOOM (BBM), and CLAVATA (CLV3) genes, as well as for most of the cyclins, starting from the 30th day of culture. Notably, WUSCHEL (WUS) transcript was detectable only on the 30th and 60th day and was not detectable in the zygotic embryos and in 90-day-old calli. View Full-Text
Keywords: arabinogalactan proteins; Brachypodium distachyon; cell wall; cyclins; extensins; pectins; ploidy instability; somatic embryogenesis; somaclonal variation; transcript accumulation arabinogalactan proteins; Brachypodium distachyon; cell wall; cyclins; extensins; pectins; ploidy instability; somatic embryogenesis; somaclonal variation; transcript accumulation
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Betekhtin, A.; Rojek, M.; Nowak, K.; Pinski, A.; Milewska-Hendel, A.; Kurczynska, E.; Doonan, J.H.; Hasterok, R. Cell Wall Epitopes and Endoploidy as Reporters of Embryogenic Potential in Brachypodium Distachyon Callus Culture. Int. J. Mol. Sci. 2018, 19, 3811.

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