Table 1.
The antibodies used in this study, the epitopes they recognise and relevant references.
Fig 1.
Morphology of selected structures in an in vitro Brachypodium culture.
(A) Immature embryos. (B) Embryogenic callus–embryogenic masses marked by yellow asterisks; vitreous and friable callus indicated by white asterisks). (C) Spontaneous somatic embryogenesis seven day after cultivation on the CIM medium.
Fig 2.
Histology of the Brachypodium embryogenic callus on the 7th day after cultivation.
(A) General view through the histological section with embryogenic masses (EM) and parenchymatous cells (PC). (B) and (C) Close-up of embryogenic masses, a black dotted square with an arrow shows examples of the cells that contain nuclei with two nucleoli; a red dotted square with an arrow indicates dividing cells. (D) Sector of callus that contains parenchymatous cells.
Fig 3.
SEM images of the Brachypodium embryogenic callus surface pattern.
A callus on the 7th (A-E) and 21st (F-H) days after cultivation. (A-C)–embryogenic masses covered by an extracellular matrix surface network (ECMSN is indicated by yellow asterisks): note the fibrillar and membranous structure. (D) Parenchymatous cells; high-magnification insert (D’) shows the contact between these cells. (E) Developing somatic embryos. (F) Developed multiple coleoptiles. (G) and (H) a callus on the 21st day after cultivation showing the absence of ECMSN on the surface.
Table 2.
Summary of the immunocytochemical detection of selected antigens in embryogenic callus.
Fig 4.
Immunolocalisation of arabinogalactans and arabinogalactan proteins in a Brachypodium embryogenic callus on the 7th day after cultivation.
(A)–(A”) and (B)–(B”) MAC207 in the cell walls of the dead cells on the callus surface (marked by red arrows) and parenchymatous cells, respectively. (C)–(C”) PC with JIM13. (D)–(D”) EM and PC with JIM16. (E)–(E”) EM and PC with LM2.
Fig 5.
Immunolocalisation of extensins in a Brachypodium embryogenic callus on the 7th day after cultivation.
(A-A”) and (B-B”)–the surface of EM and PC with JIM11, respectively. (C-C”)–PC with JIM12.
Fig 6.
Immunolocalisation of pectins in a Brachypodium embryogenic callus on the 7th day after cultivation.
(A-A”) and (B-B”)–EM and PC with LM6, respectively. (C-C”)–EM and PC with LM16.
Fig 7.
Immunolocalisation of pectins in a Brachypodium embryogenic callus on the 7th day after cultivation.
(A-A”) and (B-B”) EM and PC with LM19. (C-C”) and (D-D”)–EM with LM20.
Fig 8.
Immunolocalisation of hemicelluloses in a Brachypodium embryogenic callus on the 7th day after cultivation.
(A-A”) and (B-B”) EM and PC with LM21. (C-C”) and (D-D”)–EM and PC with LM25.
Fig 9.
Negative control for the experiment with JIM13 antibodies.
(A-A”)–no fluorescent signals were observed for these antibodies. Analogous results were obtained for all of the antibodies that were used.