Immunolocalization of Extensin and Pectin Epitopes in <i>Liparis loeselii</i> Protocorm and Protocorm-like Bodies
<i>Liparis loeselii</i> (L.) Rich, an endangered member of the Orchidaceae family, is found in alkaline fens. With the declining populations of <i>L. loeselii</i>, there is a pressing need to reintroduce this species in Central Europe. As in vitro germination is a crucial too...
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| author | Michał D. Starke Małgorzata Kapusta Bartosz J. Płachno Jerzy Bohdanowicz |
| author_facet | Michał D. Starke Małgorzata Kapusta Bartosz J. Płachno Jerzy Bohdanowicz |
| author_sort | Michał D. Starke |
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| description | <i>Liparis loeselii</i> (L.) Rich, an endangered member of the Orchidaceae family, is found in alkaline fens. With the declining populations of <i>L. loeselii</i>, there is a pressing need to reintroduce this species in Central Europe. As in vitro germination is a crucial tool for obtaining plants for introduction into the environment, we looked at the morphological changes occurring during the early stages of <i>L. loeselii</i> development in vitro. As the early stages of orchid development, especially the protocorm stage, are thought to be responsible for SAM formation and the initiation of symbiotic association, we focused on cell wall elements whose epitopes have been found in similar processes in other species: the extensin and pectin rhamnogalacturonan I (RG-I) side chain epitopes. We addressed the following questions: Does the cell wall of <i>L. loeselii</i> change its composition during the early stages of development, as noted in other species? Are there noticeable similarities in the cell wall to organs of different species whose function is to contact microorganisms? Are there regularities that allow the recognition of individual structures on this basis? Immunolocalization revealed changes in the distribution of certain extensins (JIM11 and JIM20) and RG-I (LM5 and LM6) side chain epitopes. Extensins, a type of cell wall protein, were observed during the initial stages of the formation of PLB and the shoot apical meristem of protocorms and PLBs. RG-I, on the other hand, was found to play a significant role in the development of the protocorm and PLB. In pseudobulbs, which appeared on the protocorms, extensins occurred in their storage part. However, RG-I side chains (1→4)-β-galactans (LM5), and (1→5)-α-L-arabinans (LM6) were not found in pseudobulbs. We revealed that a common feature of protocorms and PLBs was an increased amount of extensins, which were detected with the JIM11 antibody, and pectins, which were detected with the LM5 antibody, that were present together, which may prove helpful in determining the identity of the induced structures and distinguishing them from pseudobulbs. Thus, our study unveiled the role of extensins and RG-I during the growth of protocorms and PLBs. We suggest that PLBs may mimic the wall remodelling that occurs in protocorms, which indicates that using cell wall components is an invitation to be colonised by a fungal partner. However, this needs to be tested in future research. The findings of this research can help interpret future studies on the propagation, acclimatisation, and introduction of <i>L. loeselii</i> into the natural environment. |
| format | Article |
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| spelling | doaj-art-6c9d5a4ebbe9496c845b8fb744e994bd2025-08-20T01:55:31ZengMDPI AGCells2073-44092024-11-011323198510.3390/cells13231985Immunolocalization of Extensin and Pectin Epitopes in <i>Liparis loeselii</i> Protocorm and Protocorm-like BodiesMichał D. Starke0Małgorzata Kapusta1Bartosz J. Płachno2Jerzy Bohdanowicz3Laboratory of Plant Cytology and Embryology, Department of Plant Experimental Biology and Biotechnology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St., 80-308 Gdansk, PolandBioimaging Laboratory, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St., 80-308 Gdansk, PolandDepartment of Plant Cytology and Embryology, Institute of Botany, Faculty of Biology, Jagiellonian University in Kraków, 9 Gronostajowa St., 30-387 Cracow, PolandLaboratory of Plant Cytology and Embryology, Department of Plant Experimental Biology and Biotechnology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St., 80-308 Gdansk, Poland<i>Liparis loeselii</i> (L.) Rich, an endangered member of the Orchidaceae family, is found in alkaline fens. With the declining populations of <i>L. loeselii</i>, there is a pressing need to reintroduce this species in Central Europe. As in vitro germination is a crucial tool for obtaining plants for introduction into the environment, we looked at the morphological changes occurring during the early stages of <i>L. loeselii</i> development in vitro. As the early stages of orchid development, especially the protocorm stage, are thought to be responsible for SAM formation and the initiation of symbiotic association, we focused on cell wall elements whose epitopes have been found in similar processes in other species: the extensin and pectin rhamnogalacturonan I (RG-I) side chain epitopes. We addressed the following questions: Does the cell wall of <i>L. loeselii</i> change its composition during the early stages of development, as noted in other species? Are there noticeable similarities in the cell wall to organs of different species whose function is to contact microorganisms? Are there regularities that allow the recognition of individual structures on this basis? Immunolocalization revealed changes in the distribution of certain extensins (JIM11 and JIM20) and RG-I (LM5 and LM6) side chain epitopes. Extensins, a type of cell wall protein, were observed during the initial stages of the formation of PLB and the shoot apical meristem of protocorms and PLBs. RG-I, on the other hand, was found to play a significant role in the development of the protocorm and PLB. In pseudobulbs, which appeared on the protocorms, extensins occurred in their storage part. However, RG-I side chains (1→4)-β-galactans (LM5), and (1→5)-α-L-arabinans (LM6) were not found in pseudobulbs. We revealed that a common feature of protocorms and PLBs was an increased amount of extensins, which were detected with the JIM11 antibody, and pectins, which were detected with the LM5 antibody, that were present together, which may prove helpful in determining the identity of the induced structures and distinguishing them from pseudobulbs. Thus, our study unveiled the role of extensins and RG-I during the growth of protocorms and PLBs. We suggest that PLBs may mimic the wall remodelling that occurs in protocorms, which indicates that using cell wall components is an invitation to be colonised by a fungal partner. However, this needs to be tested in future research. The findings of this research can help interpret future studies on the propagation, acclimatisation, and introduction of <i>L. loeselii</i> into the natural environment.https://www.mdpi.com/2073-4409/13/23/1985cell wallcell wall remodellingendangered speciesextensinsOrchidaceaeorchid germination |
| spellingShingle | Michał D. Starke Małgorzata Kapusta Bartosz J. Płachno Jerzy Bohdanowicz Immunolocalization of Extensin and Pectin Epitopes in <i>Liparis loeselii</i> Protocorm and Protocorm-like Bodies Cells cell wall cell wall remodelling endangered species extensins Orchidaceae orchid germination |
| title | Immunolocalization of Extensin and Pectin Epitopes in <i>Liparis loeselii</i> Protocorm and Protocorm-like Bodies |
| title_full | Immunolocalization of Extensin and Pectin Epitopes in <i>Liparis loeselii</i> Protocorm and Protocorm-like Bodies |
| title_fullStr | Immunolocalization of Extensin and Pectin Epitopes in <i>Liparis loeselii</i> Protocorm and Protocorm-like Bodies |
| title_full_unstemmed | Immunolocalization of Extensin and Pectin Epitopes in <i>Liparis loeselii</i> Protocorm and Protocorm-like Bodies |
| title_short | Immunolocalization of Extensin and Pectin Epitopes in <i>Liparis loeselii</i> Protocorm and Protocorm-like Bodies |
| title_sort | immunolocalization of extensin and pectin epitopes in i liparis loeselii i protocorm and protocorm like bodies |
| topic | cell wall cell wall remodelling endangered species extensins Orchidaceae orchid germination |
| url | https://www.mdpi.com/2073-4409/13/23/1985 |
| work_keys_str_mv | AT michałdstarke immunolocalizationofextensinandpectinepitopesiniliparisloeseliiiprotocormandprotocormlikebodies AT małgorzatakapusta immunolocalizationofextensinandpectinepitopesiniliparisloeseliiiprotocormandprotocormlikebodies AT bartoszjpłachno immunolocalizationofextensinandpectinepitopesiniliparisloeseliiiprotocormandprotocormlikebodies AT jerzybohdanowicz immunolocalizationofextensinandpectinepitopesiniliparisloeseliiiprotocormandprotocormlikebodies |