Phylogenetic and structural analysis of annexins in pea (<i>Pisum sativum</i> L.) and their role in legume-rhizobial symbiosis development
Annexins as Ca2+/phospholipid-binding proteins are involved in the control of many biological processes essential for plant growth and development. In a previous study, we had shown, using a proteomic approach, that the synthesis of two annexins is induced in pea roots in response to rhizobial inocu...
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Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders
2021-09-01
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| Series: | Вавиловский журнал генетики и селекции |
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| Online Access: | https://vavilov.elpub.ru/jour/article/view/3108 |
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| author | O. A. Pavlova I. V. Leppyanen D. V. Kustova A. D. Bovin E. A. Dolgikh |
| author_facet | O. A. Pavlova I. V. Leppyanen D. V. Kustova A. D. Bovin E. A. Dolgikh |
| author_sort | O. A. Pavlova |
| collection | DOAJ |
| description | Annexins as Ca2+/phospholipid-binding proteins are involved in the control of many biological processes essential for plant growth and development. In a previous study, we had shown, using a proteomic approach, that the synthesis of two annexins is induced in pea roots in response to rhizobial inoculation. In this study, phylogenetic analysis identified these annexins as PsAnn4 and PsAnn8 based on their homology with annexins from other legumes. The modeling approach allowed us to estimate the structural features of these annexins that might influence their functional activity. To verify the functions of these annexins, we performed comparative proteomic analysis, experiments with calcium influx inhibitors, and localization of labeled proteins. Essential down-regulation of PsAnn4 synthesis in a non-nodulating pea mutant P56 (sym10) suggests an involvement of this annexin in the rhizobial symbiosis. Quantitative RT-PCR analysis showed that PsAnn4 was upregulated at the early stages of symbiosis development, starting from 1–3 days after inoculation to up to 5 days after inoculation, while experiments with the Ca2+ channel blocker LaCl3 revealed its negative influence on this expression. To follow the PsAnn4 protein localization in plant cells, it was fused to the fluorophores such as red fluorescent protein (RFP) and yellow fluorescent protein (YFP) and expressed under the transcriptional regulation of the 35S promoter in Nicotiana benthamiana leaves by infiltration with Agrobacterium tumefaciens. The localization of PsAnn4 in the cell wall or plasma membrane of plant cells may indicate its participation in membrane modification or ion transport. Our results suggest that PsAnn4 may play an important role during the early stages of pea-rhizobial symbiosis development. |
| format | Article |
| id | doaj-art-3e594091c75f45d6bf3567ce36e4412e |
| institution | Kabale University |
| issn | 2500-3259 |
| language | English |
| publishDate | 2021-09-01 |
| publisher | Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders |
| record_format | Article |
| series | Вавиловский журнал генетики и селекции |
| spelling | doaj-art-3e594091c75f45d6bf3567ce36e4412e2025-02-01T09:58:10ZengSiberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and BreedersВавиловский журнал генетики и селекции2500-32592021-09-0125550251310.18699/VJ21.0571184Phylogenetic and structural analysis of annexins in pea (<i>Pisum sativum</i> L.) and their role in legume-rhizobial symbiosis developmentO. A. Pavlova0I. V. Leppyanen1D. V. Kustova2A. D. Bovin3E. A. Dolgikh4All-Russia Research Institute for Agricultural MicrobiologyAll-Russia Research Institute for Agricultural MicrobiologyAll-Russia Research Institute for Agricultural MicrobiologyAll-Russia Research Institute for Agricultural MicrobiologyAll-Russia Research Institute for Agricultural MicrobiologyAnnexins as Ca2+/phospholipid-binding proteins are involved in the control of many biological processes essential for plant growth and development. In a previous study, we had shown, using a proteomic approach, that the synthesis of two annexins is induced in pea roots in response to rhizobial inoculation. In this study, phylogenetic analysis identified these annexins as PsAnn4 and PsAnn8 based on their homology with annexins from other legumes. The modeling approach allowed us to estimate the structural features of these annexins that might influence their functional activity. To verify the functions of these annexins, we performed comparative proteomic analysis, experiments with calcium influx inhibitors, and localization of labeled proteins. Essential down-regulation of PsAnn4 synthesis in a non-nodulating pea mutant P56 (sym10) suggests an involvement of this annexin in the rhizobial symbiosis. Quantitative RT-PCR analysis showed that PsAnn4 was upregulated at the early stages of symbiosis development, starting from 1–3 days after inoculation to up to 5 days after inoculation, while experiments with the Ca2+ channel blocker LaCl3 revealed its negative influence on this expression. To follow the PsAnn4 protein localization in plant cells, it was fused to the fluorophores such as red fluorescent protein (RFP) and yellow fluorescent protein (YFP) and expressed under the transcriptional regulation of the 35S promoter in Nicotiana benthamiana leaves by infiltration with Agrobacterium tumefaciens. The localization of PsAnn4 in the cell wall or plasma membrane of plant cells may indicate its participation in membrane modification or ion transport. Our results suggest that PsAnn4 may play an important role during the early stages of pea-rhizobial symbiosis development.https://vavilov.elpub.ru/jour/article/view/3108legume-rhizobial symbiosispea annexinsthree-dimensional modelingproteomicscalcium inhibitorslocalization |
| spellingShingle | O. A. Pavlova I. V. Leppyanen D. V. Kustova A. D. Bovin E. A. Dolgikh Phylogenetic and structural analysis of annexins in pea (<i>Pisum sativum</i> L.) and their role in legume-rhizobial symbiosis development Вавиловский журнал генетики и селекции legume-rhizobial symbiosis pea annexins three-dimensional modeling proteomics calcium inhibitors localization |
| title | Phylogenetic and structural analysis of annexins in pea (<i>Pisum sativum</i> L.) and their role in legume-rhizobial symbiosis development |
| title_full | Phylogenetic and structural analysis of annexins in pea (<i>Pisum sativum</i> L.) and their role in legume-rhizobial symbiosis development |
| title_fullStr | Phylogenetic and structural analysis of annexins in pea (<i>Pisum sativum</i> L.) and their role in legume-rhizobial symbiosis development |
| title_full_unstemmed | Phylogenetic and structural analysis of annexins in pea (<i>Pisum sativum</i> L.) and their role in legume-rhizobial symbiosis development |
| title_short | Phylogenetic and structural analysis of annexins in pea (<i>Pisum sativum</i> L.) and their role in legume-rhizobial symbiosis development |
| title_sort | phylogenetic and structural analysis of annexins in pea i pisum sativum i l and their role in legume rhizobial symbiosis development |
| topic | legume-rhizobial symbiosis pea annexins three-dimensional modeling proteomics calcium inhibitors localization |
| url | https://vavilov.elpub.ru/jour/article/view/3108 |
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