CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation
54 Pags.- 19 Figs.- 2 Tabls. The definitive version is available at: https://nph.onlinelibrary.wiley.com/journal/14698137
| Autores: | , , , , , , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión aceptada para publicación |
| Fecha de publicación: | 2019 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:dnet:digitalcsic_::c197693f2877407c7b8fb2ff4ad6777c |
| Acceso en línea: | http://hdl.handle.net/10261/191637 |
| Access Level: | acceso abierto |
| Palabra clave: | CRISPR/Cas9 leghemoglobins nitrogen fixation nodule senescence phytoglobins reactive oxygen species RNA-seq. |
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CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation Wang, Longlong Rubio Luna, María Carmen Xin, Xian Zhang, Baoli Fan, Qiuling Wang, Qiang Ning, Guogui Becana Ausejo, Manuel Duanmu, Deqiang CRISPR/Cas9 leghemoglobins nitrogen fixation nodule senescence phytoglobins reactive oxygen species RNA-seq. 54 Pags.- 19 Figs.- 2 Tabls. The definitive version is available at: https://nph.onlinelibrary.wiley.com/journal/14698137 Legume nodules contain high concentrations of leghemoglobins (Lbs) encoded by several genes. The reason for this multiplicity is unknown. CRISPR/Cas9 technology was used to generate stable mutants of the three Lbs of Lotus japonicus. The phenotypes were characterized at the physiological, biochemical and molecular levels. Nodules of the triple mutants were examined by electron microscopy and subjected to RNA‐sequencing (RNA‐seq) analysis. Complementation studies revealed that Lbs function synergistically to maintain optimal N2 fixation. The nodules of the triple mutants overproduced superoxide radicals and hydrogen peroxide, which was probably linked to activation of NADPH oxidases and changes in superoxide dismutase isoforms expression. The mutant nodules showed major ultrastructural alterations, including vacuolization, accumulation of poly‐β‐hydroxybutyrate and disruption of mitochondria. RNA‐seq of c. 20 000 genes revealed significant changes in expression of carbon and nitrogen metabolism genes, transcription factors, and proteinases. Lb‐deficient nodules had c. 30–50‐fold less heme but similar transcript levels of heme biosynthetic genes, suggesting a post‐translational regulatory mechanism of heme synthesis. We conclude that Lbs act additively in nodules and that the lack of Lbs results in early nodule senescence. Our observations also provide insight into the reprogramming of the gene expression network associated with Lb deficiency, probably as a result of uncontrolled intracellular free O2 concentration. This work was supported by grants from the National Natural Science Foundation of China (31870220 and 31570233), the Fundamental Research Funds for the Central Universities (Program No. 2662015PY171), and Spanish Ministry of Economy and Competitiveness‐European Regional Development Fund (AGL2017‐85775‐R). Peer reviewed John Wiley & Sons http://hdl.handle.net/10261/191637 |
| title |
CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation |
| spellingShingle |
CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation Wang, Longlong CRISPR/Cas9 leghemoglobins nitrogen fixation nodule senescence phytoglobins reactive oxygen species RNA-seq. |
| title_short |
CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation |
| title_full |
CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation |
| title_fullStr |
CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation |
| title_full_unstemmed |
CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation |
| title_sort |
CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation |
| author |
Wang, Longlong |
| author_facet |
Wang, Longlong Rubio Luna, María Carmen Xin, Xian Zhang, Baoli Fan, Qiuling Wang, Qiang Ning, Guogui Becana Ausejo, Manuel Duanmu, Deqiang |
| author_role |
author |
| author2 |
Rubio Luna, María Carmen Xin, Xian Zhang, Baoli Fan, Qiuling Wang, Qiang Ning, Guogui Becana Ausejo, Manuel Duanmu, Deqiang |
| author2_role |
author author author author author author author author |
| topic |
CRISPR/Cas9 leghemoglobins nitrogen fixation nodule senescence phytoglobins reactive oxygen species RNA-seq. |
| topic_facet |
CRISPR/Cas9 leghemoglobins nitrogen fixation nodule senescence phytoglobins reactive oxygen species RNA-seq. |
| description |
54 Pags.- 19 Figs.- 2 Tabls. The definitive version is available at: https://nph.onlinelibrary.wiley.com/journal/14698137 |
| publishDate |
2019 |
| format |
article |
| status_str |
acceptedVersion |
| url |
http://hdl.handle.net/10261/191637 |
| eu_rights_str_mv |
openAccess |
| publisher |
John Wiley & Sons |
| institution |
Consejo Superior de Investigaciones Científicas (CSIC) |
| collection |
DIGITAL.CSIC. Repositorio Institucional del CSIC |
| reponame_str |
DIGITAL.CSIC. Repositorio Institucional del CSIC |
| instname_str |
Consejo Superior de Investigaciones Científicas (CSIC) |
| _version_ |
1878440798641455105 |
| publishDateSort |
2019 |
| author_browse |
Becana Ausejo, Manuel Duanmu, Deqiang Fan, Qiuling Ning, Guogui Rubio Luna, María Carmen Wang, Longlong Wang, Qiang Xin, Xian Zhang, Baoli |
| publisherStr |
John Wiley & Sons |
| score |
6,924472 |