Simultaneous CAS9 editing of cpSRP43, LHCA6, and LHCA7 in Picochlorum celeri lowers chlorophyll levels and improves biomass productivity
Abstract High cellular pigment levels in dense microalgal cultures contribute to excess light absorption. To improve photosynthetic yields in the marine microalga Picochlorum celeri, CAS9 gene editing was used to target the molecular chaperone cpSRP43. Depigmented strains (>50% lower chlorophyll)...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-09-01
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| Series: | Plant Direct |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/pld3.530 |
| _version_ | 1797629635916201984 |
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| author | Anagha Krishnan Melissa Cano Devin A. Karns Tyson A. Burch Maria Likhogrud Moena Aqui Shaun Bailey John Verruto William Lambert Fedor Kuzminov Mahva Naghipor Yingjun Wang Christopher C. Ebmeier Joseph C. Weissman Matthew C. Posewitz |
| author_facet | Anagha Krishnan Melissa Cano Devin A. Karns Tyson A. Burch Maria Likhogrud Moena Aqui Shaun Bailey John Verruto William Lambert Fedor Kuzminov Mahva Naghipor Yingjun Wang Christopher C. Ebmeier Joseph C. Weissman Matthew C. Posewitz |
| author_sort | Anagha Krishnan |
| collection | DOAJ |
| description | Abstract High cellular pigment levels in dense microalgal cultures contribute to excess light absorption. To improve photosynthetic yields in the marine microalga Picochlorum celeri, CAS9 gene editing was used to target the molecular chaperone cpSRP43. Depigmented strains (>50% lower chlorophyll) were generated, with proteomics showing attenuated levels of most light harvesting complex (LHC) proteins. Gene editing generated two types of cpSRP43 transformants with distinct lower pigment phenotypes: (i) a transformant (Δsrp43) with both cpSRP43 diploid alleles modified to encode non‐functional polypeptides and (ii) a transformant (STR30309) with a 3 nt in‐frame insertion in one allele at the CAS9 cut site (non‐functional second allele), leading to expression of a modified cpSRP43. STR30309 has more chlorophyll than Δsrp43 but substantially less than wild type. To further decrease light absorption by photosystem I in STR30309, CAS9 editing was used to stack in disruptions of both LHCA6 and LHCA7 to generate STR30843, which has higher (5–24%) productivities relative to wild type in solar‐simulating bioreactors. Maximal productivities required frequent partial harvests throughout the day. For STR30843, exemplary diel bioreactor yields of ~50 g m−2 day−1 were attained. Our results demonstrate diel productivity gains in P. celeri by lowering pigment levels. |
| first_indexed | 2024-03-11T10:55:54Z |
| format | Article |
| id | doaj.art-16c0af3cc9f64dcc8aea3d57de705432 |
| institution | Directory Open Access Journal |
| issn | 2475-4455 |
| language | English |
| last_indexed | 2024-03-11T10:55:54Z |
| publishDate | 2023-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Plant Direct |
| spelling | doaj.art-16c0af3cc9f64dcc8aea3d57de7054322023-11-13T07:55:56ZengWileyPlant Direct2475-44552023-09-0179n/an/a10.1002/pld3.530Simultaneous CAS9 editing of cpSRP43, LHCA6, and LHCA7 in Picochlorum celeri lowers chlorophyll levels and improves biomass productivityAnagha Krishnan0Melissa Cano1Devin A. Karns2Tyson A. Burch3Maria Likhogrud4Moena Aqui5Shaun Bailey6John Verruto7William Lambert8Fedor Kuzminov9Mahva Naghipor10Yingjun Wang11Christopher C. Ebmeier12Joseph C. Weissman13Matthew C. Posewitz14Department of Chemistry Colorado School of Mines Golden Colorado USADepartment of Chemistry Colorado School of Mines Golden Colorado USADepartment of Chemistry Colorado School of Mines Golden Colorado USADepartment of Chemistry Colorado School of Mines Golden Colorado USAExxonMobil Technology and Engineering Company Annandale New Jersey USAViridos La Jolla California USAViridos La Jolla California USAViridos La Jolla California USAViridos La Jolla California USAViridos La Jolla California USAViridos La Jolla California USAViridos La Jolla California USADepartment of Chemistry and Biochemistry University of Colorado Boulder Colorado USAExxonMobil Technology and Engineering Company Annandale New Jersey USADepartment of Chemistry Colorado School of Mines Golden Colorado USAAbstract High cellular pigment levels in dense microalgal cultures contribute to excess light absorption. To improve photosynthetic yields in the marine microalga Picochlorum celeri, CAS9 gene editing was used to target the molecular chaperone cpSRP43. Depigmented strains (>50% lower chlorophyll) were generated, with proteomics showing attenuated levels of most light harvesting complex (LHC) proteins. Gene editing generated two types of cpSRP43 transformants with distinct lower pigment phenotypes: (i) a transformant (Δsrp43) with both cpSRP43 diploid alleles modified to encode non‐functional polypeptides and (ii) a transformant (STR30309) with a 3 nt in‐frame insertion in one allele at the CAS9 cut site (non‐functional second allele), leading to expression of a modified cpSRP43. STR30309 has more chlorophyll than Δsrp43 but substantially less than wild type. To further decrease light absorption by photosystem I in STR30309, CAS9 editing was used to stack in disruptions of both LHCA6 and LHCA7 to generate STR30843, which has higher (5–24%) productivities relative to wild type in solar‐simulating bioreactors. Maximal productivities required frequent partial harvests throughout the day. For STR30843, exemplary diel bioreactor yields of ~50 g m−2 day−1 were attained. Our results demonstrate diel productivity gains in P. celeri by lowering pigment levels.https://doi.org/10.1002/pld3.530biomass productivityCAS9 editingcpSRP43photosynthetic efficiencyPicochlorum celeri |
| spellingShingle | Anagha Krishnan Melissa Cano Devin A. Karns Tyson A. Burch Maria Likhogrud Moena Aqui Shaun Bailey John Verruto William Lambert Fedor Kuzminov Mahva Naghipor Yingjun Wang Christopher C. Ebmeier Joseph C. Weissman Matthew C. Posewitz Simultaneous CAS9 editing of cpSRP43, LHCA6, and LHCA7 in Picochlorum celeri lowers chlorophyll levels and improves biomass productivity Plant Direct biomass productivity CAS9 editing cpSRP43 photosynthetic efficiency Picochlorum celeri |
| title | Simultaneous CAS9 editing of cpSRP43, LHCA6, and LHCA7 in Picochlorum celeri lowers chlorophyll levels and improves biomass productivity |
| title_full | Simultaneous CAS9 editing of cpSRP43, LHCA6, and LHCA7 in Picochlorum celeri lowers chlorophyll levels and improves biomass productivity |
| title_fullStr | Simultaneous CAS9 editing of cpSRP43, LHCA6, and LHCA7 in Picochlorum celeri lowers chlorophyll levels and improves biomass productivity |
| title_full_unstemmed | Simultaneous CAS9 editing of cpSRP43, LHCA6, and LHCA7 in Picochlorum celeri lowers chlorophyll levels and improves biomass productivity |
| title_short | Simultaneous CAS9 editing of cpSRP43, LHCA6, and LHCA7 in Picochlorum celeri lowers chlorophyll levels and improves biomass productivity |
| title_sort | simultaneous cas9 editing of cpsrp43 lhca6 and lhca7 in picochlorum celeri lowers chlorophyll levels and improves biomass productivity |
| topic | biomass productivity CAS9 editing cpSRP43 photosynthetic efficiency Picochlorum celeri |
| url | https://doi.org/10.1002/pld3.530 |
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