High productivity algal mutants having reduced photosynthetic antenna
Disclosed herein are mutant photosynthetic microorganisms having an attenuated SGI1 gene. The mutants have reduced chlorophyll and increased productivity with respect to wild type cells. Also disclosed are methods of using such mutants for producing biomass or bioproducts, and methods of screening for such mutants.
1. A method of isolating a mutant Chlorophyte alga having higher biomass productivity with respect to the progenitor alga from which it is derived, comprising:
culturing a Chlorophyte algal strain under photoautotrophic semi-continuous or continuous conditions having a disruption in an SGI1 gene encoding a polypeptide having at least 90% sequence identity to SEQ ID NO: 3 or SEQ ID NO: 14-16; and
isolating at least one strain from the culture, wherein the at least one isolated strain has a higher biomass productivity than the progenitor alga;
thereby isolating a mutant Chlorophyte alga having higher biomass productivity with respect to the progenitor alga from which it is derived.
2. The method according to claim 1 , wherein the photoautotrophic semicontinuous or continuous culture conditions include exposure to constant light of an intensity of at least 600 uE.
3. The method according to claim 1 , wherein the photoautotrophic semicontinuous or continuous culture conditions include exposure to light of an intensity of at least 800 uE.
4. The method according to claim 1 , wherein the photoautotrophic semicontinuous or continuous culture conditions include exposure to light of an intensity of at least 1000 uE.
5. The method according to claim 1 , wherein the photoautotrophic semicontinuous or continuous culture conditions include exposure to light of an intensity of at least 1200 uE.
6. The method according to claim 1 , wherein the photoautotrophic semicontinuous or continuous culture conditions include exposure to light of an intensity of at least 1500 uE.
7. The method according to claim 6 , wherein the photoautotrophic semicontinuous or continuous culture conditions include exposure to light of an intensity of at least 2000 uE.
8. The method according to claim 7 , wherein the photoautotrophic semicontinuous or continuous culture conditions include exposure to light of an intensity of at least 2500 uE.
9. The method according to claim 1 , wherein the photoautotrophic semicontinuous or continuous culture conditions include maintaining the algal culture at a density of less than or equal to about 1.5×10 6 cells/mL.
10. The method according to claim 1 , wherein the photoautotrophic semicontinuous or continuous culture conditions include maintaining the algal culture at a density of less than or equal to about 1×10 6 cells/mL.
11. The method according to claim 1 , wherein the algal-strain mutant Chlorophyte alga is mutagenized using UV, chemical mutagenesis, insertional mutagenesis, or targeted genome editing.
12. The method according to claim 1 , further comprising performing PCR to amplify at least a portion of the SGI1 gene locus of the isolated mutant alga.
13. The method according to claim 1 , further comprising sequencing at least a portion of the SGI1 gene locus of the isolated mutant alga.
14. The method according to claim 1 wherein the Chlorophyte algal strain is a member of the genus Parachlorella or Tetraselmis.
15. The method according to claim 1 , wherein the isolated mutant Chlorophyte alga has reduced total chlorophyll/total organic carbon with respect to the control photosynthetic microorganism.
16. The method according to claim 1 , wherein the isolated mutant Chlorophyte alga has an increased chlorophyll a:b ratio with respect to the control photosynthetic organism.
17. The method according to claim 1 , wherein the isolated mutant Chlorophyte alga has a reduced photosystem II (PSII) antenna size with respect to the control photosynthetic organism.
18. The method according to claim 1 , wherein the isolated mutant Chlorophyte alga has a higher 14C Pmax with respect to the control photosynthetic organism.
19. The method according to claim 1 , wherein the isolated mutant Chlorophyte alga has a higher Fv/Fm than the control organism.