Transgenic algae engineered for higher performance

Abstract

The present disclosure relates to transgenic algae having increased growth characteristics, and methods of increasing growth characteristics of algae. In particular, the disclosure relates to transgenic algae comprising a glutamine phenylpyruvate transaminase transgene and to transgenic algae comprising a glutamine phenylpyruvate transaminase transgene and a glutamine synthetase

1 . A transgenic alga comprising a plant-derived or algal-derived glutamine phenylpyruvate transaminase (GPT) transgene and a glutamine synthetase transgene, wherein each of said GPT transgene and said GS transgene is operably linked to a promoter. 
     
     
         2 . The transgenic alga according to  claim 1 , wherein the alga is a green alga. 
     
     
         3 . The transgenic alga according to  claim 2 , wherein the green alga is a  Chlorella  species. 
     
     
         4 . The transgenic alga according to  claim 1 , wherein the GPT transgene is a plant-derived GPT. 
     
     
         5 . The transgenic alga according to  claim 1 , wherein the GPT is an algal-derived GPT. 
     
     
         6 . The transgenic alga of  claim 4 , wherein
 the GPT transgene encodes a polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 9, SEQ ID NO: 15, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO 24, SEQ ID NO: 30, SEQ ID NO:31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 44, and SEQ ID NO: 47, an amino acid sequence that is at least 75% identical to SEQ ID NO: 2, an amino acid sequence that is at least 75% identical to SEQ ID NO: 9, an amino acid sequence that is at least 75% identical to SEQ ID NO: 15, an amino acid sequence that is at least 75% identical to SEQ ID NO: 19, an amino acid sequence that is at least 75% identical to SEQ ID NO: 21, an amino acid sequence that is at least 75% identical to SEQ ID NO 24, an amino acid sequence that is at least 75% identical to SEQ ID NO: 30, an amino acid sequence that is at least 75% identical to SEQ ID N0:31, an amino acid sequence that is at least 75% identical to SEQ ID NO: 32, an amino acid sequence that is at least 75% identical to SEQ ID NO: 33, an amino acid sequence that is at least 75% identical to SEQ ID NO: 34, an amino acid sequence that is at least 75% identical to SEQ ID NO: 35, an amino acid sequence that is at least 75% identical to SEQ ID NO: ID NO: 36, an amino acid sequence that is at least 75% identical to SEQ ID NO: 44, and an amino acid sequence that is at least 75% identical to SEQ ID NO: 47.   
     
     
         7 . The transgenic alga according to  claim 5 , wherein the GPT transgene encodes a polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NO: 48, and an amino acid sequence that is at least 75% identical to SEQ ID NO: 48. 
     
     
         8 . The transgenic alga according to  claim 1 , wherein the GS transgene encodes a polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NO: 4, SEQ ID NO: 7, SEQ ID NO: 39, an amino acid sequence that is at least 75% identical to SEQ ID NO: 4, an amino acid sequence that is at least 75% identical to SEQ ID NO: 7, and an amino acid sequence that is at least 75% identical to SEQ ID NO: 39. 
     
     
         9 . The transgenic alga according to  claim 1 , wherein the GPT and GS transgenes are incorporated into the genome of the alga. 
     
     
         10 . A progeny of any generation of the transgenic alga of  claim 9 , wherein said progeny comprises said GPT transgene and said GS transgene. 
     
     
         11 . The transgenic alga of  claim 9 , which displays a faster growth rate, increased chlorophyll production, increased ribulose bisphosphate carboxylase (RUBISCO) level, increased nitrogen use efficiency, increased biomass yield, increased GPT activity, increased GS activity, increased 2-oxoglutaramate levels, and/or increased tolerance to salt or saline conditions when compared to a control wild-type or untransformed alga. 
     
     
         12 . A method for increasing growth characteristics of an alga relative to an wild type or progenitor alga of the same species, comprising:
 (a) introducing a plant-derived or algal-derived GPT transgene into the alga;   (b) introducing a GS trans gene into the alga or a progeny of the alga;   (c) expressing the GPT transgene and the GS transgene in the alga or the progeny of the alga; and,   (d) selecting an alga having an increased growth characteristic relative to an alga of the same species that does not comprise the GPT transgene or the GS transgene.   
     
     
         13 . The method according to  claim 12 , wherein the increased growth characteristic is selected from the group consisting of: faster growth rate, increased chlorophyll production, increased ribulose bisphosphate carboxylase (RUBISCO) level, increased nitrogen use efficiency, increased biomass yield, increased GPT activity, increased GS activity, increased 2-oxoglutaramate levels, and/or increased tolerance to salt or saline conditions. 
     
     
         14 . A transgenic alga comprising a plant-derived or algal-derived GPT transgene, wherein said GPT transgene is operably linked to a promoter. 
     
     
         15 . The transgenic alga according to  claim 14 , wherein the alga is a green algae. 
     
     
         16 . The transgenic alga according to  claim 15 , wherein the green alga is a  Chlorella  species. 
     
     
         17 . The transgenic alga according to  claim 14 , wherein the GPT transgene is a plant-derived GPT. 
     
     
         18 . The transgenic alga according to  claim 14 , wherein the GPT is an algal-derived GPT. 
     
     
         19 . The transgenic alga of  claim 17 , wherein the GPT transgene encodes a polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 9, SEQ ID NO: 15, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO 24, SEQ ID NO: 30, SEQ ID NO:31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 44, and SEQ ID NO: 47, an amino acid sequence that is at least 75% identical to SEQ ID NO: 2, an amino acid sequence that is at least 75% identical to SEQ ID NO: 9, an amino acid sequence that is at least 75% identical to SEQ ID NO: 15, an amino acid sequence that is at least 75% identical to SEQ ID NO: 19, an amino acid sequence that is at least 75% identical to SEQ ID NO: 21, an amino acid sequence that is at least 75% identical to SEQ ID NO 24, an amino acid sequence that is at least 75% identical to SEQ ID NO: 30, an amino acid sequence that is at least 75% identical to SEQ ID N0:31, an amino acid sequence that is at least 75% identical to SEQ ID NO: 32, an amino acid sequence that is at least 75% identical to SEQ ID NO: 33, an amino acid sequence that is at least 75% identical to SEQ ID NO: 34, an amino acid sequence that is at least 75% identical to SEQ ID NO: 35, an amino acid sequence that is at least 75% identical to SEQ ID NO: 36, an amino acid sequence that is at least 75% identical to SEQ ID NO: 44, and an amino acid sequence that is at least 75% identical to SEQ ID NO: 47. 
     
     
         20 . The transgenic alga according to  claim 18 , wherein the GPT transgene encodes a polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NO: 48, and an amino acid sequence that is at least 75% identical to SEQ ID NO: 48. 
     
     
         21 . The transgenic alga according to  claim 14 , wherein the GPT transgene is incorporated into the genome of the alga. 
     
     
         22 . A progeny of any generation of the transgenic alga of  claim 21 , wherein said progeny comprises said GPT transgene. 
     
     
         23 . The transgenic alga of  claim 21 , which displays a faster growth rate, increased chlorophyll production, increased ribulose bisphosphate carboxylase (RUBISCO) level, increased nitrogen use efficiency, increased biomass yield, increased GPT activity, increased GS activity, increased 2-oxoglutaramate levels, and/or increased tolerance to salt or saline conditions when compared to a control wild-type or untransformed alga. 
     
     
         24 . A method for increasing growth characteristics of an alga relative to an wild type or progenitor alga of the same species, comprising:
 (a) introducing a plant-derived or algal-derived GPT transgene into the alga;   (b) expressing the GPT transgene in the algae or the progeny of the alga; and,   (c) selecting an alga having an increased growth characteristic relative to an alga of the same species that does not comprise the GPT transgene.   
     
     
         25 . The method according to  claim 24 , wherein the increased growth characteristic is selected from the group consisting of: faster growth rate, increased chlorophyll production, increased ribulose bisphosphate carboxylase (RUBISCO) level, increased nitrogen use efficiency, increased biomass yield, increased GPT activity, increased GS activity, increased 2-oxoglutaramate levels, and/or increased tolerance to salt or saline conditions.