Lettuce with increased shelf life
Inventors
Boddupalli, Sekhar • TREMBLAY, Arianne • Aasen, Eric • Rout, Jyoti R. • Li, Zhongsen
Assignees
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Publication Number
US-12203086-B2
Publication Date
2025-01-21
Expiration Date
Abstract
The present application is directed to lettuce plants comprising a mutation in each of at least two different PPO genes, where said mutations reduce the activity of PPO protein compared to a wild type lettuce plant. The present application is also directed to methods for making such lettuce plants.
Core Innovation
The invention relates to a genetically modified mutant lettuce plant in which non-naturally occurring mutations are introduced into each of at least a PPO-E gene and a PPO-S gene. The PPO-E protein and PPO-S protein in the mutant plant have at least 99% and 98% amino acid sequence identity, respectively, to the sequences set forth in SEQ ID NO:10 and SEQ ID NO:28, and the mutations eliminate or reduce activity of the PPO-E and PPO-S proteins by at least 30% compared to a wild type lettuce plant of the same variety.
The mutations comprise deletions or insertions of nucleotides in the coding sequences of SEQ ID NO:9 (PPO-E) and SEQ ID NO:27 (PPO-S). These mutations are directed to reduce PPO-E and PPO-S activity to achieve a mutant phenotype at harvest.
At harvest, the genetically modified mutant lettuce plant comprises one or more characteristics compared to the wild type lettuce plant of the same variety lacking the mutations. The characteristics include reduced tip burn and/or higher levels of polyphenolics, vitamin A, beta-carotene, vitamin C, and vitamin K, and also reduced browning, higher shelf life, lower CO2 production, higher carbohydrate levels, and/or less yellowing measured by midvein scoring.
The patent content also describes making the genetically modified mutant lettuce plant by introducing the non-naturally occurring mutations into each of the PPO-E and PPO-S genes of a lettuce plant. The resulting genetically modified mutant lettuce plant has PPO-E and PPO-S activity reduced by at least 30% relative to a wild type lettuce plant of the same variety, and the plant at harvest may include the same set of quality and nutritional characteristics as compared to the wild type.
Claims Coverage
The provided claim set includes two independent claims: one for a genetically modified mutant lettuce plant and one for a method of making the plant. Across these claims, the inventive subject matter centers on dual PPO-E and PPO-S targets with specified sequence identity requirements and deletions or insertions in their coding sequences to reduce PPO activity by at least 30% and yield harvest characteristics.
Dual PPO-E and PPO-S gene mutations for reduced PPO activity in lettuce
A genetically modified mutant lettuce plant comprising non-naturally occurring mutations in each of at least a PPO-E gene and a PPO-S gene, wherein the mutations eliminate or reduce activity of the PPO-E and PPO-S proteins by at least 30% compared to a wild type lettuce plant of the same variety.
Specific coding-sequence indels in SEQ ID NO:9 and SEQ ID NO:27
The mutations comprise deletions or insertions of nucleotides in the coding sequences of SEQ ID NO:9 (PPO-E) and SEQ ID NO:27 (PPO-S), with the PPO-E protein and PPO-S protein having at least 99% and 98% amino acid sequence identity, respectively, to SEQ ID NO:10 and SEQ ID NO:28.
Harvest characteristics associated with reduced tip burn and increased nutritional components
The genetically modified mutant lettuce plant at harvest comprises one or more characteristics including reduced tip burn; higher levels of polyphenolics; higher levels of vitamin A; higher levels of beta-carotene; higher levels of vitamin C; or higher levels of vitamin K compared to the wild type lettuce plant of the same variety lacking the mutations.
Method of making mutant lettuce by introducing dual PPO-E and PPO-S coding-sequence indels
A method of making a genetically modified mutant lettuce plant comprising introducing non-naturally occurring mutations into each of at least a PPO-E gene and a PPO-S gene of a lettuce plant, wherein the mutations reduce the activity of the PPO-E and PPO-S proteins by at least 30% compared to a wild type lettuce plant of the same variety.
Resulting mutant lettuce with harvest characteristics after dual PPO-E/PPO-S mutation introduction
The method results in a genetically modified mutant lettuce plant at harvest comprising one or more characteristics including reduced tip burn; higher levels of polyphenolics; higher levels of vitamin A; higher levels of beta-carotene; higher levels of vitamin C; or higher levels of vitamin K compared to a wild type lettuce plant of the same variety lacking the mutations.
The independent claims focus on a genetically modified mutant lettuce plant and a method of making it by introducing non-naturally occurring insertions or deletions in PPO-E and PPO-S coding sequences under specified sequence identity constraints, reducing PPO-E and PPO-S activity by at least 30% relative to wild type and yielding harvest-associated reductions in tip burn and improvements in nutritional and quality characteristics.
Stated Advantages
Reduced tip burn
Higher levels of polyphenolics
Higher levels of vitamin A
Higher levels of beta-carotene
Higher levels of vitamin C
Higher levels of vitamin K
Reduced browning
Higher shelf life
Lower CO2 production
Higher carbohydrate levels
Less yellowing (midvein scoring)
Documented Applications
Genetically modified mutant lettuce plants with reduced PPO activity and harvest characteristics such as reduced tip burn, reduced browning, increased shelf life, and increased nutritional components compared to wild type lettuce.
A method for making genetically modified mutant lettuce plants with reduced PPO activity by introducing non-naturally occurring mutations into PPO-E and PPO-S genes to produce harvest characteristics compared to wild type.
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