Plant body of genus Nicotiana with low alkaloid content and production method thereof
Inventors
TAKEUCHI, Takanori • Arai, Masao • UDAGAWA, Hisashi • Magome, Hiroshi • NISHIGUCHI, Ryo • Takakura, Yoshimitsu
Assignees
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Publication Number
US-12239090-B2
Publication Date
2025-03-04
Expiration Date
Abstract
An embodiment of the present invention provides a tobacco plant having a low alkaloid content. In the tobacco plant, a function of a gene encoding aspartate oxidase AO2 is suppressed.
Core Innovation
The invention relates to a tobacco plant in which a mutation specifically causes suppression of a function of an endogenous gene in the genome. The suppressed endogenous gene is selected from endogenous genes whose coding regions encode polypeptides having sequence identity of 95% or higher with amino acid sequences represented by SEQ ID NO: 2 or SEQ ID NO: 5, and/or with an amino acid sequence represented by SEQ ID NO: 6.
The background problem is that tobacco alkaloids, notably nicotine and nornicotine, are present in tobacco plants and the document aims at reducing alkaloid content while maintaining other essential endogenous gene functions. Suppression is configured to be specific, so that suppression of the selected endogenous genes does not suppress the respective functions of endogenous genes containing coding regions encoding polypeptides with sequence identity of 95% or higher to amino acid sequences represented by SEQ ID NO: 62, SEQ ID NO: 63, and SEQ ID NO: 64.
The described core concept is to engineer a tobacco plant and/or a cured leaf produced from the tobacco plant by introducing mutations in endogenous genes such that suppression is achieved for selected endogenous genes based on the stated sequence identity thresholds relative to SEQ ID NO: 2/5 and SEQ ID NO: 6, while the functions of the endogenous genes corresponding to SEQ ID NO: 62/63 and SEQ ID NO: 64 are not suppressed. The document further distinguishes suppression outcomes at the levels of mRNA amount, mRNA degradation, transcription to mRNA, or translation into a polypeptide, and relates the resulting plants or cured leaves to lower alkaloid content compared with cured leaves from wild-type tobacco plants.
Claims Coverage
The document contains three independent claims: a tobacco plant, a method of producing the tobacco plant, and a cured leaf. Across the independent claims, the coverage centers on specifically introducing a mutation that suppresses the function of selected endogenous genes while ensuring functions of other specified endogenous genes are not suppressed.
A tobacco plant with specific suppression of selected endogenous gene functions while keeping other endogenous gene functions not suppressed
A tobacco plant in which a mutation that specifically causes suppression of a function of an endogenous gene is introduced in an endogenous gene in a genome, wherein the endogenous gene is at least one of genes encoding polypeptides with sequence identity of 95% or higher with amino acid sequences represented by SEQ ID NO: 2 or SEQ ID NO: 5 and genes encoding polypeptides with sequence identity of 95% or higher with an amino acid sequence represented by SEQ ID NO: 6, and wherein respective functions of other endogenous genes are not suppressed: genes encoding polypeptides with sequence identity of 95% or higher with amino acid sequences represented by SEQ ID NO: 62 or SEQ ID NO: 63 and genes encoding polypeptides with sequence identity of 95% or higher with an amino acid sequence represented by SEQ ID NO: 64.
A method of producing a tobacco plant by introducing a mutation that specifically causes suppression while keeping other endogenous gene functions not suppressed
A method of producing a tobacco plant comprising introducing a mutation in a genome of a tobacco plant, wherein the mutation specifically causes suppression of a function of at least one endogenous gene encoding a polypeptide with sequence identity of 95% or higher with amino acid sequences represented by SEQ ID NO: 2 or SEQ ID NO: 5 and/or an amino acid sequence represented by SEQ ID NO: 6, and wherein respective functions of other endogenous genes are not suppressed, including genes encoding polypeptides with sequence identity of 95% or higher with amino acid sequences represented by SEQ ID NO: 62 or SEQ ID NO: 63 and an amino acid sequence represented by SEQ ID NO: 64.
A cured leaf with mutations causing lower alkaloid content while keeping specified other endogenous gene functions not suppressed
A cured leaf having a mutation in an endogenous gene encoding a polypeptide with sequence identity of 95% or higher with amino acid sequences represented by SEQ ID NO: 2 or SEQ ID NO: 5 and/or a mutation in an endogenous gene encoding a polypeptide with sequence identity of 95% or higher with an amino acid sequence represented by SEQ ID NO: 6, wherein the mutations suppress functions of the endogenous genes, the cured leaf has lower alkaloid content as compared with a cured leaf produced from a tobacco leaf harvested from a wild-type tobacco plant, and respective functions of other endogenous genes are not suppressed, including genes encoding polypeptides with sequence identity of 95% or higher with amino acid sequences represented by SEQ ID NO: 62 or SEQ ID NO: 63 and an amino acid sequence represented by SEQ ID NO: 64.
Across the independent claims, the essential inventive coverage is the introduction of a mutation that specifically suppresses functions of endogenous genes defined by polypeptide sequence identity to SEQ ID NO: 2/5 and/or SEQ ID NO: 6, while explicitly maintaining the functions of other endogenous genes defined by sequence identity to SEQ ID NO: 62/63 and SEQ ID NO: 64; for the cured leaf claim, the maintained-function requirement is combined with a lower alkaloid content comparison versus wild-type cured leaves.
Stated Advantages
Lower alkaloid content compared with a cured leaf produced from a tobacco leaf harvested from a wild-type tobacco plant.
Documented Applications
A cured leaf included in a tobacco product.
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