Method for modulating the alkaloid content of a tobacco plant

A plant or part thereof or cell, or plant propagation material obtained therefrom, which has been modified to achieve a modulation in alkaloid content in comparison to an unmodified plant, wherein the modification is the modulation of the expression or decrease the activity of at least one gene encoding a BTB/POZ NPH3 domain-containing protein from tobacco,

• wherein
• i) the content of one or more alkaloids selected from nicotine, nornicotine, pseudooxynicotine (PON), anabasine, and anatabine is modulated;
• ii) the plant or part thereof, or cell, or plant propagation material obtained therefrom is from the genus Nicotiana;
• iii) modulation of the expression of said at least one gene is achieved by overexpression, mutation, knock out, gene silencing or RNA silencing of said at least one gene;
• iv) the BTB/POZ NPH3 domain-containing protein comprises an amino acid sequence:
• a) as set out in SEQ ID NO:
• 1, or
• b) a functional variant of SEQ ID NO:
• 1 comprising a BTB/POZ domain with a sequence set forth in SEQ ID NO:
• 31 or a sequence with at least 98% identity thereto, and a NPH3 domain with a sequence as set forth in SEQ ID NO:
• 32 or a sequence with at least 98% identity thereof,
• or
• c) a sequence which has at least 98% identity to SEQ ID NO:
• 1;
• or
• the at least one gene encoding the BTB/POZ NPH3 domain-containing protein encodes the BTB/POZ NPH3 domain-containing protein as defined in a), b) or c) above and comprises a nucleotide sequence:
• a) as set out in SEQ ID NO:
• 2 or SEQ ID NO:
• 3,
• b) a functional variant of SEQ ID NO:
• 2 or SEQ ID NO:
• 3 comprising a nucleotide sequence encoding a BTB/POZ domain with a sequence set forth in SEQ ID NO:
• 31 or a sequence with at least 98% identity thereto, and a NPH3 domain with a sequence as set forth in SEQ ID NO:
• 32 or a sequence with at least 98% identity thereof, or
• c) a sequence which has at least 98% identity to SEQ ID NO:
• 2 or 3;
• and
• v) when the plant or part thereof or cell, or plant propagation material obtained therefrom is modified to increase the expression of the at least one gene encoding a BTB/POZ NPH3 domain-containing protein, the plant or part thereof or cell, or plant propagation material obtained therefrom exhibits increased alkaloid content in comparison to a plant or part thereof or cell, or plant propagation material obtained therefrom which has not been modified to modulate the expression of the at least one gene encoding a BTB/POZ NPH3 domain-containing protein, or
• when the plant or part thereof or cell, or plant propagation material obtained therefrom is modified to decrease the activity or expression of the at least one gene encoding a BTB/POZ NPH3 domain-containing protein, the plant or part thereof or cell, or plant propagation material obtained therefrom exhibits decreased alkaloid content in comparison to a plant or part thereof or cell, or plant propagation material obtained therefrom which has not been modified to modulate the expression or decrease the activity of the at least one gene encoding a BTB/POZ NPH3 domain-containing protein.

A mutant of a plant carrying a heritable mutation in a nucleotide sequence of at least one gene encoding the BTB/POZ NPH3 domain-containing protein, wherein the BTB/POZ NPH3 domain-containing protein comprises an amino acid sequence:

• a) as set out in SEQ ID NO:
• 1, or
• b) a functional variant of SEQ ID NO:
• 1 comprising a BTB/POZ domain with a sequence set forth in SEQ ID NO:31 or a sequence with at least 98% identity thereto, and a NPH3 domain with a sequence as set forth in SEQ ID NO:32 or a sequence with at least 98% identity thereof,
• or
• c) a sequence which has at least 98% identity to SEQ ID NO:
• 1;
• or
• the at least one gene encoding the BTB/POZ NPH3 domain-containing protein encodes the BTB/POZ NPH3 domain-containing protein as defined in a), b) or c) above and comprises a nucleotide sequence:
• a) as set out in SEQ ID NO:
• 2 or SEQ ID NO:
• 3,
• b) a functional variant of SEQ ID NO:
• 2 or SEQ ID NO:
• 3 comprising a nucleotide sequence encoding a BTB/POZ domain with a sequence set forth in SEQ ID NO:31 or a sequence with at least 98% identity thereto, and a NPH3 domain with a sequence as set forth in SEQ ID NO:32 or a sequence with at least 98% identity thereof, or
• c) a sequence which has at least 98% identity to SEQ ID NO:
• 2 or 3;
• wherein said heritable mutation modulates the expression or decrease the activity of the at least one gene encoding the BTB/POZ NPH3 domain-containing protein by overexpression, mutation, knock out, gene silencing or RNA silencing of said at least one gene;
• and wherein the mutant plant has modulated alkaloid content relative to a comparable plant which does not carry said heritable mutation, wherein
• i) the content of one or more alkaloids selected from nicotine, nornicotine, pseudooxynicotine (PON), anabasine, and anatabine is modulated;
• ii) the plant is from the genus Nicotiana, and
• iii) when said heritable mutation increases the expression of the at least one gene encoding a BTB/POZ NPH3 domain-containing protein the plant exhibits increased alkaloid content in comparison to a plant which does not carry said heritable mutation, or when said heritable mutation decreases the activity or expression of the at least one gene encoding a BTB/POZ NPH3 domain-containing protein the plant exhibits decreased alkaloid content in comparison to a plant which does not carry said heritable mutation.