Regulatory Nucleic Acid Molecules for Reliable Gene Expression in Plants

A method for producing a population of plants with reduced percentage of low-expressing plants comprising the steps of:

• a) providing a recombinant nucleic acid construct comprising at least one RENA molecule selected from the group consisting of:
• i) a nucleic acid molecule having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• ii) a nucleic acid molecule having a sequence with an identity of at least 80% to a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iii) a fragment of at least 100 consecutive bases of a nucleic acid molecule of i) or ii) which has reliability enhancing activity as the corresponding nucleic acid molecule having the sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iv a nucleic acid molecule hybridizing under conditions equivalent to hybridization in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 2×SSC, 0.1% SDS at 50° C. to a nucleic acid molecule comprising at least 50 consecutive nucleotides of the complement of a reliability enhancing nucleic acid having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• and
• v) a nucleic acid molecule which is the complement or reverse complement of any of the previously mentioned nucleic acid molecules under i) to iv);
• functionally linked to a promoter and/or a nucleic acid molecule of interest wherein at least the promoter is heterologous to the RENA molecule;
• b) integrating said recombinant nucleic acid construct in the genome of plant cells, plants or parts thereof;
• and
• c) regenerating a population of independent primary transformant plants from the transgenic plant cells, plants or parts thereof produced in step b);
• wherein the population of plants has an at least 5% reduced percentage of plants exhibiting low expression of the nucleic acid of interest compared to a population of plants comprising in their genome a respective recombinant nucleic acid construct not comprising the respective RENA molecule as defined in a) i) to v).

A method for reducing the number of low expressing plants in a population of independent primary transformant plants, comprising the steps of

• a) providing a recombinant nucleic acid construct comprising at least one RENA molecule selected from the group consisting of:
• i) a nucleic acid molecule having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• ii) a nucleic acid molecule having a sequence with an identity of at least 80% to a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iii) a fragment of at least 100 consecutive bases of a nucleic acid molecule of i) or ii) which has reliability enhancing activity as the corresponding nucleic acid molecule having the sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iv) a nucleic acid molecule hybridizing under conditions equivalent to hybridization in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 2×SSC, 0.1% SDS at 50° C. to a nucleic acid molecule comprising at least 50 consecutive nucleotides of the complement of a reliability enhancing nucleic acid having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• and
• v) a nucleic acid molecule which is the complement or reverse complement of any of the previously mentioned nucleic acid molecules under i) to iv);
• functionally linked to a promoter and/or a nucleic acid molecule of interest wherein at least the promoter is heterologous to the RENA molecule;
• b) integrating said recombinant nucleic acid construct in the genome of plant cells, plants or parts thereof;
• and
• c) regenerating independent primary transformant plants from the transgenic plant cells, plants or parts thereof produced in step b);
• wherein the number of low expressing independent primary transformant plants is reduced by at least 5% compared to a population of independent primary transformant plants comprising in their genome a respective recombinant nucleic acid not comprising the respective RENA molecule as defined in a) i) to v).

A recombinant expression construct comprising at least one RENA molecule selected from the group consisting of:

• i) a nucleic acid molecule having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• ii) a nucleic acid molecule having a sequence with an identity of at least 80% to a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iii) a fragment of at least 100 consecutive bases of a nucleic acid molecule of i) or ii) which has reliability enhancing activity as the corresponding nucleic acid molecule having the sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iv) a nucleic acid molecule hybridizing under conditions equivalent to hybridization in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 2×SSC, 0.1% SDS at 50° C. to a nucleic acid molecule comprising at least 50 consecutive nucleotides of the complement of a reliability enhancing nucleic acid having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• and
• v) a nucleic acid molecule which is the complement or reverse complement of any of the previously mentioned nucleic acid molecules under i) to iv);
• functionally linked to a promoter and/or a nucleic acid molecule of interest wherein at least the promoter is heterologous to the RENA molecule.

A transgenic cell or transgenic plant or part thereof comprising:

• at least one RENA molecule selected from the group consisting of:
• i) a nucleic acid molecule having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• ii) a nucleic acid molecule having a sequence with an identity of at least 80% to a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iii) a fragment of at least 100 consecutive bases of a nucleic acid molecule of i) or ii) which has reliability enhancing activity as the corresponding nucleic acid molecule having the sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iv) a nucleic acid molecule hybridizing under conditions equivalent to hybridization in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 2×SSC, 0.1% SDS at 50° C. to a nucleic acid molecule comprising at least 50 consecutive nucleotides of the complement of a reliability enhancing nucleic acid having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• and
• v) a nucleic acid molecule which is the complement or reverse complement of any of the previously mentioned nucleic acid molecules under i) to iv);
• functionally linked to a heterologous promoter and/or a heterologous nucleic acid of interest to be expressed.

A method for the production of an agricultural product by introducing a RENA molecule selected from the group consisting of:

• i) a nucleic acid molecule having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• ii) a nucleic acid molecule having a sequence with an identity of at least 80% to a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iii) a fragment of at least 100 consecutive bases of a nucleic acid molecule of i) or ii) which has reliability enhancing activity as the corresponding nucleic acid molecule having the sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• iv) a nucleic acid molecule hybridizing under conditions equivalent to hybridization in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 2×SSC, 0.1% SDS at 50° C. to a nucleic acid molecule comprising at least 50 consecutive nucleotides of the complement of a reliability enhancing nucleic acid having a sequence selected from SEQ ID NOS:
• 1 to 16 or 94 to 116666;
• and
• v) a nucleic acid molecule which is the complement or reverse complement of any of the previously mentioned nucleic acid molecules under i) to iv);
• into a plant, growing the plant, harvesting and processing the plant or parts thereof.