Methods and compositions for identifying NBB1 mutations
Two genes, A622 and NBB1, can be influenced to achieve a decrease of nicotinic alkaloid levels in plants. In particular, suppression of one or both of A622 and NBB1 may be used to decrease nicotine in tobacco plants.
1. A primer or probe pair capable of amplifying or detecting a nucleic acid molecule as set forth in SEQ ID NO: 3.
2. The primer or probe pair of claim 1 , wherein the probe pair comprises isolated nucleic acid molecules comprising:
(a) a first nucleic acid molecule comprising 15-1680 contiguous nucleotides of SEQ ID NO: 3; and
(b) and a second nucleic acid molecule comprising 15-1680 contiguous nucleotides of SEQ ID NO: 3,
wherein the first and second nucleic acid molecules function together as primers or probes for a nucleic acid sequence as set forth in SEQ ID NO: 3.
3. A method for identifying a tobacco plant comprising a mutant allele of the polynucleotide sequence as set forth in SEQ ID NO: 3, comprising:
(a) obtaining a nucleic acid sample from one or more tobacco plants; and
(b) contacting the sample with a nucleic acid primer comprising 15-1680 contiguous nucleotides of SEQ ID NO: 3 or its complement; and
(c) identifying a sample having a nucleic acid sequence comprising the mutant allele of the polynucleotide as set forth in SEQ ID NO: 3.
4. The method of claim 3 , wherein the identifying method step comprises detecting formation of a heteroduplex, wherein the heteroduplex detects the mutant allele of the nucleic acid sequence as set forth in SEQ ID NO: 3.
5. A method for identifying a tobacco plant with reduced expression of the polynucleotide sequence as set forth in SEQ ID NO: 3, comprising:
(a) measuring expression of the polynucleotide sequence as set forth in SEQ ID NO: 3 in a population of tobacco plants; and
(b) selecting a tobacco plant with reduced expression of the polynucleotide sequence as set forth in SEQ ID NO: 3 relative to wild-type levels of expression of the polynucleotide sequence as set forth in SEQ ID NO: 3.
6. The method of claim 5 , wherein the measuring of expression of the polynucleotide sequence as set forth in SEQ ID NO: 3 uses a primer or probe pair capable of amplifying or detecting a nucleic acid molecule as set forth in SEQ ID NO: 3.
7. A tobacco plant identified with the method of claim 3 , wherein the plant comprises the mutant allele of the polynucleotide sequence as set forth in SEQ ID NO: 3 and has reduced expression of the polynucleotide sequence as set forth in SEQ ID NO: 3 as compared to wild-type levels of expression of the polynucleotide sequence as set forth in SEQ ID NO: 3.
8. A product produced from the tobacco plant of claim 7 , wherein the product comprises the mutant allele of the polynucleotide sequence as set forth in SEQ ID NO: 3.
9. The product of claim 8 , wherein the product is selected from the group consisting of smoking cessation products, cigarettes, cigarette tobacco, cigars, cigar tobacco, snus, pipe tobacco and chewing tobacco.
10. The product of claim 8 , wherein the product is selected from the group consisting of a food product, food ingredient, feed product, feed ingredient, nutritional supplement, and biofuels.