Novel plant genes and uses thereof

Abstract

Homologues of the Arabidopsis NIM1 gene, which is involved in the signal transduction cascade leading to systemic acquired resistance (SAR), are isolated from Nicotiana tabacum (tobacco), Lycopersicon esculentum (tomato), Brassica napus (oilseed rape), Arabidopsis thaliana, Beta vulgaris (sugarbeet), Helianthus annuus (sunflower), and Solanum tuberosum (potato). The invention further concerns transformation vectors and processes for expressing the NIM1 homologues in transgenic plants to increase SAR gene expression and enhance broad spectrum disease resistance.

What is claimed is:  
     
         1 . An isolated nucleic acid molecule comprising: 
 (a) a nucleotide sequence that encodes SEQ ID NO:2, 4, 6, 8, 16, 18, 20, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 62, 64, 66, 68, 70, 72, or 74;    (b) SEQ ID NO:1, 3, 5, 7, 15, 17, 19, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 61, 63, 65, 67, 69, 71, or 73;    (c) a nucleotide sequence that comprises an at least 20 consecutive base pair portion identical in sequence to an at least 20 consecutive base pair portion of SEQ ID NO:1, 3, 5, 7, 15, 17, 19, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 61, 63, 65, 67, 69, 71, or 73;    (d) a nucleotide sequence that can be amplified from a  Lycopersicon esculentum  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:9 and 10, SEQ ID NO:21 and 24, SEQ ID NO:22 and 24, SEQ ID NO:25 and 28, SEQ ID NO:26 and 28, or SEQ ID NO:59 and 60;    (e) a nucleotide sequence that can be amplified from a  Beta vulgaris  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:22 and 24 or SEQ ID NO:26 and 28;    (f) a nucleotide sequence that can be amplified from a  Helianthus annuus  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:26 and 28;    (g) a nucleotide sequence that can be amplified from a  Solanum tuberosum  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:21 and 24, SEQ ID NO:21 and 23, SEQ ID NO:22 and 24, SEQ ID NO:25 and 28, or SEQ ID NO:26 and 28;    (h) a nucleotide sequence that can be amplified from a  Brassica napus  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:9 and 10 or SEQ ID NO:26 and 28;    (i) a nucleotide sequence that can be amplified from an  Arabidopsis thaliana  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:13 and 14, SEQ ID NO:21 and 24, or SEQ ID NO:22 and 24;    (j) a nucleotide sequence that can be amplified from an  Nicotiana tabacum  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:9 and 10, SEQ ID NO:11 and 12, SEQ ID NO:21 and 24, SEQ ID NO:22 and 24, SEQ ID NO:25 and 28, or SEQ ID NO:26 and 28; or    (k) a nucleotide sequence that can be amplified from an plant DNA library using the polymerase chain reaction with a pair of primers comprising the first 20 nucleotides and the reverse complement of the last 20 nucleotides of the coding sequence (CDS) of SEQ ID NO:1, 3, 5, 7, 15, 17, 19, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 61, 63, 65, 67, 69, 71, or 73.    
     
     
         2 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that encodes SEQ ID NO:2, 4, 6, 8, 16, 18, 20, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 62, 64, 66, 68, 70, 72, or 74.  
     
     
         3 . An isolated nucleic acid molecule according to  claim 1 , comprising SEQ ID NO:1, 3, 5, 7, 15, 17, 19, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 61, 63, 65, 67, 69, 71, or 73.  
     
     
         4 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that comprises an at least 20 consecutive base pair portion identical in sequence to an at least 20 consecutive base pair portion of SEQ ID NO:1, 3, 5, 7, 15, 17, 19, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 61, 63, 65, 67, 69, 71, or 73.  
     
     
         5 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that can be amplified from a  Lycopersicon esculentum  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:9 and 10, SEQ ID NO:21 and 24, SEQ ID NO:22 and 24, SEQ ID NO:25 and 28, SEQ ID NO:26 and 28, or SEQ ID NO:59 and 60.  
     
     
         6 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that can be amplified from a  Beta vulgaris  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:22 and 24 or SEQ ID NO:26 and 28.  
     
     
         7 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that can be amplified from a  Helianthus annuus  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:26 and 28.  
     
     
         8 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that can be amplified from a  Solanum tuberosum  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:21 and 24, SEQ ID NO:21 and 23, SEQ ID NO:22 and 24, SEQ ID NO:25 and 28, or SEQ ID NO:26 and 28.  
     
     
         9 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that can be amplified from a  Brassica napus  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:9 and 10 or SEQ ID NO:26 and 28.  
     
     
         10 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that can be amplified from an  Arabidopsis thaliana  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:13 and 14, SEQ ID NO:21 and 24, or SEQ ID NO:22 and 24.  
     
     
         11 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that can be amplified from an  Nicotiana tabacum  DNA library using the polymerase chain reaction with the pair of primers set forth as SEQ ID NO:9 and 10, SEQ ID NO:11 and 12, SEQ ID NO:21 and 24, SEQ ID NO:22 and 24, SEQ ID NO:25 and 28, or SEQ ID NO:26 and 28.  
     
     
         12 . An isolated nucleic acid molecule according to  claim 1 , comprising a nucleotide sequence that can be amplified from a plant DNA library using the polymerase chain reaction with a pair of primers corresponding to the first 20 nucleotides and the reverse complement of the last 20 nucleotides of the coding sequence (CDS) of SEQ ID NO:1, 3, 5, 7, 15, 17, 19, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 61, 63, 65, 67, 69, 71, or 73.  
     
     
         13 . A chimeric gene comprising a promoter active in plants operatively linked to the nucleic acid molecule of  claim 1 .  
     
     
         14 . A recombinant vector comprising the chimeric gene of  claim 13 .  
     
     
         15 . A host cell comprising the chimeric gene of  claim 13 .  
     
     
         16 . A plant comprising the chimeric gene of  claim 13 .  
     
     
         17 . The plant of  claim 16 , which is selected from the following: rice, wheat, barley, rye, corn, potato, canola, sunflower, carrot, sweet potato, sugarbeet, bean, pea, chicory, lettuce, cabbage, cauliflower, broccoli, turnip, radish, spinach, asparagus, onion, garlic, eggplant, pepper, celery, squash, pumpkin, cucumber, apple, pear, quince, melon, plum, cherry, peach, nectarine, apricot, strawberry, grape, raspberry, blackberry, pineapple, avocado, papaya, mango, banana, soybean, tobacco, tomato, sorghum and sugarcane.  
     
     
         18 . Seed from the plant of  claim 16 .  
     
     
         19 . A method of increasing SAR gene expression in a plant, comprising expressing the chimeric gene of  claim 13  in said plant.  
     
     
         20 . A method of enhancing disease resistance in a plant, comprising expressing the chimeric gene of  claim 13  in said plant.  
     
     
         21 . A PCR primer selected from the group consisting of SEQ ID NO:9-14, 21-28, 59, and 60.  
     
     
         22 . A method for isolating a NIM1 homologue involved in the signal transduction cascade leading to systemic acquired resistance in plants comprising amplifying a DNA molecule from a plant DNA library using the polymerase chain reaction with a pair of primers corresponding to the first 20 nucleotides and the reverse complement of the last 20 nucleotides of the coding sequence (CDS) of SEQ ID NO:1, 3, 5, 7, 15, 17, 19, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 61, 63, 65, 67, 69, 71, or 73 or with the pair of primers set forth as SEQ ID NO:9 and 10, SEQ ID NO:11 and 12, SEQ ID NO:13 and 14, SEQ ID NO:21 and 24, SEQ ID NO:22 and 24, SEQ ID NO:21 and 23, SEQ ID NO:25 and 28, SEQ ID NO:26 and 28, or SEQ ID NO:59 and 60.  
     
     
         23 . The method of  claim 22 , wherein said plant DNA library is a  Nicotiana tabacum  (tobacco),  Lycopersicon esculentum  (tomato),  Brassica napus  (oilseed rape),  Arabidopsis thaliana, Beta vulgaris  (sugarbeet),  Helianthus annuus  (sunflower), or  Solanum tuberosum  (potato) DNA library.