Pesticidal Nucleic Acids and Proteins and Uses Thereof

Abstract

The invention provides compositions comprising polynucleotide molecules encoding certain pesticidal polypeptides which exhibit plant parasitic nematode and/or insect control properties, and are particularly directed to controlling plant parasitic pest species of nematodes and insects known to infest crop plant species. Methods for controlling pests are disclosed in which the toxic proteins are provided in the diet of the targeted plant pests. The invention also provides compositions such as nucleic acids, proteins, and plant and bacterial cells, plants, and seeds containing the nucleic acid and protein compositions, as well as methods and kits for identifying, detecting, and isolating the compositions of the present invention. The invention further provides a method of producing crops from recombinant seeds which contain the polynucleotide molecules encoding the pesticidal polypeptides of the present invention.

1 - 9 . (canceled) 
     
     
         10 . A DNA construct comprising a polynucleotide operably linked to a heterologous promoter, wherein said polynucleotide encodes a pesticidal polypeptide that comprises an amino acid sequence at least 95% identical to SEQ ID NO:42. 
     
     
         11 . The DNA construct of  claim 10 , wherein said polynucleotide encodes a pesticidal polypeptide that comprises an amino acid sequence at least 99% identical to SEQ ID NO:42. 
     
     
         12 . The DNA construct of  claim 10 , wherein said polynucleotide encodes a pesticidal polypeptide that comprises the amino acid sequence as set forth in SEQ ID NO:42. 
     
     
         13 . The DNA construct of  claim 10 , wherein said polynucleotide is codon-optimized for expression in a plant. 
     
     
         14 . The DNA construct of  claim 13 , wherein said polynucleotide comprises the nucleic acid sequence as set forth in SEQ ID NO:43. 
     
     
         15 . A host cell comprising a DNA construct that comprises a polynucleotide operably linked to a heterologous promoter, wherein said polynucleotide encodes a pesticidal polypeptide that comprises an amino acid sequence at least 95% identical to SEQ NO:42. 
     
     
         16 . The host cell of  claim 15 , wherein said polynucleotide encodes a pesticidal polypeptide that comprises an amino acid sequence at least 99% identical to SEQ ID NO:42. 
     
     
         17 . The host cell of  claim 15 , wherein said polynucleotide encodes a pesticidal polypeptide that comprises the amino acid sequence as set forth in SEQ ID NO:42. 
     
     
         18 . The host cell of  claim 15 , wherein said host cell is a bacterial cell or a plant cell. 
     
     
         19 . The host cell of  claim 18 , wherein said bacterial cell is selected from the group consisting of an  Agrobacterium , a  Bacillus , an  Escherichia , a  Salmonella , a  Pseudomonas , and a  Rhizohium  cell, and wherein said plant cell is selected from the group consisting of a alfalfa, banana, barley, bean, broccoli, cabbage, canola, carrot, cassava, castor, cauliflower, celery, chickpea, Chinese cabbage, citrus, coconut, coffee, corn, clover, cotton, a cucurbit, cucumber, Douglas fir, eggplant, eucalyptus, flax, garlic, grape, hops, leek, lettuce, Loblolly pine, millets, melons, nut, oat, olive, onion, ornamental, palm, pasture grass, pea, peanut, pepper, pigeonpea, pine, potato, poplar, pumpkin, Radiata pine, radish, rapeseed, rice, rootstocks, rye, safflower, shrub, sorghum, Southern pine, soybean, spinach, squash, strawberry, sugar beet, sugarcane, sunflower, sweet corn, sweet gum, sweet potato, switchgrass, tea, tobacco, tomato, triticale, turf grass, watermelon, and a wheat plant cell. 
     
     
         20 . A plant, or part thereof, comprising a polynucleotide encoding a pesticidal polypeptide that comprises an amino acid sequence at least 95% identical to SEQ ID NO:42. 
     
     
         21 . The plant, or part thereof, of  claim 20 , wherein said polynucleotide comprises the nucleic acid sequence as set forth in SEQ ID NO:43. 
     
     
         22 . The plant, or part thereof, of  claim 20 , wherein said plant is elected from the group consisting of a alfalfa, banana, barley, bean, broccoli, cabbage, canola, carrot, cassava, castor, cauliflower, celery, chickpea, Chinese cabbage, citrus, coconut, coffee, corn, clover, cotton, a cucurbit, cucumber, Douglas fir, eggplant, eucalyptus, flax, garlic, grape, hops, leek, lettuce, Loblolly pine, millets, melons, nut, oat, olive, onion, ornamental, palm, pasture grass, pea, peanut, pepper, pigeonpea, pine, potato, poplar, pumpkin, Radiata pine, radish, rapeseed, rice, rootstocks, rye, safflower, shrub, sorghum, Southern pine, soybean, spinach, squash, strawberry, sugar beet, sugarcane, sunflower, sweet corn, sweet gum, sweet potato, switchgrass, tea, tobacco, tomato, triticale, turf grass, watermelon, and a wheat plant, and wherein said part is selected from the group consisting of a leaf, a stem a flower, a sepal, a fruit, a root, or a seed. 
     
     
         23 . A method of controlling a pest infection of a plant, said method comprising providing in a diet of said pest a plant, or part thereof, said plant or part comprising a polynucleotide encoding a pesticidal polypeptide that comprises an amino acid sequence at least 95% identical to SEQ ID NO:42. 
     
     
         24 . The method of  claim 23 , wherein said pest is an insect or a nematode. 
     
     
         25 . The method of  claim 24 , wherein said insect is an insect from the insect order selected from the group consisting of Coleoptera, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, and Trichoptera, and wherein said nematode is selected from the group consisting of  Acontylus, Anguina, Aorolaimus, Aphasmatylenchus, Aphelenchoides, Aphelenchus, Atalodera, Atylenchus, Bakernema Belonolaimus, Brachydorus, Bursaphelenchus, Cacopaurus, Caloosia, Carphadorus, Criconema, Criconemella, Cryphodera, Ditylenchus, Dolichodorus, Eutylenchus, Globodera, Gracilacus, Helicotylenchus, Hemicriconemoides, Hemicycliophora, Heterodera, Hirschmanniella, Histotylenchus, Hoplolaiinus, Hoplotylus, Longidorus, liacrotrophurus, Meloidodera, Meloidogyne, Merlinius, Morulaimus, Nacobbus, Nothanguina, Nothotylenchus, Paralongidorus, Paratrichodorus, Paratrophurus, Paratylenchus, Peltamigratus, Pratylenchoides, Pratylenchus, Psilenchus, Radopholoides, Radopholus, Rhadinaphelenchus, Rototylenchus, Rotylenchoides, Rotylenchus, Sarisodera, Scutellonema, Sphaeronema, Subanguina, Telotylenchoides, Telotylenchus, Trichotylenchus, Trophonerna, Trophotylenculus, Trophurus, Tylenchorhynchus, Tylenchulus, Tylenchus, Tylodorus, Xiphinema , and  Zygotylenchus nematode.    
     
     
         26 . The method of  claim 23 , said method further comprising providing in the diet of said pest a pesticidally effective amount of one or more other toxic agents selected from the group consisting of methylketone synthase, dsRNA, a Cry protein, a VIP protein, and a chemical nematicide. 
     
     
         27 . The method of  claim 23 , said method further comprising providing in a diet of said pest a pesticidally effective amount of one or more pesticidal polypeptides, wherein said one or more pesticidal polypeptides comprise SEQ ID NO:2, SEQ ID NO:6, SEQ ID NO:10, SEQ ID NO:14, SEQ II) NO:18, SEQ ID NO:22, SEQ ID NO:26, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:38, SEQ ID NO:46, SEQ ID NO:50, SEQ ID NO:54, SEQ ID NO:58, or SEQ ID NO:60, or a pesticidal fragment thereof. 
     
     
         28 . The method of  claim 27 , wherein the pesticidally effective amount of said one or more pesticidal polypeptides is provided by the plant, which is a recombinant plant, a part of the plant, or a product of the plant or the plant part. 
     
     
         29 . The method of  claim 27 , wherein the pesticidally effective amount of said one or more pesticidal polypeptides is provided in one or more formulations topically applied on the plant or a part of the plant, said one or more formulations comprising bacterial cells, spores, or parasporal crystals that comprise said one or more pesticidal polypeptides.