Pesticidal nucleic acids and proteins and uses thereof
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. A DNA construct comprising a polynucleotide operably linked to a heterologous promoter, wherein said polynucleotide encodes a pesticidal polypeptide that comprises the amino acid sequence as set forth in SEQ ID NO:50.
2. The DNA construct of claim 1 , wherein said polynucleotide is codon-optimized for expression in a plant.
3. The DNA construct of claim 2 , wherein said polynucleotide comprises the nucleic acid sequence as set forth in SEQ ID NO:51.
4. 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 the amino acid sequence as set forth in SEQ ID NO:50.
5. The host cell of claim 4 , wherein said host cell is a bacterial cell or a plant cell.
6. The host cell of claim 5 , wherein said bacterial cell is selected from the group consisting of an Agrobacterium , a Bacillus , an Escherichia , a Salmonella , a Pseudomonas , and a Rhizobium 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.
7. A plant, or part thereof, comprising a polynucleotide encoding a pesticidal polypeptide that comprises the amino acid sequence as set forth in SEQ ID NO:50.
8. The plant, or part thereof, of claim 7 , 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.
9. 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 the amino acid sequence as set forth in SEQ ID NO:50.
10. The method of claim 9 , wherein said pest is an insect or a nematode.
11. The method of claim 10 , 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, Carphodorus, Criconema, Criconemella, Cryphodera, Ditylenchus, Dolichodorus, Eutylenchus, Globodera, Gracilacus, Helicotylenchus, Hemicriconemoides, Hemicycliophora, Heterodera, Hirschmanniella, Histotylenchus, Hoplolaimus, Hoplotylus, Longidorus, Macrotrophurus, 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, Trophonema, Trophotylenculus, Trophurus, Tylenchorhynchus, Tylenchulus, Tylenchus, Tylodorus, Xiphinema , and Zygotylenchus nematode.
12. The method of claim 9 , 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, a Cry protein, a VIP protein, and a chemical nematicide.
13. The method of claim 9 , 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 ID 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:42, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:58, or SEQ ID NO:60, or a pesticidal fragment thereof.
14. The method of claim 13 , 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.
15. The method of claim 13 , 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.