Compositions and methods of nematode control

Abstract

The present invention provides compositions and methods for identifying agents that stimulate or inhibit nematode reproduction, especially related to oocyte maturation, sheath cell contraction, and ovulation. It is disclosed that the major sperm protein (MSP) is acts in signal transduction of female sexual maturation in nematodes. Provided are compositions and methods for identifying anti-nematode agents with MSP as a target and for controlling nematode populations. MSP is an excellent target for identification of anti-nematode factors because it is highly conserved among members of the phylum Nematoda and is not known to exist in other organisms, especially crops, livestock, pets, and humans. Thus, anti-nematode agents that target MSP are less likely to induce severe side effects when administered to a host and the nematode is unlikely to develop resistance to a highly conserved molecule involved in sexual reproduction.

What is claimed is:  
     
         1 . A method of identifying an anti-nematode agent, comprising: 
 contacting a test compound to a nematode; and    monitoring a female sexual maturation of the nematode, wherein inhibition of the female sexual maturation indicates that the test compound includes the anti-nematode agent.    
     
     
         2 . The method of  claim 1 , wherein the female sexual maturation comprises an oocyte maturation or a gonadal sheath cell contraction.  
     
     
         3 . The method of  claim 1 , wherein the female sexual maturation comprises an ovulation.  
     
     
         4 . The method of  claim 1 , wherein the monitoring step includes optical detection.  
     
     
         5 . The method of  claim 1 , wherein the monitoring step includes optical detection of more than one female sexual maturation episode at the same time.  
     
     
         6 . The method of  claim 1 , wherein the nematode is an animal parasitic nematode.  
     
     
         7 . The method of  claim 1 , wherein the nematode is a plant parasitic nematode.  
     
     
         8 . The method of  claim 1 , wherein the nematode is of the genus Caenorhabditis.  
     
     
         9 . The method of  claim 1 , wherein the nematode is  Caenorhabditis elegans.    
     
     
         10 . The method of  claim 1 , wherein the nematode is of the genus Ascaris.  
     
     
         11 . The method of  claim 1 , wherein the nematode comprises a roundworm.  
     
     
         12 . The method of  claim 1 , wherein the nematode comprises a member of a genus selected from a group consisting of: Ascaris, Heterodera, Globodera, Meloidogyne, Ditylenchus, Anguina, Pratylenchus, Radopholus, Hirschmanniella, Hoplolaimus, Rotylenchulus, Tylenchulus, Helicotylenchus, Criconemella, Xiphinema, Longidorus, Trichodorus, Paratrichodorus, Aphelenchs, Onchocerca, Brugia, and Wuchereria.  
     
     
         13 . The method of  claim 1 , wherein the nematode includes a defect in a production of a sperm.  
     
     
         14 . The method of  claim 1 , wherein the nematode includes a genetic mutation.  
     
     
         15 . The method of  claim 1 , wherein the nematode is selected from a group consisting of: a fog-1 nematode, a fog-2 nematode, a fog-3 nematode, a fem-1 nematode, a fem-2 nematode, a fem-3 nematode, and a gld-1 nematode.  
     
     
         16 . The method of  claim 1 , wherein the nematode comprises a transgenic nematode including a transgenic expression of a major sperm protein, or a biologically active fragment thereof.  
     
     
         17 . The method of  claim 1 , wherein the nematode comprises a transgenic nematode including an ectopic expression of a major sperm protein, or a biologically active fragment thereof.  
     
     
         18 . The method of  claim 1 , wherein the nematode comprises a transgenic nematode including a transgenic expression of a major sperm protein from a nematode of a genus Caenorhabditis or a genus Ascaris, or a biologically active fragment thereof.  
     
     
         19 . A method of inhibiting a reproduction of the nematode of  claim 1 , comprising: administering the anti-nematode agent identified in  claim 1  to the nematode.  
     
     
         20 . The method of  claim 19 , wherein the anti-nematode agent comprises an antibody that binds specifically to the major sperm protein, or a biologically active fragment thereof.  
     
     
         21 . A method of controlling a population of nematodes, comprising: administering the anti-nematode agent identified in  claim 1  to the nematodes.  
     
     
         22 . A method of controlling a population of nematodes in an animal in need thereof, comprising: administering the anti-nematode agent identified in  claim 1  to the animal in an amount effective to control the population of nematodes.  
     
     
         23 . A method of increasing a host resistance to a nematode infection in an animal in need thereof, comprising: administering the anti-nematode agent identified in  claim 1  to the animal in an amount effective to increase the host resistance.  
     
     
         24 . A method of increasing a host resistance to a nematode infection in a non-human animal in need thereof, comprising: expressing the anti-nematode agent identified in  claim 1  in the animal, wherein the agent comprises a polypeptide which binds a major sperm protein.  
     
     
         25 . A method of controlling a population of nematodes in a plant in need thereof, comprising: administering the anti-nematode agent identified in  claim 1  to the plant in an amount effective to control the population of nematodes.  
     
     
         26 . A method of increasing a host resistance to a nematode infection in a plant in need thereof, comprising: administering the anti-nematode agent identified in  claim 1  to the plant in an amount effective to increase the host resistance.  
     
     
         27 . A method of increasing a host resistance to a nematode infection in an plant in need thereof, comprising: expressing the anti-nematode agent identified in  claim 1  in the plant, wherein the agent comprises a polypeptide which binds a major sperm protein.  
     
     
         28 . A process of making a factor that inhibits a female sexual maturation of a nematode, the method comprising: 
 contacting a test compound to a nematode;    monitoring the female sexual maturation, wherein inhibition of the female sexual maturation indicates that the test compound comprises the factor; and    manufacturing the factor.    
     
     
         29 . A method of identifying an anti-nematode agent, comprising: 
 contacting a test compound to a polypeptide comprising a major sperm protein or a portion thereof capable of stimulating a female sexual maturation; and    detecting a composition including the test compound and the polypeptide, wherein the presence of the composition indicates that the test compound is the anti-nematode agent.    
     
     
         30 . The method of  claim 29 , further comprising screening the anti-nematode agent for inhibition of a female sexual maturation.  
     
     
         31 . The method of  claim 29 , wherein the MSP comprises the amino acid sequence set forth in SEQ ID NO:2: or nematode major sperm protein alignment variants thereof, or a portion thereof capable of stimulating a female sexual maturation.  
     
     
         32 . A method of inhibiting a reproduction of a nematode, comprising: inhibiting a stimulation of a female reproductive cell by a major sperm protein.  
     
     
         33 . A method of identifying an agent that binds to a biologically active domain of a nematode major sperm protein, comprising: 
 contacting a test compound to the domain; and    detecting a composition including the test compound and the domain, wherein the presence of the composition indicates that the agent binds to the domain.    
     
     
         34 . An isolated nematode major sperm protein domain, comprising: a polypeptide including 125 or fewer consecutive amino acids of a nematode major sperm protein, wherein the domain is capable of stimulating a female sexual maturation.  
     
     
         35 . The major sperm protein domain of  claim 34 , wherein the polypeptide includes 20 or fewer amino acids of the nematode major sperm protein, wherein the domain is capable of stimulating the female sexual maturation.  
     
     
         36 . The major sperm protein domain of  claim 34 , wherein the polypeptide includes 10 to 30 amino acids of the nematode major sperm protein, wherein the domain is capable of stimulating the female sexual maturation.  
     
     
         37 . An isolated nematode major sperm protein domain, comprising: a polypeptide including 125 or fewer consecutive amino acids as set forth in SEQ ID NO:2 or nematode major sperm protein alignment variants thereof, wherein the domain is capable of stimulating a female sexual maturation.  
     
     
         38 . The major sperm protein domain of  claim 37 , wherein the polypeptide includes 20 or fewer amino acids, wherein the domain is capable of stimulating a female sexual maturation.  
     
     
         39 . The major sperm protein domain of  claim 37 , wherein the polypeptide includes 10 to 30 amino acids, wherein the domain is capable of stimulating a female sexual maturation.  
     
     
         40 . An antibody that selectively binds to a biologically active domain of a nematode major sperm protein, wherein the biologically active domain comprises a sheath cell contraction domain.  
     
     
         41 . The antibody of  claim 40 , wherein the biologically active domain comprises 20 or fewer amino acids of a carboxyl-terminus of the domain.  
     
     
         42 . An antibody that selectively binds to a biologically active domain of a nematode major sperm protein, wherein the biologically active domain comprises an oocyte maturation domain.  
     
     
         43 . The antibody of  claim 42 , wherein the biologically active domain includes an amino acid sequence having a group of consecutive residues in the range of 96-110 of SEQ ID NO:2, or nematode major sperm protein alignment variants thereof; and wherein the amino acid sequence includes 20 or fewer consecutive residues of SEQ ID NO:2, or nematode major sperm protein alignment variants thereof.  
     
     
         44 . The antibody of  claim 42 , wherein the biologically active domain includes an amino acid sequence having a group of consecutive residues in the range of 1-106 of SEQ ID NO:2, or nematode major sperm protein alignment variants thereof; and wherein the amino acid sequence includes 20 or fewer consecutive residues of SEQ ID NO:2, or nematode major sperm protein alignment variants thereof.  
     
     
         45 . A process of making an antibody, comprising immunizing a non-human animal with an immunogenic fragment of a nematode major sperm protein domain including a polypeptide having 20 or fewer consecutive amino acids as set forth in SEQ ID NO:2 or nematode major sperm protein alignment variants thereof, wherein the domain is capable of stimulating a female sexual maturation.