GENETICALLY MODIFIED PSEUDOMONAS STRAINS WITH ENHANCED BIOCONTROL ACTIVITY

Abstract: Strains of Pseudomonas have been genetically engineered to have enhanced biocontrol properties. The strains of the invention are particularly effective against plant pathogenic fungi such as species of Rhizoctonia and Pythium, because the strains produce enhanced amounts of antifungal metabolites such as pyrrolnitrin that are active against these fungal pathogens. Both the genetically modified biocontrol strains and the antifungal metabolites can be used as active agents for biocontrol compositions.

Description: GENETICALLY MODIFIED PSEUDOMONAS STRAINS WITH ENHANCED BIOCONTROL ACTIVITY The present invention relates to genetically modified strains of Pseudomonas that have improved biocontrol properties. More specifically it relates to strains that are effective against plant pathogenic fungi. It has been recognized that crops grown in some soils are naturally resistant to certain fungal pathogens. Furthermore, soils that are conducive to the development of these diseases can be rendered suppressive or resistant to the pathogen by the addition of small quantities of soil from a suppressive field ( Scherarx. Baker (1980) Conversely, suppressive soils can be made conducive to fungal disease susceptibility by autoclaving, indicating that the factors responsible for disease control are biological. Subsequent research has demonstrated that root colonizing bacteria are responsible for this phenomenon, which is known as biological disease control ( Cook and Baker (1983), The Nature and Practice of Biological Control of Plant Palhogens; Amer. Phytopalhol. Soα, St Paul, MN). In many cases, the most efficient strains of biological disease controlling bacteria are fluorescent pseudomonads ( WeBereta/. (1983) Phytopathology, 73: 463469). These bacteria have also been shown to promote plant growth in the absence of a specific fungal pathogen by the suppression of detrimental rhizosphere microflora present in most soils ( Woepperet a/.(1981) Phytopatholog 71: 1020-1024). Important plant pathogens that have been effectively controlled by seed inoculation with these bacteria include Gaemannomyces graminis, the causative agent of take-all in wheat ( Cook era/.(1976) Sol Bbl. B chem 8: 269-273) and Pythium and Rhizoctonia, pathogens that cause damping off of cotton ( Howeflefa/. (1979)F 5 hytopa1nology 69:480-482). Rhizoctonia is a particularly problematic plant pathogen for several reasons. First, it is capable of infecting a wide range of crop plants, and second, there are no commercially available chemical fungicides that are effective in controlling the fungus. Many biological disease controlling Pseudomonas strains produce antibiotics that inhibit the growth of fungal pathogens ( Howelefa/. (1979) FrrytDpathofog 69:480-482; Howelefa/. (1980) Ftτytopatho gy70:712-715). These antibiotics have been implicated in the control of fungal pathogens in the rhizosphere. For example, Howell et al. (Rτytopatholog 69: 480482; 1979) disclose a strain of Pseudomonas fluorescens that produces an antibiotic substance antagonistic to Rhizoctonia solani. In addition, other strains of Pseudomonas fluorescens having enhanced biocontrol activity against plant pathogenic fungi such as Rhizoctonia and Pythium are disclosed in U.S. Patent Nos. 5,348,742 and 5,496,547, both of which are hereby incorporated by reference in their entireties. Several other past studies have focused on the effects of mutations that result in the inability of the disease control bacterium to synthesize these antibiotics ( Kloepperef a/. (1981) FtiytDpatho g 71: 1020-1024; HoweS etal. (1983) Can. J. Mcfobiol.29: 321-324). In these cases, the ability of the organism to control the pathogen is reduced, but not eliminated. A particularly effective antibiotic against fungal p athogens is pyrrolnitrin, which is biosynthesized from tryptophan (Cnangefa/. J.Anlbiot 34: 555-566). Pyrrolnitrin is a phenylpyrrole derivative with strong antibiotic activity that has been shown to inhibit a broad range of fungi (Hommaera/.,So_ Bid. Biochem.21: 723-728(1989); r ύdae*a/.,J.Arιtt ±,ser. 18: 211-219(1965)). Pyrrolnitrin was originally isolated from Pseudomonas pyrrocinia (Arimaetal, J. Aπtb t,ser.Ai8:201-204(1965)), but has since been isolated from Myxococcus species, Burkholdaria species, and several other Pseudomonas species such as Ps fluorescens (Gerth eta/. J. Arttbbt 35: 1101-11CG (1982); J.N. Floitma^ MαotøologyarriBbtedΥrølogy 34:381-386 (1990)). The compound has been reported to inhibit fungal respiratory electron transport ( T athi & Gottfieb, J. Bacteriol.100: 310-318 (1969)) and uncouple oxidative phosphorylation ( l_ambcwitz&Slayman,J.Baιcterft It has also been proposed that pyrrolnitrin causes generalized lipoprotein membrane damage ( Nose & Arirra, J. AritiDbt, ser A, 22 135-143 (1969); Ca^^ AppBcata53: 111 -123 (1974)). U.S. Patent No. 5,639,949 and Application Serial No. 08/729,214, both of which are hereby incorporated by reference in their entireties, describe the cloning and characterization of the pyrrolnitrin biosynthetic genes from Ps. fluorescens and Ps. pyrrocinia. An important factor in biological control is the ability of a biocontrol organism to compete in a given environment (Baker etal. (1982) Biological (Itoritid of PlartPatix^ Phytopathobgcal Society, St Paul, Mnn., pages 61-106). Thus, it is desirable to obtain strains of biocontrol agents that effectively control the growth of fungal pathogens such as Rhizoctonia and Pythium and that are also able to aggressively compete with indigenous bacteria and microflora existing in the rhizosphere of the plant. The present invention is drawn to genetically engineered biocontrol strains of Pseudomonas that are able to effectively control pathogenic attack on crop plants. Preferred biocontrol strains include the following strains of Pseudomonas fluorescens, which are described in detail in the examples below: CGA376146, CGA364473, CGA375258, CGA376148, CGA364476, CGA375260, CGA375259, CGA378584, CGA267pPhz, CGA364474, CGA364475, CGA366259, CGA376150, NOA402208, NOA402212, NOA402214, NOA402216, CGA267356/Phl, NOA409068, NOA413174, NOA413175, NOA413176, NOA413177, and NOA413178. The biocontrol strains of the invention produce at least one antifungal substance that is capable of inhibiting a broad spectrum of plant pathogens such as Rhizoctonia and Pythium. In a preferred embodiment, the biocontrol strains of the invention produce enhanced quantities of pyrrolnitrin; see, e.g., Table 1. As shown in Table 2, such strains have increased biocontrol properties and are able to aggressively compete in the plant rhizosphere. The genotypes of the biocontrol strains of the invention are summarized in Table 3 and deposit information for the biocontrol strains of the invention is given in Table 4. The present invention is also intended to encompass pyrrolnitrin producing strains derived from the above-listed strains. The present invention is also drawn to biocontrol compositions comprising the biocontrol strains of the invention in combination with a chemical fungicide such as a metalaxyl compound. In addition, methods of making the biocontrol strains as well as methods of using the strains and biocontrol compositions for control of pathogenic attack on crops are described. SEQ ID NO:1 is the nucleotide sequence of the 11 kb EcoRI fragment containing the gacA gene and derived from the chromosome of Pseudomonas fluorescens strain CGA267356. The coding sequences in this sequence include genes encoding: a methyltransferase (bases 210-1688) with homology to the cheR and frzF genes from E. coli and Myxococcus xanthus, respectively; a sensor kinase (bases 1906-3633) with homology to the rcsC, frzE and bvgS genes of E. coli, M. xanthus, and Borditella pertussis, respectively; a tRNA (bases 4616-4691 , complementary DNA strand) with homology to glyWirom E. coli ; CDP-diacylglycerol-glycerol-3-phosphate-3-phosphatidyltransferase (bases 4731-5318, complementary DNA strand) with homology to pgsA; UVR exonuclease subunit C (bases 5574-7397, complementary DNA strand) with homology to uvrC; and a response regulator/transcription activator (gacA; bases 7400-8041 , complementary DNA strand) with homology to the uvrY and GACA genes of E. coli and P. fluorescens, respectively. SEQ ID NO:2 is the nucleotide sequence of the native gacA regulatory gene. SEQ ID NO:3 is the protein sequence encoded by the native gacA regulatory gene. SEQ ID NO:4 is the nucleotide sequence of the ATG- gacA regulatory gene, wherein the first base in the coding sequence has been changed from the native thymidine (T) to an adenine (A) to create the more efficient ATG translation initiation codon. SEQ ID NO:5 is the protein sequence encoded by the altered ATG- gacA regulatory gene. SEQ ID NO:6 is the nucleotide sequence of the pyrrolnitrin gene cluster. SEQ ID NO:7 is the nucleotide sequence of the tac promoter/rmB transcription terminator cassette. SEQ ID NO:8 is the nucleotide sequence of the lemA gene. SEQ ID NO:9 is the nucleotide sequence of the gac*3 regulatory gene, wherein the adenine (A) base at position 395 has been changed from the native adenine (A) to a guanine (G) so that codon 132 encodes an arginine residue instead of the usual glutamine. SEQ ID NO:10 is the protein sequence encoded by the altered gac * 3 regulatory gene. SEQ ID NO:11 is the nucleotide sequence of the phenazine gene cluster. The present invention provides improved biocontrol strains of Pseudomonas that can be used to control pathogenic attack on crop plants. Such strains are able to aggressively compete in the plant rhizosphere as well as produce one or more antifungal substances that are effective against a broad spectrum of plant pathogenic fungi such as Rhizoctonia and Pythium. The biocontrol strains of the present invention are important for several reasons. First, Rhizoctonia such as Rhizoctonia solani are particularly pernicious plant pathogens. The affected plants include beans, wheat, tomato and potato, in addition to cotton. Second, there are few environmentally safe and effective fungicide treatments available for the protection of crops from Rhizoctonia. Therefore, the use of the disclosed biocontrol strains to control or prevent Rhizoctonia infections in crop plants provides an environmentally safe and effective method of controlling this and other plant pathogens. Pseudomonas fluorescens strain CGA267356 (also known as both MOCG134 and BL915) has been shown to be effective in controlling plant pathogenic fungi such as Rhizoctonia and Pythium. Strain CGA267356 is one of the subjects of U.S. Patent No. 5,348,742. Two mutants of CGA267356, strain s CGA321730 (a.k.a. MOCG134-8392) and CGA319115, have been constructed and shown to demonstrate even better biological control (biocontrol) of these phytopathogens. CGA321730 and CGA319115 are the subject of U.S. Patent No. 5,496,547. Two genes have been isolated from strain CGA267356 that encode proteins that regulate the synthesis of several antifungal compounds produced by the strain , including the antifungal metabolite pyrrolnitrin (prn). These are the /e A gene and gafA (a.k.a. gacA) gene that encode sensor kinase and response regulator proteins, respectively, which function as a typical two-component bacterial regulatory system. These genes and their use to activate biocontrol activity in biocontrol strains are described in U.S. Patent No. 5,670,350, which is hereby incorporated by reference in its entirety. In addition, U.S. Patent No. 5,639,949 describe a four gene cluster isolated from strain CGA267356 that encodes proteins that direct the biosynthesis of pyrrolnitrin. In the present invention, the lemA and gacA regulatory genes and the pyrrolnitrin biosynthetic genes have been utilized to genetically modify parent Ps. fluorescens strains such as CGA267356 and also mutant strains such as, for example , strains CGA321730 and CGA267356, to construct altered strains that demonstrate enhanced production of antifungal metabolites, i.e. pyrrolnitrin, and accordingly enhanced biocontrol activity. In addition, genes from Pseudomonas aureofaciens strain 30-84 that are involved in the synthesis of the antifungal metabolite phenazine-1-carboxylic acid (PCA) have been utilized to genetically modify parent Ps. fluorescens strain CGA267356 to produce PCA, thereby improving the biocontrol activity of strain CGA267356. In particular, a plasmid comprising the gacA and lemA regulatory genes, such as plasmid pE11 is transformed into a transposon mutant of a wild-type P. fluorescens strain that has enhanced biocontrol activity over the wild-type strain resulting in a biocontrol strain according to the invention . In a further embodiment of the invention the first base in the coding sequence of the gacA gene is changed from a thymidine (T) to an adenine (A) to create the more efficient ATG translation initiation codon . This change can be created in vitro by PCR technology and the native gacA gene in the chromosome of a wild-type P. fluorescens strain can be replaced with the ATG/gacA gene by homologous gene replacement resulting in a biocontrol strain according to the invention . In a still further embodiment of the invention a plasmid is introduced into a wild-type or mutant Pseudomonas strain, but preferably a P. fluorescens strain, comprising the pmABCD gene cluster with a strong constitutive bacterial promoter such as, for example, the tac promoter, upstream of the gene cluster and a suitable transcription terminator such as the trnB transcription terminator downstream of the pmABCD gene cluster. The use of the tac promoter with the pm genes causes constitutive, high-level expression of these genes in the transformed Pseudomonas strain resulting in a biocontrol strain according to the invention. The invention further provides a Pseudomonas strain transformed with a plasmid with the lemA and gacA genes derived from the chromosome of a wild-type strain. In a preferred embodiment of the invention a Pseudomonas strain is provided which has been transformed to contain both the the pmABCD gene cluster and a plasmid with the lemA and gacA genes. In a further embodiment of the invention a single base change within the coding sequence of the native gacA gene is accomplished resulting in gacA genes that encode a GacA protein exhibiting a single amino acid change in the encoded GacA protein such that do no longer require phosphorylation by LemA in order to be active as a transcriptional activator. Said gacA genes are isolated and used to replace the native gacA gene in wild- type and mutant Pseudomonas strains by perfect site replacement mediated through homologous recombination resulting in a biocontrol strain according to the invention . Preferred within the scope of the invention is a mutant gacA gene comprising a single base change in codon 132, which is CAG and encodes a glutamine residue in the native GacA protein. The adenine base in this codon is changed to guanine to create a codon that encodes an arginine residue (CGG) in the altered strain. In the normal regulatory system and under the proper conditions, the LemA protein phosphorylates GacA and in the phosphorylated state it activates transcription of genes involved in the synthesis of antifungal compounds. This single base change in the GacA protein renders it active irrespective of the kinase activity of the LemA protein. In a further embodiment of the invention the native promoter controlling expression of the chromosomal gacA gene is replaced with a strong constitutive bacterial promoter such as, for example, the tac promoter from E. coli and the thus modified gacA gene is introduction into a wild-type or mutant Peudomonas strain, optionally together with a plasmid comprising the pmABCD gene cluster. The newly inserted tac/gacA gene resulted in a higher level of expression of the gacA gene, compared to the low level of expression from the native gacA gene promoter. The invention further provides a no-phenazine, pyrrolnitrin producing Pseudomonas strain which is transformed with a plasmid carrying DNA that contains 5 genes known to encode the pathway for the biosynthesis of the antifungal metabolite phenazine-1- carboxylic acid (PCA). The resulting strain is shown to produce PCA, an antifungal metabolite not normally produced by the parent strain in addition to the metabolites normally produced by the parent strain, including pyrrolnitrin. In a further embodiment of the invention a plasmid is constructed comprising the lac promoter element from £. coli with the gacA gene such that expression of the gacA gene is regulated by the lac promoter. This plasmid is transformed into a wild-type or mutant Pseudomonas strain resulting in a biocontrol strain according to the invention. In a still futher embodiment of the invention the tac promoter element from £. coli is operably linked with the pmABCD gene cluster and introduced in the chromosome of a wild- type or mutant Pseudomonas strain such that expression of the pmABCD genes is from a strong constitutive bacterial promoter such as, for example, the tac promoter. In additon, a plasmid, but preferably plasmid pE11 , containing the native gacA gene derived from P. fluorescens strain CGA267356 is introduced in said strain by conjugation. The new strain is shown to produce higher amounts of the antifungal metabolite pyrrolnitrin and to provide greater biocontrol activity than the parent strain. The invention also provides a 2-hexyl-5-propyl-resorcinol non-producing deletion mutant of a P. fluorescens strain. Said strain is transformed with a plasmid comprising the pmABCD gene cluster under the control of the tac promoter resulting in a biocontrol strain according to the invention. A further embodiment of the invention pro vides a method for controlling or inhibiting the growth of a plant pathogenic fungus by applying the genetically engineered biocontrol strains of the invention to an environment in which the plant pathogenic fungus may grow. This can be to the plant/s or parts of the plant s (before or after harvest) or to the seeds (prior to planting) of the plant/s to be protected, or alternatively to soil in which the plant/s to be protected are growing or will grow. The biocontrol strains are applied in an effective amount; that is, in an amount sufficient to control or inhibit the pathogen. The rate of application may vary according to the crop to be protected, the efficacy of the biocontrol strain, the pathogen to be controlled, and the severity of the disease pressure. Generally, the rate of application is about 1.3 x 10 5 cfu/cm to about 1.3 x 10 10 cfu/cm, specifically about 1.3 x 10 6 cfu/cm to about 1.3 x 10 9 cfu/cm, more specifically about 1.3 x 10 7 cfu/cm to about 1.3 x 10 8 cfu/cm. A more particular embodiment of the present invention provides methods of inhibiting the growth of Rhizoctonia and Pythium by applying the biocontrol strains of the invention to environments in which the plant pathogenic fungi may grow. This can be to the plant/s or parts of the plant/s (before or after harvest) or to the seeds (prior to planting) of the plant/s to be protected, or alternatively to soil in which the plant/s to be protected are growing or will grow. As noted above, the rate of application varies depending on various factors. However, the general rate of application is about 1.3 x 10 5 cfu/cm to about 5 x 10 9 cfu/cm, specifically about 1.3 x 10 6 cfu/cm to about 1.3 x 10 9 cfu/cm more specifically about 1.3 x 10 7 cfu/cm to about 1.3 x 10 8 cfu/cm. The recombinant biocontrol strains of the present invention may be used in any manner known in the art, including coating seeds with an effective amount of the biocontrol strains, in furrow application of the biocontrol strains directly into the soil, in foliar application, and in post-harvest disease control. Such methods are well known in the art and are described, for example, in U.S. Patent No. 5,348,742 and in the published European Application EP 0 472 494 A2, which is hereby incorporated by reference. Furthermore, the strains of this application can also be mixed in formulation with known pesticides in a manner described in WO 94/10845, which disclosure is herein incorporated by reference. EXAMPLES The invention is illustrated in further detail by the following detailed procedures, preparations, and examples. The examples are for illustration only, and are not to be construed as limiting the scope of the present invention. Standard recombinant DNA and molecular cloning techniques used here are well known in the art and are described by andbyTJ.Slτavy. M.LBeιτnan.arxJLW. Erκ^ Laboratory, Coϋ Spring Harbor, NY (1984) and by Ausubel, F.M etal., Current Protocols in Molecular Biology, pub. by Greene PubBshing Asscc and WBey-lnterscience (1987). EXAMPLE 1 Construction of Strain CGA376146 (MON0571) (CGA321730, pE11) Strain CGA376146 (a.k.a. MON0571) was constructed by introducing plasmid pE11 into P. fluorescens strain CGA321730 (a.k.a. MOCG-0134-8392) by conjugation. Strain CGA321730 is described in U.S. Patent No. 5,496,547 and is a transposon mutant of wild- type P. fluorescens strain CGA267356 (U.S. Pat. No. 5,348,742) that has enhanced biocontrol activity over the wild-type strain. Strain CGA267356 has been deposited with the ATCC and assigned ATCC accession number 55169. Strain CGA321730 has been deposited with the NRRL and assigned accession number NRRL B-21173. Plasmid pE11 (Patent No. 5,670,350) was constructed by ligating the 1 1 kilobase (kb) EcoRI fragment derived from the chromosome of strain CGA267356 into the broad host range plasmid vector pRK290 (Dittaera/.,P/oc. Naϋ.Acad. Sά. USA 77:7347-7351 (1980). Plasmid pE11 has been deposited with ATCC and has been assigned ATCC accession number 40869. The 11-kb EcoRI fragment contains the gacA (gafA) gene that encodes a response regulator protein known to regulate the synthesis of antifungal compounds. This 11-kb EcoRI fragment is described in U.S. Patent No.5,670,350 and is set forth herein as SEQ ID NO. . The plasmid vector, pRK290, used in constructing pE11 , is derived from native Pseudomonas plasmids. It is mobilizable but not self-transmissible by conjugation and it carries a tetracycline resistance gene (DittaetaL, 1980). New strain CGA376146 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to have higher biocontrol activity (Table 2) compared to the related strains CGA267356 and CGA321730. New strain CGA376146 was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B- 21811. EXAMPLE 2 Construction of Strain CGA364473 (MON0517) (ATG-gacA) P. fluorescens strain CGA364473 (a.k.a. MON0517) was derived from parent P. fluorescens strain CGA267356 by changing a single base in the chromosome of the parent strain. The native gacA regulatory gene begins with the unusual TTG translation initiation codon (SEQ ID NO:2). All proteins in nature are synthesized with methionine as the first amino acid on the amino terminus and ATG is the only codon that encodes methionine. Therefore, the normal translation initiation codon for most genes is ATG. Alternate translation initiation codons GTG and TTG will also result in the incorporation of methionine as the first amino acid since methionine must always be the first amino acid in a newly synthesized protein, but they usually cause a reduction in the efficiency of translation. As a result, fewer protein molecules are made from the same amount of messenger RNA. To create strain CGA364473, the first base in the coding sequence of the gacA gene was changed from a thymidine (T) to an adenine (A) to create the more efficient ATG translation initiation codon (SEQ ID NO:4). This change was created in vitro by PCR technology and the native gacA gene in the chromosome of strain CGA267356 was replaced with the ATG/gacA gene by homologous gene replacement. The amino acid sequence of the GacA protein encoded by this altered gacA gene (SEQ ID NO:5) is identical to that encoded by the native gene (SEQ ID NO:3), but translation should be more efficient, resulting in synthesis of higher amounts of the protein. New strain CGA364473 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to have higher biocontrol activity (Table 2) compared to strain CGA267356. New strain CGA364473 was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B-21812. EXAMPLE 3 Construction of Strain CGA375258 (MON0568) (pPrn) Strain CGA375258 (a.k.a. MON0568) was created by introducing plasmid pPrn into the parent P. fluorescens strain CGA267356. Plasmid pPrn was constructed by cloning a 6.2 kb Xba\/Not gene fragment from plasmid pCIB169 (which was derived from P. fluorescens strain CGA267356) into the expression vector pKK223-3 , as described in Examples 7-11 of U.S. Patent No. 5,639,949 and Application Serial No. 08/729,214. (See SEQ ID NO:6). Plasmid pCIB169 has been deposited with the NRRL and assigned accession number NRRL B-21256. The 6.2 kb XbaUNott gene fragment of SEQ ID NO:6 contains the pmABCD gene cluster that encodes genes for the biosynthesis of pyrrolnitrin as described in Patent No. 5,639,949 and Serial No. 08/729,214. A 6.9 kb βg/ll fragment containing the pmABCD gene cluster with the tac promoter (see SEQ ID NO:7) upstream of the gene cluster and the rrnB transcription terminator (SEQ ID NO:7) derived from plasmid pKK223-3 downstream of the cluster was subsequently cloned into the Sg/ll site of plasmid pRK290 to create plasmid pPrn. The tac promoter is a small DNA fragment (less than 100 bases) derived from E. coli, which is known to be a regulatory e iement or promoter (Amann, etal, Gene 25:167-178 (1983)) that does not itself encode a protein product. SEQ ID NO:7 presents the sequence of the SssHII DNA fragment containing the tac promoter and the rrnB transcription terminator derived from plasmid pKK223-3. The tac promoter is known to be highly expressed in a constitutive manner in Pseudomonas. Its use with the pm genes causes constitutive, high-level expression of these genes. On the other hand, in the native strain, the promoter for the pyrrolnitrin genes is regulated by the gacA gene product; accordingly, the pyrrolnitrin genes are expressed in the native strain only in the stationary phase of growth. New strain CGA375258 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to have higher biocontrol activity (Table 2) compared to the related strain CGA267356. New strain CGA375258 was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B-21813. EXAMPLE 4 Construction of Strain CGA376148 (MON0573) (CGA321730, pPrn) The chromosomal background of strain CGA376148 (a.k.a. MON0573) is the same as strain CGA376146 (Example 1). In addition, strain CGA376148 contains the plasmid pPrn (Example 3) that carries the tac promoter fused to the pmABCD gene cluster. New strain CGA376148 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to have higher biocontrol activity (Table 2) compared to the related strain CGA267356. New strain CGA376148 was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B-21814. EXAMPLE 5 Construction of Strain CGA364476 (MONO520) (pLem/Gac) Strain CGA364476 (a.k.a. MONO520) is the same as wild-type strain CGA267356, except that strain CGA364476 also contains a plasmid with the lemA and gacA genes derived from the chromosome of the wild-type strain CGA267356. The plasmid containing the lemA and gacA genes, pLem/Gac, was constructed as follows: The plasmid pCIB146 (Examples 20 and 21 and Figure 4 of Patent No. 5,670,350) contains about 25-kb of chromosomal DNA from strain CGA267356. The lemA gene (see SEQ ID NO:8) has been described and shown to be located in this DNA. The subcloned DNA in pCIB146 is flanked on each side by Not\ and EcoRI sites. An approximately 16-kb Hind\\\INot\ fragment from pCIB146, which contains the lemA gene, was excised from pCIB146. The Not\ end was converted to a Hind\\\ site to facilitate cloning of the fragment into the unique Hind\\\ fragment of plasmid pE11 (Example 1) , which contains the gacA gene. Insertion of the 16-kb /emA-containing Hind\\\ fragment into the Hind \\ site of pE11 did not disrupt the function of the gacA gene, because the H/ndlll site is not within its coding sequence. New strain CGA364476 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to have higher biocontrol activity (Table 2) compared to the related strain CGA267356. New strain CGA364476 was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B-21815. EXAMPLE 6 Construction of Strain CGA375260 (MONO570) (gac*3) Strain CGA375260 (a.k.a. MONO570) differs from the parent strain CGA267356 by a single base change within the coding sequence of the native gacA gene (SEQ ID NO:2). This modification was generated by introducing the native gacA gene into the hypermutagenic E. coli strain XL1-Red (from Stratagene, Inc.). The plasmid was recovered and introduced into a lemA mutant of strain CGA267356 that also contained lacZY genes inserted into an unknown chromosomal gene whose expression is regulated by LemA and GacA. Clones containing randomly mutated gacA genes that resulted in expression of the lacZ genes, as indicated by the formation of blue colonies on agar containing X-Gal , were further analyzed. These clones contained gacA genes that did not require phosphorylation by LemA in order to be active as a transcriptional activator. Three such gacA genes were isolated in this manner and the nucleotide sequence of each was determined. In each, there was a different single base change that resulted in a different single amino acid change in the encoded GacA protein. Each of the three modified gacA genes were used to replace the native gacA gene in strain CGA267356 by perfect site replacement mediated through homologous recombination. Of the three, only one clone with a LemA-independent gacA gene (gac * 3) was shown to have enhanced pyrrolnitrin synthesis and biocontrol activity (Tables 1 and 2). The nucleotide sequence of the gac*3 gene of this clone, CGA375260, was determined (SEQ ID NO:9) and it was found that a single base change occurs in codon 132, which is CAG and encodes a glutamine residue in the native GacA protein. The adenine base in this codon was changed to guanine to create a codon that encodes an arginine residue (CGG) in the altered strain. Therefore, the GacA protein (SEQ ID NO: 10) in this strain has an arginine at amino acid 132 instead of the usual glutamine. In all other respects , this strain is identical to the parent strain. In the normal regulatory system and under the proper conditions, the LemA protein phosphorylates GacA and in the phosphorylated state it activates transcription of genes involved in the synthesis of antifungal compounds. This single base change in the GacA protein renders it active irrespective of the kinase activity of the LemA protein. New strain CGA375260 was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B-21816. EXAMPLE 7 Construction of Strain CGA375259 (MON0569) (tac/gacA, pPrn) Strain CGA375259 (a.k.a. MON0569) was derived from strain CGA267356 by replacement of the native promoter controlling expression of the chromosomal gacA gene with the tac promoter from E. coli (Example 3) and introduction of plasmid pPrn (Example 3). The promoter of the gacA gene was replaced with the tac promoter as follows: A unique Nru\ site in the 2-kb Xho\ gacA gene-containing fragment of pCIB137 (Examples 6 and 7 of Patent No. 5,670,350) located 12-bp upstream of the translation start site of the gacA gene was modified by PCR to change it to a SamHI site. Plasmid pCIB137 has been deposited with the NRRL and assigned accession number NRRL B-18981. A second SamHI site was inserted immediately 5' to the gacA translation start site using PCR. This created a small SamHI fragment immediately preceding the gacA coding sequence. This short SamHI fragment was excised and the DNA was religated to create a new SamHI site. The tac promoter was excised from plasmid pKK223-3 (SEQ ID NO:7) as a Sg/ll/SamHI fragment and cloned in the appropriate orientation into the new SamHI site 5' to the beginning of the gacA gene, which was created by the excision of the above short SamHI fragment. This tac promoter/gacA gene (tac/gacA) fragment was excised as an Xho\ fragment and was used to replace the native gacA gene on the 2-kb Xho\ site in a plasmid containing the Hind\WEcoR\ fragment from pE11. This plasmid was introduced into a gacA deletion mutant of strain CGA267356 (Example 9 of Patent No. 5,670,350). Perfect replacement clones were generated (Example 9 of Patent No. 5,670,350) and selected by restoration of the wild-type colony morphology. The newly inserted tac/gacA gene resulted in a higher level of expression of the gacA gene, compared to the low level of expression from the native gacA gene promoter. Plasmid pPrn described in Example 3 above was subsequently introduced into the strain containing the tac/gacA gene to make strain CGA375259. New strain CGA375259 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to have higher biocontrol activity (Table 2) compared to the related strain CGA267356. New strain CGA375259 was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B-21817. EXAMPLE 8 Construction of Strain CGA378584 (MON0591) (cPrn, pLem/Gac) Strain CGA378584 (a.k.a. MON0591 ) contains the tac/pmABCD gene cluster described in Example 3 in the chromosome and it also contains plasmid pLem/Gac described in Example 5. The tac/pmABCD genes were inserted in the chromosome of strain CGA267356 by modification of the £. coli cloning vector pKK223-3 by adding a kanamycin resistance gene derived from plasmid pUC4K (Pharmacea) into the Pst\ site of the multiple cloning site and by removing the EcoRI, Nott, and SamHI sites of pKK223-3. A 16-kb Kpn\ gene fragment derived from pCIB169 (Figure 4 of Patent No. 5,639,949) was cloned into the modified pKK223-3. Plasmid pCIB169 has been deposited with the NRRL and assigned accession number NRRL B-21256. The pmABCD genes were deleted by digestion of the plasmid with EcoRI and Noti, conversion of the EcoRI and Λ/ofl ends to SamHI by fill-in, linkering, and religation. The 6.9- kb Sg/ll fragment (Example 3), which contains the tac promoter/ pmABCD/ rmB terminator construction, was ligated into the SamHI site, thus introducing these modified genes into the plasmid. This plasmid was introduced into strain CGA267356 by conjugation and the native pmABCD gene cluster was replaced with the tac promoter/pmABCD/tmB terminator construct by homologous recombination. Plasmid pLem/Gac (Example 5) was introduced into the strain with the chromosomally located tac promoter/pmABCD/tmB terminator to create strain CGA378584. New strain CGA378584 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to have higher biocontrol activity (Table 2) compared to the related strain CGA267356 . New strain CGA378584 was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B-21818. EXAMPLE 9 Construction of Strain CGA267pPhz (MON0597) (pPhz) This strain is the same as Pseudomonas strain CGA267356, except that strain CGA267pPhz (a.k.a. MON0597) contains a plasmid carrying DNA from Pseudomonas aureofaciens strain 30-84 that contains 5 genes known to encode the pathway for the biosynthesis of the antifungal metabolite phenazine-1-carboxylic acid (PCA). A 5.7-kb EcoRI/H/>7dlll gene fragment (SEQ ID NO:11) containing the pftzFABCD gene cluster from P. aureofaciens (Pieπson, etal., FEMS Mctob l. Lett 134299-307 (1995)) was ligated into DNA of plasmid pUCP26 ( West βt al., Gene 128:81-86(1994)) that had been restricted with EcoRI and Hind\\\ to create plasmid pPhz. Plasmid pUCP26 is an E. coli/Pseudomonas shuttle plasmid that has a lac promoter flanking the multiple cloning site. The EcoRI and Hind\\\ sites are oriented with the lac promoter such that the EcoRI site is closer to the promoter. Therefore, cloning of the EcoRI/H/πdlll pήzFABCD gene fragment, in which the phz genes are cotranscribed on a single operon oriented in the EcoRI to HindlW direction into plasmid pUCP26, results in the proper juxtaposition of the lac promoter and phz gene cluster to cause expression of these genes from that promoter. New strain CGA267pPhz has been shown to produce PCA, an antifungal metabolite not normally produced by parent strain CGA267356. It also produces the metabolites normally produced by the parent strain, including pyrrolnitrin. New strain CGA267pPhz was deposited with the NRRL on September 5, 1997, and assigned accession no. NRRL B- 21819. EXAMPLE 10 Construction of Strain CGA364474 (MON0518) (lac/gacA) Strain CGA364474 (a.k.a. MON0518) was constructed by juxtaposition of the lac promoter element from E. coli (de BoeretaL, Free. Nat. Acad Set. USA 8021-25 (1983)) with the gacA gene (SEQ ID NO:2) residing in the chromosome of P. fluorescens strain CGA267356 (U.S. Patent No. 5,348,742) such that expression of the gacA gene is regulated by the lac promoter. The lac promoter is a small DNA fragment that is known to promote gene expression and does not itself encode a protein product. It provides strong, constitutive expression of genes in Pseudomonas. The lac promoter and the gacA gene were fused precisely by overlapping PCR and the lac/gacA promoter/gene DNA fragment replaced the native gacA gene in the chromosome of strain CGA267356 by homologous recombination. Strain CGA364474 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to provide higher biocontrol activity (Table 2) than parent strain CGA267356. New strain CGA364474 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21887. EXAMPLE 11 Construction of Strain CGA364475 (MON0519) (tac/pmABCD) P. fluorescens strain CGA364475 (a.k.a. MON0519) was derived from the parent P. fluorescens strain CGA267356 by juxtaposition of the tac promoter element from E. coli (Example 3, SEQ ID NO:7) with the pmABCD gene cluster (U.S. Patent No. 5,639,949) in the chromosome such that expression of the pmABCD genes is from the tac promoter. In strain CGA364475, the pmABCD genes are expressed from the strong, constitutive tac promoter rather than the weaker, regulated native pm gene promoter. As a result, this strain produces more of the antifungal metabolite pyrrolnitrin (Table 1 ) and provides greater biocontrol activity (Table 2) than parent strain CGA267356. New strain CGA364475 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B- 21888. EXAMPLE 12 Construction of Strain CGA366259 (MON0524) (tac/pmABCD, pE11 ) Strain CGA366259 (a.k.a. MON0524) was created by introducing plasmid pE11 (Example 1 ) containing the native gacA gene derived from P. fluorescens strain CGA267356 into P. fluorescens strain CGA364475 (Example 11) by conjugation. New strain CGA366259 has been shown to produce higher amounts of the antifungal metabolite pyrrolnitrin (Table 1 ) and to provide greater biocontrol activity (Table 2) than parent strain CGA267356. New strain CGA366259 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21889. EXAMPLE 13 Construction of Strain CGA376150 (MON0575) (CGA267355, pE11 ) Strain CGA376150 (a.k.a. MON0575) was constructed by introducing plasmid pE11 (Example 1 ) into the wild-type P. fluorescens strain CGA267355 by conjugation. Strain CGA267355 was isolated from soil in Texas. It normally does not produce the antifungal metabolites pyrrolnitrin and 2-hexyl-5-propyl-resorcinol , or the hydrolytic enzyme chitinase. However, upon the introduction of plasmid pE11 containing the gacA gene, strain CGA376150 was demonstrated to produce pyrrolnitrin (Table 1), 2-hexyl-5-propyl- resorcinol, and chitinase and was shown to be an effective biocontrol agent (Table 2). New strain CGA376150 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21890. EXAMPLE 14 Construction of Strain NOA402208 (MONO630) (gac*3, pPrn) Strain NOA402208 (a.k.a. MONO630) was constructed by introducing the plasmid pPrn containing the pmABCD gene cluster under the control of the tac promoter (Example 3) into P. fluorescens strain CGA375260 (Example 6) by conjugation. Strain CGA375260 differs from the parent strain CGA267356 by a single base change in the gacA coding sequence (SEQ ID NO:9) that renders GacA (SEQ ID NO:10), the protein product of the modified gacA gene and an activator of transcription, active irrespective of phosphorylation by the LemA protein. As a result, strain NOA402208 produces more pyrrolnitrin (Table 1) and has greater biocontrol activity (Table 2) than parent strain CGA267356. New strain NOA402208 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21891. EXAMPLE 15 Construction of Pseudomonas fluorescens Strain NOA402210 (MON0632) (res, pPrn) Strain NOA402210 (a.k.a. MON0632) was constructed from P. fluorescens strain CGA267356 (U.S. Patent No. 5,348,742) by deletion of a region of the chromosome that resulted in no production of the antimicrobial metabolite 2-hexyl-5-propyl-resorcinol and introduction of the plasmid pPrn by conjugation. P. fluorescens strain CGA319115 is a transposon mutant of the parent strain CGA267356 that is incapable of the production of 2-hexyl-5-propyl-resorcinol (U.S. Patent No. 5,496,547) and that provides greater biocontrol activity compared to strain CGA267356. A cosmid clone, BL3610, from a gene library of DNA from strain CGA267356 was found that restores production of 2-hexyl-5-propyl-resorcinol to strain CGA319115. An in vivo marker exchange was performed with cosmid clone BL3610 in strain CGA319115 in order to rescue the transposon and the flanking DNA. The cosmid clone BL3610Tn containing the mutagenized genomic DNA from strain CGA319115 with the transposon was thus isolated. A 6.5 kilobase pair (kb) EcoRI DNA fragment from cosmid BL3610 that corresponded to the region in cosmid BL3610Tn that contained the transposon insertion was cloned into plasmid pBluescript II (Pharmacea, Inc.) to create plasmid pBL3632. The DNA sequence of this fragment and the DNA flanking the transposon in strain CGA319115 were determined and compared to reveal the precise location of the transposon insertion in the 6.5 kb DNA fragment. Two unique Bc/I restriction sites approximately 200 base pairs apart and flanking the transposon insertion site were identified within this region , and the DNA between these sites was deleted by restriction with Sc l and religation. A 4.0 kb Xho\ DNA fragment derived from the 6.5 kb EcoRI fragment of pBL3632 and containing the deletion of the 200 base pair Sc/l fragment was cloned into a plasmid vector to facilitate homologous ex change in strain CGA267356. Homologous exchange between this plasmid and the chromosome of strain CGA267356 resulted in replacement of the wild-type region in the chromosome with the deleted DNA of the plasmid, thereby rendering the strain incapable of produc ing 2-hexyl-5-propyl-resorcinol. Southern hybridization was performed to confirm that the Sc l fragment was absent. This 2-hexyl-5-propyl-resorcinol non-producing deletion mutant of P. fluorescens strain CGA267356 was given the strain name NOA402209. Strain NOA402210 was created by introducing plasmid pPrn containing the pmABCD gene cluster under the control of the tac promoter into strain NOA402209. Strain NOA402210 produces more pyrrolnitrin (Table 1) than the parent strain CGA267356 and it provides greater biocontrol activity (Table 2) compared to the parent strain. Strain NOA402210 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21901. EXAMPLE 16 Construction of Strain NOA402212 (MON0634) (gac*3, res ' , pPrn) Strain NOA402212 (a.k.a. MON0634) was constructed in a manner identical to that described for strain NOA40221 0 in Example 15 above, except that the strain used as the starting strain for NOA402212 was P. fluorescens strain CGA375260 (Example 6) instead of strain CGA267356. Strain NOA402212 was created by introducing the deletion of the 200 bp Sc/l fragment into the chromosome of P. fluorescens strain CGA375260 to create strain NOA402211 and by the subsequent introduction of the plasmid pPrn containing the pmABCD gene cluster under the control of the tac promoter. This strain produces more pyrrolnitrin (Table 1) than either the parent strain CGA267356 or strain NOA402208 and it provides greater biocontrol activity (Table 2) compared to the parent strain. Strain NOA402212 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21892. EXAMPLE 17 Construction of Strain NOA402214 (MON0636) (ATG-gacA, res , pPrn) Strain NOA402214 (a.k.a. MON0636) was constructed in a manner identical to that described for strain NOA40221 0 in Example 15 above, except that the strain used as the starting strain for NOA402214 was P. fluorescens strain CGA364473 (Example 2) instead of strain CGA267356. Strain CGA364473 is identical to strain CGA267356 except for a single base change in the coding sequence of the gacA gene. The gacA gene was noted to have the unusual TTG translation start codon. In order to improve translational efficiency of the gacA-specific mRNA, the first base in the coding sequence of the gacA gene was changed to an adenine (A), thus creating the normal ATG translation start codon ( SEQ ID NO:4). Strain CGA364473 was further modified in the same manner as strain NOA402212 to contain the deletion of the 200 base pair Sc/l fragment, creating strain NOA402213 , which does not produce 2-hexyl-5-propyl-resorcinol. Strain NOA402214 was created by introduction of the plasmid pPrn containing the pmABCD genes under the control of the tac promoter by conjugation into strain NOA402213. Strain NOA402214 produces higher amounts of pyrrolnitrin (Table 1) and provides greater biocontrol activity (Table 2) than parent strain CGA267356. Strain NOA402214 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21893. EXAMPLE 18 Construction of Strain NOA402216 (MON0638) (lac/ATG-gacA, res ' , pPrn) Strain NOA402216 (a.k.a. M0N0638) contains a chromosomal gacA gene with an ATG translation start codon as in Example 17 that is expressed from the lac promoter of E. coli. It also contains a chromosomal deletion of the 200 base pair Sc/l fragment that results in a 2-hexyl-5-propyl-resorcinol non-producing phenotype. Finally, it contains the plasmid pPrn described in Example 14, which contains the pmABCD gene cluster under regulation of the tac promoter. The ATG-gacA gene was fused to the lac promoter and integrated into the chromosome of P. fluorescens strain CGA267356 essentially as described in Example 7, except that the lac promoter was used instead of the tac promoter. The resulting strain containing the lac/ATG-gacA gene in its chromosome was converted to a 2-hexyl-5-propyl- resorcinol non-producing phenotype by the same method described for the construction of strain NOA402212 (Example 15) to create strain NOA402215. Finally, strain NOA402216 was constructed by introduction of the plasmid pPrn into strain NOA402215. Strain NOA402216 produces more pyrrolnitrin (Table 1) and provides greater biocontrol activity (Table 2) than parent strain CGA267356. Strain NOA402216 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B- 21894. EXAMPLE 19 Construction of Strain NOA409063 (MONO686) (pGac*3/Pm) Strain NOA409063 (a.k.a. MON0686) was constructed from the parent P. fluorescens strain CGA267356 by the introduction of a plasmid containing both the gac*3 and pmABCD genes. Plasmid pGac * 3/Pm was constructed by cloning the pmABCD gene cluster under the control of the tac promoter as a 6.9 kb Sg ll fragment (Example 3) into the unique Sg/ll site of the broad host-range plasmid pVK101 (Knauf, V. and Nester, E. Plasmid 8:45-54 (1982)) followed by cloning the gac * 3 gene (Example 6) as an Xho\ DNA fragment into the unique Xho\ site. This plasmid was introduced into the parent P. fluorescens strain CGA267356 to create strain NOA409063. This strain produces more pyrrolnitrin (Table 1 ) and provides greater biocontrol activity (Table 2) than the parent strain CGA267356. Strain NOA409063 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21902. EXAMPLE 20 Construction of Strain NOA409068 (MON0691) (pKT-Prn) Strain NOA409068 (a.k.a. MON0691) was constructed in a manner identical to that described for strain CGA375258 in Example 3 above, except that plasmid pKT231 was used in strain NOA409068 to maintain the exogenous DNA instead of the plasmid pRK290 used in strain CGA375258. Strain NOA409068 was created by introducing plasmid pKT-Prn into parent P. fluorescens strain CGA267356 by conjugation. Plasmid pKT-Prn was constructed by cloning the 6.9 kb Sg/ll fragment described in Example 3 containing the pmABCD gene cluster from strain CGA267356 with the tac promoter and rmB transcription terminator, into the broad host-range plasmid pKT231 (Pϋhler, Vectors for Gram-negative Bacteria. 1985. Elsevier Scientific Publishers). Strain NOA409068 produces more pyrrolnitrin (Table 1 ) and provides greater biocontrol activity (Table 2) than parent strain CGA267356. Strain NOA409068 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B- 21895. EXAMPLE 21 Construction of Strain NOA413174 (MONO706) (res ' , pKT-Prn) Strain NOA413174 (a.k.a. MONO706) was created by introduction of plasmid pKT- Prn (Example 20) by conjugation into the P. fluorescens 2-hexyl-5-propyl-resorcinol non- producing strain NOA402209 described in Example 15. Strain NOA413174 produces more pyrrolnitrin (Table 1 ) and provides greater biocontrol activity (Table 2) compared to parent strain CGA267356. Strain NOA413174 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21896. EXAMPLE 22 Construction of Strain NOA413175 (MONO707) (gac*3, pKT-Prn) Strain NOA413175 (a.k.a. MONO707) was created by introduction of plasmid pKT- Prn (Example 20) by conjugation into the P. fluorescens strain CGA375260 (Example 6). Strain NOA413175 produces more pyrrolnitrin (Table 1) and provides greater biocontrol activity (Table 2) than parent strain CGA267356. Strain NOA413175 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21897. EXAMPLE 23 Construction of Strain NOA413176 (MONO708) (gac * 3, res ' , pKT-Prn) Strain NOA413176 (a.k.a. MONO708) was created by introduction of plasmid pKT- Prn (Example 20) by conjugation into P. fluorescens strain NOA402211 (described in Example 16). Strain NOA413176 produces more pyrrolnitrin (Table 1) and provides greater biocontrol activity (Table 2) than parent strain CGA267356. Strain NOA413176 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B- 21898. EXAMPLE 24 Construction of Strain NOA413177 (MONO709) (ATG-gacA, res ' , pKT-Prn) Strain NOA413177 (a.k.a. MONO709) was created by introduction of plasmid pKT- Prn by conjugation into P. fluorescens strain NOA402213 (described in Example 17). Strain NOA413177 produces more pyrrolnitrin (Table 1) and provides greater biocontrol activity (Table 2) than parent strain CGA267356. Strain NOA413177 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21899. EXAMPLE 25 Construction of Strain NOA413178 (MONO710) (lac/ATG-gacA, res ' , pKT- Prn) Strain NOA413178 (a.k.a. MONO710) was created by introduction of plasmid pKT- Prn by conjugation into P. fluorescens strain NOA402215 (described in Example 18). Strain NOA413178 produces more pyrrolnitrin (Table 1) and provides greater biocontrol activity (Table 2) than parent strain CGA267356. Strain NOA413178 was deposited with the NRRL on November 20, 1997, and assigned accession no. NRRL B-21900. EXAMPLE 26 Cultivation of Bacteria and Fungi for Screening Assays a. Cultivation of bacteria The bacterial strains are stored in 20% glycerol at -80 9 C prior to use. One loop from the stored culture is suspended in 5 ml Luria Broth (LB: 10 g Bacto-Tryptone, Difco; 5 g yeast extract, Oxoid; 0.25 g MgS0 4 H 2 0; 8 g NaCI; and 1 L distilled water; pH 7) and shaken at 150 rpm and 25 9 C overnight. 100 ml LB is inoculated with 1 ml of the preculture and incubated under the same conditions. 10 ml of the last culture are centrifuged (10 min at 10,000 rpm), and the pellet is resuspended in 200 ml saline (0.8% NaCI) giving a concentration of approximately 10 8 cfu/ml. For exact determination, a dilution series (10° to 10 "8 , 20 μl in 180 μl) is prepared in microtiter plate and drops of 10 μl are spotted onto Luria Agar (LB with 1.5% Bacto-Agar, Difco) with an Eppendorf pipette. The cfu are counted after 24 hrs incubation at 28 9 C. Antibiotics may be added if required for selection of bacteria: tetracycline - 15 μg/ml; kanamycin ~ 50 μg/ml. b. Cultivation of Rhizoctonia solani Rhizoctonia solani is grown on Potato Dextrose Agar (PDA, Difco) pH 5.6 in a petri dish. A 300 ml Erlenmeyer flask with 25 g millet and 50 ml distilled water is autoclaved and incubated with one agar plug (5mm diameter) from a PDA culture of R. solani. After incubation at 20 9 C in the dark for 3 weeks, the overgrown millet is air-dried and ground in a Culatti mill (1mm sieve, 6000 rpm). c. Cultivation of Pythium aphanidermatum Pythium aphanidermatum is grown on Malt Agar (Oxoid), pH 5.6 in a petri dish. One agar plug (6mm diameter) from this culture is transferred to a petri dish with 8 ml oatmeal agar (50 g Oatmeal, 3 ml 1.5% cholestrin in ethanol, and 1 L distilled water) with a slant surface. Two hrs later, 13 ml of sterile distilled water are added, and the plates are incubated for 10 to 14 days in the dark. The mycelium that grows from the agar surface into the water is transferred to a mixer and cut into small pieces. The concentration of oospores is counted in a Thoma chamber and adjusted with distilled water to 2 x 10 4 /ml. EXAMPLE 27 Assays For Biocontrol Activity Preparation of bacterial cultures: All bacteria cultures are cultured in Luria broth for 2 days at 28° C. Bacterial cells are collected by centrifugation and resuspended in water to 10 9 or 10 8 bacterial cells/ml. 10 ml of each suspension is used to drench a pot containing 50 ml soil, resulting in 2 x 10 8 (high rate, CGA267356 only) or 2 x 10 7 cells/ml soil (all strains). Preparation of fungal inocula: Rhizoctonia solani is grown on twice-autoclaved millet seed until fully colonized, then air dried for several days. Dried inoculum is ground to a fine powder for use in all assays except the poinsettia assays, in which whole colonized millet seeds are used. A large supply of inoculum is stored at room temperature and used for several months. Pythium aphanidermatum is inoculated on twice-autoclaved millet seed and grown for one week. The colonized millet seed is air dried for 2 hours, then used immediately. Pym/ ' um-infested millet seed is prepared weekly. a. Pathosystem Rhizoctonia so/an/ ' -cucumber One cucumber seed is planted per pot containing a standard commercially available peat/bark type potting soil. The bacterial suspension is drenched on each pot and Rhizoctonia inoculum is broadcast over the surface of the soil. Each treatment in an experiment contains 5 sets of 12 plants which are randomized and placed in greenhouse under automatic sprinklers. Each experiment is repeated a minimum of three times. Stand counts are recorded at 1 and 2 weeks after planting and compared to uninfested and untreated healthy controls and infested and untreated diseased controls. b. Pathosystem Rhizoctonia so/aπ/ ' -impatiens A single hole is drilled in the center of each pot and Rhizoctonia inoculum is broadcast over the surface of the soil and into the hole. One commercially purchased impatiens seedling plug is transplanted into the hole of each pot and bacteria suspension is applied as a drench. Each treatment in an experiment contains 5 sets of 12 plants which are randomized and placed in greenhouse under automatic sprinklers. Each experiment is repeated a minimum of three times. Stand counts are recorded at 1 and 2 weeks. c. Pathosystem Rhizoctonia so/an/ ' -poinsettia The bacterial suspension is drenched on strips of ten Oasis rooting cubes (the cell suspensions were adjusted so that each 50ml cube was drenched with 40ml of suspension), resulting in 2 x 10 8 or 2 x 10 7 cells/ml of cube, and one commercially purchased poinsettia cutting is inserted into each cube in the normal method for rooting. Five Rhizoctonia -infested millet seeds are placed in the middle of the strip (between plants 5 and 6), and the strips are placed in the greenhouse under automatic sprinklers. Each treatment in an experiment contains 4 rooting strips with 10 cuttings each and each experiment is repeated a minimum of three times. Stand counts are recorded at 10 days and 21 days after planting. d. Pathosystem Pythium aphanidermatum-c c mber One cucumber seed is planted per pot and the bacterial suspension is drenched on top of each pot. Pythium inoculum is broadcast over the surface of the soil and replicates are randomized and placed in the greenhouse under automatic sprinklers. Each treatment in an experiment contains 5 sets of 12 plants and each experiment is repeated a minimum of three times. Stand counts are recorded at 1 and 2 weeks . EXAMPLE 28 Extraction of Antifungal Metabolites Active antifungal metabolites such as pyrrolnitrin (prn) can be extracted from the growth medium of bacterial strains that produce inhibitory antibiotics. For example, using strain CGA376146, this can be accomplished by extraction of the growth medium with 80% acetone followed by removal of the acetone by evaporation and a second extraction with diethyl ether. The diethyl ether is removed by evaporation and the dried extract is resuspended in a small volume of methanol. Alternately, the antifungal metabolites can be extracted with methanol using conventional methods. Small aliquots of the antibiotic extract applied to small sterile filter paper discs placed on an agar plate will inhibit the growth of Rhizoctonia solani, indicating the presence of the active antibiotic compound. EXAMPLE 29 Combination of Biocontrol Strain With Fungicides The biocontrol strains of the invention are each applied to non-sterile soil as a drench at 2 x 10 8 cfu/ml soil, while metalaxyl fungicide is either drenched (Ridomil at 0.02, 0.5, or 2 ppm) or coated onto seeds (Apron at 35 g a.i./100 kg seed). Pythium aphanidermatum is introduced as an oospore suspension (1400 spores/ml soil). Rhizoctonia solani is introduced as a pelleted millet powder (5 mg in the center of each pot). After incubation for 19 days in the greenhouse, the hypocotyls of cotton seedlings are rated for disease on an observation scale. Almost complete control of damping-off may be achieved when one of the biocontrol strains is applied together with Ridomil at 2 ppm. Using Apron instead of Ridomil results in the same level of control. Each of the biocontrol strains alone still gives significant suppression of both pathogens. Metalaxyl fungicide alone, however, fails to control the disease complex. 0.2 ppm Ridomil alone gives approximately 40% suppression of Pythium aphanidermatum. However, the combination of 0.02 ppm Ridomil with one of the biocontrol strains significantly increases the level of control. Thus, the combined application of the biocontrol bacterial strains of the invention with a reduced rate of metalaxyl fungicide achieves almost complete control of the seedling disease complex in cotton caused by Rhizoctonia solani and Pythium aphanidermatum. EXAMPLE 30 Antifungal Compositions Formulations of antifungal compositions containing as the active ingredient th e antifungal metabolites that are produced by the biocontrol strains of the invention and that are inhibitory to the growth of Rhizoctonia and Pythium are produced according to Examples 10 and 11 in U.S. Patent No. 5,348,742. These formulations include emulsifiable concentrates, solutions, granulates, dusts, wettable powders, extruder granulates, coated granulates, and suspension concentrates. The antifungal compositions may be used to control or inhibit the growth of a plant pathogenic fungus by applying an effective amount of the biocontrol composition to an environment in which the fungus may grow, to a plant or plant part, and/or to seed. EXAMPLE 31 Field Trial of Biocontrol Strains Cultures of the biocontrol strains are stored in 20% glycerol at -80 s C. One loop from the stored culture is suspended in 5 ml Luria Broth (LB: 10 g Bacto-Typtone, Difco; 5 g yeast extract, Oxold; 0.25 g MgS0 4 H 2 0; 8 g NaCI; and 1 L distilled water; pH 7) and shaken at 150 rpm and 25 s C for 24 hrs. 100 ml LB is inoculated with 1 ml of the preculture and incubated under the same conditions. After 16 hrs, the culture is centhfuged for 10 minutes at 10,000 rpm, and the pellet is resuspended in saiine (0.8% NaCI) and adjusted to 3 x 109 cfu/ml (OD2). Thus, 100 ml of culture will give approximately 200-300 ml drench of OD2. A hemocytomerter and/or spectrophotometer is used to adjust the concentration of bacteria in the drench. Otherwise, a standard salt solution of a known OD (e.g., Phillips' Milk of Magnesia TM = Mg(OH) 2 ) can be used to adjust the OD of the drench. If a centrifuge is not available, the whole culture broth has to be applied; a hemocytometer is then used to determine the cfu/ml. For exact determination, a dilution series (10 9 to 10 " ^ 20 μl in 180 μl) is prepared in a microtiter plate and drops of 10 μl are spotted onto Luria Agar (LB with 1.5% Bacto-Agar, Difco) with an Eppendorf pipette. The cfu are counted after 24 hrs incubation at 28 s C. 250 mL of the bacterial suspension per 10' (=1 rep) are drenc hed onto the covered seeds (200 seeds per rep). A handheld sprayer or watering can free of pesticide residues is used to apply the drench in a narrow band of approximately 1.5 inches width. Rhizoctonia and Pythium are prepared for inoculation as in Examples 10 and 11 above. Emergence is recorded at 10 days after planting to assess pre-emergence damping off. Stands are recorded at 21 days and 28 days after planting to assess post-emergence damping-off. While the present invention has been described wit h reference to specific embodiments thereof, it will be appreciated that numerous variations, modifications, and embodiments are possible, and accordingly, all such variations, modifications and embodiments are to be regarded as being within the scope of the present invention. Furthermore, all publications and patent applications mentioned in this specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are therefore hereby incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incoφorated by reference. TABLE 1 Pyrrolnitrin production by P. fluorescens strain CGA267356 and genetically modified strains derived from it. Pyrrolnitrin was isolated from 3 day old cultures of the strains grown in CMMMAD medium by extraction with methanol and quantified by HPLC analysis. TABLE 2 TABLE 3 a Plasmid pRK290 carrying an 11 kb EcoRI fragment with the native gacA gene. 0 Plasmid pRK290 carrying the pmABCD gene cluster under control of the tac promoter. c 2-hexyl-5-propyl-resorcinol non-producing phenotype from a chromosomal deletion. d Plasmid pKT231 carrying the pmABCD gene cluster under control of the tac promoter. TABLE 4 SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: NOVARTTS AG (B) STREET: Schwarzwaldallee 215 (C) CITY: Basel (E) COUNTRY: Switzerland (F) POSTAL CODE (ZIP) : 4058 (G) TELEPHONE: +41613241111 (H) TELEFAX: + 41613227532 (ii) TITLE OF INVENTION: Genetically Modified Pseudomonas Strains with Enhanced Biocontrol Activity (iii) NUMBER OF SEQUENCES: 11 (iv) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) CCMHjrER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOSMS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.30 (2) INFORMATION FOR SEQ ID N0:1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10763 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Pseudomonas fluorescens (B) STRAIN: CGA267356 (aka M0CG134 and aka BL915) (vii) IMMEDIATE SOURCE: (B) CLONE: Plasmid pELl (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 210..1688 (D) OTHER INFORMATION: /product= "msthyltransferase" /note= "Coding sequence for methyltransferase has homology to the cheR and frzF genes from E. coli and Myxococcus xanthus, respectively. " (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 1906..3633 (D) OTHER ΓNPORMATTCN: /product= "sensor kinase" /note= "Coding sequence for sensor kinase has homology to the rcsC, frzE, and bvgS genes of E. coli, M. Xanthus, and Borditella pertussis, respectively." (ix) FEATURE: (A) NAME/KEY: misc_RNA (B) LOCATION: complement (4616..4691) (D) OTHER INFORMATION: /product= "tRNA" /note= " (complementary DNA strand) Homology to gly from E. Coli." (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: complement (4731..5318) (D) OTHER INFORMATION: /product= "CDP-diacylglycerol-glycerOl-3-phosphate-3-phosph atidyltrans. " /note= "Coding sequence for CDP-diacylglycerol-glycerol-3-phosphate-3-phosphatidyltransfere se has homology to pgsA. " (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: complement (5574..7397) (D) OTHER INFORMATION: /product= "UVR exonuclease subunit C" /note= "Coding sequence for UVR exonuclease subunit C has homology to uvrC. " (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: complement (7400..8041) (D) OTHER INFORMATION: /function= "response regulator/transcription activator" /product= "gacA (aka gafA) " /note= "Coding sequence for gacA (aka gafA) has homology to the uvrY and gacA genes of E. coli and Ps. fluorescens, respectively. " (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: GAATΓCGATG ACATGCCGCG CGCCGGCATC GACACGCAAA TGGTCGACCT GGTSCTOCCG 60 GTOGTCGAAA TGCCGCAGAA GCGCTGGAG CTGTGGOGCA ACTCTCAGCT CATCACCCTG 120 CCGACCGCCA AOGATCCGCA AATCAAGGTC TCGGCGCCGG TCTCCAAACG CGATGCCGCG 180 QCGGCGAACA GCAGCTGCAA GACATCCTGA TGCIGTIGCG CACCQGCACC GGCCATGACT 240 TCAAGCATTA CAAGCGCGCC ACGGTGCTGC GGCGGATCGA GCGCCCGCIG CAGGTCACCG 300 CCCAGCCGGA CC CGCCGCC TATCACGATT ACCIGCAGAT QCACCCIGAA GAAACCAAGG 360 CQCTGCTGGG CGACATGCTG ATCGGCGTGA CCAATI CTT TCGCGACCGC GAGQCCTICG 420 AAGCCCTGGA GCGCAAT3TC ATTCCTGCCC TGGTGAAGTC CTTGCAQGAC AGCCAACCGC 480 AOCOTGAAGA CX3IGCGCATC TGGTCCGCCG GCTGCTCCAC QOGTGAAGAG GCCTATAGCC 540 TGGCAATCGT CGCCAGCGAG CAGATG3CCC TGGAGGCCTG CAACGCCAAG C GCAGGTAT 600 TOGCGACCGA TATCGACGAT CGTGCCATCG OCCAGGGACG (-AAGGGGGTC TATOCCGAAG 660 CGATCGTTAC OGATGTGCCT CO3CAGOGCA TCCGCCAGTA CTITTC0CG3 GAAAACCAGC 720 ATTACCGGGT GOGCAAGGAG ATTCGCGAAA AGGTGCTGTT CGCCAAGCAC AGCCTGCTGG 780 cGGATCcσσc ATΓΓΓCGCAG ATCGACTΓGA TOGTCTGCCG TAACCTGCTG ATCTACCIΌG 840 ACCGCGACGT GCAACGGGAG ATCCIGCAGA TGTICCACTT CGCCCTGCGT CCTGGAQQCT 900 ACCTGT CCT CGGΓTCCΓCC GAATCCGCGG ACGGCTGCCA GGATCTGTΓC GTOCCGGTOG 960 ACAAGCGCAA C03CATTTIC CCΞGGTAOGQC CCAACTCGGC CACGGTTCGC CGCGCGCCCA 1020 CCATGCCGCG AOCGCGTACA TGOGCACCAT CGGCAGCCCC CACCCCGT3G AAACCAAGTG 1080 TCTCGcαcAA AACCTCGTΓC GCCGACATCC AOCTΓCGCGC CCΓGGAAAAG TGCGCGCCGC 1140 CGAGCATGAT CGTCGATGCC AACσCCGACA CCTGCACAT GAGCGAAQQC GCCGGCCOGT 1200 TOCTOCGCTA TGTCGCGGGG GAAATCACCC GCΆACCTGCT G&CCCΓGATC CAGCCCGAGC 1260 TGCGCCTTGA ACΓGCGCACC A03CIGT CC AGGTGCAACA σrCCGGT3TT GOGGTGACCG 1320 CCGCCGGGTG 03 -ATCGAGC GGGAAAAGAA GCCTTGTTTC ATCGACCTCA CAGCCCGCCC 1380 CTΓCAAQGAC GAGGAAACCG ACAACGAATA TσrαCTGGTG GTGTΓCGAGG AGACCGAOaC 1440 OGACCCACGG GAGCTGCGCG AGACCAGCGC CAGCCAGACG GAAAACCAGA TOCTGGCCAA 1500 CCTCGAGCGG GAGTPGCAGC GGACCAAATT GCACCTGCAG GACACCATCG AGCAAICOGA 1560 AGTCTCCAQC GAGGAGCTCA AGGCGTCGAA OGAAGAAATG CAGGCGCTCA ATGAAGAGCT 1620 GCGCTOGGCC ACCGAAGAGC T3GAAACCAG CAAQGAAGAG TTGCAGTCGA TCAATGAAGA 1680 GCTGCTGACG GTCAATTACG AGCTGAAAAC CAAGGTCGAG GAAACCGACA AGATCAACGA 1740 CTACCΓGACC AACCTGATCG CCTCCACCGA CATCGCCACG GTGTTCGTCG ACCGCAACAT 1800 GCGCATCCGC TOGTTCACCC OGCGCGCCAC OGACAΪTTTC AGCATGCTGC CGGTGGACAC 1860 OGACGCTCAT TACTGGACAT CACCCACCGC CTGAACTACC CGGAAATGGC CGAGGACQCC 1920 GCGACCGTGT TCGAGTCGTT GAGCATGATC GAGCGTGAAG TCAACAGCGA CGATCAQCGC 1980 TGGTACATCG CAOGCCTGTT GCCCTATCGC TCCAQCGAAG ACCATATCGA CGGCACCGTG 2040 CTGACCITCA TCGATATCAC CAAGCGCCGG CTGGCCGAGG AGGAACTGCG CCK3GQCGAA 2100 GAAOGCATGC GCCT3GTO3C CGAAAQCACC CATGATITOG O-ATCATCAT OCTCGACAAC 2160 CAQGGCCTCA TCACCGACTG GAACACCQQG GCGCAACTGA TC TOGGCTA TACCAAGGAC 2220 GAAGTGCTGG GCGCCTATTA CGACCTGATT TICGCGCCTG AGGACCGCGC CGGCαGCGTG 2280 COGGAAAGCG AGCTGCTCAC CGCCCGCGAA CACGGCCGCA GCGACGATGA ACXΞCTGGCAT 2340 ATACGCAAGG ACGGCGAGCG CTITITCIGC AGCGGCGAAG TCACGCGGCT CAAQGGTGAC 2400 AQCCTGCAAG G IACGTGAA AATAGCCCGC GACCTGACGG GCCACAAACG CATGCAGGAC 2460 GAQCAGAACC AGAAGCTGAT GGAGACCCAG ACCCACAGCC ACCTCAAGGA TGAGUTI IC 2520 GCGGTGATGT CCCATGAACT CAAGCATCCG CTCAACCTGA TCCAGCTCAA CGCCGAGTTG 2580 CTGCGTOGCC T3CCGACGAC CAAGGCGQCC GCCCCTGCCC TCAAQQCGGT CAATACCATT 2640 TGCGAGGCTG TCTCCAGCCA GGCGCGGATC ATCGACGACC TGCTGGATGT QCGGCGΠTG 2700 CGCACCGGCA AGCTCAAGCT GAAGAAACAG CCGGTGGATC TTGGCCGGAT CCIGCAGGAC 2760 ATCCATACCG T3GTGCTCAG CGAAGGGCAT CGCTGCCAGG TGACGCIGCA AGTGCCGT G 2820 CCACCGCAAC CGCCG TAAT GATCGATGCC GATGCGACGC GGCK3GAGCA QGTGATCTOG 2880 ACAACGCCCT GAAATTCACC CCQGCCAATG GCTIGGTCCA GTTGATGGCC 2940 CAGCGGGTCG AGGATAAGGC GCACGTGGAT GTCATCGACA GCGGCG GαG CCTGGCCGAG 3000 GAAGACCAGA ACAAQGTGTT OGAO T T1C GGCCAGGCGG CCAACCAGCA CGGCACTCAT 3060 CAACGCGACG QQCTGGQCAT CGGCCTGTCA CTGGTGCGCC AGCIGGTGGA AGCCCACGGC 3120 G3CTCGGTCA GCGTGCAGTC GAAGGGGCTG QGCCAGGGAT QOCCT TAC CGTGCTCTTG 3180 CCCCIGAGCC ACCCCAACGA CAGCGCTCCC AAACAGCCCG CGTCGCGGGG T3TCGAACGC 3240 OTGCCGGCA TCAAQGTGCT GCTGGTGGAC GACTCGCGQG AAGTCATGGA AGTCCIGCAA 3300 CTOCT3CT3G AGATGGAQ33 CGCGCAAGTC GAGGCCITCC ACGACCCGCT GCAGQCCTTG 3360 GQCAATOCCA GGAACAACAG TTACGACCTG ATCATTTCAG ACATCQQCAT Q.CGATTATG 3420 AACGGCTACG AACTGATGCA GAACCTQCGC CAGATCGCTC ACCTGCACCA TACGCCAGCG 3480 ATTGCGCTGA CXX3GITACGG CGCCAQCAQC GACCAGAAGA AG CCCAQCA TQCQGGATTC 3540 GATCGGCATG TGAGCAAACC OGrrGGCTCAG GiACCCGCPGA TCGACCTGAT CAGGGAGCTG 3600 T Q CAQCCAQG QCTIGCGCTC GGCTGAGCAC TGATGGTCTA GACCCGQCGA ACCCACCTCG 3660 TCX3GCCTTGA G XSCQGCGAG CQCCATTGCC TGCΓGGGCAG CTATTCACGC TTQCGGATCG 3720 TCQCGCCTGC QGGCCACCGC CTC - GATG GCTTGCTCAT AQGCGQCGTT QGCCTQ3TCC 3780 TTGAGCTTSA GCCAATCGTC CCAATCGATC ACGCCGTIGC QCAGCAACTC CTCGGCCGCG 3840 CTIAACAGCG CCTGATQCCA QGCGTCCQGC GAQCCGGAAC GGTAGTCACG GTCTTCCAGC 3900 AQGCCTTGCC AQ3CGTCCAG TTCCQGTGTC TIQCGTTCAT TQACCATGQC AGCCACQ3CC 3960 TΓΓGTΓCATT GCCGATAAAT CQGCQAG GG GTOGΓGOGTΓ TCTCGGATAT GCQCCCTG C 4020 CTGCTCGAGA ACQGCCAGGC CGGGACATIG CRCAACOGTC AGCGACCQGA TGGAGCTCGA 4080 GCGQCATGCC ATCGACCAGC σrCAAGGTCA GGTTCTCGAT G3TGCCGGCG ATCCGGTCCT 4140 TGAATACCQG TTCGCCGTCC QGATCCAACT CATCGTAGAA AAAQCαCGTG CXTTTCGAQCC 4200 AGCCAATGGT OUI IUCAGG TCCGGCCCCA QGTAATACTT GCCGTCAAQG AAAAACCCGG 4260 TAAAQQGCTC CA CCGCTCG OGATTCTCAA TGACATAACG TATΓCCAGCG TGCATACCTG 4320 TOGAΠTATC GAGCATQGCG TCGATCTCCC AQCAGATGAA TCCGGTAGAC CGCGTGGCTT 4380 TITCACTGTΓ CCTΠTGATT GCCCQCCCGA CCCTQQCGAG CCTΓGCTCQC QCGTCCΓGGC 4440 CQCAT QCQC QGCGAATGQG CGACGTCGAA TODGATCTGC AAGTGCCCAG CTAGCQGCCC 4500 QGCCACGGCA ATACGGGCTT CAGGTACQGC TTAGAAAGAA GAATGACGAT TGGCTCGACA 4560 TATΠTΠGG CQCAAAAAAA AATGGACCTC TΠTCAGAGG TCCATΓΠTA ATATΓTGGAG 4620 CGQGAAACGA GACTCGAACT CQCGACCCCG ACCTIGGCAA GGTCGTGCTC TACCAACTGA 4680 GCTATTCCCG GGTCTT3GTG GTQTGCATTT TATAGAAATΓ CX3AAACTQCG TCAACCCCTT 4740 GATTCAAAAA QITTTATITC TTITCTACCA TOGGTCTTCA GGTGCGGCCA QGCAGCQCGC 4800 AQGTACTQCA ACATCGACCA CAQGGTCAGC CCTCCGGCGA TCAQCAQGAA QGCATAACCC 4860 AGCAGCACCC AGAAGGTGAA QQCCOGCGGA TIGQCCAGCA OGATCACCAG CGCCAGCATC 4920 TGCGCGGCAG TT-TCCGATT TGCCCATGTT GGACACCGGC CACCIGGGCG CGTGCGOCCG 4980 AGCTCGGCCA TCCACTCGCG AAGGGCGGAC ACCACGATIT CaCGCCCGAT GATCACCGCT 5040 GCCGGCAQGG TCAGCCACAG GTTGCCGTGC TCT GCACCA GCAGCACCAG QGCCACCGCC 5100 ACCATCAACT TGTCGQCCAC CGGATCGAGG AAGGCCCCGA ACGGCGTGCT CIQCTCCAGA 5160 CGCCGCGCCA QGTAGCCATC AAGCCAOTCG GT3GCCGCGG CGAACGCAAA GACGGAACTG 5220 GCGGCCAT3T AGCTCCAGTT GTAAQGCAGG TAAAACAQCA AAATGAAGAT CGGGATGAGC 5280 AGAACGCGTA GAACGGTGAT CAGATTAQGG ATATTCATCG GCACAACTQΞ CTACGAQGTG 5340 AGIGGCAATC TACTCGGAAA AGACAGCAGA TGAGGTAQCA CGGCCATTCT A033GCTTCT 5400 QCCACAGCGT GTCTAACACT GTTCCAAGAC TICGGGCCGC TCGAAAGAGC AACTTCAGAA 5460 GGTCTACACG CGCAAAATAA GACATTCAGT TCTTCTGTAA GIAOOGTGTA GATCGOGATC 5520 TATCAGOGGT GO CCGCCAA AAAQGAAGCC TTGAAGCTTC CTTGAGCαCT CCCCTACTCG 5580 CTATGCAAGT TCGCATAAAT CAGCTCAGCG AGCTITTTAC TGATCCCCGG CGCTTTGGCG 5640 ATCTCCTCAA TGCTGGCGCG AGACAGTTCC TGCAACCCAC CAAAGTGTIT CAACAQGTCG 5700 CGGCGGCGCT TGGGGCCGAC GCCOGCCACG TCTTCGAQGG TCGAAGTGCG GCGGGTCTTT 5760 CCGCGACGGG CGCG3TGGCC AGTGATGGCG AAACGGTGAG CCTCGTCGCG GATCTOCTGG 5820 ATCAGGTGCA GCGCCGGCGA GTCGCCCI 3 AGGGTGAACT CATGGGCGGC ATCGTΓGAGG 5880 TAGAGGGTCT CGAAACCCGT CITGCGCGTC GCACCCTTOG CCACACCCAG CAGGATCAQG 5940 TCAGGCACOG CCAACTCGTT GAGCACGTCG CGGGCCATGG ACAGCT3GCC CTTGCOQCCG 6000 TCCACCAGCA O.ATGTCCGG CAACITGCCC TCGCCGTCCT TGAGTTTGCT GAAGCGTCGT 6060 GTCAGGGCCT GGTGCATCGC CGCATAGTCA TCαCCGGCGG TGACGCCITC GATGTTGTAG 6120 CGCCGATAGT CGGACT CAG CGGCCCTTCC QGACCGAACA CCACGCAGGA CGCCACGGTC 6180 GCCTCGcσσc TGGAGTGGCT GATGTCG AG CACTCCAGGC GΠ ΓGCGGΓGG CDCGTCCAGG 6240 TTCAGCACTT CGGCCAGGGC CTCGAAACGC GCCGCCACAT GCTGCCGGTΓ GGCCAGQCGC 6300 GCACTCAGCG σCTGTTCGGC GTTGGTCACT GCCAATTGCT GCCAGCGCαC CCGCGTACCG 6360 CGCACCCGGT GGCTGATQCT CAQCTCGCGG CCACGCAGCT CCTGGATCGC CGCGATCAGG 6420 GCCGGGAAAT CCTCATGGAC CA03TTGACG ATCAGCTCGC TGGGCAGGTC GCGCTCCGGG 6480 CTGCΓGAGAA AGΓACTOGCC GAGGAAGGCC GACATGACTT CGGCCACCTC TΓCCΓCGATG 6540 CCCACCT3GG GAAAGAAGTT CITGCTGCCC AGCACCCGCC CX_CCCCGCAC GCTGATCAGG 6600 TGCACACAGG CGCOGCCCGG GTTGACGAAG GCCGCGACCA CGTCGACGTC GCX_ACIOCCG 6660 CCTTCCATGC TCIGCTGGTC CTGGACCCGT CGCAGCAGGG AAATCTGGTC GCGCAGCTCA 6720 GCGGCCΠTT CGAAGTCCAG GGTGCTGGCC GCCTGCTCCA TGCCσσCCGA CAGTΓCGTCG 6780 GTCAGGGCAT T3CTQCGGCC TTCGAGGAAC ATCACCGAGT GGCGCACATC CTCQGCGTAC 6840 ACCTCGGCCT CCACCAQGCC GAOGCACGGC GCCTTGCAGC GCTTGATCTG ATATTGCAGA 6900 CATGGCCGGG TGCGGTTCTT GTAGTAGCTG TCCTCGCACT QGCGGACCAT GAAGU1C 1 6960 TGCAGCAQGC TGAQGCTCTC GCGAATGQCC CCOGCGCTGG QGTACαGGCC GAAATACITG 7020 cccTTCTGCT TCTΓCGCCCC ACGATGGATG CΓGAAACGCG GAAACTCGCC GTCCGAGAGA 7080 AACACATAGG GATAGGACTT ATCGTCGCGC AGCAGGATGT TGTACGGCGG CCGCCATTCC 7140 TIGATCAQCG TCTGCTCGAG CAGCAGGGCT TCGGTCTCGT TGGCQGTGAT GGTGGITTCG 7200 ATCTGCGCGA T333CCCCAC CAQGGCAGCG GTCTTGGGCG CCAD3CCGGT CTTGCGAAAG 7260 TAGCT3GCCA GGCGGCTCTT CAGGITCTIG GCTTTACCGA CGTACAGCAG ACGCGTATCG 7320 CT3TCGAACA T3CGATACAC GCCAGGACGG CCACTGCAGG TGGAAAGAAA AGCACTGGGA 7380 TCAAA0333T CG3TCATT3T CAGGCACTGG C^TCGACCAT GCCGTGQCGA ACCGCCAGCA 7440 ATGTCAGT C AACAT GCTG CTGATCGAGA GCTT-TCGAA GATGCGGTAA 033TAQGTAT 7500 TAACGGTTTT CGGAGACAGG CACAGCTIUI' CGGAGATGAT CTGCAC-TTC TGGCAGCCGA 7560 CG GATCTGG ATTTCCCGCT CGGACAAAGC ATCGAACGGT G TCACTGG 7620 AAGGCTGGAA TGACTTGAAC ACCAACTGCT GGGCAATTIG CGGGCTGATG TAACGCTGGC 7680 CσαCAAACAC CAGOCGAATG GOCTGCACCA TTTCAΪTGAG GCCCGCCCCC TIO3TCAQGT 7740 AACCCGCCGC GCCGGCTΓGC AGCAAGCGGG TCGGGAACGG ATCΓΓCTΓCA CACACGGTGA 78θo CCGCCACGAC TTIGATATCC GGGTGACTGC GCAACAATTT CCGCGTGGCT TCAAGACCGC 7860 CGATCCCGGG CATCTTGACG TCCATGAGGA CX-ACATCCOG TΓΓCAACICC CGGGCCTΓGA 7920 GCAGGGATTC CTCCCCTGAC TCGGCCTGGC CGACCACTTG ( .AGGCCATCG ATGTCAGCCA 7980 GCATTCOTGT AATACCTCTA CGAACGAGAT CAT3GTCATC GACTACTAGC ACCCTAATCA 8040 AQCAGACACC T030GATTTG QGTCITATAG GITGCCGGAC ACCTTAGCAA AAAAGCAGCG 8100 TGCIGACCTA ATGACAAACA CCATATAAAA AGCACTΪGTT CATCAGGGGT ATCCGGTGGA 8160 TGGTTGATGC GCTGCGAACG CCCTGCCCTA AGGCTCTCOG GCGTCCGCCT TTCTTTTCAT 8220 GAQGCTGGAA ACCGAAAGCT CGGCAAGGGT ATGGGTCAGG TGCCGGATCG CCTOCTGATC 8280 TTCCT 3TAC AAGGCCCGGA TAACTGACGA GGC TTCTC TICGCTCTGG CIGAGATTCT 8340 CAGOCTGOT CGGGGTCCGC CTGCCGGTCA CTATATAAAG GACATCGACC CCTTTTGTCG 8400 CCACCCGGGA CAAGTAATCC GCCTTGGGCA CACGGTCTCC AC ITCATAT CTGCCTTGGG 8460 C-ATTGGCTTC CACGCOGCCA ACTTCACCAA ATTITTTCTG CGACAACCCC AGQCGTTCCC 8520 TTTCCTGTCG TCAACCGCGA ACCGATTCCA CTCATTTGGA TGTATGATCC TTTTTTATGC 8580 ACσCCTAGGG GTGTTACACC CTTCAAGCAT TGAACAAATT TGAACGGTTT TGAACTATGC 8640 CCG3TTATCC CCACTGCCGC ACAAGCCAAG GCCTGGCITG AACATCAAGG TAAATCGGTT 8700 CAACAGTTCG C CGTGATCA CGOGGTCGAT CCAGCCACCA CTTATCAGGT ATIQGCIGGC 8760 CGCAAGAAAG GACGGCGCGG CGAGGCGCAC AAQGTGACCG TATTGCTGGG CATGAAAGAC 8820 GGCGTCATCC TGGCCGAACC OGAGGGTCCC GACCAGACGC C03CCTGATC TPCGAATGCC 8880 ATCATCCTGC CGGGAAGAAT CAACTGGCGG CTACGCCTCC ACGGCGCTQC TICGCTCCAT 8940 CCGCAGAAAA CACTCGTCTT CCCCGACCAC TTTCAGCCCC ATCCGCCCAT AAAGCGCCTG 9000 CGCCGGATTG TCCTTGAACA CCGTCAGCCG CAGCAQCCCA CGCCGCTCGT CATGOGCCAT 9060 CGCACGCACC TGTTCGATGG TCCAGGCCCC GADGCCTTGC CCGCGCGACG CCTCGAGCAC 9120 ATGCAATTCA OGAATGTACA AAGCCTTGGC ATOGOGACTC AGGCTGACGA ACCCCAGCAC 9180 CXπOGCGCCC TQGCAGATCA ACAQGTTCTG CCGACCGGCC CAGGCCACAT CGAAQQCCΓC 9240 ATCCAGCCAC AACAQSTCGT GACGAATGTG ATAACCCAGC ATGGTGCTGC GGGTCAGGTC 9300 GOGGGCGAAC ACCAGATCCT CGTGGCTGCC GGCCGGACGT AGCTGCAQGC CGTTCAAQGC 9360 GCGTCAACCG GCAATGACTG GCCACTCCAG CGO GGCAT TGCGCCTGGC AATCAGCAGT 9420 TCσrCGCCTG TACCGGCCGA AGCCATGATC AGCCCGCCGC CGQCσCCCCA GATCGCGCTA 9480 CGCCCCσCCG ACACCCAGCC CCCGGTCGGC CCQCCGT3GT TGGCCATCAG CACCAQCATT 9540 Q 03TGCTCGG CQGCATACCC CTGCAACAAG GCACTGTCCG CGGCATAGOC TGTITCGCCG 9600 ATCAAAACGC CAGCGGCATA GATGCCQQCA CCGGAGTGCG CCGCCGCACG CGCGTGGCTG 9660 GCCTGGGAGA AGTCGGCGCA CACCGCCAGG GCCACCTGGT CTTCQGCGAA CCTCAGGTTC 9720 GCαCCACCGG TGCCGQGGCT GAATACCCGC TCCTCGCCAG CATGCAGATG CTQCTTQCTG 9780 TACACOGCCA GCGAACCATC GGCGQCCAGC ACCAGGGCGC CGATCAACAG CX3QCCCCTCG 9840 ACCGACAQGC GQACQ3GCAT GCCCACCACC GCTQ TACQC CC03CTCCCG GGCCAGGTCG 9900 CGCAACGGTT GCAACAGCGG GCTCTGCGGC AGTATCGCCA GCTCGGCCGC CAQCGCGGQC 9960 TCATAACOQG TCAGGGACAG CTCGGGAAAT ACCAGCAATT GCADQCCCTG CTCCGCCGCG 10020 ACGCGGATAA AGGCCTGGTG CCGGGCGATA TTGCCCGGCA QGTCTCCGGC AACGGAAATC 10080 GACTGQGCGG CQGCAAQQ3T CAGCATQGTC ATQGTTCAAC CTGAATCQGC ATTCQGGAGG 10140 GCGTGGCGAG TGTGTCATAA AAAACTCAAA GCQCTTCACT CATAGACAQC GACTGAAAAC 10200 GCAATAGGAT TITCTGATIG AACCGCGCCC OCQGCCTCTA G^AAGCTCGG CO.ACTTCAC 10260 GGAGAAACAG CATQTCGTCC CTCACCCTTA CCATGCATΌG TCACACΓGCC AGCGCCGCαC 10320 QCTCCGGTGC QQC GCCIGG GTGAAAAACG OnOCGCTCC GGCGGQCTTT TAΠTTOGGT 10380 ATTGGTTTAG CCACTGGCGC QCCTGATACC CAAACGGOGC CCACITGAAC GGGTCGCCTA 10440 CCAGAGAAAA TCTCACCCCC QGTCQGCCTC CCGACCGGGG GTTTΓGTTTT TCTQGGCCGA 10500 AGAT ΠTAA ccms C TΓAAGCAACA CAOOGAACTT ACCGAQGATT GAAOCATGAA 10560 CTACQCCACC TATTACCGTT ACGACACT Q CACCACCTGG CGAΠTAQCA QOTCCGTTC 10620 QGGACAQOCT QCCCCCTCCG ATOGGTCACC TACTGGTGGC AAACATACCT GOi AGCCAA 10680 TCCGSCCAAT TGTCGAACAC CCCAGTAGGG CCGCGCGCGG GAAATCACCC GCCGCCTGCC 10740 CAQGAAGCCT TGAACATGAA TTC 10763 (2 ) INFORMATION FOR SEQ ID NO:2 : (i) SEQUENCE CHARACTERISTICS : (A) LENGTH: 642 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Pseudomonas fluorescens (B) STRAIN: CGA267356 (aka MOCG134 and aka BL915) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..639 (D) OTHER INFORMATION: /transl_except= (pos: 1 .. 3, aa: Met (xi) SEQUENCE DESCRIPTICN: SEQ ID NO:2: TTG ATT AGG GTG CTA GTA GTC GAT GAC CAT GAT CTC GTT COT ACA GOT 48 Met He Arg Val Leu Val Val Asp Asp His Asp Leu Val Arg Thr Gly 1 5 10 15 ATT ACA CGA ATG CTG GCT GAC ATC GAT QGC CTG CAA GTG GTC GGC CAG 96 He Thr Arg Met Leu Ala Asp He Asp Gly Leu Gin Val Val Gly Gn 20 25 30 QCC GAG TCA GOG GAG GAA TCC CTG CTC AAG GCC 033 GAG TTG AAA CCC 144 Ala Glu Ser Gly Glu Glu Ser Leu Leu Lys Ala Arg Glu Leu Lys Pro 35 40 45 GAT GTG GTC CTC ATG GAC GTC AAG ATG CCC GOG ATC GGC Q3T CTT GAA 192 Asp Val Val Leu Met Asp Val Lys Met Pro Gly He Gly Gly Leu Glu 50 55 60 GCC ACG CQC AAA TTG TTG CGC AGT CAC CCG GAT ATC AAA GTC GTG GCC 240 Ala Thr Arg Lys Leu Leu Arg Ser His Pro Asp He Lys Val Val Ala 65 70 75 80 GTC ACC GTG TGT GAA GAA GAT CCG TTC CCG ACC CGC TTG CTG CAA QCC 288 Val Thr Val Cys Glu Glu Asp Pro Phe Pro Thr Arg Leu Leu Gin Ala 85 90 95 QGC CCG GOG GST TAC CTG ACC AAG GOG GOG QGC CTC AAT GAA ATG GTG 336 Gly Ala Ala Gly Tyr Leu Thr Lys Gly Ala Gly Leu Asn Glu Met Val 100 105 110 CAG GCC ATT CGC CTG GTG TTT GCC GGC CAG CGT TAC ATC AGC CCG CAA 384 Gin Ala He Arg Leu Val Phe Ala Gly Gin Arg Tyr He Ser Pro Gin 115 120 125 ATT GCC CAG CAG TTG GTG TTC AAG TCA TTC CAG CCT TCC AGT GAT TCA 432 He Ala Gin Gin Leu Val Phe Lys Ser Phe Gin Pro Ser Ser Asp Ser 130 135 140 CCG TTC GAT GCT TTG TCC GAG COG GAA ATC CAG ATC GCG CTG ATG ATT 480 Pro Phe Asp Ala Leu Ser Glu Arg Glu He Gin He Ala Leu Met He 145 150 155 160 GTC QGC TOC CAG AAA GTG CAG ATC ATC TCC GAC AAG CTG TGC CTG TCT 528 Val Gly Cys Gin Lys Val Gin He He Ser Asp Lys Leu Cys Leu Ser 165 170 175 CCG AAA ACC GTT AAT ACC TAC GOT TAC CGC ATC TTC GAA AAG CTC TOG 576 Pro Lys Thr Val Asn Thr Tyr Arg Tyr Arg He Phe Glu Lys Leu Ser 180 185 190 ATC AGC AGC GAT GTT GAA CTG ACA TTG CTG QCG GTT CGC CAC QGC ATG 624 He Ser Ser Asp Val Glu Leu Thr Leu Leu Ala Val Arg His Gly Met 195 200 205 GTC GAT GCC AGT GCC TGA 642 Val Asp Ala Ser Ala 210 (2) INPORMATTCN FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 213 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: Met He Arg Val Leu Val Val Asp Asp His Asp Leu Val Arg Thr Gly 1 5 10 15 Ile Thr Arg Met Leu Ala Asp He Asp Gly Leu Gin Val Val Gly Gin 20 25 30 Ala Glu Ser Gly Glu Glu Ser Leu Leu Lys Ala Arg Glu Leu Lys Pro 35 40 45 Asp Val Val Leu Met Asp Val Lys Met Pro Gly He Gly Gly Leu Glu 50 55 60 Ala Thr Arg Lys Leu Leu Arg Ser His Pro Asp He Lys Val Val Ala 65 70 75 80 Val Thr Val Cys Glu Glu Asp Pro Phe Pro Thr Arg Leu Leu Gin Ala 85 90 95 Gly Ala Ala Gly Tyr Leu Thr Lys Gly Ala Gly Leu Asn Glu Met Val 100 105 110 Gin Ala He Arg Leu Val Phe Ala Gly Gin Arg Tyr He Ser Pro Gin 115 120 125 He Ala Gin Gin Leu Val Phe Lys Ser Phe Gin Pro Ser Ser Asp Ser 130 135 140 Pro Phe Asp Ala Leu Ser Glu Arg Glu He Gin He Ala Leu Met He 145 150 155 160 Val Gly Cys Gin Lys Val Gin He He Ser Asp Lys Leu Cys Leu Ser 165 170 175 Pro Lys Thr Val Asn Thr Tyr Arg Tyr Arg He Phe Glu Lys Leu Ser 180 185 190 He Ser Ser Asp Val Glu Leu Thr Leu Leu Ala Val Arg His Gly Met 195 200 205 Val Asp Ala Ser Ala 210 (2) INFORMATTON FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 642 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Pseudomonas fluorescens (B) STRAIN: CGA267356 (aka M0CG134 and aka BL915) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 1..3 (D) OTHER INFORMATION: /note= "TTG initiation codon in native sequence modified to ATG initiation codon. " (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..639 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: ATG ATT AQG GTG CTA GTA GTC GAT GAC CAT GAT CTC GTT COT ACA GGT 48 Met He Arg Val Leu Val Val Asp Asp His Asp Leu Val Arg Thr Gly 1 5 10 15 ATT ACA CGA ATG CTG GCT GAC ATC GAT GGC CTG CAA GTG GTC GGC CAG 96 He Thr Arg Met Leu Ala Asp He Asp Gly Leu Gin Val Val Gly Gin 20 25 30 GCC GAG TCA QCG GAG GAA TCC CTG CTC AAG GCC COG GAG TTG AAA CCC 144 Ala Glu Ser Gly Glu Glu Ser Leu Leu Lys Ala Arg Glu Leu Lys Pro 35 40 45 GAT GTG CTC CTC ATG GAC GTC AAG ATG CCC QGG ATC GGC GGT CTT GAA 192 Asp Val Val Leu Met Asp Val Lys Met Pro Gly He Gly Gly Leu Glu 50 55 60 GCC ACG CGC AAA TTG TIG CGC AGT CAC CCG GAT ATC AAA GTC GTG GCC 240 Ala Thr Arg Lys Leu Leu Arg Ser His Pro Asp He Lys Val Val Ala 65 70 75 80 GTC ACC GTG TGT GAA GAA GAT CCG TTC CCG ACC CGC TTG CTG CAA QCC 288 Val Thr Val Cys Glu Glu Asp Pro Phe Pro Thr Arg Leu Leu Gin Ala 85 90 95 GGC QCG QCG GGT TAC CTG ACC AAG QGG QCG GGC CTC AAT GAA ATG CTG 336 Gly Ala Ala Gly Tyr Leu Thr Lys Gly Ala Gly Leu Asn Glu Met Val 100 105 110 CAG GCC ATT CGC CTG CTG TTT QCC QGC CAG CCT TAC ATC AGC CCG CAA 384 Gin Ala He Arg Leu Val Phe Ala Gly Gin Arg Tyr He Ser Pro Gin 115 120 125 ATT GCC CAG CAG TTG GTG TTC AAG TCA TTC CAG CCT TCC ACT GAT TCA 432 He Ala Gin Gin Leu Val Phe Lys Ser Phe Gin Pro Ser Ser Asp Ser 130 135 140 CCG TTC GAT GCT TTG TCC GAG COG GAA ATC CAG ATC GCG CTG ATG ATT 480 Pro Phe Asp Ala Leu Ser Glu Arg Glu He Gn He Ala Leu Met He 145 150 155 160 GTC GGC TGC CAG AAA GTG CAG ATC ATC TCC GAC AAG CTG TOC CTG TOT 528 Val Gly Cys Gin Lys Val Gin He He Ser Asp Lys Leu Cys Leu Ser 165 170 175 CCG AAA ACC GTT AAT ACC TAC CCT TAC CGC ATC TTC GAA AAG CTC TOG 576 Pro Lys Thr Val Asn Thr Tyr Arg Tyr Arg He Phe Glu Lys Leu Ser 180 185 190 ATC AGC AGC GAT GTT GAA CTG ACA TTG CTG GCG GTT CGC CAC QGC ATG 624 He Ser Ser Asp Val Glu Leu Thr Leu Leu Ala Val Arg His Gly Met 195 200 205 GTC GAT GCC AGT GCC TGA 642 Val Asp Ala Ser Ala 210 (2) INFORMAnON FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 213 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: Met He Arg Val Leu Val Val Asp Asp His Asp Leu Val Arg Thr Gly 1 5 10 15 He Thr Arg Met Leu Ala Asp He Asp Gly Leu Gin Val Val Gly Gin 20 25 30 Ala Glu Ser Gly Glu Glu Ser Leu Leu Lys Ala Arg Glu Leu Lys Pro 35 40 45 Asp Val Val Leu Met Asp Val Lys Met Pro Gly He Gly Gly Leu Glu 50 55 60 Ala Thr Arg Lys Leu Leu Arg Ser His Pro Asp He Lys Val Val Ala 65 70 75 80 Val Thr Val Cys Glu Glu Asp Pro Phe Pro Thr Arg Leu Leu Gin Ala 85 90 95 Gly Ala Ala Gly Tyr Leu Thr Lys Gly Ala Gly Leu Asn Glu Met Val 100 105 110 Gin Ala He Arg Leu Val Phe Ala Gly Gin Arg Tyr He Ser Pro Gin 115 120 125 He Ala Gin Gin Leu Val Phe Lys Ser Phe Gin Pro Ser Ser Asp Ser 130 135 140 Pro Phe Asp Ala Leu Ser Glu Arg Glu He Gin He Ala Leu Met He 145 150 155 160 Val Gly Cys Gin Lys Val Gin He He Ser Asp Lys Leu Cys Leu Ser 165 170 175 Pro Lys Thr Val Asn Thr Tyr Arg Tyr Arg He Phe Glu Lys Leu Ser 180 185 190 He Ser Ser Asp Val Glu Leu Thr Leu Leu Ala Val Arg His Gly Met 195 200 205 Val Asp Ala Ser Ala 210 (2) INFORMATION FOR SEQ ID N0:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7001 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Pseudomonas fluorescens (B) STRAIN: CGA267356 (aka MCCG134 and aka BL915) (vii) IMMEDIATE SOURCE: (B) CLONE: pCIB169 (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 423..2036 (D) OTHER ΓNFORMAΠON: /product= "PrnA" /note= "ORF1" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 2039..3121 (D) OTHER INFORMATION: /product= "PrnB" /note= "ORF2" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 3167..4867 (D) OTHER INFORMATION: /product= "PmC" /note= "ORF3" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 4895..5983 (D) OTHER INFORMATION: /product= "PrnD" /note= "ORF4" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6 : GAATTCCGAC AACGCCGAAG AAGCG03GAA CCGCTGAAAG AGGAGCAQGA ACTGGAGCAA 60 ACQCTCTCCC AQGTGATCGA CMCCTGCCA CTGCGCATCG AGGGCCGATG AACAGCATIG 120 GCAAAAGCTG QCGGTGCGCA GTGCGCGACT GATCCGATCA TTTTTGATCG QCTCGCCTCT 180 TCAAAATCGG OGGTGGATGA AGTCGACGGC GGACTGATCA GGCGCAAAAG AACATGCGCC 240 AAAACCTTCT TTTATAGOGA ATACCTTTGC ACTTCAGAAT GTTAATTCOG AAACGGAA T 300 TGCATCGCTT TTCCGGCACT CTAGAGTCTC TAACAGCACA TIGATCTGCC TCTTGCAT3G 360 ATGCACGAAG ACTQGCQGCC TCCCCTCGTC ACAGGCQGCC OGCCTTTGAA ACAAGGAGTG 420 TTATGAACAA GCCGATCAAG AATATCGTCA TOGTGGGCQG OCCTACTGCG Q3CTGGATGG 480 COGOCTCCTA CCTCGTCCGG GCCCTCCAAC AGCAGQCGAA CATTACGCTC ATCGAATCTG 540 CGQCGATCCC TCQGATCGGC GTQGGCGAAG CGACCATCCC AAGTΠGCAG AAGGTGTTCT 600 TCGATTTCCT CGGGPOACCG GAQCG33AAT GGATGCCCCA AGTGAAOGGC GCGTTCAAGG 660 CCGCGATCAA GTTCGTGAAT TGGAGAAACT CTCCαGACCC CTCGCGCGAC GATCACTTCT 720 ACCATTTGTT 03GCAACGTG CCGAACTGCG ACGGCGTGCC GCTTACCCAC TACTGGCTGC 780 GCAAGCGCGA ACAGGGCTTC CAGCAGCOGA TGGAGTACQC GTGCTACCCG CAGCC03QG3 840 CACTCGACGG CAAGCTQGCA O TGCCTCT CCX.ACGGCAC CCGCCAGATG TCCCACGCCT 900 GQCACTTCGA CGCGCACCTG GTGGCCGACT TCTTGAAQCG CTGQGCCGTC GAC<X_CGGGG 960 TGAACCG03T GCTCGATGAG GTGGTGGACG TICGCCTGAA CAACCG033C TACATCTCCA 1020 ACCTGCTCAC CAAG3AG33G CGGACGCTGG AGGCGGACCT GITCATCGAC TGCTCCGQCA 1080 TGCGGGQGCT CCTGATCAAT CAGGOGCTGA AGGAACCCTT CATCGACATG TOCGACTACC 1140 TGCTGTGCGA CAGCGCGGTC GCCAGCGCCG TGCCCAACGA CGACGCGCGC GATGGGGTCG 1200 AGCCCTACAC CTCCTCGATC GCCATGAACT CGGGATGGAC CTGGAAGATT CCGATGCTGG 1260 GCCGGTTCQG CAGCGGCTAC GTCTTCTCGA GCCATTTCAC CTCGCGCGAC CAGGCCACCG 1320 C03ACTTCCT OΛACTCTGG GGCCTCICOG ACAATCAGCC QCTCAACCAG ATCAAGTTCC 1380 QGGTCQQQCG CAACAAQCGG GOGTGGGTCA ACAACTGCCT CTCGATC Q QG CTGTCGTCCT 1440 GCTITCTGGA GCCOCTGGAA T0GACG33GA TCTACTTCAT CTACGCGGOG CTTTACCAQC 1500 TCGTGAAGCA CTTCCCCGAC ACCTCGTTCG AOCCGCQGCT GAQOGACGCT TTCAACGCCG 1560 AGATOGTCCA CATGTICGAC GACTGCCGGG ATTTCGTCCA AGCGCACTAT TICACCACGT 1620 CQOGCGATGA CAOGCCGTTC TGGCTCGCGA ACCGGCACGA CCTGCGGCTC TCQGACGCCA 1680 TCAAAGAGAA QGTTCAGCGC TACAAGG033 GGCTQCCQCT GACCACCACG TCGTΓCGACG 1740 ATTCCACCTA CTACGAGACC TTCGACTACG AATTCAAGAA TTIT?IQGTTG AACGGCAACT 1800 ACTACTGCAT CTTTQCOQGC TT33GCATGC TGCCCGACCG GTCGCTGCCG CTGTTGCAQC 1860 ACCGACOGGA GTCGATCGAG AAAQCCGAQG CGATGTTCGC CAGCΆTCCGG CσCGAGGCCG 1920 AGCGTCTGCG CACCAGCCTG CCGACAAACT ACGACTACCT QCGSTCGCTG CGTGACGGCG 1980 ACC<33GGGCT GTCGCGCGGC CAQCGTQGQC CGAAGCTCGC AGCGCAQGAA AGCCTGTACT 2040 GGAACGCACC TTCGACCGGG TAGGCGTATT CQCG3CCACC CACGCTGCCG TGGCQQCCTG 2100 ∞ATCCGCTG CAG3CGCGCG CGCTCGTTCT GCAACTGCCG GGCCTGAACC GTAACAAGGA 2160 CGTΌCCCGCT ATCGTCGQCC TGCTGCGCGA GTTCCTTCCG GTGCGCGGCC TGCCCTGCGG 2220 CTC 3G3TTTC GTCGAAGCCG CCGCCGCGAT GCGGGACATC GGGITCTTCC TGGGGTCGCT 2280 CAAGCGCCAC GGACATGAGC CCGCQGAGGT QGTGCCCGGG CTTGAGCCGG TGCTGCTCGA 2340 CCTGGCACGC GCGACCAACC TGCCGCCGCG CGAGACGCTC CTGCATGTGA 03GTCTGGAA 2400 CπXACGGCG GCCGACGCGC AGCQCAGCTA CACCGQGCTG CCCGACGAAG 03CACCTGCT 2460 CGAGAGCGTG CGCATCTCGA TGQCGQCCCT CCAG3CQ3CC ATCQCGTTGA COCTCGAQCT 2520 GTTCGATCTG TCCCTGCGCT CGCCCGAGTT CGCGCAAAGG TGCGACGAGC TO3AAGCCTA 2580 TCTGCAGAAA ATQGTCGAAT CX_ATCGTCTA CGCCTACCGC TTCATCTCGC OQCAQGTCTT 2640 CTACGATGAG CIGCGCCCCT TCTACGAACC GATTCGAGTC QGαGGCCAGA GCTACCTCQG 2700 CCDC Q CTGCC GTAGAGATGC Cπ TCTTCCT GCTG3AGCAC GTCCTCTGGG GCTCGCAATC 2760 GGACGACCAA ACTTATCGAG AATTCAAAGA GACGTAOCTG CCCTATGTQC TTCCCGCGTA 2820 CAGGGCGGTC TACGCTCGCT TCTCOQQGGA GCCGGCGCTC ATGGACCGCG CGCTCGACGA 2880 GGCQCGAGCG GTOGGTADGC GGGACGAGCA CGTCCGGGCT GGGCTGACAG CCCTCGAGCG 2940 GGTCTTCAAG GTCCTGCTGC GCTICCGGGC GCCTCACCTC AAATTGQCCG AG033GCGTA 3000 CGAAGTCGGG CAAAGCGGCC CCGAAATCGG CAGCGGSGQG TACGCGCCCA GCATGCTCQG 3060 TGAGCTGCTC ACQCTGACGT ATGCCQCGCG GTCCCGOGTC CGCGCCQCGC TCGACGAATC 3120 CIGATGCGCG CGACCCAGTG TTATCTCACA AGGAGAGTTT OCCCCCATGA CTCAGAAGAG 3180 CCCCGCGAAC GAACACGATA GCAATCACTT CGACGTAATC ATOCTCGGCT CGGGCATGTC 3240 033CACCCAG ATGG333CCA TCTTGGCCAA ACAACACTTT CGCGTGCTGA TCATCGAGGA 3300 GTCGTCGCAC CCGCGGTTCA OGATCGQCGA ATCGTCGATC CCOGAGACCT CTCTTATGAA 3360 CCGCATCATC GCTGATCGCT AOQGCATTCC GGAGCTCGAC CACATCACGT CGT-'i'lATTC 3420 GAOSCAACCT TACGTCGCGT CGAGCA03G3 C.ATTAAQCGC AACTTCGGCT TCGTGTTCCA 3480 CAAGCCCG3C CAGGAGCACG ACCCGAAGGA GTTCACCCAG TGCGTCATTC CCGAGCTGCC 3540 GTQGGGGCCG GAGAGCCATT ATTACCGGCA AGACGTCGAC GCCTACTTGT TOCAAGCCGC 3600 CATTAAATAC QGCTQCAAQG TCCACCAGAA AACTACCGTG ACCGAATACC ACGCCGATAA 3660 AGACQGCGTC Q CGGTGACCA CCGCCCAQGG OGAACQGTTC ACCGGCCQCT ACATGATCGA 3720 CTGCGGAGGA CCTCGCQCGC CGCTCQCGAC CAAGTTCAAG CTCCGCGAAG AACCGTGTCG 3780 CTTCAAGACG CACTCGCGCA GCCTCTACAC GCACMGCTC G3GGTCAAQC OGTΓCGACGA 3840 C^TCTTCAAG GTCAAQGGGC ACOXTGGCG CTGGCACGAG GQGACCTTGC ACCACATGTT 3900 CGAGGGCGGC TGGCTCTGGG TGATTCCGTT CAACAACCAC CCGCGGTCGA CCAACAACCT 3960 QGTGAGOCTC GGOCTGCAGC TCGACCCGCG TGTCTACCCG AAAACCGACA TCTCCGCACA 4020 GCAGGAATTC GATGAGTTCC TO30GCGGTT CCCGAGCATC QGGQCTCAGT TCCQGGAOGC 4080 CGTGCCGGTG CGCGACTGGG TCAAGACCGA ( GCCTGCAA TTCTCGTCGA AGGCCTGCCT 4140 (-GGCGACCGC TACTGCCTGA TGCTGCACGC GAACQQCTTC ATCGACCCGC TCTTCTCCCG 4200 QGGGCTQGAA AACADCGCQG TGACCATCCA OGCOCTCGCG GCGCGCCTCA TCAAGGCGCT 4260 GCGCGACGAC GACTTCTCCC OCGAGCGCTT CGAGTACATC GAGCGCCTGC AGCAAAAGCT 4320 TTTGGACCAC AACGACGACT TCCTCAGCTG CIGCTACACG GCGTTCTCGG ACTTCCGCCT 4380 ATαGGACGCG TTCCACAGGC TGTO3GCGCT (OGQCACCATC CIO333CACT TCCCGCTCGT 4440 O.AGGCCCAC QCGAGGTTCC GCGCGTCGCG (-AACGAGGGC GACCTCGATC ACCTCGACAA 4500 (-GACCCTCCG TATCTCQGAT ACCT3TGCGC QGACATGGAG GAGTACTAOC AGTICTTCAA 4560 CGACGCCAAA QCOGAGGTCG AGGCCGTGAG TGCCαQGCGC AAGCCGGCCG ATGAGSCCQC 4620 GGCGCGGATT CACGCCCTCA TTGACGAAOG AGACTTCGCC AAGCCGATCT TCQQCTTCGG 4680 GTACTGCATC ACCGQGGACA AGOCGCAGCT CAACAACTCG AACTACAGCC TGCTGCOGGC 4740 GATG033CTG ATGTACIGGA CGCAAACCOG CGCGCCGGCA GAG3TGAAAA AGTACTTCGA 4800 CTACAACCCG ATGTTCGCGC TGCTCAAQGC GTACATCACG ACOCQCATCG GCCTGSCGCT 4860 GAAGAAGTAG CO3CTCGA0G ACGACATAAA AACGATGAAC GACATTCAAT TGGATCAAGC 4920 GAGCGTCAAG AAGCGTCCCT CG33CGCCTA CGACGCAACC ACQCGCCTGG _GCGAGCTG 4980 GTACGΓΓCGCG AIGOGCICCA ACGAGCΓCAA GGACAAGCCG ACOGAGTTGA CQCTCTTCGG 5040 CCGTCCCTGC GIGGCGIGGC GCGGAGCCAC GGGGCGGGCC GT3GTGATGG ACCGCCACIG 5100 CTCGCACCIG GGCGCGAACC TGGCTGACGG GOGGATCAAG GACGQGTGCA TCCAGTGCCC 5160 GTTTCACCAC TGGCQGTACG ACGAACAG3G CCAGTGOGTT CACATCCCOG QCCAIAACCA 5220 GG03GTG03C CAGCTQGAGC CGGTGCCGCG OGGGGCGCGT CAGCCGACGT TGGTCACCGC 5280 03AGCGATAC QGCTAOGTGT GGGTCTQGTA OQGCTCCCCG CTGCCGCTGC ACCCGCTGCC 5340 CGAAATCTCC GCGGCCGAIG TCGACAAOGG CGACTTTAIG CACCTGCACT TOQCGTTCGA 5400 GACGACCACG GOQGICTTGC QGATCGTCGA GAACTTCTAC GAOGCGCAGC ACGCaACCCC 5460 GGTGCACGCA CTCCCGATCT CGGCCTTCGA ACICAAGCTC TTCGACGATT G30GCCAGTG 5520 GCCGGAGCTT GAGTCGCTGG CCCTGGCGGG CGCGTGGTTC Q3TGCCGGGA TCGACTTCAC 5580 α-TGGACCGG TACTTCGQCC CCCTOOGCAT GCTGTCAOGC QCQCTCGGCC TGAACATGTC 5640 GCAGATGAAC CTGCACTTCG ATGQCTACCC CGGCGGGTQC GTCATGACCG TCGCCCTQGA 5700 CGGAGACGTC AAATACAAGC TGCTCCAGTG TGTGACGCCG GIGAGCGAAG QCAAGAACGT 5760 CATGCACATG CTCATCTCGA TCAAGAAGGT QGGCGGCATC CTGCGCCGCG CGACCGACTT 5820 CGTGCTGTTC GG3CTGCAGA CCAGGCAQGC CGCGQQGTAC GACGTCAAAA TCTGGAACGG 5880 AATGAAGOCG GACGGCGGCG GCGCGTACAG CAAGTAOGAC AAGCTOCTGC TCAAGTACCG 5940 GGCGTTCTAT CGAGGCTGGG TCGACCGCCT OGCAAGTGAG CGGTGATQCG TGAAGCCGAG 6000 COGCTCTCGA CCGCGTCGCT GOGCCAGGCG CTCGCGAACC TQGCGAGCGG CGTGACGATC 6060 ACGGCCTACG GCGCGCCGGG CCCGCTTGGG CTCGCGGCCA CX-AGCTTCCT CTCGGAGTCG 6120 CTCTITGCGA QGTATTCATG ACTATCTGGC TGTTGCAACT CGTQCTGGTG ATCQCGCTCT 6180 GCAACGTCTG CQGCCQCATT GCCGAACGGC TCGQCCAGTG CGCGGTCATC GGCGAGATCG 6240 C Q GCCG3TTT GCTGTTGGGG COCTCGCTGT TCQGCGTGAT CQCACCGAGT TICIACGACC 6300 TGTTGTTCGG CCCCCAGCTG CTGTCAGCGA T ΞCGCAACT ( .aGCGAAGTC QGCCTGGTAC 6360 TGCTGATGTT CCAGGTCGGC CTGCATATGG AGTTGGGCGA GAOGCTGCQC GACAAGCGCT 6420 GGCGCATGCC CGTCGCGATC GCAQCGGGCG GGCTCGTCGC ADCGGCOGCG ATCGGCATGA 6480 T03TCQCCAT CGTTTCGAAA GGCAOQCTCG CCAGCGACGC QCCGGCGCTG CCCTATGTGC 6540 TCTTCTGCGG TGTCGCACTT GCGCTATCGG CGGTGCCGCT GATGGCGCGC ATCATCGACG 6600 ACCTGGAGCT CAGCGCCATG GTGGGCGCGC GGCACGCAAT GTCTGCCGCG ATGCTGACGG 6660 ATGOGCTCGG ATGGATQCTG CTTGCAACGA TIGCCTCGCT ATCGAQCG33 CΠDSGCIGGG 6720 CATTIGCGCG CATGCTCGTC AGCCTGCTCG CGTATCTGGT G TGTGCGCG 6780 TCGACCGACC CTTGCQCQGC TOGCGTCGAC CDCGCATGCG ACGOGCGACC 6840 GCTIGGCCCT TTCGTAATGT TGTCGGCACT CGCGACGTCG CTGATCQGAT 6900 TCCATAGCGC TTITGGCGCA CTT3CCGCGG CGCTGTTCCT GCGCCGGGTG 6960 CGAAGGACTG GOGCGACAAC GTCGAAGGTT TCGTCAAQCT T 7001 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1097 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genαtlic) (vii) IMMEDIATE SOURCE: (B) CLONE: pKK223-3 (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 1 (D) OTHER INFORMAΠON: /note= "BssHII site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "Bglll site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 265 (D) OTHER INFORMATTON: /note= "EcoRI site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 270 (D) OTHER INFORMATTON: /note= "Smal site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 275 (D) OTHER INPORMATION: /note= "BamHI site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 281 (D) OTHER INFORMATTON: /note= "Sail site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 287 (D) OTHER INFORMATTON: /note= "PstI site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 293 (D) OTHER INFORMATTON: /note= "Xbal site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 299 (D) OTHER INFORMATTON: /note= "Xhol site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 305 (D) OTHER INFORMATION: /note= "Kpnl site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 311 (D) OTHER INFORMATTON: /note= "N tl site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 319 (D) OTHER INFORMATTON: /note= "Hindlll site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 1086 (D) OTHER INFORMATTON: /note= "Bgll site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 1092 (D) OTHER INFORMATTON: /note= "BssHII site" (ix) FEATURE: (A) NAME/KEY: promoter (B) LOCATION: 185..264 (D) OTHER INFORMATION: /standard_name= "tac" (ix) FEATURE: (A) NAME/KEY: terminator (B) LOCATION: 327..752 (D) OTHER INFORMATTON: /standard_name= "rmB" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7 : GCGOGCAGAT CTGGGCTTAT CGACTGCACG GTGCACCAAT GCTTCIGGCG TCAQQCAGCC 60 ATCΑGAAQCT GTGCTATGGC TGTGCAGQTC GTAAATCACT QCATAATTCG TGTCQCTCAA 120 QQCGCACTCC CGTTCTGGAT AATGTTTTTT GCGCCGACAT C-ATAACQGTT CTGQCAAATA 180 TTCTGAAATG AGCTGTTGAC AATTAATCAT CXXXTCGTAT AATCTGTGGA AITCTGAQCG 240 GATAACAATT TCACACAQGA AACAGAATTC CCX3QGGATCC GTOGACCTGC AGTCTAGACT 300 CGAGGGTACC GOGQCCQCAA GCTIGACTGT TTTQSCGGAT GAGAGAAGAT TTICAGCCTG 360 ATACAGATTA AATCAGAACG CAGAAGCGGT CTGATAAAAC AGAATTTGCC T -CGGCAGT 420 AGOGCQGTGG TCCCACCTGA CCCCATGCCG AACTCAGAAG TGAAACGCCG TAQOGCOGAT 480 GCTAGTGTGG GGTCTCCCCA TGCGAGAGTA G3GAACTGCC AGQCAICAAA TAAAACGAAA 540 GQCΓCAGTCG AAAGACTGGG CCTITCCTTT TATCTGTTCT TTCTCGGTGA ACGCTCTCCT 600 GAGRΠAQGACA AATCCQCCGG GAQCQGATTT GAACX-TTGCG AAGCAACGGC CCGGAGQCTG 660 GCGGGCAGGA CGCCCGCCAT AAACTGCCAG C ΑTCAAATT AAQCAGAAGG CTATCCTGAC 720 QGATGGCCTT TITGσGTTTC TACAAACTCT TTIGTTTATT TTTCTAAATA CATTCAAAIA 780 TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTCA ATAATATTGA AAAAQGAAGA 840 GTATGAGTAT TCAACATTTC OGTGTCGCCC TTATTCCCTT TTITGCGGCA TITIGCCITC 900 CTGTTTTTGC TCACCCAGAA ACGCTGGTGA AACTAAAAGA TGCTGAAGAT CAGTTGGGTG 960 CAOGAGTGGG TTAC-ATCGAA CTGGATCTCA ACAGCQGTAA GATCCTTGAG AGITTTCGCC 1020 CCGAAGAAOG TTITCCAATG ATGAGCACTT TTAAAGTTCT GCTATOTGGC GCGGTATTAT 1080 CCCGTAGATC TGCQCGC 1097 (2 ) INPORMATTON FOR SEQ ID NO: 8 : (i) SEQUENCE CHARACTERISTTCS : (A) LENGTH: 3186 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANT -SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Pseudomonas fluorescens (B) STRAIN: CGA267356 (aka MCCG134 and aka BL915) (vii) IMMEDIATE SOURCE: (B) CLONE: pCIB146 (ix) FEATURE: (A) NAME/KEY: RBS (B) LOCATION: 245..251 (D) OTHER INFORMATTON: /note= "potential ribosome binding site" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 256..3006 (D) OTHER INFORMATTON: /product= "LemA" /note= "LemA coding sequence. " (x±) SEQUENCE DESCRIPTION: SEQ ID NO: 8 : GATCCGGGGA TGGCCGGCAG ATACQGGATT CATIGGCTTC TGCAAGTAAT TCTCACTTGC 60 GCGATTATTC AAGATTGTCC GCGGCCαQGC AACCGACACC OJrCGACAAA ACQCTGGCCG 120 «3α3COGAGA CATCCGAGCC ATIGCGCQCT CAATTTTGCG AAGAATGCCG TCAAQCAAAT 180 GGCTACACTG CCCAGGTGGT GCQCACCGGA CGTGCGCAQG GTTCATTCAA AATGGCGTGG 240 TAGCAQGAGA GTTGCGTGCT TAAGAAACTG GGAATCAAAG GCCGCGTGCT GTTACTGACC 300 TTGCTGCCAA CCAGCCTGAT GGCGTTGGTA CTGG3CGGTT ATITCACCTG GATGCAGCAA 360 T03GACCTGC AAAOCCAGCT TCTGCACCGC GQ03AAATGA TO3CCGAGCA ACTGGCGCCC 420 CTGGT3GCTC CCQCCCTGQC CCAOCAGGAC ACTICCCTGC TGGAGCGCAT COXaCCCAA 480 TCCCTGGAAC AGCAGGACCT GCGCGCAGTG ACITTCCTCG OGCCCGACCG CACQCCGCTG 540 σσccATGCCG GCCCGAGCAT GCTCAACCAG GCGCCGACCG GCΆPCGGCΆG CCAGCΓGCTG 600 CAACGCACOG GCAGCGACGC CAOCCGCTAT CTC [QCCQG TATTCG3CCG CCACOGCAAC 660 CTQGCCGQCG ACCTGATTCC CGACGACTCC GACCQCCTGC TCQGCTGGCT CX-AGCTGGAA 720 CTCTCCCATA ACAQCATGCT GCTGCGCGGC TAC03CAGCC TGITCGCCAG OCTGCTGCTG 780 ATTACCGCCG GGCTGATCTG CACCQGCCTG CTCO.ACTGC QCAIQ33GCG AACCATCAAC 840 GACCCGCTGA QCCAGATCAA ACAGGCOGTC ACO-AGCTCA AQGACGGCAA CCTSGAAACC 900 CGCCT3CCCT TGCTOOGCAG OCAGGAACTG GACGAGCTGG CCTO3GGCAT CAACCGCATG 960 GCOQGCACCC TGCAGAATGC CCAGGAAGAA CTGCAGCACA GCATCGACCA GGCCAOCGAG 1020 GACGTCCGGC AAAACCTGGA GACCATCGAG ATCCAGAACA TCGAGCTGGA CXTIQGCQCGC 1080 AAQGAG3CCC TO3AGGCCAG C033ATCAAG TCCGAATTCC TGGCCAACAT GAGCCATGAA 1140 ATCCGCAC Q C CGCTCAACGG CATOCTCGGC TTCACTCATT TGTTGCAGAA AAGOGAGCTG 1200 ACCCOGCGCC AQCTGGATTA CCT3GGCACC ATCGAAAAAT CCGCCGACAG CCTGCTGGGA 1260 ATCATCAACG AAATTCTCGA CTTCTCGAAA ATCGAAGCCG QCAAQCTGCT QCTCGACAGC 1320 ATICCGTTCA ADCTGCGCGA CCTGTTGCAG GACACCCTGA CCATICTCGC TCCQGCCGCC 1380 CACGCCAAGC AGCTGGAACT CCTCAGCCTG GTGTATCGCG ATAGCCCGCT GTCQCTGCTG 1440 QGCGACOCGC TGCGCCTCAA GCAGATCCTC ACCAATCTGG TGAGCAACQC CATCAAGTTC 1500 ACCCGCGAAG QCACCATCGT CGOCOGGGCC AT3CTTGAAG AGGAGCACGA AGACAQCGTG 1560 CAACTGCGCA TCAGCATTCA QGACAC03GC ATCGGCCTCT CGAACCAGGA CGTGCQ03CC 1620 CIGTTOCAGG CGTTCAGCCA GGCCGACAAT TCGCTGTCGC GACAACCCGG CGQGACTGGC 1680 CIGGQQCTGG TGATTTCCAA GCGCCTGATC GAACAGATGG QCGGCGAGAT CQ3CGTCGAC 1740 AGCACGCCCG GCGAAQGTTC QGAGTTCTGG ATCAGCCTGC GCCTGCCGAA AACCCGOGAC 1800 GACGCCGAAG ACCTGC0Q3C CCCGCOGCTG CTCGGCAG3C QIGTCGCGGT CC GGAAAAC 1860 CATGAGCTGG 03CGCCAGGC CCTQCAGCAT CAACTCGAGG ACTCΩQGCCT GGAAGTCACT 1920 CCGITCAACA CCCTQGAAGC CCTGACCAAC «_αjPGACCG GCGTGCACCA GACCGACCAG 1980 C<3GATCGATC TGGCGGTCCT CGGCATCACC ACCAACGACA TGCTGCOGGA AD3CCTCAAC 2040 CAGCACATCT QGGACCTCGA GCACCTGGGC TGCAAAGTCC TGGTGCTCTG CCCGAOCACA 2100 GAACAGACAC TCTTCCACCT GTCGGTGCCC AACXXTCACA QCCAGTTGCA GQCCAAACCG 2160 GCGTGCACGC GCAAACTGCG GCGCQCCCTG GCCGACCTGG TCAACCCCAA OTIGGTGCGC 2220 AGCGAGCCGA GOGAACCGAT CGCCAGCCQC CO3CCACG3G TCCTGTGTGT CGATGACAAC 2280 CCGGCCAACC TGCTQCTGCT GC-AGACCCTG CTCGAAGACA TC«3CGCCAA AGTGCTOQCG 2340 GTCCACAGCG GCTATGCGGC QGTCAAGGCG GTGCAGAGOG AGTOCTTCGA CCTGGTGATG 2400 AΪGGACGTGC AGATGCCCQG CATGGACGGT CGCCAGAGCA CCGAGGCGAT TCGCX-ACTGG 2460 GAAAGCQGGC GCAACTGCTC GCCGCTGCCG GTGATCGCCC TCAC03CCCA CGCCATGGCC 2520 AACGAAAAAC GO-CGCTGCT GCAAAGCGGC ATGGACGATT ACCTGAOCAA ACCCATCACT 2580 CAGCGGCAAC TGGCCCAGCT QGTGCTGAAG TGGACCGGCC TGGCCCTGCG CAACCAAGCT 2640 CCQGAACGCT CTGGCGAACT GTCTGGCAAC GQCCTCGAGC TGCAAGTGCT GGATCACGAC 2700 GAAC<3CTTGC TCCTGGCCGC CGGCAAGGCG GACCTGGCGG CCGACATGCT QGCCATGCTC 2760 CT3GCCTCGC TαGAAGCCGA TOSCGAAGOG ATTCGCGCCG (CCCGTGCCGC CAACGATCAC 2820 AATGCGTTGA TCGAGCQGGT CCATCGCCTG CACGGGGCGA CCCGCTATTG TGQOGTGCCG 2880 CAGTTGCGCG CCGCCTQCCA GCGCAGCGAA ACCCTGCTCA AGCAGGAAGA GGTCAAGGCC 2940 TTΌGCCGCCC TCGAOGAGCT CGAACGQGCC ATTAGTCGCC TGGCCACGGA GGCCCGCATC 3000 AAOGCCTGAT TCAAGGCAAC GACACGTCAG CCCCGCAGCT TCATGCTCGG GGCAACTTTC 3060 ACAAQGACGA CGCCAIGCGC AOGATTCTCT TCAGCAGCCA GAACTATGAC OGCGACAGCT 3120 TCCTCGGCGC CGCCCTGCCG CCGGGCATCG AGCTGCAATT CCAGGCGGCG CGCCTGAGOC 3180 TGGACA 3186 (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 642 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETTCAL: NO (iv) ANTT-SENSE: NO (vii) IMMEDIATE SOURCE: (B) CLONE: CGA375260 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..639 (D) OTHER ΓNFORMAΠQN: /product= "gac*3 gene" (D) OTHER INFORMATTON: /transl_except= (pos: 1 .. 3, aa: Met (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: TTG ATT AGG GTG CTA CTA GTC GAT GAC CAT GAT CTC CTT CCT ACA GCT 48 Met He Arg Val Leu Val Val Asp Asp His Asp Leu Val Arg Thr Gly 1 5 10 15 ATT ACA CGA ATG CTG GCT GAC ATC GAT QGC CTG CAA GTG GTC GGC CAG 96 He Thr Arg Met Leu Ala Asp He Asp Gly Leu Gin Val Val Gly Gin 20 25 30 GCC GAG TCA GOG GAG GAA TCC CTG CTC AAG GCC 003 GAG TTG AAA CCC 144 Ala Glu Ser Gly Glu Glu Ser Leu Leu Lys Ala Arg Glu Leu Lys Pro 35 40 45 GAT GTG GTC CTC ATG GAC GTC AAG ATG CCC QGG ATC QGC GCT CTT GAA 192 Asp Val Val Leu Met Asp Val Lys Met Pro Gly He Gly Gly Leu Glu 50 55 60 GCC ACG CGC AAA TTG TTG CGC AGT CAC CCG GAT ATC AAA GTC GTG GCC 240 Ala Thr Arg Lys Leu Leu Arg Ser His Pro Asp He Lys Val Val Ala 65 70 75 80 GTC ACC GTG TCT GAA GAA GAT CCG TTC COG ACC CGC TTG CTG CAA GCC 288 Val Thr Val Cys Glu Glu Asp Pro Phe Pro Thr Arg Leu Leu Gin Ala 85 90 95 QGC GCG GCG GCT TAC CTG ACC AAG QGG QCG QGC CTC AAT GAA ATG GTG 336 Gly Ala Ala Gly Tyr Leu Thr Lys Gly Ala Gly Leu Asn Glu Met Val 100 105 110 CAG GCC ATT CGC CTG CTG TTT GCC QGC CAG CCT TAC ATC AGC CCG CAA 384 Gin Ala He Arg Leu Val Phe Ala Gly Gin Arg Tyr He Ser Pro Gin 115 120 125 ATT GCC CAG 033 TTG GTG TTC AAG TCA TTC CAG CCT TCC ACT GAT TCA 432 He Ala Gin Arg Leu Val Phe Lys Ser Phe Gin Pro Ser Ser Asp Ser 130 135 140 CCG TTC GAT GCT TTG TCC GAG COG GAA ATC CAG ATC GCG CTG ATG ATT 480 Pro Phe Asp Ala Leu Ser Glu Arg Glu He Gin He Ala Leu Met He 145 150 155 160 GTC QGC T3C CAG AAA GTG CAG ATC ATC TCC GAC AAG CTG TGC CTG TOT 528 Val Gly Cys Gin Lys Val Gin He He Ser Asp Lys Leu Cys Leu Ser 165 170 175 CCG AAA ACC GTT AAT ACC TAC CCT TAC CGC ATC TTC GAA AAG CTC TOG 576 Pro Lys Thr Val Asn Thr Tyr Arg Tyr Arg He Phe Glu Lys Leu Ser 180 185 190 ATC AGC AGC GAT CTT GAA CTG ACA TTG CTG GCG CTT CGC CAC GGC ATG 624 He Ser Ser Asp Val Glu Leu Thr Leu Leu Ala Val Arg His Gly Met 195 200 205 GTC GAT GCC AGT GCC TGA 642 Val Asp Ala Ser Ala 210 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 213 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xL) SEQUENCE DESCRIPTION: SEQ ID NO:10: Met He Arg Val Leu Val Val Asp Asp His Asp Leu Val Arg Thr Gly 1 5 10 15 He Thr Arg Met Leu Ala Asp He Asp Gly Leu Gin Val Val Gly Gin 20 25 30 Ala Glu Ser Gly Glu Glu Ser Leu Leu Lys Ala Arg Glu Leu Lys Pro 35 40 45 Asp Val Val Leu Met Asp Val Lys Met Pro Gly He Gly Gly Leu Glu 50 55 60 Ala Thr Arg Lys Leu Leu Arg Ser His Pro Asp He Lys Val Val Ala 65 70 75 80 Val Thr Val Cys Glu Glu Asp Pro Phe Pro Thr Arg Leu Leu Gin Ala 85 90 95 Gly Ala Ala Gly Tyr Leu Thr Lys Gly Ala Gly leu Asn Glu Met Val 100 105 110 Gin Ala He Arg Leu Val Phe Ala Gly Gin Arg Tyr He Ser Pro Gin 115 120 125 He Ala Gin Arg Leu Val Phe Lys Ser Phe Gin Pro Ser Ser Asp Ser 130 135 140 Pro Phe Asp Ala Leu Ser Glu Arg Glu He Gin He Ala Leu Met He 145 150 155 160 Val Gly Cys Gin Lys Val Gin He He Ser Asp Lys Leu Cys Leu Ser 165 170 175 Pro Lys Thr Val Asn Thr Tyr Arg Tyr Arg He Phe Glu Lys Leu Ser 180 185 190 He Ser Ser Asp Val Glu Leu Thr Leu Leu Ala Val Arg His Gly Met 195 200 205 Val Asp Ala Ser Ala 210 (2) INFORMATTCN FOR SEQ ID N0:11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5698 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTT-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Pseudomonas aureofaciens (B) STRAIN: 30-84 (vii) IMMEDIATE SOURCE: (B) CLONE: phzFABCD (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 105..1307 (D) OTHER INFORMATTON: /prcduct= "phzF" /note= "ORFl" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LCCATTON: 1323..1946 (D) OTHER INFORMATTON: /product= "phzA" /note= "ORF2" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 1943..3856 (D) OTHER INFORMATTON: /product= "phzB" /note= "ORF3" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 3859..4695 (D) OTHER TN ΌRMAΠCN: /product= "phzC" /note= "ORF4" (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATTON: 4692 . .5360 (D) OTHER BJFCW ION: /product= "phzD" /note= "ORF5" (xi) SEQUENCE DESCRIPTTCN: SEQ ID NO: 11 : GAATTCATGA AGCπCTTTCA GCAACTGCGC GCCCTGGGTA TTCCAGTACC GCAAATCAAG 60 OGOGAAQGCA TTCCAACTTA ATOCCTCGTG AGACTGATCG CATCATGGAA GACTTACTGA 120 AA033CTTTT AAGITGTGAA GCX3TTCCAQC AGCCTCAATG GAGCGAGCCC TCACAATTGC 180 ATC4AO30SCA GGCCTACCIC AQGGACAGCG CCTCATTGAT ACGAGTGGAA GACATCCTGG 240 TGCTGCGCGC CACGCTOQCG (-CTCTAGCGG CXX3QCGAAGC AATGGTCATC CAGTCCGGTG 300 ACTGCGCOGA QGACATGGAT GAAAGCACTC CCGACCATGT GGCCCGCAAA GCCGCQGTAC 360 TGGACATCCT Q3C03CTA0G TTCCQGCTGG TGACCCAACA 420 GAATIGCCGG GCAGTTTGCC AAC CQCGTT CCAACAACAA CGAACGCATC GGCGATGTCG 480 AATTACCQCT GTATOGCGGC GACATGGTCA ACO3TCGCGA GGTCATCGCC 540 AGCACGATGC C<^A03CCTG GTΓCGAGGCT ATAGCGCCGC GCQGGACATC ATGCAACACC 600 TO33CT3GAA AGCCTCGGCA AGCCAQGAAC AACTCAQOQG TTCACCGGOC TGGACCAACC 660 ATGAAATGCT CGTACTCGAC TA03AACTGC CACAACTGCG CCAG3A0GAA CAQGGCCGGG 720 TATTICTCGG TTCTACCCAC TGGCCGTGGA TCGQOGAGOG TACCOCTCAG TTAACGGGCG 780 CTCACCTGAC GCTGCTCAGC GAAGTGCTCA ATCCGGTGGC GTGCAAGCTC QGCCCGGACA 840 TTACCCAAGA CCAGTTACTG AGOCTGTGTG AACGCCTGGA COα-AAGCGC GAACCCGGCC 900 GQCTGACCCT GAIT3COCGC ATGGGCGCGC AAAAGGTCGC CGAGCGCCTG (XG∞QCTGG 960 TCGAAGCGCT GCGCCAQGCC QGCCACAAGA TCATCTGGCT GAGCGACCCG ATGCACGGCA 1020 ACACCATOCT CGCGOCCTGC GGCAACAAGA CCCGCATGCT QCAGGCCATC ACCGAGGAAA 1080 TOGCCGCCTT (.AACCAΪGCC GIGACCTCCG CCGGIGGCCT GGCCGCCQQC CTGCNXTTGC 1140 AAACCACCCC TGACGACGTC AGCGAGTGCG CTTCCGATGC CGCCGGCCTG CATCAGGTCG 1200 OCAGCCGCTA CAAAAGCCTG TGCGACCCGC GCCTGAAOCC CTGGCAAQCC ATTACT3CGG 1260 TGATGGCCTG GAAAAACCAG CπTICTTCAA CX TIGCCTC CTTTIGACTG GAGTTTGTCG 1320 TCATGACCGG CATTCCATCG ATCGTCCCTT ACGCCTTGCC TACCAAOCGC GAOCTGCCCG 1380 TCAACCTCGC GCAATGGAGC AIOGACCCCG AGCGTGCCCT GCTGCTGCTG CATGACATGC 1440 AGCQCTACTT CCTGCGGCCC TTGCCCGACG CCCTGCGTGA CGAAGTOCTG AGCAATGCCG 1500 03CGCΑTTCG CCAGTGQGCT GCCGACAACG GCGTTCCGCT QGCCTACACC GCOCAGCCCG 1560 GCAGCATGAG CGAGGAGCAA CGCGGGCTGC TCAAQGACTT CTGOGGCOCG GGCATGAAGG 1620 O-AQCCCCGC CGACCGCGAG GTQGTCGGCG CCCTGACGCC CAAGCCCGGC GACTGQCTGC 1680 TGACCAAGTG GCGCTACAGC GCGTICTTCA ACIC03ACCT QCTGGAACGC ATGOGCGCCA 1740 AOGGGCGOGA TCAGTTGATC CTGTGCG333 TGTACGCCCA TGTCG33GTA CTGATTTCCA 1800 CCGTGGATGC CTACTOCAAC GATATCCAGC CGTICCTCCT TC^XGACQCG ATCGCOGACT 1860 TCAGCAAAGA GCACCACTGG AIGGCCATCG AATACQCCGC CAGCCGTTGC GCCAIGGTCA 1920 TCACOiCCGA CGAC ?IGGTG CTATGAQCCA GACCQCAGCC CACCTCATGG AACGCATCCT 1980 GCAACCGGCT OCCGAGCOCT TIGCCOTCTT CTA∞GCCCG GAATCCAGTG GOOCCGGCCT 2040 GCTGGACGTG CTGAXCQGCG AAATGTCGGA ACOGCAGGTC CTGGCCGATA TCGACTTGCC 2100 TGCCACCTCG ATCGGCGCGC CICGCCTGGA TGTACTGGCG CTGATCCCCT ACCQCCAGAT 2160 CGCCGAACGC GGTTTCGAGG CGGTGGACGA TGAGTCGCCG CTGCTGGCGA TGAACATCAC 2220 CGAGCAGCAA TCCATCAGCA TCGAQCGCTT GCTGGGAATG CTGCCCAACG TGCCGATCCA 2280 GΓTGAACAGC GAACGCTTCG ACCTCAQOGA OGCGAQCTAC GCCGAGATCG TCAGCCAGCT 2340 GATCGCCAAT GAAATCQGCT CCGGGGAAGG OGCCAACTTC GTCATCAAAC GCAOCTTCCT 2400 G3CCGAGATC AGCGAATACG GCCCGGOCAG TGCGCTCTCG TTCITICQOC ATCTGCTGGA 2460 ACGGGAGAAA QGCGCCTACT GGACGTTCAT CATCCACACC GGCAQCCCTA CCTTCGTGGG 2520 TGCGTCCCCC GAGCGCCACA TCAGCATCAA GGATGGGCTC TCGGTGATGA ACCCCATCAG 2580 CασCACTTAC CGCTATCCGC CCGCOGSCCC CAACCTGTCG GAAGTCATGG ACTTCCTG3C 2640 CGAT03CAAG GAAGCCGACG AQCTCTACAT QGTGQTGGAT GAAGAGCTGA AAATGATGGC 2700 GCGCATTTCT GAGGACGGCG GCCACCTCCT CG3CCX_TTAC CTCAAQGAAA TGGOGCACCT 2760 Q GCCCACACC GACTACTTCA TCGAAGGCAA GACCCATCGC GATGTACGGG AAATCCTGCG 2820 CGAAAOCCTG TITGCαCCCA CCGTCACCGG CAGCCCACTG GAAAGCGCCT C«_GQ3TCAT 2880 CCAGCGCTAT GAGCCGCAAG QCD3CG0CTA CTACAGCGGC ATGQCTGCGC TGAT033CAG 2940 CGATGGCAAG QGCQGGOCTT CCCTQGACTC CGCX_ATCCTG ATTCCTAOCG COGACATCGA 3000 TAACAGOQGC GAGGTGCGGA TCAGOGTGGG CTCGACCATC GTGCGCCATT COGACCCGAT 3060 GADOGAGGCT GCCGAAAGCC QG3CCAAGGC CACTGGCCTG AICAGCGCAC TGAAAAACCA 3120 GOCGCCCTCG ∞CTTCGQCA ATCACCΓGCA AGTGCGCGCC GCATTOGCCA GCCGCAATGC 3180 CTAOGTCTOG GACTTCTGGC TGATGGACAG CCAGCAGCGG GAGCAGATOC AGGCOGACTT 3240 CAGTGGGCGC CAGGTGCTGA TCGTCGAD3C CGAAGACACC TTCACCTCGA TGATCGCCAA 3300 GCAACTGCGG QCCCTGGGCC TCGTAGTGAC GGTCriGCAGC TTCAGCGACG AAIACAGCTT 3360 TGAAGGCTAC GACCTGGTCA TCATGGQCCC OGGCOOCGGC AAOCCGAGCG AACTCCAACA 3420 GCCGAAAATC AACCACCTGC ACGTGGCCAT CCGCTCCTTG CTCAGCCAQC AC03GCCATT 3480 CCTCGOGGTG TGCCT3AGCC ATCAQCTGCT GAGCCT3TGC CTOGGCCTQG AACIGCAGCG 3540 CAAAQCCATT CCCAACCAGG GCGTGCAAAA ACAGATCGAC CTGITTGGCA ATGTCGAACG 3600 GGTGQGTT C TACAACACCT TCGCCGCCCA GAGCTCGAGT GACCGCCTGG ACATOGACGG 3660 CATCGQCACC GICGAAATCA GOCQCGACAG C3AGACCGGC GAGGTGCATG CCCTGCGTQG 3720 CCCCTO3TTC GCCTCCATGC AGTTTCATGC CGAGTCGCTG CTGACCCAGG AAO-TCCGCG 3780 CATCATCGCC C XTGCTGC GGCACGCCCT GATCCACACA CCT3TCGAGA ACAAD3CTTC 3840 Q3CCGCCGGG AGATAACCAT GGAGCATTAC GTCATCATCG ACGCOTTTGC CAGOSTCCCG 3900 CTGGAAQGCA ATOCGGTCGC G3TGTTCTTT GACQCCGATG ACTIGTCGGC CGAGCAAATG 3960 CAAOGCATTG CCCG3GAGAT GAACCIGTCG GAAACCACTT TCGTGCTCAA GCX-AOGTAAC 4020 TGCGQCGATG CXXTGATCCG GATCTTCACC (3CGGTCAACG AACTGCCCTT CXKX0333CAC 4080 C0GTT3CTGG GCACGGACAT TGCCCTGQCT GCGCGCAOCG ACAATCACCG GCTGTTCCTG 4140 GAAACCCAGA TGGGCACCAT CQCOTTTGAG CTGGAGCGCC AGAACGGCAG OGTCATCGCC 4200 GOCAGCATGG ACCAGCCGAT ACCGACCTGG ACGGCCCTGG GQCGCGAOGC CGAGTTGCTC 4260 AAGGCCCTGG GCATCAGOGA CTCGACCTTT CCCATCGAGA TCTATCACAA CGCOCCGCCT 4320 CATGTUmU TCQ3CCTGCC AAGCATCGCC OMCTCTCGG CCCTGCACOC CGACCAOCCT 4380 QCCCTGTACA QCTICCACGA CATGGCCATC AACTGTTTTG CCGGTG03GG ACGOCGCTGG 4440 CGCAQCCGGA TGTICTOGCC GGCCTATGGG GIGGICGAGG ATGαSGCCAC GGGCTCOGCT 4500 GCX3QGGCCCT TC^XGATCCA TCTGGCGCGG CATGGCCAGA TCCiAGITCOG CCAGCAGATC 4560 GAAATTCTTC AGGQCGTGGA AATCQGCCGC CXXTCACTCA TGTIO3CCCG GGCCGAGGQC 4620 σSCGCOGATC AACTGACGCG QGTCGAACTA TCAQGCAATG GCATCACCTT CGGAOGGGGG 4680 AC OOGTTC TATGAACACT TCAGTACTAG GCAXSCCGCT GTTGGGTAAA GGCATGTGQG 4740 AATOGCIGAC CX33CACACTG GATCCGCCGT TCCCCGAGTA O-AGAAGCCG CCTGCCGATC 4800 CCAIGAGCGT GCTGCACAAC TC*_CTCGAAC GCGCACGCCG OCTG3QCATC CGCGAACCCC 4860 GTGCGCTGGC GCTGGCCACG GCTGACAGCC AGGGCCGGCC TIOGACAOGC ATCGTGGTGA 4920 TCAGTGAGAT CAGTGACACC GGGGTGCTCT TCAGCACCCA TGCCGGAAGC CAGAAAGGOC 4980 GCGAACTGAC AGAGAACCCC TGGGCCTCQG QGA03CICTA TIOGCGCGAA ACCAGOCAGC 5040 AGATCATCCT CAATGGCCAG GCCGTGOGCA TGCOQGATGC CAAGGCTGAC GAGGCCTGCT 5100 TGAAGOQCCC TTATGCCACG CATCCGATGT CATCGGTGTC TCGCCAGACT GAAGAACTCA 5160 AQGATGTTCA AGCCATGCQC AACGCCGOCA QGGAACTGGC α_AQ3TTCAA GGTCCGCTCC 5220 CGCGTCCCGA GGGTTATTGC CTUlTi ACT TAOQQCTTGA ATOGCTGGAG TTCTGG33TA 5280 A0GGCGAG3A CC Q CCTGCAT GAACQCTTGC GCTATGACCG CAQOGCTGAA GGCTGGAAAC 5340 ATO3C03GTT ACAGCCATAG QGTC003CGA TAAACATGCT TTGAAGTGCC TGGCTGCTCC 5400 AGCTTCGAAC TCATTGCGCA AACTTCAACA CTTATGACAC CCGGTCAACA TGAGAAAACT 5460 CCAGATGCGA AAGAACGCCT ATTCGAAATA CCAAACAGAG AGTCCGGATC ACCAAAGTGT 5520 GTAACGACAT TAACTCCTAT CTGAATTTTA TAGTTGCTCT RGΑΑCGTTGT CCTTGACCCA 5580 GCGATAGACA TCGQQCCAGA ACCTACATAA ACAAAGTCAG ACATTACTGA G3CTGCTA0C 5640 AIGCTAGATT TTCAAAACAA CCGTAAATAT CTGAAAAGTG CAGAATCCTT CAAAGCTT 5698

Claims: What Is Claimed Is: 1. A genetically engineered biocontrol strain of Pseudomonas that is able to effectively control pathogenic attack on crop plants by pathogenic fungi such as Rhizoctonia and Pythium and that is also able to aggressively compete with indigenous bacteria and microflora existing in the rhizosphere of the plant wherein the lemA and gacA regulatory genes and/or the genes involved in the synthesis of the antifungal metabolite phenazine-1- carboxylic acid (PCA) and/or pyrrolnitrin have been utilized to genetically modify a parent Ps. fluorescens strain to construct altered strains that demonstrate enhanced production of antifungal metabolites, i.e. pyrrolnitrin, and accordingly enhanced biocontrol activity. 2. A biocontrol strain of Pseudomonas fluorescens selected from the group consisting of: CGA376146, CGA364473, CGA375258, CGA376148, CGA364476, CGA375260, CGA375259, CGA378584, CGA267pPhz , CGA364474, CGA364475, CGA366259, CGA376150, NOA402208, NOA402210, NOA402212, NOA402214, NOA402216, NOA409063, NOA409068, NOA413174, NOA413175, NOA413176, NOA413177, and NOA413178; or a pyrrolnitrin producing strain derived therefrom. 3 A biocontrol strain of claims 1 or 2, wherein the gacA and lemA regulatory genes are transformed into a transposon mutant of a wild-type P. fluorescens strain. 4. A biocontrol strain of claims 1 or 2, wherein the first base in the coding sequence of the gacA gene is changed from a thymidine (T) to an adenine (A) to create the more efficient ATG translation initiation codon . 5. A biocontrol strain of claims 1 or 2, wherein the genes involved in the synthesis of the antifungal metabolite pyrrolnitrin are operably linked to a strong constitutive bacterial promoter that causes high-level expression of these genes in the transformed Pseudomonas strain. 6. A biocontrol strain of claims 1 or 2, wherein said strain is transformed with a plasmid comprising the lemA and gacA genes derived from the chromosome of a wild-type strain. 7. A biocontrol strain of claims 1 or 2, wherein said strain is transformed to contain both the the pmABCD gene cluster and a plasmid comprising the lemA and gacA genes. 8. A biocontrol strain of claims 1 or 2, wherein said strain is transformed to contain a gacA gene that encodes a GacA protein exhibiting a single amino acid change in the encoded GacA protein owing to a single base exchange in the correpsonding gacA gene such that the modified GacA protein does no longer require phosphorylation by LemA in order to be active as a transcriptional activator. 9. The biocontrol strain of claim 8, wherein said modified gacA gene comprises a single base change in codon 132, which is CAG and encodes a glutamine residue in the native GacA protein. 10. The biocontrol strain of any one of claims 1 , 2, 8 and 9, wherein the gacA gene is operably linked to a strong constitutive bacterial promoter that causes high-level expression of the gene in the transformed Pseudomonas strain. 11. A biocontrol strain of claims 1 or 2, wherein a no-phenazine, pyrrolnitrin producing Pseudomonas strain is transformed with a plasmid carrying DNA that contains 5 genes known to encode the pathway for the biosynthesis of the antifungal metabolite phenazine-1-carboxylic acid (PCA). 12. A biocontrol strain of claims 1 or 2, wherein the gacA gene is operably linked to a lac promoter element from E. coli such that expression of the gacA gene is regulated by the lac promoter. 13. A biocontrol strain of claims 1 or 2 comprising the pmABCD gene cluster operably linked to a tac promoter element from E. coli such that expression of the pmABCD genes is from the tac promoter, together with the gacA gene. 14. A biocontrol strain of any one of claims 1 or 13, wherein the parent strain is 2- hexyl-5-propyl-resorcinol non-producing deletion mutant of P. fluorescens. 15. A biocontrol composition comprising the biocontrol strain of any one of claims 1 to 14 together with an agronomically acceptable carrier. 16. A biocontrol composition of claim 15 comprising the biocontrol strain of any one of claims 1 to 14 in combination with a chemical fungicide. 17. The biocontrol composition of claim 16, wherein said chemical fungicide is a metalaxyl compound. 18. A method for controlling or inhibiting the growth of a plant pathogenic fungus by applying an effective amount of the biocontrol strain of of any one of claims 1 to 14 to an environment in which the plant pathogenic fungus may grow. 19. A method for controlling or inhibiting the growth of a plant pathogenic fungus by applying an effective amount of the biocontrol strain of any one of claims 1 to 14 to a plant or plant part in order to protect said plant or plant part from a plant pathogenic fungus. 20. A method for controlling or inhibiting the growth of a plant pathogenic fungus by applying an effective amount of the biocontrol strain of any one of claims 1 to 14 to seed in order to protect a plant that develops from said seed from a plant pathogenic fungus. 21. The method of either claim 18, 19 or 20, wherein said plant pathogenic fungus is Rhizoctonia or Pythium. 22. A method for controlling or inhibiting the growth of a plant pathogenic fungus by applying an effective amount of the biocontrol composition of any one of claims 15 to 17 to an environment in which the plant pathogenic fungus may grow. 23. A method for controlling o r inhibiting the growth of a plant pathogenic fungus by applying an effective amount of the biocontrol composition of any one of claims 15 to 17 to a plant or plant part in order to protect said plant or plant part from a plant pathogenic fungus. 24. A method for controlling or inhibiting the growth of a plant pathogenic fungus by applying an effective amount of the biocontrol composition of any one of claims 15 to 17 to seed in order to protect a plant that develops from said seed from a plant pathogenic fungus.

Inventors: GAFFNEY THOMAS DEANE, HILL DWIGHT STEVEN, LAM STEPHEN TING, LIGON JAMES MADISON, STAFFORD JILL MICHELLE, TORKEWITZ NANCY REEVES, GAFFNEY, THOMAS, DEANE, HILL, DWIGHT, STEVEN, LAM, STEPHEN, TING, LIGON, JAMES, MADISON, STAFFORD, JILL, MICHELLE, TORKEWITZ, NANCY, REEVES
Applicants: CIBA GEIGY AG, NOVARTIS AG
Application year: 1997
Main classification: C12N15
IPC codes: A01N63, C07K14