Progress 10/01/02 to 09/30/07

Outputs
OUTPUTS: Trichoderma spp. are very diverse their abilities to make large quantities of extracellular enzymes, to control diseases and to enhance plant growth. Some strains are plant symbionts. Recent developments indicate that they infect plant roots, establish asymptomatic infections, and create a zone of chemical communication between the fungus and the host. This communication frequently results in better plant growth and yield, and in systemic resistance to plant diseases. Most of this understanding is the result of informal international collaboration over the past few years. It has permitted an increase in abilities to screen for better biocontrol systems both through direct effects on plants such as for systemic resistance to pathogens such as powdery mildew, and also through molecular signatures, which is direct screening for combinations of plant gene products that are indicative of positive plant-microbe interactions. The enzymes also are important; in this project we have identified better methods for large-scale production of chitinolytic enzymes from Trichoderma. These improvements have been through increased understanding of factors that control enzyme production and through processes such as response surface modeling. We also have identified chitinolytic enzymes with highly desirable characteristics from plant, bacterial and plant sources. We have identified critical sites of catalytic activity in a bacterial enzyme. PARTICIPANTS: Postdocs: Shiping Deng, Bruno Donzelli TARGET AUDIENCES: The target audiences are both the academic community, where increased understanding of the science is critical and for commercial use of fungi and enzymes. There are several companies interested in the enzymes and Trichoderma strains for commerical use. Some of the Trichoderma strains are being tested in the field by companies.

Impacts
Work in this project has been extremely helpful in developing better methods to screen Trichoderma strains with better potential in commercial agriculture than those currently available. They are currently being evaluated for commercial use. On the enzyme side, we continue to find enzymes that are more and more active both in degradation of chitin and in their antifungal activity. We are developing synergistic additives to the antifungal materials. We also have made basic progress in understanding further how to enhance both enzyme production and in the protein structures that provide catalytic activities.

Publications

• Park, S. K., Kim, C. W., Kim, H., Jung, J. S. and Harman, G. E. 2007. Cloning and high-level production of a chitinase from Chromobacterium sp. And the role of conserved or nonconserved residues on its catalytic activity. Appl. Microb. Biotechnol. 74:791-804
• Harman, G. E. and Shoresh, M. 2007. The mechanisms and applications of symbiotic opportunistic plant symbionts. pages Pages 131-155 in Vurro, M. and Gressel, J. (eds.). Novel Biotechnologies for Biocontrol Agent Enhancement and Management.
• Deng, S., Pentilla, M., Lorito, M. and Harman, G. E. 2007. Overexpression of an endochitinase gene (ThEn-42) in Trichoderma atroviride for increased production of antifungal enzymes and enhanced antagonist action against phytopathogenic fungi. Appl. Biochem. Biotechnol. 142: 81-94.

Progress 01/01/06 to 12/31/06

Outputs
Certain microorganisms function as plant symbionts including fungi in the genus Trichoderma; the international community has determined that these organisms infect the outer layers of root cells and secrete bioactive compounds that serve as plant communication factors. These materials, designated symbiont associated molecular patterns (SAMPs) are diverse and different SAMPs cause different reactions in the plant. Among the most useful properties of these organisms are their abilities to increase plant growth and induce systemic resistance. The latter capability induces resistance in the foliage even when the fungi are localized on roots. These responses must be associated with significant changes in the plant transcriptome and proteome. We have investigated the changes in the plant proteome as induced by root colonization with Trichoderma harzianum strain T22. There are about 300 proteins in shoots whose expression is changed in maize seedlings by T22. Many of the shoot proteins upregulated by T22 are those involved in disease resistance, which is expected given the fact that T22 does induce systemic resistance, and basic metabolism and phtotosynthesis, which is reasonable given the fact that T22 increases growth and, frequently leaf greenness. Our data suggests that T22 induces the jasmonate/ethylene pathway, as is definitely the case with T. asperellum/cucumber. However, by changing the SAMPs to include glucose oxidase probably results in increased activity of the salicylate pathway. In addition to this basic approach, we also have been developing improved general methods for formulation of biocontrol microbes and have developed a universal, inexpensive process that results in good shelf life. A patent has been filed on this process but it has not yet been published. Further, we have developed high-throughput plant screens for induced resistance, increased plant growth and for testing efficacy of spray applications of biocontrol agents. These are being used to screen for strains with improved characteristics and a number of fungal and bacterial agents with very good characteristics already have been identified. Finally, in cooperative work with Korean scientists, a chitinase gene was modified to give increased activity of the protein. This protein may have several uses in agriculture for plant pest control and in other processes.

Impacts
The basic discoveries in this project have demonstrated that our understanding of the nature of biocontrol interactions was, at best, limited. The identification of specific proteins whose expression is altered by the presence of biocontrol organisms gives insights into plant metabolic pathways that are altered by the organisms. These can be used to develop molecular signatures that can provide rapid screening systems for successful biocontrol interactions and that can be used, for example, for screening of superior biocontrol agents, or for identification of effective interactions in the laboratory or even in the field. Better formulations and better strains, plus the molecular approaches identified here, are expected to result in more robust and effective biocontrol systems and for unique methods to increase plant growth and productivity. These will enhance viability of biocontrol companies and commercial agricultural practices.

Publications

• Harman, G. E. 2006. Overview of mechanisms and uses of Trichoderma spp. Phytopathology 96:190-194.
• Park, S. K., Kim, C. W., Kim, H., Jung, J. S. and Harman, G. E. 2007. Cloning and high-level production of a chtinase from Chromobacterium sp. And the role of conserved or nonconserved residues on its catalytic activity. Appl. Microb. Biotechnol. (in press).
• Harman, G. E. and Shoresh, M. 2007. The mechanisms and applications of symbiotic opportunistic plant symbionts. Vurro, M. and Gressel, J. (eds.). NATO Workshop Proceedings (in press).
• Deng, S., Pentilla, M., Lorito, M. and Harman, G. E. 2007. Overexpression of an endochitinase gene (ThEn-42) in Trichoderma atroviride for increased production. Appl. Biochem. Biotechnol. (In press).

Progress 01/01/05 to 12/31/05

Outputs
Trichoderma spp. are now known to be opportunistic plant symbionts. As such, they infect the cortical tissues of plants and induce in many cases increased growth responses and systemic resistance to plant diseases. Even though they colonize and are restricted to roots, they induce an ISR-type response on plant leaves when pathogens attack. The interaction of Trichoderma spp. with plants induces large changes in the plant proteome; in the proteome of at least bean and maize, there are several hundred proteins detected that are not visualized on 2D gels in the absence of the organism. The induced systemic resistance, like that caused by PGPR, frequently occurs in the absence of pathogenesis related proteins or phytoalexin production but when pathogens attack, the resistance is more rapid and more massive than if the Trichoderma is not present. Trichoderma spp., at least some strains, are both biocontrol agents and plant growth stimulants. They are present in high numbers when large numbers of healthy plant roots are present.

Impacts
The agricultural applications of Trichoderma strains are large and growing. By understanding the organism, we can develop systems that will improve plant agriculture both by reducing pesticide use and by increasing plant yields.

Publications

• Donzelli, B. D. D., Seibert, K. J. and Harman, G. E. 2005. Response surface modeling of factors influencing the production of chitinolytic and b-1, 3-glucanolytic enzymes in Trichoderma atroviride strain P1. Enz. Microbial. Technol. 38:1823-1833.
• Chen, J., Harman, G. E., Comis, A., Cheng, G-W. 2005. Proteins related to the biocontrol of Pythium damping-off in maize with Trichoderma harzianum Rifai. J. Integ. Plant Biol. 47: 988-997.

Progress 01/01/04 to 12/31/04

Outputs
Extracellular enzymes from fungi in the genus are highly diverse and are produced abundantly. Among the uses of the chitinolytic complex are those that, when combined with enzymes with similar function, degrade chitin efficiently. This mixture is probably the one known that is capable of complete release of the chitin monomer, N-acetyglucosamine (NAG), from crustacean chitin. We are developing large-scale methods for the enzymatic release of NAG from chitin for pharmaceutical purposes. Further, similar fungal enzymes release certain small molecular substances from fungal cell walls that are potent elicitors of fungal growth and enzyme production. These elicitors are being purified and their uses determined. For T. atroviride, we used response surface modeling techniques to optimize production of glucanolytic and chitinolytic enzyme production.

Impacts
Inflammatory bowel disease (IBD) affects large numbers of people. N-acetyglucosamine has been shown to reduce IBD symptoms. We are developing systems for large scale production of NAG and, in cooperative work, futher developing it as a pharmaceutical for treatment of the disease.

Publications

• Donzelli, B. D. D., Seibert, K. J. and Harman, G. E. 2005. Response surface modeling of factors influencing the production of chitinolytic and b-1, 3-glucanolytic enzymes in Trichoderma atroviride strain P1. Enz. Microbial. Technol. In press.

Progress 01/01/03 to 12/31/03

Outputs
In the past year, we have focused on uses of enzymes from Trichoderma for useful purposes. In one study we have optimized chitinolytic enzyme production from T. atroviride strain P1, using response surface modeling. This has resulted in substantial improvements in enzyme production efficiency. Moreover, we have also discovered enzyme mixtures that efficiently degrade native crustacean chitin to N-acetlglucosamine (NAG). No single organism produces enzymes that totally degrade this native chitin to its monomer but we have discovered that a mixture of fungal enzymes (from Trichoderma strains) and bacterial enzymes (from Steptomyces albidoflavus or Serratia marcescens) are capable of essentially total degradation of chitin to NAG. This is of strong interest since glucosamine, a degraded form of NAG, is widely sold as a nutriceutical. The processes we have developed are much more environmentally friendly than the acid based systems used to produce glucosamine from native chitin. Moreover, NAG is expected to be more effective than glucosamine for diseases such as irritable bowel syndrome. This new method of producing NAG is expected to be commercialized starting in 2004 by Biomarinex, a company with locations in Newfoundland and Geneva, NY.

Impacts
The findings of this research are expected to provide a major new use for shrimp chitin, a resource currently dumped back into the ocean. It also will provide a high quality and quite valuable nutriceutical.

Publications

• Donzelli, B. D. D., Ostroff, G. and Harman, G. E. 2003. Enhanced enzymatic hydrolysis of langostino shell chitin with mixtures of enzymes from bacterial and fungal sources. Carboh. Res. 338:1823-1833.

Progress 01/01/02 to 12/31/02

Outputs
Significant progress was made on commercial uses of microbial enzymes. Response surface modeling approaches were used to optimize chitinolytic and glucanolytic enzyme production from Trichoderma atroviride strain P1. In addition, we have pursued the enzymatic degradation of crustacean shells to produce N-acetylglucosamine as a neutriceutical product. Chitin in this difficult material could be totally degraded to its monomers (N-acetylglucosamine plus a small quantity of glucosamine) by a mixture of enzymes from bacterial and Trichoderma sources. This synergistic mixture was highly effective in this application. In addition, we are developing production systems for hemicellulases and proteases for animal feed additives. Patents have been filed on the N-acetylglucosamine system by Cornell University and commercial development is proceeding.

Impacts
The impact of these developments are expected to be considerable and include the following: 1. The development of large scale commercial enzyme production for a variety of purposes. 2. A significant new industry that will enzymatically produce a very high quality N-acetylglucosamine product. 3. This, in turn, provides more incentives for genome sequencing in important fungi in the genus Trichoderma.

Publications

• No publications reported this period

Progress 01/01/01 to 12/31/01

Outputs
Research on this objective has focused primarily on enzyme production technologies and gene regulation of fungitoxic chitinolytic and glucanolytic enzymes. The genes in general are regulated primarily by the presence or absence of both simple carbon compounds and ammonia levels of the medium. Under permissive nutritional conditions, the presence of chitin may have a secondary inducing role. Exochitinases are also regulated and specifically induced by particular chito-oligomers. In addition, response surface modeling has provided a means to substantially improve levels of enzymes produced by Trichoderma spp. Research also has demonstrated that mixtures of bacterial source and fungal source enzymes are highly synergistic in their ability to degrade native chitin and in release of N-acetylglucosamine from chitin.

Impacts
The enzymes studied have substantial potential for control of plant diseases and for nonagricutural applications. The work in this project is essential to develop systems and enzymes permitting production of high levels of enzymes.

Publications

• Donzelli, B. G. G., Lorito, M., Scala, F. and Harman, G. E. 2001. Cloning, sequence and structure of an gene encoding an antifungal glucan 1,3-b-glucosidase from Trichoderma atroviride (T. harzianum). Gene 277: 199-208.
• Bolar, J. P., Norelli, J. L., Harman, G. E., Brown, S. K. and Aldwinckle, H. S. 2001. Synergistic activity of endochitinase and exochitinase from Trichoderma atroviride (T. harzianum) against the pathogenic fungus Venturia inaequalis in transgenic apple plants. Transgen. Res. 10:533-543.
• Donzelli, B. G. G. and Harman, G. E. 2001. Interaction of ammonium, glucose and chitin regulates the expression of cell wall-degrading enzymes in Trichoderma atroviride strain P1. Appl. Environ. Microbiol. 67:5463-5647.
• Harman, G. E., Broadway, R. M., Tronsmo, A., Lorito, M., Hayes, C. K. and DiPietro, A. Purified chitinases and use thereof. US Patent 6,251,390, issued June 26, 2001.

Progress 01/01/00 to 12/31/00

Outputs
Trichoderma spp. produce numerous enzymes that degrade fungal cell walls. These enzymes are fungitoxic but expected to have little effects on mammals or plants since the substrates for these enzymes largely are lacking in these organisms. We have examined in detail the regulation of genes encoding these enzymes in Trichoderma. We have discovered that regulation is complex. High levels of either ammonia or glucose in the medium are repressive to expression of the enzymes. The presence of inducers such as chitin may have no effect, a small effect or be required for expression of specific genes depending on the nitrogen or carbon status of the medium and the enzyme or gene being measured. No doubt this control of enzyme expression is of ecological importance. For example, in soils the level of available organic carbon and nitrogen source and inorganic nitrogen usually are less than those that permit enzyme expression. In other previous research, we have identified genes that encode specific enzymes that are synergistic in their control of fungi. Recently, we have shown that the enzymes also are synergistic in transgenic plants that produce two of these enzymes. Useful levels of resistance are present in field grown transgenic plants.

Impacts
Cell wall degrading enzymes (CWDEs) are likely to be important in soil ecology and in production of enzymes for use in agriculture and other industries. An understanding of specific regulatory elements permits more intelligent use of the producing fungi in biocontrol and enzyme production. Genes for production of disease-resistant plants also are important to reduce pesticide use in agriculture. Genes from Trichoderma appear to be among the most promising for production of plants resistant to fungal diseases.

Publications

• Bolar, J. P., Norelli, J. L., Wong, K-W, Hayes, C. K., Harman, G. E. and Aldwinckle, H. S. 2000. Expression of endochitinase from Trichoderma harzianum increases resistance to apple scab and reduces vigor. Phytopathology 90: 72-77.
• Harman, G. E., Hayes, C. K., Lorito, M.and Tronsmo, A and Klemsdhal, S. 6,020,540. Gene encoding for endochitinase, CIP to US Patent 5,378,821, Issued Feb. 1, 2000.
• Broadway, R. M. and Harman, G. E. 2000. 6,029, 299. Fungus and Insect Control with Chitinolytic Enzymes, issued May 30, 2000.

Progress 01/01/99 to 12/31/99

Outputs
Genes from biocontrol fungi in the genus Trichoderma have shown promise as transgenes to produce plants highly resistant to fungal diseases. The genes thus far identified encode antifungal proteins with high levels of synergy between themselves and also with membrane active compounds. Two of these, an endochitinase and an exochitinase, have been inserted into apple singly and in combination. Transgenic apple lines with either gene singly are more resistant to Venturia inaequalis than nontransformed lines. The level of resistance was strongly correlated with levels of expression in individual lines. Surprisingly, however, the endochitinase gene reduced vigor of transformed lines, also in a dose-dependent manner. The exochitinase was less effective than the endochitinase in reducing disease but did not affect plant growth. More importantly, expression of both enzymes demonstrated in planta synergy. Thus, it was possible to select lines with moderate levels of exochitinase expression and low levels of endochitinase expression that were strongly resistant to the pathogen. This is the first report of disease resistance enhanced by an exochitinase or of in planta synergy from the combined effects of two different classes of chitinases. Further, a gene encoding another antifungal protein, a beta 1,3 glucosidase was cloned and sequenced and is being mobilized for plant transformation. The control of these antifungal proteins in the host fungus also has been studied. This regulatory elements of the beta 1,3 glucanase are unusual and may suggest regulation to provide more than one physiological function in the fungus.

Impacts
Genes to confer resistance to plant pests, as well as other useful traits, are desirable both to reduce agriculture's dependence upon pesticides, to reduce farmer's production costs and to mitigate worker safety issues. In spite of the current furor over GMOs, transgenic crops with benefit to agriculture and consumers will be important components of future agricultural practices.

Publications

• Woo, S. L., B. Donzelli, B., Scala, F., Mach, R., Harman, G. E., Kubicek ,C. P., Del Sorbo, G., and Lorito, M. 1999. Disruption of ech42 (endochitinase-encoding) gene affects biocontrol activity in Trichoderma harzianum strain P1. Molec. Plant Microbe Interact. 12:419-429.
• Bolar, J. P., Norelli, J. L., Wong, K-W, Hayes, C. K., Harman, G. E. and Aldwinckle, H. S. 2000. Expression of endochitinase from Trichoderma harzianum increases resistance to apple scab and reduces vigor. Phytopathology 90: 72-77.

Progress 01/01/98 to 12/31/98

Outputs
Research on the project has focused on the cloning of the beta-1,3-glucosidase gene from Trichoderma, and this was recently completed. Other genes and gene products with strong antifungal activity have been identified in the past and additional ones are being identified now. There are several uses for the genes and gene products including direct topical application of the proteins to control diseases, including postharvest diseases of fruits and vegetables. Methods for production of useful material using transgenic and nontransgenic fungi have been developed. Further, the genes have been expressed in plants where they confer a high level of disease resistance. "Knockout" mutants of Trichoderma lacking ability to espress endochitinase genes have been produced; they have reduced ability to protect plants against Botrytis cinerea but have enhanced ability to control Rhizoctonia solani.

Impacts
(N/A)

Publications

• Kubicek, C. P. and Harman, G. E. 1998. Trichoderma and Gliocladium, Vol. 1. Taylor & Francis, London. 278 pg.
• Harman, G. E. and Kubicek, C. P. 1998. Trichoderma and Gliocladium, Vol. 2. Taylor & Francis, London. 393 pg.
• Lorito, M., Woo, S. L., Fernandez, I. G., Colucci, G., Harman, G. E., Pintor-Toro, J. A., Fillipone, E., Muccifora, S., Lawrence, C. B., Zoina, A., Tuzun, S., and Scala, F. 1998. Genes from mycoparasitic fungi as a souce for improving plant resistance to fungal pathogens. Proc. Natl. Acad. Sci., USA 95:7860-7865.
• Woo, S. L., B. Donzelli, B., Scala, F., Mach, R., Harman, G. E., Kubicek ,C. P., Del Sorbo, G., and Lorito, M. 1999. Disruption of ech42 (endochitinase-encoding) gene affects biocontrol activity in Trichoderma harzianum strain P1. Molec. Plant Microbe Interact. (in press).

Progress 01/01/97 to 12/31/97

Outputs
Research on this project is designed to discover, identify, and characterize and use antifungal genes and gene products to control plant diseases. Research has focused primarily on chitinases and beta-1,3-glucosidases. Previous research has indicated that some forms of these enzymes are strongly antifungal. There are various methods whereby these genes and enzymes may be used. The enzymes themselves may be effective as topical applications for various purposes, including control of post-harvest diseases of fruits. The genes may be used in producing transgenic plants that are resistant to plant diseases. Finally, the genes may be used to produce transgenic microbes that are better biocontrol agents. All approaches have been successful and some applications will be described in the report to project NYG6323555. In 1997, we have cloned and sequenced an additional strongly antifungal chitinase from the bacterium Chromobacterium violaceum. Further, we have produced transgenic plants of a number of crop species using single and multiple genes from the biocontrol fungus Trichoderma harzianum. Fungal genes impart substantial resistance to a number of plant pathogenic fungi, including species of Alternaria, Botrytis, and Rhizoctonia.

Impacts
(N/A)

Publications

• Kubicek, C. P. and Harman, G. E. 1998. Trichoderma and Gliocladium, Vol. 1. Taylor & Francis, London, in press.
• Harman, G. E., Hayes, C. K., and Ondik, K. L. 1998. Asexual genetics in Trichoderma and Gliocladium: Mechanisms and implications. p. 243-270. In Kubicek, C. P. and Harman, G. E., eds, Trichoderma and Gliocladium. Vol. 1. Taylor & Francis, London, in press.

Progress 01/01/96 to 12/30/96

Outputs
Research on this topic is designed to discover, identify, and characterize antifungal gene proteins, primarily from biocontrol fungi, to clone and sequence the genes encoding the proteins, and to use these genes in biorational and genetic approaches to plant pest control. In the past year, genes were sequenced and used for various purposes. One of these was to place the genes into various plants to attempt to produce disease-resistant crops. Research was conducted in cooperation with a number of other labs, and involved work with grapes, apples, tomatoes, tobacco, Brassica spp. and other crops. The genes inserted were from the biocontrol fungus Trichoderma harzianum, and these genes express good levels of antifungal protein in plants. Some of these plants possess enhanced levels of resistance to fungal diseases relative to nontransformed plants of the same varieties.

Impacts
(N/A)

Publications

• Lorito, M., D Ambrosia, M., Woo, S. L., Harman, G. E., Hayes, C. K., and Scala, F. 1996. Synergistic interaction between cell wall-degrading enzymes and membrane-affecting compounds. Molec. Plant Microbe Interact. 3:206-213.
• Klemsdahl, S. S., Hayes, C. K., Hjeljord, L., Harman, G. E., and Tronsmo, A. 1996. Isolation and characterization of of a cDNA from Trichoderma harzianum P1 encoding a 14.3.3 protein homologue. Gene 171:123-127.
• Margolles-Clark, E., Harman, G. E., and Pentilla, M. 1996. Enhanced expression of endochitinase in T. harzianum using the cbh 1 promoter of T. reesei. Appl. Env.. Micro. 62:2152-2155.
• Margolles-Clark, E., Harman, G. E., Hayes, C. K. and Pentilla, M. 1996. Improvedproduction of Trichoderma harzianum endochitinase by expression in T. reesei. Appl. Environ. Microbiol. 62:2145-2151.
• Peterbauer, C., Lorito, M., Hayes, C. K., Harman, G. E., and Kubicek, C. K. 1996. Molecular cloning and expression of nag 1 (N-acetyl-beta-D-glucosaminidase-encoding) gene from Trichoderma harzianum P1. Curr. Genet. 30:325-331.

Progress 01/01/95 to 12/30/95

Outputs
Research in 1995 dealt with discovery and use of genes and gene products for thecontrol of plant diseases. Cooperative research has demonstrated a number of synergistic combinations of proteins that are likely to be highly useful in plant disease control. Genes coding for a number of these proteins have been cloned and sequenced. These genes are being used for plant transformation and for preparing large quantities of enzymes in transgenic microorganisms. Publication of details of these discoveries await patent filings. Patent/Application No. 5378821, 01/03/95; 5433947, 07/17/95.

Impacts
(N/A)

Publications

Progress 01/01/94 to 12/30/94

Outputs
Primary activity on this project has been to characterize proteins with antifungal activity, to clone and sequence genes coding for highly active proteins, and to examine synergy between these proteins and also between other factors. In addition, we wished to develop large-scale production methods for these antifungal proteins in order to obtain sufficient enzyme for large-scale in situ tests. The antifungal proteins of primary interest are enzymes that degrade chitin and #-1,3-glucosidases. A gene coding for the endochitinase of T. harzianum strain P1 was cloned and sequenced and portions of genes coding for several other enzymes have been cloned and sequenced. In addition, strongly antifungal enzymes from the bacterium Streptomyces albidoflavus have been identified. In cooperative work, methods for large-scale enzyme production largely have been developed. Synergies between chitinolytic enzymes and antibiotics produced by T. harzianum have been identified. Patent/Invention No.: 5360608. Date: July 5, 1994.

Impacts
(N/A)

Publications

Progress 01/01/93 to 12/30/93

Outputs
Enzymes that catalyze the degradation of cell walls of pathogenic fungi were purified and characterized from the biocontrol fungi TRICHODERMA HARZIANUM and GLIOCLADIUM VIRENS. The enzymes were shown to be complex mixtures. Each enzyme has activity against a wide range of fungi, and more importantly, they are strongly synergistic. Mixtures of enzymes provide provide strong antifungal activity at 1-10 ppm, while each individual enzyme requires 50 to 150 ppm to achieve the same level of control. These synergistic combinations probably effective at lower concentrations and control a wider range of pathogenic fungi than similar enzymes from any other source. Further, they are synergistic with fungicides and biocontrol bacteria. The gene coding for one enzyme, and endochitinase, has been cloned and sequenced. These findings permit the development of new plant pest strategies using the enzymes themselves, either alone or in combination with fungicides, and also for the development of superior transgenic biocontrol agents and transgenic plants resistant to plant diseases.

Impacts
(N/A)

Publications