Expression of organophosphate hydrolase in the filamentous fungus Gliocladium virens

Dave, Kirti I.; Lauriano, Crystal M.; Xu, Ben; Wild, James R.; Kenerley, Charles M.

doi:10.1007/bf00221231

Cited by 30 publications

(5 citation statements)

References 19 publications

(21 reference statements)

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“…Stable cotransformants were selected on PDA plates supplemented with hygromycin (Dave et al, 1994). Resistant colonies were screened for the presence of green fluorescence and the insertion of pTEFEGFP was further analyzed by Southern-blot analysis.…”

Section: Gfp-tagged T Virens Strain Constructionmentioning

confidence: 99%

A Proteinaceous Elicitor Sm1 from the Beneficial FungusTrichoderma virensIs Required for Induced Systemic Resistance in Maize

et al. 2007

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We have previously shown that the beneficial filamentous fungus Trichoderma virens secretes the highly effective hydrophobinlike elicitor Sm1 that induces systemic disease resistance in the dicot cotton (Gossypium hirsutum). In this study we tested whether colonization of roots by T. virens can induce systemic protection against a foliar pathogen in the monocot maize (Zea mays), and we further demonstrated the importance of Sm1 during maize-fungal interactions using a functional genomics approach. Maize seedlings were inoculated with T. virens Gv29-8 wild type and transformants in which SM1 was disrupted or constitutively overexpressed in a hydroponic system or in soil-grown maize seedlings challenged with the pathogen Colletotrichum graminicola. We show that similar to dicot plants, colonization of maize roots by T. virens induces systemic protection of the leaves inoculated with C. graminicola. This protection was associated with notable induction of jasmonic acidand green leaf volatile-biosynthetic genes. Neither deletion nor overexpression of SM1 affected normal growth or development of T. virens, conidial germination, production of gliotoxin, hyphal coiling, hydrophobicity, or the ability to colonize maize roots. Plant bioassays showed that maize grown with SM1-deletion strains exhibited the same levels of systemic protection as nonTrichoderma-treated plants. Moreover, deletion and overexpression of SM1 resulted in significantly reduced and enhanced levels of disease protection, respectively, compared to the wild type. These data together indicate that T. virens is able to effectively activate systemic disease protection in maize and that the functional Sm1 elicitor is required for this activity.

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Section: Gfp-tagged T Virens Strain Constructionmentioning

confidence: 99%

A Proteinaceous Elicitor Sm1 from the Beneficial FungusTrichoderma virensIs Required for Induced Systemic Resistance in Maize

et al. 2007

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“…The genetically engineered strain, GvT6, was constructed by transformation and heterologous integration of the plasmid pCL1, which included the hygromycin resistance gene, hygB, and a gene encoding organophosphate hydrolase (OPH), opd. GvT6 was selected based on high expression levels of the OPH and stable hygromycin resistance [43]. Mutant strain Tv10.4 was created by a single point mutation in the arg 2 gene, resulting in an inability to grow on media lacking arginine [20].…”

Section: Strains and Cultural Conditionsmentioning

confidence: 99%

“…Conidia of T. virens were incorporated at a rate of 10 8 conidia per gram of solution containing 1% sodium alginate (medium viscosity, Sigma A2033), 20% polyethylene glycol (PEG 8000) and 2% ground wheat bran (all on w/w basis) [43,45]. The solution was allowed to drip through large orifice pipette tips into a 0.25M solution of calcium chloride.…”

Section: Production Of Alginate Prillsmentioning

confidence: 99%

Competitiveness of a Genetically Engineered Strain of Trichoderma virens

Weaver¹,

Kenerley

2008

Mycopathologia

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The intraspecific competitiveness of a genetically engineered strain of Trichoderma virens was assessed relative to the non-transformed, progenitor strain and an isogenic, auxotrophic strain using a replacement series design. The transformed strain was less fit, but appeared more competitive than the wild type or the auxotroph in this assay. To validate this finding and to evaluate the ability of a strain to establish in an environment occupied by a competitor another experimental approach was developed. In various treatments the transformed strain was unaffected or only slightly inhibited by a competing strain. In contrast, the wild type and the auxotrophic strain were strongly inhibited by the presence of the transformed strain. These findings support the conclusion that this genetically engineered strain is more competitive than the wild-type strain and thus may be persistent in the environment.

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“…An opd transformant (GvT6) of T. virens was used throughout this study [31]. The fungal culture was grown on potato dextrose agar (PDA) medium containing 0.1 g l 3I of hygromycin.…”

Section: Fungal Strain and Soil Inoculationmentioning

confidence: 99%

“…The objective of our research was to design an appropriate protocol for extraction of fungal DNA from soil and to develop a QC-PCR technique that would allow us to monitor the population of a genetically modi¢ed strain of T. virens among the soil micro£ora. Dave et al [31] have transformed a wild-type strain of T. virens (Gv29-8) with a plasmid carrying the bacterial opd (organophosphate degrading) gene for potential use in the bioremediation of soil contaminated with organophosphate pesticides. One of these transformants (GvT6) was used to develop a QC-PCR technique for soilborne fungi.…”

Section: Introductionmentioning

confidence: 99%

Detection and enumeration of a genetically modified fungus in soil environments by quantitative competitive polymerase chain reaction

Baek

1998

FEMS Microbiology Ecology

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A procedure of quantitative competitive polymerase chain reaction was developed for detection and quantitation of a genetically modified strain of Trichoderma virens (GvT6) in the soil environment. A 1.0-kb coding region of the opd (organophosphate degrading) gene on GvT6 genomic DNA was co-amplified with a standard DNA. The standard DNA was a 1.5-kb fragment of the gpd (glyceraldehyde-3-phosphate dehydrogenase) gene of T. virens containing opd-primer-binding sites on the ends. Soil samples containing various types of propagules of GvT6 were treated with a combined procedure of SDSheating and bead-beating and the released nucleic acids were extracted and purified. Relative efficiency of T. virens DNA recovery from soil samples was about 85^95% compared with DNA yield from spore suspensions. Following the PCR, the product ratio between the target and the standard was plotted against a dilution series of the standard which was added into the amplification reaction. Then, the concentration of the opd gene and the GvT6 in the soil sample were estimated. Despite the differences in size and nucleotide sequences between the target and the standard, they were amplified with similar efficiency. The detection limit of the PCR was 10^1000 times lower compared to traditional dilution plating. The quantitative competitive PCR technique will enhance our ability to understand the ecology of soilborne fungi in the soil environment. z 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V.

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Expression of organophosphate hydrolase in the filamentous fungus Gliocladium virens

Cited by 30 publications

References 19 publications

A Proteinaceous Elicitor Sm1 from the Beneficial FungusTrichoderma virensIs Required for Induced Systemic Resistance in Maize

A Proteinaceous Elicitor Sm1 from the Beneficial FungusTrichoderma virensIs Required for Induced Systemic Resistance in Maize

Competitiveness of a Genetically Engineered Strain of Trichoderma virens

Detection and enumeration of a genetically modified fungus in soil environments by quantitative competitive polymerase chain reaction

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