Progress 10/01/09 to 09/30/10
Outputs
Progress Report Objectives (from AD-416) Develop a real-time PCR procedure for quantificatioin of Verticillium dahliae. Approach (from AD-416) A real-time PCR technique based on TaqMan chemistry will be developed to identify and quantify inoculum of Verticillium dahliae in the soil. This procedure will validate against a range of soilborne fungi to ensure specificity. Documents Reimbursable with CA Strawberry Commission. Log 37807. The objective of the project is to develop a molecular marker system for identification and quantification of Verticillium dahliae from soil samples. A region of the genome was identified where differences were observed between V. dahliae and related species and PCR primers were developed. Real-time PCR detection techniques (both SYBR green and TaqMan chemistries) were validated for species-specificity. Work was conducted to optimize DNA extraction techniques from soil to inhibitors that can reduce the efficiency of PCR amplification and an internal control was developed to assess PCR amplification efficiency. The technique has been validated by comparing the results with pathogen inoculum levels determined by soil plating and was found to be highly accurate down to below 2 microsclerotia/g soil (r2 = .956). Communication on the project status is done with the granting agency by e- mail, personal conversations and submission of written yearly progress reports to the CSC.
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Progress 02/01/09 to 01/31/10
Outputs
Progress Report Objectives (from AD-416) Develop a real-time PCR procedure for quantificatioin of Verticillium dahliae. Approach (from AD-416) A real-time PCR technique based on TaqMan chemistry will be developed to identify and quantify inoculum of Verticillium dahliae in the soil. This procedure will validate against a range of soilborne fungi to ensure specificity. The objective of the project is to develop a molecular marker system for identification and quantification of Verticillium dahliae from soil samples. A region of the genome was identified where differences were observed between V. dahliae and related species and polymerase chain reaction (PCR) primers were developed. Real-time PCR detection techniques (both SYBR green and TaqMan chemistries) were validated for species- specificity. DNA extraction techniques from soil were optimized to eliminate inhibitors that can reduce the efficiency of PCR amplification and an internal control was developed to assess PCR amplification efficiency. The technique has been validated by comparing the results with pathogen inoculum levels determined by soil plating and was found to be highly accurate down to below 2 microsclerotia/g soil (r2 = 0.956). Communication on the project status is done with the granting agency by e- mail, personal conversations and submission of written yearly progress reports to the CSC.
Impacts
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Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) Develop a real-time PCR procedure for quantificatioin of Verticillium dahliae. Approach (from AD-416) A real-time PCR technique based on TaqMan chemistry will be developed to identify and quantify inoculum of Verticillium dahliae in the soil. This procedure will validate against a range of soilborne fungi to ensure specificity. Documents Reimbursable with CA Strawberry Commission. Log 37807. Significant Activities that Support Special Target Populations The objective of the project is to develop a molecular marker system for identification and quantification of Verticillium dahliae from soil samples. A region of the genome was identified where differences were observed between V. dahliae and related species and PCR primers were developed. Real-time PCR detection techniques (both SYBR green and TaqMan chemistries) were validated for species-specificity. Work was conducted to optimize DNA extraction techniques from soil to inhibitors that can reduce the efficiency of PCR amplification and an internal control was developed to assess PCR amplification efficiency. Communication on the project status is done with the granting agency by e- mail, personal conversations and submission of written yearly progress reports to the CSC.
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Publications
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