The purpose of this plan first is to promote the plant pathogens detection and isolation efficiency of the laboratory. This year,regarding TAF-certification for laboratory equipment, design and layout of the laboratory has already been completed and laboratory bench and instruments have been acquired, calibrated and approved for operations.Second purpose of this plan, with reference to published literature,detection methods for Cucurbit chlorotic yellows virus, CCYV and Strawberry latent ringspot virus, SLRSV have been researched and cataloged. Current detection methods regarding Cucurbit chlorotic yellows virus, CCYV include: RT-PCR, Western blotting, ELISA, and Immunoelectron microscopy; for Strawberry latent ringspot virus, SLRSV, current detection methods include: RT-PCR, IC-RT-PCR, ELISA as well as symptom diagnosis. Utilizing various detecting methods on different samples, the most suitable detection method will be chosen as the standard detection procedure. One required index of high quality of seed/seedling is genetic purity, especially for hybrid seed. Genetic purity in a commercial hybrid of solanaceous crops is very important. This year, we setup this quality control mode by using molecular markers in seed production system of the crops of tomato and pepper, last year we did some part work in tomato. We obtain seven sets of SCAR primer for six specific DNA fragments in tomato and ten sets of SCAR primer for six specific DNA fragments in pepper,they can identify hybrid from parents. The SCAR primer can have the sharp and easy-read DNA fragment after PCR amplification and electrophoresis.We will collect these SCAR primers as members of data base for testing F1 hybrid seed purity in tomato and pepper till year 2013. Last object of this program is for the ISTA accreditation on the seed testing of Xanthomonas campestris pv. campestris (Xcc) which causes black rot of crucifers. The Xcc testing procedure which conforms to ISTA rules has been established. Artificial infested seeds were adopted to check the Xcc testing procedure. On FS and mCS20ABN selective media, Xcc showed individully pale green and pale yellow mucoid colonies. The cabbage leaves that were stab-inoculated with Xcc showed typical yellowing necrotic V-shaped symptoms in pathogenicity test. The Xcc-carried ratio of 0.02% could be detected in sensitivity test.
The purpose of this plan first is to promote the plant pathogens detection and isolation efficiency of the laboratory. This year,regarding TAF-certification for laboratory equipment, design and layout of the laboratory has already been completed and laboratory bench and instruments have been acquired, calibrated and approved for operations.Second purpose of this plan, with reference to published literature,detection methods for Cucurbit chlorotic yellows virus, CCYV and Strawberry latent ringspot virus, SLRSV have been researched and cataloged. Current detection methods regarding Cucurbit chlorotic yellows virus, CCYV include: RT-PCR, Western blotting, ELISA, and Immunoelectron microscopy; for Strawberry latent ringspot virus, SLRSV, current detection methods include: RT-PCR, IC-RT-PCR, ELISA as well as symptom diagnosis. Utilizing various detecting methods on different samples, the most suitable detection method will be chosen as the standard detection procedure. One required index of high quality of seed/seedling is genetic purity, especially for hybrid seed. Genetic purity in a commercial hybrid of solanaceous crops is very important. This year, we setup this quality control mode by using molecular markers in seed production system of the crops of tomato and pepper, last year we did some part work in tomato. We obtain seven sets of SCAR primer for six specific DNA fragments in tomato and ten sets of SCAR primer for six specific DNA fragments in pepper,they can identify hybrid from parents. The SCAR primer can have the sharp and easy-read DNA fragment after PCR amplification and electrophoresis.We will collect these SCAR primers as members of data base for testing F1 hybrid seed purity in tomato and pepper till year 2013. Last object of this program is for the ISTA accreditation on the seed testing of Xanthomonas campestris pv. campestris (Xcc) which causes black rot of crucifers. The Xcc testing procedure which conforms to ISTA rules has been established. Artificial infested seeds were adopted to check the Xcc testing procedure. On FS and mCS20ABN selective media, Xcc showed individully pale green and pale yellow mucoid colonies. The cabbage leaves that were stab-inoculated with Xcc showed typical yellowing necrotic V-shaped symptoms in pathogenicity test. The Xcc-carried ratio of 0.02% could be detected in sensitivity test.