a. 符合目標市場外銷國之種子苗驗證技術盤點與研發。By papers literature research, we selected the RT-PCR as the major tech as our TAF testing method for Tomato Mosaic Virus. At the first we design a set of RT-PCR primer for ToMV specific primer set. We also design the internal control primer set for more reliable detection result. In 2012,the stand operation procedure of ToMV detection of Taiwan Accreditation Foundation, Taiwan Accreditation Foundation (TAF) was established. ToMV Detection of Tomato leaf in-House Method(Document No.:No.WI33).。b. 外銷重要蔬菜番茄及甜椒種子處理技術之研究。The methods including electrolyzed KCl water solution, hot water and bactericides were adopted to study the seed disinfection treatments for decreasing the seed transmission and primary inoculum of bacterial spot of tomato and sweet pepper. The electrolyzed KCl water solution with fifty ppm and above of HClO concentration were effectively to disinfect the artificial infested sweet pepper and tomato seeds that were treated five and above minutes. The treatments of thirty and above minutes with 45°C hot water obviously decreased the Xv-carried ratio of artificial infested sweet pepper seeds. The treatment of ten minutes with 50°C hot water could totally disinfect the Xv on artificial infested sweet pepper seeds and, meanwhile, that did not reduce the germination rate. In the trial of varied bactericides, the treatment with 0.6% sodium chloride showed the best disinfection on artificial infested sweet pepper seeds,and the treatment with Kasugamycin and copper oxychloride could effectively reduce Xv-carried ratio and even thoroughly disinfect the artificial infested sweet pepper seeds without diminishing the germination rate.。c. 外銷蔬菜種子品質提升技術研究 (委辦)。The objective of this study was to establish a disinfection method for seeds of Crucifer vegetables for preventing black rot disease caused by Xanthomonas campestris pv. campestri (XCC). As applied the previous established protocol of ‘ 30 seconds of 3% H2O2 followed by heated dry at 70oC for 40 min’ to large scale experiment (1 Kg seeds) could completely disinfect the pathogen carried seeds; however, the germination rate dropped from 80% to 45%. Considering the seed quality per se, the seed materials were switched to high quality seeds of cauliflower and cabbage with germination rate higher than 95%. Those seeds were first contaminated with Xanthomonas campestris pv. campestri (XCC1-1) before disinfection treatments including ‘ 30 seconds of 3% H2O2 followed by heated dry at 70oC for 40 min’, ‘heated at 70oC for 7 days’, ‘hot ZnSO4’ , and the seeds were storage at electronic dry cabinet. The germination rate was maintained rather constant at 96% for 6 months of monthly check, which indicated that seed germination did not significantly influence by all the treatments. For the XCC1-1 disinfection, the treatment of ‘heated at 70oC for 7 days’ was the best for the seeds’ wash-through remained 0 detection for 6 months of monthly check. The results of the other 2 treatments were not as complete and constant. Therefore, we concluded that ‘heated the seeds at 70oC for 7 days’ was a valuable methods, which is simple and easy to apply to large amount of seeds. The morphology of the seedlings evaluated 2 weeks after germination was normal but the seedling index was smaller in cauliflower but not cabbage; therefore, further growth should be evaluated for the side effect on mature plants.。d. 建立重要蘭花病毒ORSV、CymMV新檢測技術。By phage display system, there were three kinds of phage library being selected by affinity panning selection using the phage-12 library. In the phage-12 library system, there were 20 phage populations being selected to do the indirect ELISA test for CymMV coat protein. By the ELISA test result, the specific peptides regions of these 6 phage populations were sequencing.。e. 建立苦瓜種子發芽檢查程序研究 (委辦)。Bitter gourd seed has a hard and thick seed coat which is impervious to water and gases exchange that result in un-uniform germination, low germinates rate, and fungi infection. This study evaluated the effects of warm water soaking, running water immersion, germination temperature, and media on germinating bitter gourd seeds. Results indicated that significant improvements in germination rate were observed for 60 °C warm water soaking for 10 min as compared with running water immersion. The optimum seed germination temperature was 30 ℃. The germination rate was increased to 80% when using between of paper treatment with 4 c.c. imbibition water. We sent three level vigors of bitter gourd seeds to three different laboratories. Analyse the data and results were within the tolerance range. It suggested that the pre-germination treatment of bitter gourd seed was suitable for International Seed Testing Association (ISTA) accreditation to update the germination rule for improving the germination test for bitter gourd。
a. 符合目標市場外銷國之種子苗驗證技術盤點與研發。By papers literature research, we selected the RT-PCR as the major tech as our TAF testing method for Tomato Mosaic Virus. At the first we design a set of RT-PCR primer for ToMV specific primer set. We also design the internal control primer set for more reliable detection result. In 2012,the stand operation procedure of ToMV detection of Taiwan Accreditation Foundation, Taiwan Accreditation Foundation (TAF) was established. ToMV Detection of Tomato leaf in-House Method(Document No.:No.WI33).。b. 外銷重要蔬菜番茄及甜椒種子處理技術之研究。The methods including electrolyzed KCl water solution, hot water and bactericides were adopted to study the seed disinfection treatments for decreasing the seed transmission and primary inoculum of bacterial spot of tomato and sweet pepper. The electrolyzed KCl water solution with fifty ppm and above of HClO concentration were effectively to disinfect the artificial infested sweet pepper and tomato seeds that were treated five and above minutes. The treatments of thirty and above minutes with 45°C hot water obviously decreased the Xv-carried ratio of artificial infested sweet pepper seeds. The treatment of ten minutes with 50°C hot water could totally disinfect the Xv on artificial infested sweet pepper seeds and, meanwhile, that did not reduce the germination rate. In the trial of varied bactericides, the treatment with 0.6% sodium chloride showed the best disinfection on artificial infested sweet pepper seeds,and the treatment with Kasugamycin and copper oxychloride could effectively reduce Xv-carried ratio and even thoroughly disinfect the artificial infested sweet pepper seeds without diminishing the germination rate.。c. 外銷蔬菜種子品質提升技術研究 (委辦)。The objective of this study was to establish a disinfection method for seeds of Crucifer vegetables for preventing black rot disease caused by Xanthomonas campestris pv. campestri (XCC). As applied the previous established protocol of ‘ 30 seconds of 3% H2O2 followed by heated dry at 70oC for 40 min’ to large scale experiment (1 Kg seeds) could completely disinfect the pathogen carried seeds; however, the germination rate dropped from 80% to 45%. Considering the seed quality per se, the seed materials were switched to high quality seeds of cauliflower and cabbage with germination rate higher than 95%. Those seeds were first contaminated with Xanthomonas campestris pv. campestri (XCC1-1) before disinfection treatments including ‘ 30 seconds of 3% H2O2 followed by heated dry at 70oC for 40 min’, ‘heated at 70oC for 7 days’, ‘hot ZnSO4’ , and the seeds were storage at electronic dry cabinet. The germination rate was maintained rather constant at 96% for 6 months of monthly check, which indicated that seed germination did not significantly influence by all the treatments. For the XCC1-1 disinfection, the treatment of ‘heated at 70oC for 7 days’ was the best for the seeds’ wash-through remained 0 detection for 6 months of monthly check. The results of the other 2 treatments were not as complete and constant. Therefore, we concluded that ‘heated the seeds at 70oC for 7 days’ was a valuable methods, which is simple and easy to apply to large amount of seeds. The morphology of the seedlings evaluated 2 weeks after germination was normal but the seedling index was smaller in cauliflower but not cabbage; therefore, further growth should be evaluated for the side effect on mature plants.。d. 建立重要蘭花病毒ORSV、CymMV新檢測技術。By phage display system, there were three kinds of phage library being selected by affinity panning selection using the phage-12 library. In the phage-12 library system, there were 20 phage populations being selected to do the indirect ELISA test for CymMV coat protein. By the ELISA test result, the specific peptides regions of these 6 phage populations were sequencing.。e. 建立苦瓜種子發芽檢查程序研究 (委辦)。Bitter gourd seed has a hard and thick seed coat which is impervious to water and gases exchange that result in un-uniform germination, low germinates rate, and fungi infection. This study evaluated the effects of warm water soaking, running water immersion, germination temperature, and media on germinating bitter gourd seeds. Results indicated that significant improvements in germination rate were observed for 60 °C warm water soaking for 10 min as compared with running water immersion. The optimum seed germination temperature was 30 ℃. The germination rate was increased to 80% when using between of paper treatment with 4 c.c. imbibition water. We sent three level vigors of bitter gourd seeds to three different laboratories. Analyse the data and results were within the tolerance range. It suggested that the pre-germination treatment of bitter gourd seed was suitable for International Seed Testing Association (ISTA) accreditation to update the germination rule for improving the germination test for bitter gourd。