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植物種苗團隊-動植物有害生物診斷鑑定技術及種子苗病害驗證技術之研究與應用繁殖技術課Establishment and Resea

「植物種苗團隊-動植物有害生物診斷鑑定技術及種子苗病害驗證技術之研究與應用繁殖技術課Establishment and Resea」於資料集「種苗研發成果」由單位「農業部」的陳乃華小姐所提供，聯繫電話是04-2582-5425，最近更新時間為：2023-07-13 01:04:11。欄位年度的內容是2012 ，欄位計畫名稱_中文的內容是植物種苗團隊-動植物有害生物診斷鑑定技術及種子苗病害驗證技術之研究與應用 ，欄位計畫名稱_英文的內容是Establishment and Research of Seed and Seedling Certification Techniques and Regulations ，欄位計畫編號_中文的內容是101農科-10.3.1-種-X3 ，欄位計畫編號_英文的內容是101AS -10.3.1-SS-X3 ，欄位主辦單位_中文的內容是繁殖技術課 ，欄位主辦單位_英文的內容是Propagation Technology Section ，欄位主辦人_中文的內容是邱燕欣 ，欄位主辦人_英文的內容是Yen-Hsin Chiu ，欄位中文摘要的內容是a.符合目標市場外銷國之種子苗驗證技術盤點與研發文獻搜尋並比較國際試驗室間檢測番茄嵌紋病毒(Tomato Mosaic Virus，ToMV)的檢測方式保括ELISA、RT-PCR、ICRT-PCR等檢測技術，本計畫前期合成針對ToMV專一之引子對以及植體之Internal control primer set, 可建立RT-PCR偵測受檢檢體內之ToMV，確定檢測之正確率。101年擬定檢查流程文件-番茄植體檢測番茄嵌紋病毒作業標準一式，並通過財團法人全國認證基金會(Taiwan Accreditation Foundation,TAF)技術認證。 b.外銷重要蔬菜番茄及甜椒種子處理技術之研究本計畫利用氯化鉀電解水、熱水及殺菌劑等方法，研究最適處理條件進行種子滅菌處理，以減少茄科細菌性斑點病之種傳情形與其初次接種源。以HClO濃度50 ppm以上的氯化鉀電解水處理5分鐘以上即可有效除滅甜椒種子與番茄種子上的病原菌；45°C熱水處理30分鐘以上可明顯降低甜椒種子上的帶菌情形，甚至50°C熱水僅處理10分鐘即可完全除滅甜椒種子上的病原菌，同時不降低種子發芽率。殺菌劑試驗中以0.6%漂白水滅菌效果最好，而嘉賜銅處理，除有效降低帶菌情形、甚至完全除滅病原菌外，同時也不會降低種子的發芽率。 c.外銷蔬菜種子品質提升技術研究 (委辦) 本研究的目的在開發不影響種子發芽率之種子殺菌技術，以防治十字花科黑腐病。應用100年度建立之條件『3% H2O2浸漬30秒再以70℃熱風烘乾種子30分鐘』處理1Kg天然帶黑腐病菌種子，結果顯示雖然可以完全殺滅黑腐病菌，但種子發芽率由80%降低至45%。考慮種子原先品質對試驗結果之影響，本年度改用發芽率95%以上之花椰菜及甘藍菜種子，經Xanthomonas campestris pv. Campestri (XCC1-1 strain)污染作為本試驗帶菌種子材料，殺菌處理分別為：『3% H2O2浸漬30秒再以70℃熱風烘乾種子30分鐘』、經70oC乾熱處理7天，及熱硫酸鋅，種子處理後分裝儲藏於防潮箱，每月追蹤其發芽率及帶菌情形。結果顯示所有處理對發芽率皆無顯著不良影響，發芽率穩定維持在95%以上，滅菌效果則以70oC乾熱處理7天為最佳，連續6個月種子洗出液之XCC1-1皆維持零檢出，其他處理之殺菌效果較不完全、也較不穩定。因此，本試驗的結論是『70oC乾熱處理7天』處理為最佳條件方便於大量種子處理，對發芽後兩週幼苗之外觀沒有影響，但估算其狀苗指數，對甘藍沒有影響，但對花椰菜則會降低，因此後續對成株之影響仍需評估。 d.建立重要蘭花病毒ORSV、CymMV新檢測技術建立胜肽親和性篩選系統(peptides 12)對於標地蛋白之親和性汰選技術，建立及生產與CymMV病毒鞘蛋白具親和性之噬菌體，篩選出可利用於ELIS檢測CymMV之噬菌體。 e.建立苦瓜種子發芽檢查程序研究 (委辦)苦瓜種子種皮厚且硬，會妨礙水分吸收及氣體交換，發芽過程中常有發霉的現象，造成種子發芽率不佳及不整齊等問題。本實驗以四品種苦瓜為研究材料，結果顯示苦瓜種子發芽最適溫度為30℃，最適發芽前處理為60℃溫湯浸種10分鐘，並且以紙間法加入4 c.c.發芽水促進發芽效果最佳，可提高苦瓜種子發芽率80%以上。為符合國際ISTA規範，將苦瓜種子依照發芽率高、中、低三等級，委請三個實驗室進行發芽試驗。統計分析三個實驗室發芽數據確定三批發芽活力之差異性未超過容許度，顯示所建議苦瓜種子發芽前處理之方法可推薦給ISTA之種子檢查規則使用。，欄位英文摘要的內容是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 。

年度

2012

計畫名稱_中文

植物種苗團隊-動植物有害生物診斷鑑定技術及種子苗病害驗證技術之研究與應用

計畫名稱_英文

Establishment and Research of Seed and Seedling Certification Techniques and Regulations

計畫編號_中文

101農科-10.3.1-種-X3

計畫編號_英文

101AS -10.3.1-SS-X3

主辦單位_中文

繁殖技術課

主辦單位_英文

Propagation Technology Section

主辦人_中文

邱燕欣

主辦人_英文

Yen-Hsin Chiu

中文摘要

a.符合目標市場外銷國之種子苗驗證技術盤點與研發文獻搜尋並比較國際試驗室間檢測番茄嵌紋病毒(Tomato Mosaic Virus，ToMV)的檢測方式保括ELISA、RT-PCR、ICRT-PCR等檢測技術，本計畫前期合成針對ToMV專一之引子對以及植體之Internal control primer set, 可建立RT-PCR偵測受檢檢體內之ToMV，確定檢測之正確率。101年擬定檢查流程文件-番茄植體檢測番茄嵌紋病毒作業標準一式，並通過財團法人全國認證基金會(Taiwan Accreditation Foundation,TAF)技術認證。 b.外銷重要蔬菜番茄及甜椒種子處理技術之研究本計畫利用氯化鉀電解水、熱水及殺菌劑等方法，研究最適處理條件進行種子滅菌處理，以減少茄科細菌性斑點病之種傳情形與其初次接種源。以HClO濃度50 ppm以上的氯化鉀電解水處理5分鐘以上即可有效除滅甜椒種子與番茄種子上的病原菌；45°C熱水處理30分鐘以上可明顯降低甜椒種子上的帶菌情形，甚至50°C熱水僅處理10分鐘即可完全除滅甜椒種子上的病原菌，同時不降低種子發芽率。殺菌劑試驗中以0.6%漂白水滅菌效果最好，而嘉賜銅處理，除有效降低帶菌情形、甚至完全除滅病原菌外，同時也不會降低種子的發芽率。 c.外銷蔬菜種子品質提升技術研究 (委辦) 本研究的目的在開發不影響種子發芽率之種子殺菌技術，以防治十字花科黑腐病。應用100年度建立之條件『3% H2O2浸漬30秒再以70℃熱風烘乾種子30分鐘』處理1Kg天然帶黑腐病菌種子，結果顯示雖然可以完全殺滅黑腐病菌，但種子發芽率由80%降低至45%。考慮種子原先品質對試驗結果之影響，本年度改用發芽率95%以上之花椰菜及甘藍菜種子，經Xanthomonas campestris pv. Campestri (XCC1-1 strain)污染作為本試驗帶菌種子材料，殺菌處理分別為：『3% H2O2浸漬30秒再以70℃熱風烘乾種子30分鐘』、經70oC乾熱處理7天，及熱硫酸鋅，種子處理後分裝儲藏於防潮箱，每月追蹤其發芽率及帶菌情形。結果顯示所有處理對發芽率皆無顯著不良影響，發芽率穩定維持在95%以上，滅菌效果則以70oC乾熱處理7天為最佳，連續6個月種子洗出液之XCC1-1皆維持零檢出，其他處理之殺菌效果較不完全、也較不穩定。因此，本試驗的結論是『70oC乾熱處理7天』處理為最佳條件方便於大量種子處理，對發芽後兩週幼苗之外觀沒有影響，但估算其狀苗指數，對甘藍沒有影響，但對花椰菜則會降低，因此後續對成株之影響仍需評估。 d.建立重要蘭花病毒ORSV、CymMV新檢測技術建立胜肽親和性篩選系統(peptides 12)對於標地蛋白之親和性汰選技術，建立及生產與CymMV病毒鞘蛋白具親和性之噬菌體，篩選出可利用於ELIS檢測CymMV之噬菌體。 e.建立苦瓜種子發芽檢查程序研究 (委辦)苦瓜種子種皮厚且硬，會妨礙水分吸收及氣體交換，發芽過程中常有發霉的現象，造成種子發芽率不佳及不整齊等問題。本實驗以四品種苦瓜為研究材料，結果顯示苦瓜種子發芽最適溫度為30℃，最適發芽前處理為60℃溫湯浸種10分鐘，並且以紙間法加入4 c.c.發芽水促進發芽效果最佳，可提高苦瓜種子發芽率80%以上。為符合國際ISTA規範，將苦瓜種子依照發芽率高、中、低三等級，委請三個實驗室進行發芽試驗。統計分析三個實驗室發芽數據確定三批發芽活力之差異性未超過容許度，顯示所建議苦瓜種子發芽前處理之方法可推薦給ISTA之種子檢查規則使用。

英文摘要

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