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植物種苗研究團隊-植物品種開發及種苗驗證之應用研究-植物種苗認驗證體系建構生物技術課Construct the Accredit

「植物種苗研究團隊-植物品種開發及種苗驗證之應用研究-植物種苗認驗證體系建構生物技術課Construct the Accredit」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2013 ，欄位計畫名稱_中文的內容是植物種苗研究團隊-植物品種開發及種苗驗證之應用研究-植物種苗認驗證體系建構 ，欄位計畫名稱_英文的內容是Construct the Accreditation system for plant seedlings ，欄位計畫編號_中文的內容是102農科-9.2.3-種-X2 ，欄位計畫編號_英文的內容是102AS -9.2.3-SS-X2 ，欄位主辦單位_中文的內容是生物技術課 ，欄位主辦單位_英文的內容是Biotechnology Section ，欄位主辦人_中文的內容是孫永偉 ，欄位主辦人_英文的內容是Yung-Wei Sun ，欄位中文摘要的內容是1.健康種苗之母本檢測與母本園建構及維護：健康種苗之母本檢測與母本園建構及維護為健康種苗繁殖的基礎，本計畫為落實開發的檢測技術與母本園維護，計畫檢測蒐集保存馬鈴薯、葡萄、草莓以及綠竹等作物母本植株，結果顯示葡萄、草莓以及綠竹皆未特定病原，但馬鈴薯有少數蒐集品種感染單一病毒，且未驗證種薯其帶病風險高，藉由持續的母本園維護、母本檢測及更新，期能支持健康種苗產業化之推動。 2.瓜類退綠黃化病毒及草莓潛隱輪斑病毒驗證體系建構：參酌國內外文獻，已完成瓜類退綠黃化病毒（Cucurbit chlorotic yellows virus , CCYV）與草莓潛隱輪斑病毒（Strawberry latent ringspot virus, SLRSV）之檢測技術盤點。瓜類退綠黃化病毒目前檢測技術主要有： RT-PCR、Western blotting、ELISA及Immunoelectron microscopy；草莓潛隱輪斑病毒目前檢測技術主要有：RT-PCR、IC-RT-PCR、ELISA與病徵判別。以不同的樣品進行各種檢測方法的方法測試，測試結果瓜類退綠黃化病毒以RT-PCR檢測效果較佳；草莓潛隱輪斑病毒則以ELISA檢測效果較佳，挑選最適合的檢測方法建立標準檢測流程，並通過能力試驗。 3.十字花科種子健康檢查驗證體系之建立：本計畫目的係為建立種子檢查室之十字花科種子黑腐病菌檢測作業流程。已依國際種子檢查協會規定之檢測方式建立檢測流程。以人工汙染種子進行檢測流程的測試，可於FS及mCS20ABN兩種選擇性培養基上觀察到測試菌落的標準型態，且靈敏度測試結果可達0.01%的帶菌率。經測試7批不同公司生產的十字花科種子樣品，均未測得Xcc。品質管理所需之表單亦已建置完成。 4.種子(苗)品質純度分子檢測技術研發及標準化：番椒利用F2族群及ISSR之DNA片段多型性，建立F1種子純度分子檢測識別標誌，以充實番椒雜交一代種子純度分子檢測之分子標誌資料庫。結果番椒13個多型性ISSRDNA片段成功回收12個，經定序、解序共獲得8個序列，重新設計條帶專一性引子，其中引子組合11組經最適PCR反應捻合温度及配方調整，均具條帶專一性。但以建立之番椒F2材料測試結果均未表現多型性，因此其識別性尚待進一步確認。番茄3個品種9個植體材料(雜交一代及父、母本親)，利用ISSR-PCR技術篩選共獲得5個新的識別標誌，其中3組引子表現條帶專一性。經最適PCR反應捻合温度及配方調整，標誌大小分別為385、463及626 bp，電泳圖呈現引子高度的條帶的專一性及識別性。，欄位英文摘要的內容是Virus and pathogen detection and establishment of mother-stock garden is the fundamental of healthy seedling production. In this program, we will implement the developing pathogen validation system and construction mother-stock garden to maintain specific pathogen free mother-stock such as potato, grapes, strawberry and bamboo etc. Our results indicated grapes, strawberry and bamboo kept in mother-stock garden were not infected by specific pathogens. Potato stocks were infected by potato virus S or potato virus Y and using none certification potato seed tube would face higher infection risk. We hope this effort could support healthy seedling production in Taiwan. To refer to the scientific or technical literature, we completed the detection technology inventory of Cucurbit chlorotic yellows virus, CCYV and Strawberry latent ringspot virus, SLRSV. The main detection technologies of CCYV are RT-PCR,Western blotting,ELISA and Immunoelectron microscopy. The main detection technologies of SLRSV are RT-PCR, IC-RT-PCR, ELISA and pathognomy. By taking different samples to test the detection technology, we choose RT-PCR for CCYV and ELISA for SLRSV to be the most suitable detectoion method. And we establish standard testing process (SOP), and finish the ability to test. The objective of this program is to establish the seed testing procedure on Xanthomonas campestris pv. campestris (Xcc) which causes black rot of crucifers. The procedure in coincidence with ISTA rules has been established. Artificial infested seeds were adopted to check the Xcc testing procedure. Xcc showed pale green and pale yellow mucoid colonies individually on FS and mCS20ABN selective media. And the Xcc-carried ratio of 0.01% could be detected in sensitivity test. No Xcc was detected in 7 lots of crucifer seeds from different producers. The documents for quality management had also been completed. 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 two year we did some part work in tomato and pepper. The year, We obtain six new sets of SCAR primer for three specific DNA fragments in tomato and ten new 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. 。

年度

2013

計畫名稱_中文

植物種苗研究團隊-植物品種開發及種苗驗證之應用研究-植物種苗認驗證體系建構

計畫名稱_英文

Construct the Accreditation system for plant seedlings

計畫編號_中文

102農科-9.2.3-種-X2

計畫編號_英文

102AS -9.2.3-SS-X2

主辦單位_中文

生物技術課

主辦單位_英文

Biotechnology Section

主辦人_中文

孫永偉

主辦人_英文

Yung-Wei Sun

中文摘要

1.健康種苗之母本檢測與母本園建構及維護：健康種苗之母本檢測與母本園建構及維護為健康種苗繁殖的基礎，本計畫為落實開發的檢測技術與母本園維護，計畫檢測蒐集保存馬鈴薯、葡萄、草莓以及綠竹等作物母本植株，結果顯示葡萄、草莓以及綠竹皆未特定病原，但馬鈴薯有少數蒐集品種感染單一病毒，且未驗證種薯其帶病風險高，藉由持續的母本園維護、母本檢測及更新，期能支持健康種苗產業化之推動。 2.瓜類退綠黃化病毒及草莓潛隱輪斑病毒驗證體系建構：參酌國內外文獻，已完成瓜類退綠黃化病毒（Cucurbit chlorotic yellows virus , CCYV）與草莓潛隱輪斑病毒（Strawberry latent ringspot virus, SLRSV）之檢測技術盤點。瓜類退綠黃化病毒目前檢測技術主要有： RT-PCR、Western blotting、ELISA及Immunoelectron microscopy；草莓潛隱輪斑病毒目前檢測技術主要有：RT-PCR、IC-RT-PCR、ELISA與病徵判別。以不同的樣品進行各種檢測方法的方法測試，測試結果瓜類退綠黃化病毒以RT-PCR檢測效果較佳；草莓潛隱輪斑病毒則以ELISA檢測效果較佳，挑選最適合的檢測方法建立標準檢測流程，並通過能力試驗。 3.十字花科種子健康檢查驗證體系之建立：本計畫目的係為建立種子檢查室之十字花科種子黑腐病菌檢測作業流程。已依國際種子檢查協會規定之檢測方式建立檢測流程。以人工汙染種子進行檢測流程的測試，可於FS及mCS20ABN兩種選擇性培養基上觀察到測試菌落的標準型態，且靈敏度測試結果可達0.01%的帶菌率。經測試7批不同公司生產的十字花科種子樣品，均未測得Xcc。品質管理所需之表單亦已建置完成。 4.種子(苗)品質純度分子檢測技術研發及標準化：番椒利用F2族群及ISSR之DNA片段多型性，建立F1種子純度分子檢測識別標誌，以充實番椒雜交一代種子純度分子檢測之分子標誌資料庫。結果番椒13個多型性ISSRDNA片段成功回收12個，經定序、解序共獲得8個序列，重新設計條帶專一性引子，其中引子組合11組經最適PCR反應捻合温度及配方調整，均具條帶專一性。但以建立之番椒F2材料測試結果均未表現多型性，因此其識別性尚待進一步確認。番茄3個品種9個植體材料(雜交一代及父、母本親)，利用ISSR-PCR技術篩選共獲得5個新的識別標誌，其中3組引子表現條帶專一性。經最適PCR反應捻合温度及配方調整，標誌大小分別為385、463及626 bp，電泳圖呈現引子高度的條帶的專一性及識別性。

英文摘要

Virus and pathogen detection and establishment of mother-stock garden is the fundamental of healthy seedling production. In this program, we will implement the developing pathogen validation system and construction mother-stock garden to maintain specific pathogen free mother-stock such as potato, grapes, strawberry and bamboo etc. Our results indicated grapes, strawberry and bamboo kept in mother-stock garden were not infected by specific pathogens. Potato stocks were infected by potato virus S or potato virus Y and using none certification potato seed tube would face higher infection risk. We hope this effort could support healthy seedling production in Taiwan. To refer to the scientific or technical literature, we completed the detection technology inventory of Cucurbit chlorotic yellows virus, CCYV and Strawberry latent ringspot virus, SLRSV. The main detection technologies of CCYV are RT-PCR,Western blotting,ELISA and Immunoelectron microscopy. The main detection technologies of SLRSV are RT-PCR, IC-RT-PCR, ELISA and pathognomy. By taking different samples to test the detection technology, we choose RT-PCR for CCYV and ELISA for SLRSV to be the most suitable detectoion method. And we establish standard testing process (SOP), and finish the ability to test. The objective of this program is to establish the seed testing procedure on Xanthomonas campestris pv. campestris (Xcc) which causes black rot of crucifers. The procedure in coincidence with ISTA rules has been established. Artificial infested seeds were adopted to check the Xcc testing procedure. Xcc showed pale green and pale yellow mucoid colonies individually on FS and mCS20ABN selective media. And the Xcc-carried ratio of 0.01% could be detected in sensitivity test. No Xcc was detected in 7 lots of crucifer seeds from different producers. The documents for quality management had also been completed. 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 two year we did some part work in tomato and pepper. The year, We obtain six new sets of SCAR primer for three specific DNA fragments in tomato and ten new 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.