The Study on Anther and Microspore Cultures of Coneflower

使用具優良農藝性狀且cichoric acid含量超過國外經濟栽培價值的單株為材料，進行花藥培養與小孢子培養，進行形態與染色體數鑑定，篩選雙單倍體(double haploid)個體，供後續評估F1種子生產，期能盡快推出適當品種，供日增的栽培者使用。Anthers and microspores of elite coneflowers (Echinacea purpurea) with good agronomic characteristics and higher cichoric acid contents than foreign cultivars will be cultured to get regenerated plantlets. These plantlets are going to be examined in terms of morphological features and chromosome numbers to select the double haploids (DH). The DHs will be used to hybridize with each others to get F1 seeds in the near future. After finished this project, it is possible to offer F1 seeds to the increasing growers

Studies of Mass Propagation of Coneflower (II)

紫錐菊多年來仍為歐美的重要保健植物，銷售金額超過20億美金，商業利益明顯。本計畫種苗之指標成分含量，如cichoric acid等，達商業標準150%以上，均為值得栽培之的品系。應用去年獲得密度與透氣對瓶苗品質影響之結果，持續改進瓶苗品質，尤其是 1. 國外報導亦指出發根率偏低的問題。其次 2. 為容器減重，目前國內塑膠培養盒單價下降，已漸趨可接受程度，用之可望減重50%以上，擬觀察透氣處理在此容器之培養反應。另外， 3. 去年的間歇浸潤系統(temporary immersion system)培養結果比一般固體培養效率高，發根率尤其良好，擬進一步修正流程以提高效率，評估取代固體培養，節省洋菜、簡化出瓶清洗、育苗等步驟。本年度計畫可望大幅提升紫錐菊組織培養苗發根率，增加出瓶存活率，使之更適合商業栽培，提供農民新的選擇。Coneflower is a famous herbal medicinal plant in western world for long time. Its products value at more than 2 billion US dollars. Elite plants containing effective ingredients (e.g. cichoric acid), over that of foreign commercial cultivars about 50%, had been selected for materials about three year ago in Tachung, Taiwan. The objectives of this project are (1) to improve the rooting rate of shoots derived from leaf culture of elite plant because of the low rooting response similar with the results of foreign report. (2) to replace the glass vessel with plastic vessel to reduce the heavy weight of glass vessel and to increase ventilation by using caps holed and taped with 3M adhesive tape on the holes. Furthermore, it is very hopeful to promote the rooting rate and survival rate extra vitro by modifying the protocols, please refer to final report of last year, of temporary immersion system. It will bring the saving of labors for cleaning agar and avoid hurting roots. The results may really reduce the cost of masspropagation of elite coneflower plants to fit the requirements for field cultivation or potting

Studies on Improving Somatic Embryogenesis of New Sweet Potato Cultivars (III)

甘藷為無性繁殖作物,改良現有品種易因多交(polycross)育種的遺傳分離,而將已聚集的優良性狀打散.因此,無性繁殖作物之誘變育種有其存在價值,因為大部分優良性狀未被改變.本年度擬利用前兩年已開發完成的甘藷體胚分化流程(固體與液體)做誘變處理的實際應用.主要利用nitrosourea處理癒合組織或懸浮細胞團,並篩檢再生的體胚是否為突變體.另使用低劑量nalidixic acid與hydroxyurea等DNA合成抑制劑,與誘變處理進行搭配,以干擾DNA合成過程,提高突變率.希望可增加多樣化有特色的葉用與塊根用甘藷品系,增加本地植物基因應用.Sweet potato is one of the asexual propagation crops, good agronomic characteristics and eating qualities may be separated after polycross breeding. It is indispensable to apply mutation treatments to change just one or few characteristics of good cultivars. We are going to treat sweet potato callus or cell aggregates with nitrosourea, and select possible mutants among seedlings of converted somatic embryos. In order to increase mutation rate low doses of nalidixic acid and hydroxyurea, inhibitors of DNA synthesis, are applied as supplementary treatments to mutation protocols. We look forward to find out some useful mutants to increase sweet potato diversity of eating qualities

Effects of glutamine and low pH on

封面 麩胺醯胺及低pH值對苜蓿體胚化反應的影響 Effects of glutamine and low pH on embryogenic response of alfalfa 圖目錄 表表錄 中文摘要 第一章 緒言 第二章 添加glutamine及auxin/cytokinin之比例對苜蓿懸浮培養之影響 一、前人研究 二、材料與方法 三、結果 四、討論 五、結論 第三章 低pH值對苜蓿組織培養之影響 一、前人研究 二、材料與方法 三、結果 四、討論 五、結論 第四章 克服轉變率(conversion rate)太低之方法 一、前人研究 二、材料與方法 三、結果 四、討論 五、結論 第五章 綜合討論 英文摘要 參考文獻 附

Studies on Masspropagation of Huang-Qi (Astragalus membranaceus) (II)

黃耆為大宗藥食均用之藥材。中藥材種類眾多，黃耆就佔台灣每年進口中藥總值約6%，金額約2.0億元。更重要者，品系不清、大陸產地品管不一，功效批次間向來紊亂，藥效與食用安全一直受到關切。將94年選出根部較大而優良的單株做為試驗材料，繼續改良組織培養系統。 1. 改善增殖芽體品質，擬藉由調整蔗糖、有機氮與paclobutrazol (PP333)改善之。 2. 改進試管苗發根，擬藉由滲透調節處理、透氣與添加tryptophan (auxin precursor)等改善之。 3. 組培苗田間生長的根部成分分析。本年計畫可望提高黃耆組織培養體系效率並獲得成分分析評估，極有助於保住優良單株。對高附加價值如黃耆注射液(已有台灣廠商2005獲准專利上市)生產等，極需單純且穩定材料的廠商，將多一選擇，避免完全依賴大陸產地。Root of huang-qi (Astragalus membranaceus) is one of the famous and used worldwide traditional Chinese herbal medicines. It alone accounted for about 6% of the imported many and diverse herbal medicines in Taiwan every year. For long time, people are bothering about the ingredients stability of huang-qi because of no uniform cultivars and no regular sources. We have obtained some primary and important information about tissue culture of huang-qi from the results of last year. The objectives of this year are (1) to improve further quality of induced shoots by means of modifying sucrose, organic nitrogen and paclobutrazol concentrations, (2) to optimize the conditions for rooting of shoots in vitro by amending auxin precursor, e.g. tryptophan, and ventilation, (3) to analyze the effective ingredients of field-harvested roots of plants derived from tissue culture. After this year, we may get further promotions of masspropagation and ingredients analysis of huang-qi and it will benefit to maintain elite plants. We can start on getting roots of uniform cultivars regularly to offer materials for producing high-priced product, e.g. injection of root extract of huang-qi. It will be another choice for pharmaceutical industry in Taiwan

Preservation of Elite Plants in Vitro and Hairy Root Culture of Huang-Qi

經由試管內選拔出來的優良黃耆單株的嫩芽將用以誘導胚化的癒合組織，並將進一步進行測試其分化能力。利用這些優良單株的莖誘導毛狀根，並將測試不同培養條件下，毛狀根的astragaloside IV含量。藉由上述試驗可望獲得優良的黃耆毛狀根品系，可供評估量化之可行性。The young buds of elite huang-qi (Astragalus membranaceus) selected in vitro will be used to induce embryogenic callus and trials for further regeneration will be done. Hairy roots will be induced from stems of these elite plants and the conditions for hairy root culture will be evaluated in terms of the contents of astragaloside IV. It is possible to get some valuable hairy root lines

Factors Improving Plant Regeneration from Suspension Culture of Sweet Potato

計畫目標: 由固體培養流程中挑取台農68號與70號胚化癒合組織為材料, 完成下列研究: 1.由甘藷癒合組織建立懸浮細胞增殖體系.2.確定誘導懸浮細胞( 團 )形成體胚的有利條件.3.體胚乾燥處理對體胚發芽之影響.重要工作項目: 1.懸浮培養的初期建立: ( 1 )2, 4-D用量比較、 ( 2 )NH？？濃度比較與 ( 3 )癒合組織大小.主要調查細胞堆疊體積( packed cell volume )之變化與細胞( 團 )乾鮮重變化為主.2.懸浮液內誘導體胚形成: ( 1 )ABA與Ca？？濃度比較、 ( 2 )胺基酸等用量比較與 ( 3 )篩選懸浮細胞團大小.主要調查不同時期的體胚形成量、體胚大小與出現時間快慢為主.3.體胚乾燥處理對發芽之影響: ( 1 )直接乾燥與 ( 2 )間接乾燥.主要調查處理後體胚發芽率、發芽速度與形態差異為主.預期效益懸浮細胞體胚分化系統的建立, 對無性繁殖作物的品種改良與增加遺傳變異有重大助益.尤其甘藷每年之栽培面積仍達10000公頃以上, 加以被視為有分子農場( molecular farming )潛力的作物, 建立經濟栽培甘藷品種的懸浮細胞培養體胚分化技術有農業與商業價值.The purpose of this study is to develop a suspension culture system for sweet potato.The successful culture system is supposed to be a powerful tool to sweet potato breeding and molecular farming in the future.Embryogenic callus of TN 68and TN 70sweet potato will be induced from shoot tips cultured on callus induction medium.Embryogenic callus will be used to establish a suspension culture and to induce somatic embryogenesis during the culture.Factors including 2, 4-D and NH？？ion concentrations and callus size will be taken into consideration in initiation of suspension.The other factors like abscisic acid, Ca？？ion, amino acid and cell aggregate size will be noticed in somatic embryo formation.Desiccation treatments will be estimated in improving germination rate of somatic embryos

Induction and Screening of Herbicide-Tolerant Rice

本研究計畫主要目的在建立除草劑抗性(耐性)水稻之誘導系統，分別由不同rice lines，取其發芽種子分生能力旺盛之芽點，以固殺草(glufosinate)除草劑之有效成分phosphinothricin (PPT)進行microshoot 之分化再生誘導，以產生具遺傳變異之多側芽。試驗同時配合cytokinin 及kinetin 之施用，以瞭解釐清PPT 所扮演的角色。為增加genetic variation，試驗中擬於建立適當的microshoot induction 條件之後，配合不同劑量NaN3 處理，以尋求產生具有genetic variation 之大量microshoot 所需之條件，包括rice lines，PPT dosage，cytokinin/kinetin ratio，以及適當之培養步驟與方法。經誘導產生之microshoot-derived plants 再進行PPT-tolerance 測試(包括in vitro 與in vivo tests) ，以及其對於不同作用機制除草劑glyphosate 與bentazon之耐性反應，以評估此誘導變異系統之效果

Studies on Mass Propagation of Coneflower

紫錐菊為歐美的重要藥用與保健植物，每年銷售金額約20億美金，有明顯商業利益。本計畫使用田間篩選出來高生物量、高有效成份的單株作為大量生產組培苗的材料，可以避免天然雜交種子造成單株的田間表現、成份等差異過大的缺點。本計畫可望找出大量繁殖紫錐菊組培苗的瓶內適當栽培密度與通氣孔數，大幅改善瓶苗品質；並且，探討應用間歇浸潤系統量化紫錐菊組培苗的方法。這些瓶苗都將接受馴化處理，用以建議業者最好的育苗方式。Products of coneflower are very popular, important healthy foods, its market is about 2 billion US dollars per year. The high biomass and rich of effective ingredients plants are selected from field and used as materials for mass propagation, it can avoid drawbacks that plants derived from open-pollinated seeds are uneven in field performance and ingredients. The quality of plantlets in vitro can be promoted by applicable planting density and suitable ventilation pores on cap, information will be showed in the results of this project, and the practical methods of temporary immersion system for plantlets production in vitro will be recommended as well. All the plantlets produced in vitro will be committed to acclimation treatments to find out recommendable nursing protocols

水稻穀粒大小之充實特性與遺傳之研究

穀粒為吾人種植水稻庂主要目的，而穀粒大小則為水稻產量的主要因子之一。本論文 主在藉著不同試驗方式以明瞭水稻穀粒大小遺傳行為及充實特性，希望能提供給今後 水稻育種所需的某些參考訊息。 穀粒大小遺傳研究係利用 F代全互交法（diallel cross ）， F代半互交法（ha lf dillel cross ）世代平均法（generation mean method）。穀粒充實特性則於民 國七十一年一、二期依，開花後每隔３日取樣，烘乾稱重，換算得Cubic polynomial curve ；同時於開花後每隔６天測劍葉，第二，第三之綠素含量。另做剪葉處 理，測其供源（source）對積儲（sinls ）的影響。資料來源則以國內外重要雜誌為 主。 試驗結果顯示控制殼粒大小遺傳之因子數並不多，主要遺傳變異成分為加性基因作用 （additive gene effect），具有高遺傳率，約在７５％以上，顯示早期選拔有效； 與國內外試驗結果大致結論相同。穀粒充實特性則可見千粒重大的品種具有較長的有 效充實期及較高的充實率，一期作較二期作具較長的充實期，顯示二期作對大型穀粒 較為不利。其主要原因之可能一期作時，不論穀粒大小，各品種之葉綠素含量均能維 持相當時間而不下降，二期作自開花後則不斷下降。而且由剪葉處理亦充分證明開花 後六天起，葉片存在與否決定穗實率與單株產量之高低