生化遺傳標識因子在作物改良上應用

[[sponsorship]]植物暨微生物學研究所[[note]]已出版;有審查制度;具代表

Detection of glyphosate residue in glyphosate-treated Leucaena leucocephala

銀合歡於恆春地區嚴重蔓延，根據林試所恆春研究中心以嘉磷塞注射樹幹方式獲致良好控制效果，本研究進ㄧ步進行嘉磷塞不同濃度處理效果，及嘉磷塞施用後在樹體及土壤之殘毒監測。研究結果顯示不同濃度嘉磷塞處理下，施藥後ㄧ週於注射孔上方約350 cm之銀合歡小葉快速凋萎。整株植株於處理藥劑後其傷害在施藥後1個月最為嚴重，雖然施藥後2個月受害植株開始萌新芽，但新芽多呈褐化畸形。檢測藥劑殘留量顯示藥劑隨時間逐漸消褪，施藥後五個月藥劑殘留量降至施用量的10%，根部則於施藥後三個月偵測出較高殘留量但亦隨時間迅速下降。進ㄧ步以HPLC及放射性同位素分析藥劑流向，研究結果顯示施藥初期藥劑可能於木質部藉擴散方式向周圍轉運，其中大量藥劑累積於兩注射孔中間之韌皮部，並隨時間進行雙向運輸，推測韌皮部為藥劑轉運之主要通道。施藥後45天發現注射孔上方韌皮部有較高之藥劑殘留，於60天後各部位藥劑濃度均隨時間下降，根據放射性同位素活性偵測亦顯示相似變化，然而60天後注射孔上方之放射活性降低而下方放射活性反而增加。探究原因推測由於苗木施藥後於注射孔下方逐漸萌新芽，可能因同化物質source-sink關係改變，使嘉磷塞及其代謝物經由韌皮部重新分配至植體下方，而配合TLC及放射活性分析可以確認此時嘉磷塞除了降解為少部分已知的AMPA、及較多量之sarcosine之外，尚有大量之未知代謝物。Leucaena leucocephala, an exotic species widely spread in Hengchun peninsula, Taiwan, was controlled efficiently by a herbicide of glyphosate . In this study, effects of glyphosate with three different concentrations but an equal amount of dosage were compared, and changes of glyphosate residues in plants and soil environment were also detected. Experimental results showed that rapid wilting of leaflets at height of 350 cm away from glyphosate injection site on basal trunk one week after treatment was found, and serious injury of whole plant occurred within one month. Although newly-formed buds appeared on basal trunk with time, subsequent bud growth was retarded and leaflets were small, chlorotic and deformed. Glyphosate detection showed that residues in plants dissipated rapidly in 3 months, and decreased to 10% 5 months after treatment. Further experiment to study glyphosate translocation in L. leucocephala with HPLC and 14C-glyphosate radioactivity assessment, suggested that most glyphosate residues in xylem likely diffused to surrounding tissues initially, and then temporarily accumulated in phloem located between two injection sites before translocating out subsequently. Therefore, it is implicated that phloem is a primary translocation channel of glyphosate. Glyphosate dissipation analysis revealed that more residues were found in upper phloem 45 days after treatment (DAT) and decreased significantly thereafter. Althrough glyphosate in whole plant including upper, middle and lower parts, was decreased after 45 DAT, radioactivities of 14C-glyphosate and its derivatives in lower phloem accumulated dramatically 90 DAT. The redistribution of 14C-glyphosate and its metabolites to lower phloem of basal trunk is partially resulted from an alteration of source-sink relationship of photosynthate due to the formation of newly-formed leaflets on basal trunk.目次 頁次 中文摘要 ……………………………………………………………i 英文摘要 ……………………………………………………………ii 目次 …………………………………………………………………iii 圖目次 ………………………………………………………………iv 第一章 緒言…………………………………………………………1 第二章 前人研究……………………………………………………4 第三章 材料與方法……………………………………………… 17 第四章 結果與討論…………………………………………………28 ㄧ、嘉磷塞控制銀合歡之效果…………………………… 28 二、嘉磷塞於環境中之消褪……………………………… 35 三、嘉磷塞之轉運研究…………………………………… 52 四、嘉磷塞代謝物變化…………………………………… 60 五、藥劑殘留重新分配之可能原因……………………… 64 六、結論…………………………………………………… 69 參考文獻………………………………………………………… 70 附錄……………………………………………………………… 7

電動輪椅之座椅升降結構

本創作係為一種「電動輪椅之座椅升降結構」，尤指一種電動輪椅可調整其座椅高度，以利使用者拿取高處物品者。 其主要：係設一由馬達驅動輪組之車台，而在車台上固設一承置板，該承置板上則固設有電池盒、驅達馬達、減速機及一頂撐軸桿，而驅動馬達與減速機及頂撐軸桿間則套設有時規皮帶，藉使驅達馬達可帶動頂撐軸桿作動，而該頂撐軸桿係藉由外管、內管、螺桿、皮帶輪及乾式軸承等組合而成，其另端係與座椅之座椅基板固設，而可藉由驅動馬達之傳動減速機，而由減速機連動頂撐軸桿之皮帶輪，而使頂撐軸桿內之螺桿轉動，而令內管可依螺桿之正逆轉作上、下之位移，而使座椅可為昇降，而達電動輪椅之升降可直線升降調整至任一高度，而可更具安全性及實用性者

電動輪椅之進退操控結構

本創作係為一種「電動輪椅之進退操控結構」，尤指一種電動輪椅之座椅可旋轉一百八十度，以利電動輪椅退行結構而言。 其主要：係設一由馬達驅動輪組之車台，而在車台頂端焊固一承置板，於承置板與車台間係固設一藉時規皮帶傳輸之驅動馬達及蝸桿輪，而該蝸桿輪則連動一端與座椅固結之轉軸齒輪盤，而在轉軸上係固設一組左、右固定片及焊固一反向對置之反轉桿，並左固定片上係設一感應片，而於承置板上係固設對置之高、低L狀片體，且於高L狀片體固設一感應器，藉令電動輪椅欲為旋轉或退行時，可在切換旋轉開關後，利用行進搖桿操作座椅旋轉至任一角度或轉至正後面時停止，並得在旋轉至正後面時令轉軸感應片與感應器為相對感應，而可切換驅動輪組之馬達使其反向旋轉，藉使電動輪椅之退行，對使用者而言仍係向前行進，而令電動輪椅之行進或取物更加實用及安全者

Utilization Of Molecular Markers In Vegetable Crop Improvement

生物技術中應用於作物改良者，分子標誌因子（molecular markers）是較為實際與典型的例子之一。在蔬菜作物中，番茄分子標誌因子之研究堪稱典範。在可資利用之分子標誌因子來源中，隨著分子生物技術之進步，已由早期利用蛋白質、酵素膠體電泳法，配合酵素活性染色的同功異構酶（isozyme）分析，進而利用DNA限制內切酶酶譜多形性（Restriction fragment length polymorphisms；RFLPs)，至新近應用聚合酶連鎖反應（Polymerase chain reaction；PCR）所產生的聚合酶逢機擴增多形性DNA(Random amplified polymorphic DNA；RAPD）為標誌因子。這三種方法中，同功異構酶之發展最早，在蔬菜作物之報告亦不勝枚舉，其法之優點為分析方法簡單，而缺點則為可當分子標誌因子之因子座有限；而RFLP 分析法係利用DNA 限制內切酶辨識特殊鹽基系列之特性，若不同個體之DNA序列不同時，限制酶切割後所產生片段之大小、數目亦因此而有不同，經與該段序列有某種程度相似性之探針複合（hybridization ）檢定，所產生不同之圖譜做為標誌因子，此方法之優點為所產生之因子數遍佈整個基因組，缺點為分析所用之方法較耗人力與時間；目前蔬菜作物除番茄已建立良好之RFLP分子標誌圖譜外，馬鈴薯、萵苣、甜菜、豌豆及一些十字花科蕓苔屬類蔬菜亦已有了報告。而有關利用聚合醃連鎖反應，其逢機擴增DNA片段大小不同為標誌因子之方法，其優點為簡單、快速，缺點為大部分RAPD分子標誌因子為顯性，無由分辨異型合子，且有關RAPD在作物上利用之報告目前十分有限，在發展上是否有其他受限之處有待進一步之探討。此外分子標誌因子之利用亦已由早期用以篩選一些主效因子，如抗病、抗蟲、雄不稔性等因子之選拔，進一步用來標定數量性狀之選拔。 The utilization of molecular markers for crop improvement is believed to be one of the benefits from biotechnology. Presently, at least three types of molecular markers such as isozyme, RFLP (Restriction Fragment Length Polymorphism) and RAPD (Random Amplified ploymorphic DNA) derived from AP-PCR (Arbitrary Primer Polymerase Chain Reaction) are available. Among these, isozyme markers were studied the earliest and a lot of research has been conducted. The advantages of isozyme as molecular marker are its simplicity and ease for analysis. However, the limitation on the number of polymorphic loci is the major drawback in its application. RFLP markers analyzed on the DNA level are believed to be very promising because the markers obtained can be saturated all over the linkage map. Nevertheless, the profoundity and complexity of its methodologies have slowed down its utilization. Among the vegetable crop species, RFLP markers and linkage map have been well established in tomato. Other crops such as potato, lettuce, sugar beet, pea and Brassica species also have several studies on the RFLP markers. RAPD derived from AP-PCR was most recently reported. The advantages of RAPD markers are its rapidity and efficiency during the analyzing processes. However, the predominance with dominant gene action on RAPD markers will require twice as many markers as those of codominant markers to distinguish a heterozygote. Since only very limited studies are avaliable now on RAPD in crop species, whether there is any other limitation requires further investigation. Furthermore, gene mapping is no longer limited to those of major qualitative genes such as disease, insect resistance, and male sterility, etc. Selection for quantitative traits have become possible through the aid of molecular markers

The Analysis of Isozyme Patterns in Sweet Potato [Ipomoea batatas (L.) Lam.]

有關同功異構酶的分析與利用目前在各主要作物已有不少之報告，然甘藷在這方面的研究仍極為有限。此與甘藷本身遺傳背景之複雜性及組織成份物質對酵素活性分析之干擾有或多或少之關係。近年來雖有一些新型分子標誌因子分析法，如核酸限制內切酶多形性分析及利用聚合醃連鎖反應所導來之逢機擴增多形性DNA等，然同功異構酶分析法仍不失為較簡單、易學、經濟且快速之分析法，其分析所需之組織量少，且幾乎各類型之組織均可利用來作分析，在應用上仍有其優點。本報告即針對目前有關甘藷同功異構酶已見報之資訊，及本研究小組近年來在甘藷同功異構酶分析與利用上之探討作一綜合性之報告。所討論之酵素主要以具多形性酶譜之酵素如酒精去氫酶、β一澱粉酶、磷酸化酶、過氧化酶、黃遞酶、酯酶、磷酸葡糖變位酶、蘋果酸去氫酶、蘋果酸酵素、磷酸葡糖異構酶、6-磷酸葡糖去氫酶、穀草酸轉氨酶、莽草酸去氫酶、甲基萘酶、還原酶及胰蛋白抑制酶等。 Studies on the analysis and utilization of isozyme patterns in sweet potato [Iponioca baiatas (L.)Lam.] are very limited in comparison with those of the major crop species. This might be due to the relative complexity of the sweet potato genomes and the existence of certain chemical components in tissue extracts tending to interfere the enzyme staining activity. In spite of the recent development of DNA molecular markers derived from restriction fragment length polymorphisms and random amplified polymorphic DNA derived from arbitrarily primed polymerase chain reaction, the isozyme approach is still the easiest and most economic and rapid method available. In addition, it also has the advantage of the small amount of tissue needed for the assay and no restriction to tissue type. In this communication, the current progress in isozyme analysis and utilization of sweet potato are reviewed. Emphases are laid on those polymorphic enzymes such as alcohol dehydrogenase, β-amylase, diaphorase, esterase, glutamatic oxaloacetate transaminase, malate dehydrogenase, malic enzyme, menadione reductase, peroxidase, phosphoglucomutase, 6-phosphogluconate dehydrogenase, phosphoglucose isomerase, phosphorylase, shikimate dehydrogenase, and trypsin inhibitor

Intelligent Manufacturing Cloud (II) & (III)

[[abstract]]為呼應政府推展智慧型自動化產業與雲端運算產業的政策，本整合型計畫將利用雲端運算、全自動虛擬量測、本體論、及虛擬機台等技術，發展一個可應用於製造業的雲製造系統，稱為「智慧型製造雲」，包含雲端與工廠端兩大部分。在雲端部分，我們將建置一個雲端運算服務平台及各種雲製造服務(封裝有製造資源的雲端運算服務)，包括資料擷取、預測模型建置、機台知識推論及虛擬機台等服務，將可讓許多工廠中的使用者與各式機台同時透過網際網路隨選使用。在工廠端，我們將基於全自動虛擬量測與虛擬機台技術，發展數種可插入式的智慧型機台應用模組，以提供機台具備如工件加工精度推估、機台狀態診斷、關鍵組件剩餘壽命推估，以及虛擬機台模擬等智慧性能力。「智慧型製造雲」將在總計畫主導下，搭配5個分項計畫，共同合作開發，預期具有以下優勢：(1)提供可根據需求自動調整規模的計算與儲存資源，符合製造業之經濟效益；(2)具有可擴充功能且可透過網際網路存取的各式雲製造服務，能為製造業建立新的商業模式；(3)可為工廠端各式不同機台提供可插入式的智慧型自動化能力。本整合型計畫在第1年已建構一個基於公有雲的「智慧型製造雲」雛型，並建置數個應用「智慧型製造雲」於CNC工具機的展示範例。基於第1年的研發成果，本計畫在第2、3年將進一步發展一個採用混合雲架構的「智慧型製造雲」，以利用私有雲確保企業機敏資料的安全，並以公有雲提供充沛的雲端運算資源，以利「智慧型製造雲」向業界推廣。本整合型計畫之預期研究成果具有技術的創新性、前瞻性與產業利用性，可以做為我國製造產業建置相關雲製造系統之重要參考。[[abstract]]To echo the government’s policy of developing the intelligent automation industry and cloud computing industry, this integrated project will leverage several technologies, including cloud computing, automatic virtual metrology (AVM), ontology, and virtual equipment (VE) to develop a cloud manufacturing system, called Intelligent Manufacturing Cloud (IMC), for the manufacturing industry. The IMC contains two major parts: the cloud side and the factory side. On the cloud side, we will create a cloud computing service platform and various cloud manufacturing services (i.e. cloud computing services encapsulated with manufacturing resources), including the data-acquisition service, prediction-model-creation service, equipment-knowledge-inference service, virtual equipment service, etc. Consequently, the IMC allow users and various kinds of equipment in many factories to simultaneously access these cloud manufacturing services on demand. On the factory side, we will base on the AVM and VE technologies to construct several pluggable intelligent equipment application modules for providing equipment with several intelligent capabilities, such as predicting workpiece precision, diagnosing equipment’s statues, predicting key components’ remaining useful life, and virtual equipment simulations. The IMC will be collaboratively developed by the main project with five sub-projects, and the main project will lead the overall research roadmap and direction. It is expected that the IMC will have the following advantages: (1) it can provide automatically-scalable computing and storage resources according to the demands to meet the manufacturing industry’s economic benefits； (2) it possesses various cloud manufacturing services that can be accessed through the Internet and expandable in functionality so that the manufacturing industry can leverage them to create new business models； and (3) it can provide various pluggable intelligent automation capabilities to equipment on the factory side. IThe expected research results of this integrated project possess novelty, advancement, and industrial applicability in technology and, thereby, can be an important reference for domestic manufacturing industries to build cloud manufacturing systems.[[note]]NSC102-2218-E006-009-MY

Intelligent Manufacturing Cloud

[[abstract]]為呼應政府推展智慧型自動化產業與雲端運算產業的政策，本整合型計晝將利用 雲端運算、全自動虛擬量測、本體論、及虛擬機台等技術，發展一個可應用於製造業的 雲製造系統，稱為「智慧型製造雲」，包含雲端與工廠端兩大部分。在雲端部分，我們 將建置一個雲端運算服務平台及各種雲製造服務(封裝有製造資源的雲端運算服務），包 括資料擷取、預測模型建置、機台知識推論及虛擬機台等服務，將可讓許多工廠中的使 用者與各式機台同時透過網際網路隨選使用。在工廠端，我們將基於全自動虛擬量測與 虛擬機台技術，發展數種可插入式的智慧型機台應用模組，如工件加工精度推估、機台 製造能力推估、機台關鍵組件狀態診斷與剩餘壽命推估，以及虛擬機台模擬等。「智慧 型製造雲」將在總計晝主導下，搭配4個分項計晝，共同合作開發，預期具有以下優勢： (1)提供可根據需求自動調整規模的計算與儲存資源，符合製造業之經濟效益；（2)具有 可擴充功能且可透過網際網路存取的各式雲製造服務，能為製造業建立新的商業模式； (3)可為工廠端各式不同機台提供可插入式的智慧型自動化能力。本整合型計晝之預期研 究成果具有技術的創新性、前瞻性與產業利用性，可以做為我國製造產業建置相關雲製 造系統之重要參考。[[abstract]]To echo the government’s policy of developing the intelligent automation industry and cloud computing industry, this integrated project will leverage several technologies, including cloud computing, automatic virtual metrology (AVM), ontology, and virtual equipment (VE) to develop a cloud manufacturing system, called Intelligent Manufacturing Cloud (IMC), for the manufacturing industry. The IMC contains two major parts: the cloud side and the factory side. On the cloud side, we will create a cloud computing service platform and various cloud manufacturing services, i.e. cloud computing services encapsulated with manufacturing resources, including the services of data acquisition, prediction model creation, equipment knowledge inference, virtual equipment, etc. Consequently, the IMC allow users and various kinds of equipment in many factories to simultaneously access these cloud manufacturing services on demand. On the factory side, we will base on the AVM and VE technologies to construct several pluggable intelligent equipment application modules, such as workpiece precision conjecture, equipment manufacturing capability conjecture, state diagnostics and remaining useful life (RUL) conjecture of equipment’s key components, and virtual equipment simulation. The IMC will be collaboratively developed by the main project with four sub-projects, and the main project will lead the overall research roadmap and direction. It is expected that the IMC will have the following advantages:? it can provide automatically-scalable computing and storage resources according to the demands to meet the manufacturing industry’s economic benefits; (2) it possesses various cloud manufacturing services that can be accessed through the Internet and expandable in functionality so that the manufacturing industry can leverage them to create new business models; and (3) it can provide various pluggable intelligent automation capabilities to equipment on the factory side. The expected research results of this integrated project possess novelty, advancement, and industrial applicability in technology and, thereby, can be an important reference for domestic manufacturing industries to build cloud manufacturing systems.[[note]]NSC101-2218-E006-02