植物向光性的訊息傳導
植物依靠向光性爭取最多的光線,以進行光合作用,製造食物供給所有生物。雖然在十九世紀時植物的向光性就已經被發現,並且參與植物向光性的主要荷爾蒙為植物生長素也已經熟知,但是主要是植物的哪一個組織接受光訊息以誘導向光性,以及細胞內的哪些分子參與訊息傳遞,則都不清楚。因此這個研究,以可以發射特殊波長的發光二極體為光源照射綠豆小苗以研究向光性,結果顯示藍光和綠光而不是紅和黃光可以誘導向光性。就向光性訊息傳導的組織層面的研究而言,將豆苗的葉、葉柄、生長點、子葉分別除去後,再側面照光,發現向光性要產生必須要有生長點或葉柄,並且發現莖可以誘導向光性,而葉子不能誘導向光性,因此莖是主要接受光訊息以誘導向光性的組織。就向光性訊息傳導的分子層面的研究而言,植物以鈣離子的螯合劑和鈣離子通道阻斷劑處理後發現,細胞質內鈣離子濃度的增加是藍光和綠光誘導的向光性所需要的過程,有趣的是藍光誘導向光性的訊息傳導過程中,除了經由細胞內的鈣離子濃度的增加外,還有其它鈣離子不參與的訊息傳導途徑。此外,以蛋白質磷酸?抑制劑和蛋白質去磷酸?抑制劑處理植物後發現,藍光和綠光所誘導的向光性訊息傳導,都包含蛋白質去磷酸?第1 和2a 型在細胞內的作用。因此植物的向光性需要有生長素才會表現,生長素由生長點製造後由生長點和葉柄儲存,在光刺激之下會誘導莖產生傳遞訊息,此訊息會傳遞到含有生長素的生長點和葉柄,使得生長素流向照光組織細胞,並且使得細胞內鈣離子濃度增加,活化蛋白質去磷酸?第1 和2a 型,進而造成植物的向光性。Phototropism allows plants to receive the most amount of light to perform photosynthesis, which produces food and energy for all organisms. The phenomenon of phototropism has been known since the 19th century, and auxin has been identified to be the main hormone involving in phototropism. However, the major plant tissue responsible for receiving light signal is not fully understood, and the signal transduction pathway within cells after light activation is not clear. Therefore, the phototropism of mungbean seedlings is examined by Light Emitting Diodes (LED) which produce the specific wavelength of light in this study. Results point out that blue and green lights rather than red and yellow lights induce phototropism of moonbeam. The phototropism of mungbean seedlings is further studied by plants whose leaves, petioles, apical meristem, or cotyledons were removed, showing that the presence of either apical meristem or petioles is needed for inducing phototropism. Also, stem ,not leaves, is the major tissue that receives light activation, and induces phototropism. The signal transduction of phototropism was further analyzed in the presence of calcium ion chelator and channel blockers. The signal transduction of phototropism induced by blue or green light contains the increasing concentration of calcium ion within cytosol. Interestingly, there is a calcium-independent tansduction pathway for blue light only to induce phototropism. Additionally, staurosporine (STA), a protein kinase inhibitor and okadaic acid (OKA), a protein phosphatase inhibitor, were used to study the signal transduction pathway of phototropism, and results indicated that protein phosphatase 1 and 2a is needed for both blue and green lights to induce phototropism. Conclusively, the phototropism is triggered by the reception of light by stem, and the light signal is transferred to apical meristem and petioles that reserve auxin produced from apical meristem. Auxin is then transferred to the cells that is illuminated, increases the concentration of calcium ion and activates protein phosphatase 1 and 2a in cells, and finally phototropism occurs.
銅影響大豆及田菁根部生長、細胞死亡和訊息傳遞
隨著迅速的工業化,重金屬汙染已是嚴重的環境問題。在植物中,當植物體內累積過量的重金屬,對於植物根部、葉部等器官的生長與發育有嚴重的影響或傷害。銅離子為植物生長所必須之重金屬,但是過量銅離子會導致細胞死亡,生長受到抑制。本實驗以大豆( Glycine max )及田菁( Sesbania roxburghii )為植物材料,藉由Evans blue 染色法、螢光染色、西方墨點法、反轉錄聚合?鏈鎖反應等,觀察過量銅離子影響植物根部生長、細胞死亡和細胞訊息傳遞物質變化之情形,並探討過量銅離子影響大豆根部細胞死亡的訊息傳遞路徑。過量銅離子會限制植物根部的生長及造成根部細胞死亡。以螢光染色觀察根尖(ROS, reactive oxygen species)、Ca2+累積情形,根尖細胞Ca2+、ROS 累積隨處理銅濃度的增加而上升,可能影響細胞死亡程度。以Ca2+螯合劑EGTA 和W-7(CDPK(calcium-dependent protein kinase)、Calmodulin 抑制劑)前處理發現可以降低過量銅離子對大豆根部的細胞死亡程度,推測Ca2+、CDPK 參與銅引發大豆根部細胞死亡的途徑。為檢驗MAPK 參與根部細胞死亡的途徑,以西方墨點法偵測根部細胞MAPK 的TEY 或TDY 磷酸化,實驗結果發現,隨著過量銅離子濃度的升高,田菁、大豆根部42-kDa MAPK磷酸化情形有上升之趨勢。以RT-PCR 分析大豆MAPK1 及MAPK2 基因之表現量,發現在銅處理時大豆之MAPK1 和MAPK2 基因的轉錄情形增加。大豆( Glycine max )及田菁( Sesbania roxburghii )皆屬於豆科植物,可作為綠肥植物。探討銅影響大豆、田菁細胞死亡之訊息傳遞路徑,希望進而控制生物體所受的毒害情形及訊息傳遞途徑,加強生物體對重金屬的防禦機制,未來可以以基因轉殖等基因工程技術,轉入抗重金屬基因或增強植物體對抗重金屬的能力等,作為綠肥植物、抗重金屬植物吸 附重金屬來復育土地達綠色淨化等用途。 Many heavy metals are necessary for plants, but excessive quantities directly affect plant growth and survival of organisms, cause cell death, or even affect human life indirectly. Cu (copper ion) is a heavy metal, which is one of micronutrients essential for normal growth and development of plants. The purpose of this experiment is to study the effect of excessive copper on Glycine max and Sesbania roxburghii root tips. I conducted some experiments by means of Evans blue staining (analysis of cell death), western blot analyses, and fluorescence microscope in order to examine the way copper results in plant death. Measurement of root length and analysis of cell death showed that excessive copper could bring about the inhibition of plant growth as well as cell death. With fluorescence microscope, I found that excessive copper might increase the level of copper-caused reactive oxygen species (ROS) in both the root tips of Glycine max and Sesbania roxburghii. In addition, I used Oregon Green 488 BAPTA-1 so as to assess the accumulation of calcium ions in root tips and found that the exposure of root tips to excessive copper results in the accumulation of calcium ions. To investigate whether calcium ions and calcium-dependent protein kinase (CDPK) play a role in the cell death caused by excessive copper, I tested W-7, calmodulin and CDPK inhibitors, and EGTA, Ca2+ chelating agents, before copper treatment – immersing copper in CuCl2. In this way, plant cells would be effectively prevented from copper-caused death. Besides, to find out whether copper activates MAPKs in plant cells, I took advantage of western blot analysis with Phosphor-Map kinase Antibody and Map kinase Antibody. The results revealed that excessive copper might lead to TEY or TDY motif phosphorylation of approximate 42- and 64-kDa MAPKs in the cells of Glycine max root-tip and approximate 42-kDa MAPKs in the cells of Sesbania roxburghii root-tip. Furthermore, with RT-PCR, I found that the transcription of Glycine max MAPK1 and MAPK2 mRNA happens more frequently in root cells after copper treatment. In addition, this study suggested that the MAPK cascade CDPK pathway may function in the heavy-metal-signaling pathway in plant, and that calcium ions and ROS might get involved in the copper-caused death of plant cells. By studying signal transduction against heavy-metal toxicity in the plants, we can know how the organisms protect themselves. Sesbania roxburghiivv (or Glycine max), as green manure, could be used for metal-hyper-accumulator with the help of genetic engineering in the future.
水生開花食蟲植物絲葉狸藻捕蟲囊構造及共質體輸送
水生食蟲植物絲葉狸藻 (Utricularia gibba) 是非常獨特的,它不但跟其他植物一樣能行光合作用,且具備捕蟲囊捕捉水中小生物,補充生長所必需的營養元素。捕蟲囊的構造精密卻不複雜,消化吸收主要靠囊內壁上的四爪腺毛,目前尚未有文獻實際以追蹤物質描述出整個共質體輸送路徑。我們是最先以螢光染劑 (carboxyfluorescein) 及共軛焦雷射掃描顯微鏡(confocal laser scanning microscope) 成功地描繪出捕蟲囊共質體運輸路徑。同時我們也以對細胞無害的食用色素,進行相同的實驗觀察。發現食用色素不但成本低,且較螢光染劑有更多的優點,如觀察時間較不受限制等,非常適合用來研究捕蟲囊吸收物質的路徑,因此,本實驗的模式可以應用在其他水生植物運輸路徑的研究。;The aquatic carnivorous plant Utricularia gibba is very unique. It has not only the ability to undertake photosynthesis just like other plants, but also can trap and obtain the nutrients from the freshwater zooplankton. Its trapping organ is very sophisticate but not complicate. The digestion and absorption process inside the trap are mainly accomplished by the quadrifids structure. According to our knowledge, we are the first to introduce the phloem-mobile, fluorescent probe carboxyfluorescein (CF) and confocal laser scanning microscope (CLSM) to the study of the symplastic transport in the Utricularia trap. In addition, we use edible food colorings as tracers for this transport study. Both approaches turn out to be very successful in delineating the symplastic transport of the trap. But CF quenches rapidly so the observation time is restricted. On the contrary, food colorings don’t have these disadvantages; it is inexpensive, easy to perform, and the transport process is not fast. As a result, the study is easily to be completed. These methods will be very helpful in the studies of symplastic transport in other plants.
約瑟夫數列(Josephus Series)
所謂約瑟夫數列,就是有n 個數排成一環狀,從頭開始,殺1(個數)留1(個數),求倒數第k 個留下的數會是多少?約瑟夫數列在台灣的全國中小學科學展覽出現多次(如下表)。全國科學展覽與本題類似的作品
資訊界演算法大師Donlad E. Knuth 在其著作The Art of Programing,CONCRETE MATHEMATICS,也針對該數列作詳細的說明。唯,不論是歷屆科學展覽或是大師的著作,對於該數列,都只是談及殺1 留β或是殺α留1。
筆者則在2005 年暑假,曾經提交於全國國小組比賽作品「老師無法解決的難題」討論到n 個人排成一圈經過殺α留β,最後留下來的情形。
本研究是將α、β、k 和n 作為變數,求:當有n 個數排成一環狀,從頭開始,殺α(個數) 留β(個數),則倒數第k 個留下的數會是多少?
需符合α、β、k、n 皆∈N,且n≧k
1.直觀觀察:發現在每一個循環中,當n 等差α時,Aα,β,n,k 則等差α+β、n- Aα,β,n,k 則等差β。
2.分類:將其分類為cα,n,使當中有規律可求。
3.循環觀察:發現每個循環的尾數n- Aα,β,n,k 都小於β。
4.循環尾數:設計公式求出每個循環節的尾數n、留下數Aα,β,n,k 及n-Aα,β,n,k 。
5.倒推:由與循環節中有等差的性質,則可以由循環節的尾數,推論出循環節中的任意一數。
Joseph Sequence is the problem that discussed the situation of eliminating1 and retaining1 in the circle formed by n people. Joseph Sequence has appeared a number of times in National Elementary School and Middle School Science Fair in Taiwan (as shown in the table below). Past national science fairs and researches on Joseph Sequence
The publications,The Art of Programing,CONCRETE MATHEMATICS ,by the expert of mathematical calculation in the IT industry,Donlad E. Knuth,has provided detailed explanation on it. However, all of those only discussed eliminating 1 and retaining β or eliminating α and retaining 1.
The researcher proposed “Problems unsolved by teachers” in the national competition, and discussed the situation of eliminating α and retaining β in the circle formed by n people. This study continued the summer project of 2005, and conducted research on the question of when is the last kth person eliminated in a circle formed by n people. In the paper, α, β, n and k were independent variables and the research process was as follows:
1. Direct observation: the series shows equal difference in each cycle.
2. Classification: to search the pattern of the series based on cα,n classification.
3. Use the end number of each cycle to obtain the pattern.
4. Reverse induction: use the equal difference of each cycle to induce when the kth person would be eliminated.
滴水不漏-冷氣水回收應用分析
當我們在開車享受冷氣同時,此時冷氣水正一滴一滴的滴水,造成水資源的浪費,在環保意識抬頭的今天,我們即針對此一問題進行研究,主要將冷氣水回收起來,並運用在補充雨刷水箱或者提供引擎水箱或冷凝器降溫作用,是否達到提高引擎工作性能及降低冷氣冷房效果,進而達到「資源回收」的。實驗結果證明在補充雨刷水系統最符合環保概念;另在引擎水箱噴水作用時,可縮短風扇運時間並增加停止運轉時間,可增長風扇使用壽命,對下水管溫度亦可降低,可防引擎過熱;在冷凝器噴水作用中亦能明顯提升汽車冷房效果。When we enjoyed driving with cool air from air-conditioning, the condensed water from air-conditioning system is dripping from the system drop by drop. It caused the issues of the waste of water resource. Facing the greater public awareness of environmental protection issues in Taiwan, we are focusing on this issue to have further research. The idea is to re-cycle the air-conditioning condensed water and re-fill it in the water tank of wipers, the water cooling tank of engine or the cooling system of condenser. The purpose is to improve the performance of engine and enhance the cooling efficiency of air-conditioning system. It is helpful to meet the objective of water resource recycling. The result of experiment has shown that re-filling water in the water tank of wipers meet the goal of environmental protection well. Also, the water injection in the water cooling tank of engine could reduce each operation time of cooling fan and increase the idle time of cooling fan as well. It prolong the equipment life of cooling fan and lower the operation temperature of Low water pipe which prevent the engine overheating. Meanwhile, It is proved that the water injection in the cooling system of condenser can enhance the cooling efficiency of air-conditioning system.
植物葉片自動辨識系統
在我們週遭環境中常可見到許多種類的植物,然而可以叫出名字的卻少之又少,或許我們可以查閱植物百之類的書籍,但是這類書籍通常多不在手邊,就算有了植物百科,也不易翻到顯示該種植物的正確章節。假如我們可以將想要認識的植物葉片影像取得後,透過網路將該影像傳送至植物葉片資料庫查詢,經過電腦的自動分析辨識後,再將結果傳送回來,這樣不是比查閱植物百科方便多了嗎?本研究提出一種利用輸入葉片的影像來進行植物資料庫辨識查詢的方法,藉著兩階段處理的策略及最佳權重組合式的特徵值來調校系統,以達到較佳的整體辨識效能,從實驗測試的結果得知,我們的策略與方法確實有效,有82%的查詢葉片可以被精確的辨識出來,而每次查詢的平均反應時間只要17.22 秒。In our living environment, there are many kinds of plants, but we can only name a few. We may consult an encyclopedia about plants, we always can’t find any encyclopedia with us. Besides, even if we have one, it won’t be easy to find out the proper section or the exact page immediately. How should we solve this problem? One significant improvement can be expected if the plant recognition can be carried out by a computer. First, we take a picture of the unknown plant’s leaf. Then, we transmit this image into a leaf database to recognize. After the recognition we will get the answer easily. By using a computer-aided leaf recognition system, non-professionals can also identify many plant species. Isn’t it much more convenient than checking the encyclopedia? In this study, we present an efficient method for leaf database retrieval by inputting leaf images. We use a two-stage approach and combined features with optimized weight to adjust the system to get the best system performance. The result of the experiment shows that our approach is workable and efficient. 82% leaves of the query images can be recognized accurately. And in general, the average response time only takes 17.22 sec per query.
解開蔗糖水解的秘密
本研究利用偏振片、量角器為刻度盤、雷射光為光源,及照度計為偵測器,組裝一個簡易且可靠的旋光度計。我們利用單位時間旋光度的變化量當作反應速率,來測量蔗糖的水解速率,同時求出蔗糖水解反應的反應級數、速率常數(k)。利用糖類的旋光度具有加成性之特性,找出不同混合比例時的旋光度,追蹤實際蔗糖水解的每個狀態,找出最後平衡狀態,同時將蔗糖水解平衡結果顯示,旋光度與濃度有線性關係,而蔗糖水解反應對蔗糖而言為一級反應。接著,我們在蔗糖水溶液中加入不同種類的酸,探討催化劑的種類與蔗糖水解反應速率的關係。 In this research, in order to measure the optical rotation accurately without expensive equipments or complex process, we assembled a polarimeter by ourselves. With simple materials which can be found in ordinary senior high school laboratories, including a calibrated scale, a simple Luxmeter, a laser as the photo source, and other side devices. The Polarimeter ended up operating fluently and accurately. We put the laser under a tube, which has two pieces of polar screens on the top of it and on the bottom of it, ,and put a luxmeter just above the tube. When we slowly rotate the polar screen on the top, the figure shown on the luxmeter changes. By numerical analysis, we can get information about the hydrolysis of polarized substance. Secondary, we measured the optical rotation of glucose, fructose, malt sugar, galactose, and sucrose to get their specific rotation. Then we measured the optical rotation of sucrose every five minutes. By doing this, we could keep track of the hydrolysis rate of sucrose, figure out the order of reaction, and the rate constant (k) and the equilibrium constant (K). Thirdly, we used different kinds of acids into sucrose solution as the catalyst, and observed the effect. The result showed that hydrochloric acid is a better catalyst to this reaction than sulfuric acid and nitric acid. The polarimeter of this research can be used in science education of junior and senior high school. By teaching students to assemble and operate the self-made polarimeter, students can know better about optical rotation and polarized substance. Also, the interest in this experiement will add to students’ motivation to do science research.
國民身分證相片規格驗證暨浮水印防偽系統
政府全面換發國民身分證,並訂定新式身分證之規格,以防範遭不法偽造之情事,確保民眾權益。然而其中的照片規格,有十多條規格的限定,若用傳統的辨別方式,近1876 萬張照片是否合乎規定,那將耗費多少的人力呢?於是本次研究主題「新式國民身分證相片規格驗證暨浮水印防偽系統」即產生,設計一套程式,提高換發國民身分證的工作效率及確保換證使用相片的正確度。並且延伸研究出使用內崁式的數位浮水印〈Digital Watermarks〉,將全國民眾的身分證照片統一建立資料庫,並自動加入個人資料浮水印。日後,照片只需透過本程式分析,即可知道其姓名、身分證字號、有無犯罪前科等個人資料。希望藉此達到降低偽造身分證之犯罪率,以保護民眾之權利。 The government is launching to renew national identification cards with new norms, to avoid fake ones. However, there are more than ten limits on photos, it could be wasting time to discern by people. Thus, I launched a research on "The xamination on new national ID card photos and watermark forgery-proof system". The program will help both to enhance efficient renewal process and to use correct photos. Also we developed the embeded Digital Watermark technology, which would create a database for ID cards of the nation and could add personal infomation automatically. With the help of the program, simply run the photo analysis, we could find out the names, ID number, criminal background, etc.We hope to decrease crimes via fake ID cards, and protect the national right.
Poly(ADP-ribose)polymerase-1 對細胞內DNA damage修補的調控
Poly(ADP-ribose) polymerase-1 (PARP-1)是一個細胞核內的酵素,它可以被因DNA damage\r 而形成的DNA片段活化,並將NAD(+)上的ADP-ribose轉載到結合蛋白質。這些結合蛋白質對\r 於DNA的合成、DNA的修補、以及細胞週期的調控都有關係。因此,PARP-1被認為是維持基\r 因完整性的重要角色。根據初步的研究,抑制PARP-1的活性對許多疾病的治療都可能有效,\r 其中包括癌症、心臟病、中風、糖尿病、發炎以及反轉錄病毒的感染。然而,以藥物抑制一\r 個對DNA修補這麼重要的酵素會有什麼潛在的問題呢?為了要得到解答,我們需要進一步了\r 解PARP-1在DNA damage反應的機能。在這一份報告中,我製造了一個失去活性的PARP-1突變\r 種,即E988K。經過對E988K詳細的研究,我將比較及分析PARP-1野生型與E988K之間不一樣\r 的互動蛋白質,希望能對PARP-1所控制的DNA修補有更進一步的了解。\r \r \r Poly(ADP-ribose) polymerase-1 (PARP-1) is a nuclear enzyme activated by DNA strand breaks\r during DNA damage response and catalyzes the transfer of ADP-ribose units from the substrate NAD(+)\r to acceptor proteins. These acceptor proteins involve in modulation of chromatin structure, DNA\r synthesis, DNA repair, transcription, and cell cycle control. Thus, PARP-1 is believed to play an\r important role in maintaining genome integrity through modulation of protein-protein and protein-DNA\r interactions. PARP-1 has been the target for design of inhibitors for over twenty-five years. Inhibitors of\r the activity of PARP-1 have been claimed to have applications in the treatment of many disease states,\r including cancer, cardiac infarct, stroke, diabetes, inflammation and retroviral infection. However, are\r there potential problems associated with inhibition of this DNA-repair enzyme? To answer this question,\r we need to further understand the biological function of PARP-1 during DNA damage response. In this\r report, an enzyme dead mutant (E988K) of PARP-1 was generated. Detailed studies of E988K show that\r E988K could be used in the following studies. Compare and identify the different associated proteins of\r PARP-1 wild-type and E988K will shed light into the molecular mechanism of PARP-1-mediated DNA\r repair.