http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/72 2026-03-12T06:02:38Z http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75105 標題: 黑豆子葉葉綠體色素蛋白複合體循環與類囊膜摺疊之研究; Study on the Pigment-protein Complex Cycle and Thylakoid Stacking of Black Bean Cothledon 作者: Liang-Suei Lu; 呂良穗 摘要: 本研究主要探討非葉綠色組織黑豆子葉於成熟、脫水及萌芽期間之色素體的形態變化、類囊膜摺疊、光合色素的變化以及色素蛋白複合體組成份的差異。在種子形成期間，子葉近軸面有類似氣孔發育早期的構造；萌芽復，子葉近軸面與遠軸面的氣孔形態差異大，且近軸面氣孔較發達。色素體的形態變化，在豆莢內種子的子葉，多為不規則狀含澱粉粒的色素體，有的只含單一澱粉粒、有的含有聚集型的澱粉粒，它們均有不具漿質類囊膜連結的葉綠餅疊層存在；乾種子子葉內的色素體具有層疊較緊密但不太平整的葉綠餅，浸水後變得鬆散且多方向性；萌芽照光復，出現晶格狀的原囊體(prolamellar body)，其放射出類囊膜，再層疊成葉綠餅，最後發育成完整的葉綠體。 豆莢內種子子葉的葉綠素此比值偏低，介於1.7至2.1之間，萌芽後升為2.8，與成熟葉月相似。各時期子葉的葉綠素總量與??(porphvrin)總量的變化趨勢相似，種子成熟脫水期問其含量遞減，萌芽照光後驟增。三種??莫耳百分比在各發育階段呈現週期性的起伏變動，protoporphyrin IX的變化趨勢恰與Mg-protoporphyrin IX和protochlorophyllide相反。HPLC分析的葉黃素中，各時期子葉lutein的含量均最高；豆莢內種子子葉的環氣化指數較萌芽後的子葉低。 以SDS-PAGE梯度膠體電泳分析成熟子葉和葉片的類囊膜蛋白質，兩者組成份相似。以Thomber和MARS電泳分析，黑豆子葉在萌芽照光前分別只含CPII和AB3兩種色素蛋白複合體；萌芽照光後，原本缺失的色素蛋白複合體均恢復為與成熟葉月相似，此即形成非葉綠色組織特有的色素蛋白複合體循環（pigment-protein complex cycle, PPCC）現象。此外，利用電顯免疫細胞化學定位類囊膜蛋白質LHCII b，其均標定於葉綠餅上，顯示LHCII b與類囊膜的摺疊具有某種程度的關聯。; The present study is try to describe the changes in non-leaf green tissue of cotyledons of black bean (Glycine max. L. Merrilx) during seed ripening desiccation and germination, including the changes in plastids morphology、thylakoid stacking、photosynthetic pigments and the compositions of pigment-protein complexes. During seeds ripening, guard mother cells appeared in the cotyledons. After germination, the stomatal feature and distribution of the adaxial and abaxial regions of cotyledons are very different. There are more stomata on the adaxial region. The plastids of cotyledons of developing seeds in the pods are quite different. They are plastids with irregular shape. Some of them are with only one starch grain and some with aggregate starch grains. All of them have grana stacking without stromal lamelle connecting. In dry seeds, there are more compact thylakoid stacking without flatted grana in the plastids. After imbibition, the thylakoid stacking of grana become looser and multi-direction orientated. After seed germination and seedling exposed to light, there are crystal-shape prolamellar bodies in plastid, from which thylakoids extended to growth and stackness becoming grana. The chlorophyll a/b ratio of the cotyledons of the seeds in the pods was low down between 1.7 and 2.1; after germinaiton, it became 2.8. The changing patterns of contents of total chlorophylls and porphyrins of cotyledons were similar. They decayed during seed ripening and desiccation, and enriched rapidly after germination. In each developmental stages, the mole fraction of three porphyrins revealed periodical fluctuation. The changing pattern of protoporphyrin IX was different from those of Mg-protoporphyrin IX and protochiorophyllide. HPLC profile of xanthophylls showed that the contents of lutein were the highest in cotiledons at all stages of seed. The epioxidation index of seed cotyledons in the pods is about 0.1 lower than that ones after germination. Similar thylakoid protein pattern of mature cotyledons and leaves were revealed by SDS-PAGE gradient gel electrophoresis. In Thornber system electrophoresis, the cotyledons of imbibition seed was deficient CPI pigment-protein complex, and only with CPII. In MARS system electrophoresis, it was deficient Al、AB1 and AB2, and only with AB3. After seed germination and seedling exposed to light, all pigment-protein complexes were recoveried and were similar with the pattern of leaves. This indicated that a phenomenon of pigment-protein complex cycle (PPCC) specifically belongs to non-leaf green tissues. Immunocytochemical studies revealed that the colloidal-golds of LHCII b protein were specifically located in grana of cotyledons. This indicated that there are relationship between LHCII b protein and thylakoid stacking. 2000-01-01T00:00:00Z http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76254 標題: 鴛鴦湖地區植物的花粉形態研究; Pollen Morphological Study of Vegetation Around the Yuen-yang Lake 作者: Yu-Fa Wang; 王裕發 摘要: 鴛鴦湖為台灣北部自然保護區內之一高山湖泊，本研究在鴛鴦湖附近 10 公里範圍內每月定期採集開花植物之新鮮花粉，以酸化處理後，經過固定、脫水、臨界點乾燥法和金屬離子覆膜等步驟，再以掃描電子顯微鏡觀察並照像，所得之資料與光學顯微資料相配合描述之。本研究共描述了 144 種種子植物的花粉，包括裸子植物 3 科 5 種，雙子葉植物 54 科 119 種，單子葉植物 7 科 20 種。依照花粉萌發孔的特徵，可將之歸納為下列 17 種類型： 1 ．具氣囊的花粉：台灣二葉松； 2 ．無萌發孔的花粉：如台灣天南星、線葉眼子菜； 3 ．具薄壁區的花粉：如水毛花、巒大杉、紅檜、台灣扁柏； 4 ．單孔型的花粉：如白頂早熟禾，高山芒，束亞黑三稜： 5 ．雙孔型的花粉：如短角冷水麻； 6 ．三到六孔型的花粉：如銳葉楊梅、水麻、山桔梗、台灣赤楊； 7 ．散孔型的花粉：如台灣瑞香、台灣黃蓮、箭葉蓼； 8 ．螺旋狀萌發孔型的花粉：台灣小蘗： 9 ．窗形的花粉：苦苣菜； 10 ．單溝型的花粉：如阿里山假寶鐸花、銳葉胡麻花、台灣沿階草； 11 ．三溝型的花粉：如昆欄樹、白花八角； 12 ．四到七溝型的花粉：如阿里山五味子、小葉四葉律、山酢漿草，塔花； 13 ．三溝孔型的花粉：如日本女貞、假柃木、水亞木、太平山冬青、紫金牛、乞食碗、台灣樹參、台灣龍膽； 14 ．四至七溝孔型的花粉：如深紅茵芋、伏牛花； 15 ．異溝型的花粉：如臺北附地草、蘭崁馬藍； 16 ．四面體具三孔的花粉：黑龍江柳葉菜： 17 ．四面體具三溝孔的花粉：如台灣杜鵑、台灣馬醉木。本研究亦記錄了鴛鴦湖附近地區 224 種植物的開花期，配合花粉形態描述及本研究所提供的檢索表，可做為宜蘭及附近地區研究空中花粉和化石花粉的基本資料。; The pollen morphology of the vegetation around the Yuen-yang Lake which is located in nature conservation area in northern Taiwan was investigated. Fresh pollen samples were collected every month from the area about ten km surrounding the lake, Of 144 species belonging to 64 families of spermatophyte were treated with acetolysis method and then mounted on slide for LM observations or were further dehydrated through alcohol series followed by critical point drying, and were coated with gold for SEM observations. The 17 pollen classes based on the germ aperture are found and categorized as follows: (1) saccate pollen; (2) inaperaturate pollen; (3) pollen with leptoma; (4) 1-porate pollen; (5) 2-porate pollen; (6) 3-6-porate pollen; (7) pantoporate pollen; (8) spiraperturate pollen; (9) fenestrate pollen; (10) sulcate pollen; (11) 3-colpate pollen; (12) 4-7-colpate pollen; (13) 3-colporate pollen; (14) 4-7- colporate pollen; (15) heterocolpate pollen; (16) tetrad with monads 3-porate; (17) tetrad with monads 3-colporate. The data of pollen flora provide a basic information for further studies such as aeropalynological studies and the reconstruction of paleo-ecology around Yuen-yang Lake. 1996-01-01T00:00:00Z http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75805 標題: 鳳凰木的植物相剋作用潛能之研究; The Allelopathic Potential of Delonix regia (Boj.) Raf. 作者: Lih-Ling Leu; 呂麗玲 摘要: 鳳凰木(Delonix regia (Boj) Raf.)係熱帶的豆科植物，經常在此植物下，其地被植物種類少，且生物量相當低。依田野的觀察及測定，此地被植物稀少的原因非物理因數如光、水或營養物質的競爭所致，而可能是由於鳳凰木的落葉或其代謝物質抑制地被植物的相剋作用所致。?證明此相剋作用，本研究以生物分析、色層分析、掃描式電子顯微鏡及組織切片等方法來證明鳳凰木植物相剋作用潛能。 將掉落的鳳凰木花、葉、枝條以不同百分比萃取的水溶萃取液如1％、2％、3％、4％、5％，分別對萵苣、苜蓿、白菜、油菜作生物分析以比較其植物毒性程度。其結果顯示花水溶萃取液在1％時抑制白菜根生長達81.5％，抑制油菜、萵苣根生長達77.6％及70.1％，其次是枝條及葉的水溶萃取液，而鳳凰木樹下的土壤及其根圈土或腐葉土之水溶萃取液之植物毒性不顯著，顯示植物毒性表示於花、葉、枝條等部位。一般言之，藿香薊常生長在陽光充足的棲地，偶而也在陰暗地方，但在具相當光度之鳳凰木樹下卻不常見，作者選用上述萃取液做一系列探討卻發現無明顯抑制現象，此原因有待進一步探討。對另二種陰地植物如日本柳葉箬、雷公根的毒性程度則較明顯，由此顯示陰地植物被抑制的現象與植物相剋作用有關。 ?進一步瞭解植物毒物植對植物根生長抑制之機制，作者選用白菜種子以鳳凰木水溶萃取液做生物分析處理後以掃描式電子顯微鏡觀察發現白菜種子根發育明顯受毒物質抑制，根冠細胞有嚴重剝落現象。 經HPLC之分析，本研究經鑑得鳳凰木中之植物毒物質有gallic acid, 3,4-dihydroxybenzoic acid, 3,4-dihydroxybenzaldehyde, chlorogenic acid, 4-hydroxybenzoic acid, 3,5-dinitrobenzoic acid, 3,4-dihydroxyhydrocinnamic acid等，尚有未被鑑定之類黃素及其它物質亦可能?植物相剋物質。 本研究結果證明鳳凰木抑制樹下地被植物生長與鳳凰木所產生之次階代謝物質及其中所含之植物毒物質有關。鳳凰木所產生之次階代謝物及其中所含之植物毒物質亦影響其土壤中之微生物相。; Delonix regia (Boj.) Raf. plantation in Tainan, southern Taiwan, exhibits a unique pattern of weed exclusion beneath canopy. The pattern has been particularly pronounced in the area where a substantial amount of D. regia litter has accumulated on the ground. Field data showed that the phenomenon was due primarily not to physical competition, involving light, soil moisture and nutrients, but due to an allelopathic interaction. Aqueous extracts fallen leaves, branches, flowers of D. regia showed significantly phytotoxic effects on many test species, including lettuce (Lactuca sativa), alfalfa (Medicago sativa), pickled cabbage (Brassica chinensis) and Brassica campestris L. subsp. napus Hook. f. & Anders. var. nippo-oleifera Mak. The aqueous extracts of the flower of D. regia revealed the highest phytotoxicity. In order to further understand the inhibition of phytotoxic effect, seeds of Brassica campestris were soaked in the aqueous extracts for various time period and the seedings of B. campestris were observed by scanning electron microscopy. Injury in seedings of the tested plants were shown by deteriorated epidermis and abnormally swelling of the seedings. The allelopathic substances isolated from D. regia leaves, flowers, branches and soil were identified by paper chromatography and a high performance liquid chromatography (HPLC). They are phenolic acids, namely, gallic acid, 3,4-dihydroxybenzoic acid, 3,4-dihydroxybenzaldehyde, chlorogenic acid, 4-hydroxybenzioc acid, 3,5-dinitrobenzoic acid, 3,4-dihydroxycinnamic acid, and some unidentified flavonoids and others as well. The findings of the present study concluded that the inhibition pattern beneath the trees of D. regia is due primarily to the allelopathic substances in D. regia. The responsible allelopathic substances are mostly phenolic acids, but could be flavonoids and others as well. These metabolites could possobly be degraded by soil microorganisms, such as Candida guilliermondii and Enterobacter agglomerans, Klebsiella spp. and Rhodotorula spp. 1991-01-01T00:00:00Z http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75326 標題: 鯉魚潭中葡萄藻代謝物質之生物毒性; The biotoxicity of metabolites of Botryococcus braunii from Lake Li-Yu 作者: 江殷儒 摘要: 面積約100公頃的鯉魚潭為花蓮縣最大湖泊，經複合參數指標判定水質長期屬於輕度優養狀態。鯉魚潭於1993、1996及1999年秋冬季皆曾發生嚴重的葡萄藻華，期間並伴隨吳郭魚大量死亡的現象。為評估葡萄藻華對該水域生態系統的影響，故對葡萄藻的生物毒性進行研究。以葡萄藻粗萃取物對分離自鯉魚潭的十五株藻種進行毒性測試，結果顯示除了Microcystis flos-aquae及Oocystis parva，其餘十三株藻種皆表現程度不一的敏感度。對鯉魚潭浮游藻群落的分析結果顯示，藻種對葡萄藻的敏感度與其在葡萄藻華期間所佔有的相對優勢度，呈現密切相關。本研究亦使用葡萄藻粗萃取物對鯉魚潭的優勢浮游動物進行毒性測試，結果顯示佔有浮游動物群落60％以上組成的橈足類較枝角類敏感得多。前述測試結果顯示，葡萄藻可能藉由相剋物質的作用，對鯉魚潭的浮游生物群落造成影響。 批次培養結果顯示，在磷或氮源成為限制因數而細胞進入指數生長末期時，葡萄藻能產生並釋出大量的毒性物質。此時期的培養液之營養鹽濃度與葡萄藻華時期的鯉魚潭水質條件最為接近，顯示該水域提供適當的化學環境，使葡萄藻能生長良好並藉由相剋作用與其他藻類競爭。 葡萄藻毒性物質溶於有機溶劑中，經薄層層析、高效液相層析等技術進一步分離及純化，並以核磁共振及質譜等技術鑑定結構後，確認為包括棕櫚酸（16:0）、油酸（18：1ω9）、亞麻油酸（18：2ω6）及α-亞麻脂酸（18：3ω3）在內的數種自由態脂肪酸。其中，三種18碳不飽和脂肪酸對多數測試藻株及甲殼綱浮游動物具有明顯毒性，半抑制濃度EC50（或LC50）約在10-20μg•mL-1之間。葡萄藻的總自由態脂肪酸含量佔細胞乾重的2.25％左右，遠高於其他七個分屬藍綠菌、綠藻及渦鞭毛藻門的測試藻種。 葡萄藻粗萃取物及其主要脂肪酸經由阻斷電子傳遞鏈，而能抑制藻細胞的光合作用，對呼吸作用則皆無明顯影響。三種18碳不飽和脂肪酸在低於明顯抑制光合作用的施加劑量下，即可造成藻細胞的死亡；而DCMU在完全抑制光合作用的高劑量下，卻不會在24小時內造成藻細胞死亡。顯示對光合作用的抑制並非葡萄藻對其他藻類的關鍵致毒機制。 三種18碳不飽和脂肪酸在施加三十分鐘內會造成藻細胞5-10％鉀離子滲漏，並在測試的四小時內持續增加。進一步的研究顯示，此些18碳不飽和脂肪酸會對藻細胞膜上的P型ATPase造成抑制，對V型及F型ATPase則無明顯影響。由於P型ATPase（尤其是H+-ATPase）為藻細胞維持膜電位的最重要酵素，不飽和脂肪酸造成藻細胞之鉀離子滲漏可能與P型ATPase受抑制有關。另一方面，葡萄藻粗萃取物及其主要脂肪酸亦會對枝角類及橈足類的Na+/K+-ATPase產生專一性抑制；而對F型及V型ATPase，甚至其他P型ATPase皆無任何抑制效果。顯示葡萄藻毒性物質可能經由抑制Na＋/K＋-ATPase活性，導致浮游動物因滲透壓失衡而死亡。; With a surface area of ca. 100 ha, the Lake Li-Yu is the largest lake in Hwa-Lian county, Taiwan. It is an eutrophic lake with Botryococcus braunii blooms occurred in autumn and winter in the years of 1993, 1996 and 1999. Death of fish, mainly Tilapia, were observed in the lake during B. braunii blooming. In order to elucidate the effects of algal blooms in this aquatic ecosystem, the toxicity of B. braunii was studied. Fifteen strains of phytoplankton were isolated from the lake and the cultures were tested for the toxicity of B. braunii extract on them. Results showed that except Microcystis flos-aquae and Oocystis parva, all other species exhibited various degrees of sensitivity to B. braunii. Population analysis of the tested species in the lake showed that the sensitivity to B. braunii was closely correlated with the abundance of these phytoplankton species in the lake during Botryococcus blooming. Toxic effects of B. braunii extract were also observed to the zooplankton collected from the Lake. In them, copepods showed higher sensitivity to crude extracts from B. braunii than cladocerans. All of these results suggest that B. braunii might play an allelopathic effect on other plankton. In batch culture, the toxicities of B. braunii were enhanced when P- or N-nutrient became a limitation. The maximum toxicity was measured when the growth of cells was near the end of exponential growth phase. Under this condition, the nutrient concentrations in culture medium were close to those measured during B. braunii bloom, suggesting that the chemical environment in the Lake Li-Yu might create a favorable condition for B. braunii to survive and to compete with other plankton species. The fractions containing active substances of B. braunii were soluble in organic solvents. Further isolation, purification, and identification of active substances, by employing TLC, HPLC, NMR, and GC-MS techniques, indicated that they were fatty acids, including palmitic acid (16:0), oleic acid (18:1ω9), linoleic acid (18:2ω6) and α-linolenic acid (18:3ω3). The highest toxicity was revealed in C18-unsaturated fatty acids whose EC50 (or LC50) were found between 10-20μg•mL-1 against phytoplankton as well as against zooplankton. Total free fatty acid content might be as high as 2.25% of dry weight, which was much higher than other 7 tested species of cyanophytes, chlorophytes and dinophytes. Addition of B. braunii extract inhibited the photosynthetic electron flow of tested algal cells, while the respiration rate was affected little. Similar effects were also observed when C18-unsaturated fatty acids were added to the algal cultures. It is noteworthy that the tested cells would die even when the added concentrations were lower than IC50 value of inhibition of photosynthesis. However, the photosynthetic electron flow inhibitor, DCMU, did not cause the death of tested algal cells within 24 hrs. This implicates that the inhibition of electron flow was not the only cause of death B. braunii exerted to other algal species. It was observed that addition of C18-unsaturated fatty acids to algal cultures caused a remarkable K+-leakage from tested cells within 30 minutes. The amount of released K+ might as high as 5-10% of total K+ in the cells. Enzymatic assays of ATPase showed that C18-unsaturated fatty acids did inhibit P-type, but not V- or F-type ATPase of the tested algal cells. Because P-type ATPase (especially H+-ATPase) is related to the maintenance of membrane potential, inhibition of this type of membrane enzyme might be attributed to K+-leakage from the cells. C18-unsaturated fatty acids did not exert inhibitory effect on V-, F-, or other P-type ATPase, but on Na+/K+-ATPase of copepods and cladocerans. It is assumed that toxic effects of B. braunii on crustacean zooplankton might be related to the interference of osmoregulation through inhibiting the Na+/K+-ATPase activity. 2002-01-01T00:00:00Z