Fruit Quality and Skin Freckle as Related to Nitrogen Application and Cluster Wrapping in 'Tainung No.2' Papaya (Carica papaya L.) Fruits

春、冬季的番木瓜果實表皮會出現生理性污斑(skin freckle)，嚴重影響果實的外觀，本試驗主要探討污斑的成因與減少污斑的方法。首先探討氮肥的影響，‘NTU 10號’與‘台農二號’植株於始花期後，每2週施用四種不同氮肥量作為處理，分別為每株11.9、25.0、38.2、51.2g硫酸銨。於施肥7個月後以及停止施肥後1個月採樣分析果實及葉片成分。試驗結果，以每株施用38.2g硫酸銨效果較佳，而施用高氮肥處理(51.2g/株)，果實於後熟後腐爛率明顯提高，而且‘台農二號’的果皮轉色較差，糖度也略低於其他處理。在礦物元素分析，隨著氮肥施用，葉片與果實N/Ca值增加，亦會間接影響鉀元素的吸收。每株施用11.9g硫酸銨處理，明顯有缺氮的情形，尤其是葉片較果實敏感，會誘導葉片總酚類化合物增加。另外，‘NTU 10號’與‘台農二號’番木瓜在催熟前污斑發生面積相似，但催熟後‘台農二號’會急劇上升，相對來說‘NTU 10號’在催熟後污斑的變化不大，顯示‘台農二號’為污斑較敏感的品種，而不同氮肥處理對污斑發生面積較無直接關聯。 其次在污斑指數高及低的季節，採集不同生長階段的果實，作相關的生理及生化成分分析，隨著果實生長發育，果皮中磷、鉀、鎂濃度會逐漸遞增，其中在11月份鈣、鋅濃度，至污斑出現時並無隨果實生育而遞減，反而維持平穩，顯示污斑發生並非元素缺乏所引起，推測因營養較多導致細胞壁較為堅硬，導致果實成熟時，膨壓增加，進而使細胞結構發生分裂而形成污斑。而果肉中氮、磷、鈣、鎂、鐵、錳、鋅、銅濃度皆會隨果實生長發育逐漸下降，至果肉完全轉色時維持平穩，而鉀濃度在生長期間僅少量變化，總酚類化合物與苯丙胺酸脫氨裂解酶活性亦隨著果實轉色而逐漸遞增。 污斑發生的原因可能與低溫有關，果實在套袋後因袋內溫度較為穩定而減少污斑產生。南投地區可能因為溫差較大而使污斑發生面積高於高雄地區。果實生長至一定大小，於污斑未出現前進行果房包覆96天可明顯降低污斑的發生，套袋材質以泰維克布的減少污斑效果較不織布佳。套袋後果皮較明亮且有光彩，但會降低果重，而水分含量、乾重、乳汁量、糖度則無明顯差異。果實在套袋48天後，果肉中鉀、鈣、鎂濃度皆較對照組低。套袋時間增加至96天後影響反而較小，僅鉀、錳濃度較對照組低。另外，套泰維克布處理的果皮其總酚類化合物與苯丙胺酸脫氨裂解酶活性較低。果皮污斑的發生可能與果實在逆境下所引發的褐化反應有關。Skin freckles on papaya fruits, as a rule, appear in spring and winter and thus affect fruit appearance seriously. The objectives of this study were to investigate the cause of this disorder and to reduce the occurrence of freckles. In the experiment of nitrogen treatments, four concentrations of ammonium sulphate (NH4)2SO4 were applied to ‘NTU 10’ and ‘Tainung No.2’ papaya trees (11.9、25.0、38.2、51.2 g/tree) at intervals of every two weeks. Compositions of leaves and fruits were measured seven months after fertilization and again one month after stop applying fertilizer. The result of the experiment showed that 38.2 g/tree was the best concentration used among treatments. The rate of fruit decay was a serious problem from high nitrogen fertilization. Peel color change of ‘Tainung No.2’ papaya was somewhat delayed, and the total soluble solids were also lower than other treatments. Increasing N/Ca ratio of leaves and fruits was found with increased nitrogen fertilizer, and thus indirectly affected potassium accumulation. Treatment with 11.9g ammonium sulphate per tree resulted in fruits and leaves nitrogen deficiency, especially in leaves. However, accumulation of total phenolic compound was found. Before ripening the freckle incidence area of ‘NTU 10’ and ‘Tainung No.2’ papaya fruits were similar, but ‘Tainung No.2’ was more serious after ripening. There was no significant correlation between N fertilization and the appearance of freckles. ‘Tainung No.2’ was consistently more susceptible than ‘NTU 10’ papaya fruits. In season of high and low skin freckles occurrence during fruit growth and development, chemical analysis of phosphorus, potassium, magnesium concentrations in the peel showed the tendency of gradual increase. Concentration of calcium and zinc in peel leveled off in November while freckle appeared. This seemed to indicate that freckles were not related to the mineral deficiency. It is assumed that the hardening of the cell wall and increasing in turgor pressure during the ripening of fruits could lead to the degeneration of wall structure and the occurrence of freckles. At the later stage of fruits development, the phosphorus, calcium, magnesium, iron, manganese, zinc, copper in pulp decreased progressively and stabilized until complete turning of pulp color. Potassium content change very little during entire period of fruit growth. Total phenolic compound and phenylalanine ammonia-lyase activity of fruit increased during the period of peel and pulp color change. The main cause of the skin freckle was related to the low-temperature. Wrapping fruits in bags to reduce the temperature fluctuation improved the situation. In Nantou County more skin freckles occurred than in Kaohsiung due to the differences in temperature in the two areas. Cluster wrapping of fruits with Tyvek material before the appearance of freckles, while still attached to the tree, could reduce the physiological disorder after bagging for 96 days. Tyvek material was better than Non-woven material for the purpose. Moreover, wrapping increased brightness and chroma of the peel, but reduced fruit weight at harvest. No significant differences in water content, dry matter, total soluble solids, and latex volume of fruits were found between Tyvek and non-wrapping treatments. The potassium, calcium and magnesium of pulp were less after 48 days wrapping with cluster Tyvek than non-wrapping treatment. With increasing in time of bagging, only potassium, manganese concentrations were decreased. Total phenolic compound and phenylalanine ammonia-lyase activity were less after cluster wrapping with Tyvek. In conclusion, skin freckles occurrence seems to be associated with browning of fruits and environmental stress.中文摘要 i Summary ii 圖目次 v 表目次 vii 壹、前言 1 貳、前人研究 2 一、番木瓜起源與概說 2 二、番木瓜果實生長發育與其成分之變化 2 三、葉片礦物元素及光合產物對番木瓜果實品質之影響 6 四、生理性斑點 10 五、套袋對果實品質之影響 15 參、材料與方法 18 一、氮肥施用量對‘NTU 10號’與‘台農二號’番木瓜果實品質之影響 18 二、氮肥施用量對‘NTU 10號’與‘台農二號’番木瓜葉片之影響24 三、不同地區果房包覆對‘台農二號’番木瓜果實品質之影響 25 四、‘台農二號’番木瓜果實不同生長階段礦物元素與化學成分之變化 28 五、‘台農二號’番木瓜果實8-11月乳汁礦物元素分析 30 肆、結果 31 一、氮肥施用量對‘NTU10號’與‘台農二號’番木瓜果實品質及成分之影響 31 二、不同污斑程度對‘NTU10號’與‘台農二號’番木瓜果實乳汁量與水分潛勢之影響 44 三、氮肥施用量對‘NTU10號’與‘台農二號’番木瓜葉片之影響47 四、不同地區果房包覆對‘台農二號’番木瓜果實品質之影響 52 五、‘台農二號’番木瓜果實不同生長階段礦物元素與化學成分之變化 69 六、‘台農二號’番木瓜果實8-11月乳汁礦物元素之變化 92 伍、討論 94 一、氮肥施用量對‘NTU10號’與‘台農二號’番木瓜葉片及果實品質及污斑指數之影響 94 二、斑點(污斑)發生原因之探討與果房包覆降低‘台農二號’番木瓜生理性斑點之可行性 98 三、‘台農二號’番木瓜果實在不同發育階段其組成分之變化 105 陸、參考文獻 110 附錄 1. 高雄與南投地區2006-2007年月平均雨量之變化(資料來源：中央氣象局) 120 附錄 2. 高雄與南投地區2006-2007年月平均溫度之變化(資料來源：中央氣象局) 121 附錄 3. 採樣果園施肥記錄 12

檬果炭疽病防治試驗(摘要)

檬果炭疽病(Mango Anthracnose: Colletotrichum gloeosporioides Penz.）為本省檬果重要病害之一，其罹病率幾連35% 以上，被害果實輕則發生黑色污斑，影響美觀，重則果皮凹陷，果肉腐爛而完全失卻經濟價值，故如何防治該病以增加檬果之增產，實為當務之急。 本試驗為探求果炭疽病坊效防治藥劑種類及濃度以便減低被害率而提高產量與品質，自民國52年1月起至6月止，在本分所檬果園學行試驗

Seasonal Fluctuation of Fruit Quality, Mineral Nutrient, and Skin Freckle of 'Tainung No.2' Papaya Fruits (Carica papaya. L.)

本試驗自2005年9月起至2006年11月止調查雲林縣林內鄉栽植之‘台農二號’番木瓜果實品質及生理性斑點發生情形。每月取樣一次，果實採收後於30℃下催熟，2日後調查。果皮於9月至11月皆可正常轉色，而12月至2月有轉色不良的現象，9~11月採收之果實可正常軟化，而12月以後果實硬度較高、後熟速率減緩。另外，糖度也以接近冬季者高於夏秋季。 番木瓜果實表面之生理性斑點(skin freckle)影響其外觀甚鉅，但發生機制未明且周年均會發生，以果實外側(向陽面)較為明顯，同常於採收前已出現斑點，催熟後常由綠色平面轉為更嚴重之木栓化突起狀，並遍佈果皮表面。斑點指數(freckle index, FI)以12月至隔年5月低溫生長季節最為嚴重，7-9月最輕微，果肉質地及果皮轉色較佳。 以電子顯微鏡進行顯微結構之觀察，果表均被角質層之蠟質覆蓋。綠熟果實之表面較為平坦完整，黃熟果則較有凹凸不平，其蠟質片段明顯為層疊板塊狀。於果實表面甚少觀察到氣孔，可能多被蠟質覆蓋，有斑點者周圍蠟塊更細薄且破碎。分析果實內外側元素濃度之週年變化，較嚴重之低溫季節或較嚴重部位(果實外側)分析結果顯示含有較高鈣濃度，但變化並不一致，內外側果肉、果皮間元素通常無顯著差異，與斑點之發生可能無直接關係，但斑點之發生與酚類化合物累積而導致褐化有關。番木瓜果實乳汁量隨果實發育階段而有減少之傾向，且斑點發生嚴重性逐漸增加，與乳汁管破裂有關，於蛋白質PAGE電泳分析中也顯示有斑點部位蛋白質條帶較趨近於乳汁，乳汁之流出造成果皮傷害，可能與斑點產生較有相關性。 葉片礦物元素之周年變化顯示，各元素濃度為N：3.5-3.7%；磷：0.32-0.33%；鉀：2.4-2.5%；鈣：2.2-2.5%；鎂：0.65-0.7%；鐵：80-100ppm；錳：20-25ppm；鋅：20-25ppm；銅：4-5ppm，可供葉片分析方法標準化及營養診斷、肥培管理之參考。而8至9月之變化較平緩，為採樣適期。Papaya fruit samples were collected once a month in Lennei County from Sep. 2005 to Nov. 2006, and were ripened at 30℃ for two days. Samples from September to November displayed normal color turning, and ripening, samples from Dec. 2005 to Feb 2006 showed relatively hard pulp, slower ripening and poor color change. Total soluble solids was higher in winter month than summer. Papaya skin freckle is a physiological disorder. It happens all year round especially on the abaxial portion of the fruit. The symptom appears before harvest and develops into suberized lumpy patch and spread to the whole skin layer after ripening, thus affects the appearance of fruits seriously. But the mechanism of the disorder is unknown. The freckle index (FI) was high from Dec. to next May, lower from July to Sept. Observing the ultrastracture of fruit peel by SEM and found the surface of fruits was covered with wax and cuticle. The surface of green papaya was smooth, while the yellow ripe fruits, rough and uneven. The fraction of wax was obviously in layer formation. Stomata were hardly visible, possibly because they were covered by the wax therein and wax-like fragment around freckles were rather thin and broken. Monthly mineral analysis showed higher calcium concentration in fruit abaxial side during the cooler season. There was no significant variation observed in mineral elements on abaxial or adaxial side. Freckles might not directly relate to the concentrations of elements, but was associated with the accumulation of total phenolic compounds. As fruits going through developing stages, papaya latex volume decreased, freckle index increased and proteins pattern demonstrated similar tendency as latex. The exudates of latex might possibly be one factor of fruit injury. It was suggested that latex played an important role in the case of freckle induction. The seasonal changes of mineral levels of ‘Tainung No. 2'papaya leaf was determined. The ranges of various mineral elements were as followed: N: 3.5-3.7%;P: 0.32-0.33%;K: 2.4-2.5%;Ca: 2.2-2.5%;Mg: 0.65-0.7%;Fe: 80-100ppm;Mn: 20-25ppm;Zn: 20-25ppm;Cu: 4-5ppm. The data could be used as a guide for the diagnosis of mineral disorders, soil management of papaya. It is concluded that August to September are the proper months to sample for analysis because of the stable condition of minerals in papaya leaves.目錄 摘要......................................................i Summary..................................................ii 圖目次...................................................iv 表目次....................................................v 壹、前言..................................................1 貳、前 人 研 究...........................................2 一、番木瓜之概說..........................................2 二、番木瓜果實生長發育與後熟之變化........................3 三、影響果實品質之因子....................................7 四、生理性斑點（Skin freckle)............................14 五、葉片礦物元素.........................................19 參、材料與方法...........................................21 ㄧ、番木瓜果實品質季節性變化之調查.......................21 二、番木瓜果實礦物元素及褐化相關成分季節性變化之調查.....23 三、番木瓜葉片礦物元素變化之調查.........................26 四、番木瓜果實不同生長期礦物元素及化學成分之變化.........27 五、番木瓜果皮蛋白質及過氧化酵素(POD)電泳分析............29 六、番木瓜果實生理性斑點(污斑)掃描式電子顯微鏡之分析.....31 肆、結 果................................................32 ㄧ、‘台農二號’番木瓜果實品質週年變化與污斑發生比例之調查.......................................................32 二、‘台農二號’番木瓜葉片及果實礦物元素之周年變化.......35 三、‘台農二號’番木瓜不同果實發育階段礦物元素之變化.....41 四、‘台農二號’番木瓜果皮有無污斑蛋白質電泳圖之比較.....43 五、‘台農二號’番木瓜污斑之顯微構造之觀察...............43 伍、討論.................................................82 一、果實之品質之周年性變化...............................82 二、生理性斑點週年性變化.................................83 三、生理性斑點發生原因之探討.............................84 參 考 文 獻..............................................91 附圖1、雲林地區月平均溫度及雨量圖(資料來源：中央氣象局).....................................................100 附表1、蛋白質電泳膠片之製作.............................101 附表2、過氧化酵素電泳膠片之製作.........................102 附表3、採樣果園整園施肥記要.............................103 圖目次 圖 1. 番木瓜生理性斑點(污斑)。...........................44 圖 2. 番木瓜生理性斑點(污斑)以外側較為嚴重。.............44 圖 3. ‘台農二號’番木瓜果實催熟前後污斑發生率之季節性變化。.....................................................45 圖 4. ‘台農二號’番木瓜果實催熟前後污斑嚴重程度之季節性變化。.....................................................45 圖 5. ‘台農二號’番木瓜果實外側催熟後污斑之變化。.......46 圖 6. ‘台農二號’番木瓜果實硬度催熟前後之季節性變化。...47 圖 7. ‘台農二號’番木瓜果實糖度催熟前後之季節性變化。...47 圖 8. ‘台農二號’番木瓜果皮催熟前後L、a*、b*值之季節性變化。.....................................................48 圖 9. ‘台農二號’番木瓜果皮催熟前後彩度與色相角之季節性變化。.....................................................49 圖 10. ‘台農二號’番木瓜果肉催熟前後L、a*、b*值之季節性變化。.....................................................50 圖 11. ‘台農二號’番木瓜果肉催熟前後彩度與色相角之季節性變化。.....................................................51 圖 12. ‘台農二號’番木瓜果肉與果皮氮濃度催熟前後之季節性變化。.....................................................52 圖 13. ‘台農二號’番木瓜果肉與果皮磷濃度催熟前後之季節性變化。.....................................................53 圖 14. ‘台農二號’番木瓜果肉與果皮鉀濃度催熟前後之季節性變化。.....................................................54 圖 15. ‘台農二號’番木瓜果肉與果皮鈣濃度催熟前後之季節性變化。.....................................................55 圖 16. ‘台農二號’番木瓜果肉與果皮鎂濃度催熟前後之季節性變化。.....................................................56 圖 17. ‘台農二號’番木瓜果肉與果皮鐵濃度催熟前後之季節性變化。.....................................................57 圖 18. ‘台農二號’番木瓜果肉與果皮錳濃度催熟前後之季節性變化。.....................................................58 圖 19. ‘台農二號’番木瓜果肉與果皮鋅濃度催熟前後之季節性變化。.....................................................59 圖 20. ‘台農二號’番木瓜果肉與果皮銅濃度催熟前後之季節性變化。.....................................................60 圖 21. ‘台農二號’番木瓜葉片大量元素之季節性變化。......61 圖 22. ‘台農二號’番木瓜葉片微量元素之季節性變化。......62 圖 23. 不同生長階段之番木瓜果實。........................63 圖 24. ‘台農二號’番木瓜乳汁、果皮無斑點及斑點樣品之蛋白質電泳圖譜。...............................................64 圖 25. ‘台農二號’番木瓜乳汁、果皮無斑點及斑點樣品之過氧化酵素同功異構.............................................65 圖 26. ‘台農二號’番木瓜有斑點及無斑點樣品之表皮結構。.....................................................66 圖 27. ‘台農二號’番木瓜黃熟有斑果皮表皮上氣孔與角質層結構。.....................................................6

(33(3):325-330)Study on the Quarantine Treatments of Insect Pests on Chrysanthemum Cut Flowers-The Application of Protection Bags and the Improved Dipping Methods

銷日菊花害蟲檢疫問題可以田間花蕾套袋與切花後藥液浸漬莖葉配合應用而獲得解決。田間自花蕾尚未展開時即予以套袋，可杜絕為害花部害蟲之發生，並減少田間之施用殺蟲藥劑。切花後浸漬切花之莖葉於殺蟲劑與殺蟎劑之混合稀釋液中，可毒殺附著莖葉上之害蟲及害蟎。此種方式處理之菊花無害蟲存在，且任何品種之菊花均可應用。以往白色花浸藥後因污斑而品質降低之現象將可消除。經處理之切花其保鮮期不受影響。 Transparent PE bags (13 x 17cm) fully covered with holes (dia. 0.2-0.5 mm) were used to protect chrysanthemum blooms in field. Young buds were covered and protected with these bags prior to the appearance of any petals, and these buds growing and blooming inside the bags. The change of attack of insect pests was hence significantly eliminated or avoided depending on the size of holes on bags. After cutting, dipping the stem below the flower head in diluted insecticides and acaricides would kill the aphids and mites stayed on leaves. Acricide 25 WP and Decis 2.8 EC were the most effective chemicals screened in this experiment. The application of the proper protection bags on buds and the improvement of dipping method offered insect-free cut flowers and eliminated the amount of insecticides used in field. Blooms, especially those in white color were not polluted with insecticides. And the keeping period of cut flowers was not influenced

Studies on the scars of humpback dolphins, Sousa chinensis, in Taiwan

台灣西海岸的中華白海豚族群被列為極度瀕危，亟需保育。海豚身上的傷疤可以反映海豚在過去所面臨到的環境或掠食者壓力。本研究利用海上調查照片與擱淺中華白海豚解剖資料來檢視中華白海豚的體表傷疤，並探討其所面臨的人為與自然環境壓力。本研究將傷疤分為兩大類，包括六種人為肇因傷疤以及一種自然的鯊魚咬痕。研究發現，在71已辨識的海豚中有42%的海豚身上帶有人為相關傷疤，年增加率為2±0.707(傷/年)。帶傷比例隨年齡增長而增加，尤以青年期進入成年期階段增加幅度為劇。雖然台灣白海豚的擱淺率相當低，但與香港和南非的白海豚族群比較，台灣帶傷之比例較高，值得關注。人為肇因最相關的兩種傷痕：纏繞傷與船槳傷，身上有此類傷疤的海豚個體多出沒於苗栗—台中以及台中—北彰化一帶，與漁業以及船隻作業繁忙地區重疊，可能受衝擊機率大。 自然肇因的傷疤皆來自鯊魚咬傷，檢視的71隻海豚中發現25.4%的海豚身上帶有鯊魚咬痕，年增加率為0.8±0.98(傷/年)。此比例與南非Natal地區白海豚族群結果相仿，推測與海豚和鯊魚棲息環境的鹽度相同有關。比較四個年齡群中各群內帶傷個體比例發現，帶傷比例增加幅度同樣以青年期進入成年期階段最劇。另外自性別比例來看，在受傷的海豚中有43.75% 為雌性，推測與海豚懷孕以及保護幼體相關。58%的鯊魚咬痕集中於海豚身體後半處(包含尾幹)，與前人推論的海豚迴聲定位盲點假說一致。致傷的鯊魚種類則根據相關生物資料推論，可能為污斑白眼鮫(Carcharhinuslongimanus)或鼬鯊(Galeocerdocuvieri)。 綜合結果發現，不論是來自人為還是自然，台灣的中華白海豚族群都面臨到相當大的環境壓力。雖然我們難以改善白海豚所面臨的自然環境威脅，但仍應盡速採取相關行動降低牠們所面臨人為環境的壓力，以達保護這個脆弱族群的目的。The humpback dolphins (Sousa chinensis) off the west coast of Taiwan are listed as a critically endangered species in the IUCN list that call for instant protection. Scars on the dolphins can be studied as records of the environmental or predatory threats these cetaceans encountered. In this study, I used photos obtained from boat surveys and necropsy reports of stranded/ by-caught humpback dolphins to examine the scars on them and to elucidate the anthropogenic and natural threats they may have encountered. There scars are classified into six types of anthropogenic scars and one type of natural predatory scars caused by shark bites. Of the 71examined Sousa, 42% carried anthropogenic scars, and the annual increase rate of anthropogenic scars was 2±0.707 (scars/year) in this population. The scarring ratio increased with age, and it increased sharply from the speckled stage to spotted adults. Although the stranding rate was very low in this population, the scarring rate on the other hand was relatively higher than those of the Sousa populations in Hong Kong and South Africa. The dolphins bearing impression marks and propeller wounds, the two types of scars most likely related to anthropogenic activities, were most frequently sighted in coastal waters from Miaoli to Taichung and between Taichung and north Changhua, overlapping with the areas characteristic of heavy vessel traffic and fishery activities. With regard to natural predatory scars, all the scars we examined were determined to be shark bites. Of the 71 dolphins 25.4% displayed shark bites. The annual increase rate of shark bites was 0.8±0.98, which was similar to the result found in Natal, South Africa. The consistency might be related to the similar salinity of the dolphins’ and sharks’ habitats. The predatory scarring ratio also increased fast during the period bwtween the speckled-stage and spotted adults. Among the sexually mature, scarred dolphins, 43.75% were females. This result may be associated with the adult dolphins’ protecting behaviors of the infants and calves. Based on the fish fauna records in Taiwan, the predatory scars are probably inflicted by the oceanic whitetip sharks (Carcharhinuslongimanus) and tiger sharks (Galeocerdocuvieri). This study reveals that humpback dolphins in Taiwan face both specific anthropogenic and environmental threats. Although it is difficult to reduce the natural predatory pressure on the dolphins of this vulnerable population, reducing the anthropogenic threats must be a priority task for their conservation

Species Identification of Shark Fins in Taiwan Markets by Using DNA Barcode: the Application to Fishery Management and Species Conservation

魚翅是整尾鯊魚個體中經濟價值最高的部位，長久以來在東亞以及東南亞國家中食用魚翅的風氣相當盛行，在中國更是一項流傳了數百年的珍貴菜餚；本研究基於對軟骨魚類保育的觀念，欲探討台灣市售魚翅種類之組成。自國內各大漁港與都會區採樣或購買的魚翅，包括新鮮魚翅、乾製魚翅、水發魚翅、罐頭和調理包魚翅等，由於沒有完整的個體，難以靠形態鑑定種類，故採取分子生物學的方式，利用生命條碼鑑定所收集到的樣本；選用的 DNA 片段為粒線體 COI 基因。研究結果顯示，新鮮魚翅、乾製魚翅與水發魚翅皆能以此方法成功鑑定物種；但罐頭及調理包魚翅則未能成功放大所需之 DNA 片段。種類分析方面，在 1170 個樣本中，成功鑑定了 686 個，總共發現 46 個物種，分屬 8 目 14 科 21 屬。分析的樣本當中，數量最多的物種是鋸峰齒鮫，其次依序是平滑白眼鮫、灰鯖鮫、深海狐鮫、紅肉丫髻鮫和淺海狐鮫。保育狀況方面，本研究鑑定出之物種，根據國際自然保護聯盟的評估與分類，極危的物種佔了 2.2 %，瀕危的物種佔了 4.3 %，易危的物種佔了 32.6 %，近危的物種佔了 39.1 %，無危的物種佔了 15.2 %，數據缺乏的物種佔了 4.3 %，未評估的物種佔了 2.2 %。 除了種類鑑定之外，本研究亦分析了各 COI 序列之間的演化距離，結果顯示，各物種內的演化距離分布為 0.002-0.035，平均 0.007；屬內為 0.003-0.102，平均 0.029；科內為 0.003-0.105，平均 0.054。之後將本研究所採集而來的單倍型序列用以建構親緣關係樹，發現各物種在親緣關係樹中均能被分離開，因此證實了 COI 序列對於軟骨魚類的種類鑑定具有很好的效果。 最後，在針對八個受區域漁業管理組織規範的物種之親緣關係分析結果顯示，淺海狐鮫、狐鮫、平滑白眼鮫、紅肉丫髻鮫、丫髻鮫等五個物種在不同的洋區有遺傳分化之現象；深海狐鮫、污斑白眼鮫、八鰭丫髻鮫此三個物種則無。 經由本研究可得知，利用粒線體 COI 基因當作生命條碼分析市售魚翅為一快速、有效且準確的方法；此方法將在漁業管理以及保育工作上能夠起到很好的輔助效果。Shark fin is the most valuable part in a whole shark body, the custom of eating shark fin is very popular in East and South-East Asia, and it has been counted a traditional noble delicacy in China for centuries. This study aims to analyze constitution of species in Taiwan market fins. We collected or purchased shark fin samples from fishing ports or metropolis in Taiwan, including fresh fin, dried fin, soaked fin, canned fin, and pack fin. Because of lacking key features, it is impossible to identify species of the samples by morphological method. We used the molecular tool- DNA barcode (the gene marker is mitochondrial COI gene) to accomplish it. The results showed that this method is appropriate in species identification of fresh fin, dried fin, and soaked fin, but inappropriate in canned fin and pack fin. In the results of species analysis, we found 46 species in the 686 successfully indentified samples of the all 1170 samples. The 46 species belong to 8 orders, 14 families, and 21 genera. In all the samples we analyzed, the most plentiful species is Blue shark, subsequently are Silky shark, Shortfin mako, Bigeye thresher, Scalloped hammerhead, and Pelagic thresher. In status of conservation, according to the evaluation and classification by IUCN, critically endangered species account for 2.2 %, endangered species account for 4.3 %, vulnerable species account for 32.6 %, near threatened species account for 39.1 %, least concern species account for 15.2 %, data deficient species account for 4.3 %, not evaluated species account for 2.2 % in all the species we found in this study. In addition to species identification, we also calculated evolutionary distances of the COI sequences. The results showed that within-species distances ranged from 0.002 to 0.035, with the mean value on 0.007; within-genus distances ranged from 0.003 to 0.102, with the mean value on 0.029; and within-family distances ranged from 0.003 to 0.105, with the mean value on 0.054. Furthermore, all the species can be perfectly separated in phylogenetic trees which are constructed by the haplotypes, it indicated that capability of COI gene in species identification of chondrichthians is very good. Moreover, phylogenetic analyses showed regional genetic divergences in Pelagic thresher, Thresher, Silky shark, Scalloped hammerhead, and Smooth hammerhead; but no divergence in Bigeye thresher, Oceanic whitetip shark, and Great hammerhead. We affirmed that using mitochondrial COI gene as a DNA barcode to identify chondrichthian species is a fast, effective, and accurate method, which will be helpful for fishery management and species conservation.致謝 .............................................................................................................................. i 摘要 ............................................................................................................................. ii Abstract ....................................................................................................................... iv 目錄 ............................................................................................................................ vi 圖目錄 ...................................................................................................................... viii 表目錄 ......................................................................................................................... x 附錄目錄 .................................................................................................................... xi 一、前言 ..................................................................................................................... 1 二、材料與方法 ......................................................................................................... 5 2.1 樣本採集 ...................................................................................................... 5 2.2 種類鑑定 ...................................................................................................... 5 2.3 演化距離之探討 .......................................................................................... 8 2.4 粒線體 COI 基因鑑別力之檢驗 ............................................................... 8 2.5 八個受區域漁業管理組織規範的物種之地域性遺傳分化情形檢驗 ...... 9 三、結果 ................................................................................................................... 10 3.1 樣本收集狀況與種類鑑定成功率 ............................................................ 10 3.2 鑑定結果 .................................................................................................... 10 3.3 各物種保育現況分析 ................................................................................ 11 3.4 演化距離之分析 ........................................................................................ 13 3.5 粒線體 COI 基因鑑別力檢驗之結果 ..................................................... 14 3.6 受規範物種之地域性遺傳分化情形檢驗結果 ........................................ 14 四、討論 ................................................................................................................... 18 4.1 利用 DNA 條碼鑑定魚翅物種之實用性 ................................................ 18 4.2 鑑定結果與保育狀況之討論 .................................................................... 19 4.3 演化距離之討論 ........................................................................................ 25 4.4 粒線體 COI 基因對於軟骨魚類物種的鑑別力與實用性之討論 ......... 27 4.5 受規範物種的親緣關係與地域性遺傳分化之討論 ................................ 28 五、結論 ................................................................................................................... 32 六、參考文獻 ........................................................................................................... 33 七、圖 ....................................................................................................................... 39 八、表 ....................................................................................................................... 65 九、附錄 .................................................................................................................... 7

Turing model for generating biological patterns

In 1952, A. M. Turing[1] proposed that the reaction-diffusion system could explain the main phenomena of biological morphogenesis. Unfortunately, he died in 1954. However, Turing's intriguing ideas influenced the thinking of theoretical biologists and scientists of many fields. The Turing mechanism has been successfully used for generating patterns in mammals[2,3], fish[4-6], bacterial colonies[7-9], phyllotaxis[10,11] and many others. We simulate the patterns on the elytras of the lady beetles using a reaction-diffusion equation with two types of morphogens based on the Turing model[12]. A part of a half spherical surface is used to approximate the geometry of the hard wings.Various patterns common to lady beetles in Taiwan can be produced on this curved surface. A complex system like the leopard's skin marking still offers an optimal level of challenge for generating it, even though previous simulations using a one-stage Turing model[13-16] might have produced final rosette patterns similar the patterns of real leopards. Based on the results of phylogenetic analysis, which showed that flecks are the primitive pattern of the felid family and all other patterns including rosettes and blotches develop from it[17], we construct a two-stage Turing reaction-diffusion model[18] which generates spot patterns initially. In the first stage, spots are generated in a similar manner for both the leopard cub and the jaguar cub. In the second stage, we tune model paraments to generate, separately, the sequence of patterns transformation during the growth of the animals.英國數學家涂林於1952年[1]提出「解釋生物形態發生的反應擴散模型」的創見，不幸卻於1954年早逝，但他的想法已經引起理論生物學家與許多領域的科學家的注意與興趣，開始從事於涂林機制的研究。至今在生物圖案形成的應用累積了許多豐碩的成果，例如成功模擬出哺乳動物毛皮圖案[2,3]、魚類的圖紋[4-6]、細菌群聚的圖樣[7-9]、與植物的葉序[10,11]…等等。 台灣特有的瓢蟲身上豔麗多樣的圖案吸引我們的注意，嘗試將部分的半球面視為瓢蟲的鞘翅(elytra)，以兩種成形素(morphogens)的涂林方程，給予不同的初始濃度分佈與參數，成功製造出不同種類的瓢蟲花紋[12]。 此外對於複雜的豹紋花樣，過去一些以單階段涂林模型的研究，雖然曾經製造出與花豹相類似的玫瑰斑紋[13-16]，但是斑紋隨年齡成長的變化過程的問題，仍然懸而未決。根據動植物種類史的分析(phylogenetic analysis)，顯示貓科動物身上的不同種的斑紋─包括複雜的玫瑰紋(rosettes)與擴散狀的污斑(blotches)─是從點(flecks)演化而來的[17]，我們提出了兩階段的涂林模型[18]，成功的模擬出花豹(leopard)與美洲豹(jaguar)身上複雜的斑紋結構：第一階段首先利用涂林模型製造出點的圖案，對照於剛出生的幼豹的斑紋；第二階段以點為初始條件，調變模型中部分的參數，隨著參數的變化過程，圖案的改變呈現出真實豹紋在成長中的變化。目錄 中文摘要 ⅰ 英文摘要 ⅱ 目錄 ⅲ 圖表目次 ⅵ 第一章 簡介 1 一、 引言 1 二、 涂林模型 2 三、 涂林模型的發展歷史 2 第二章 數學分析 6 一、 反應動力學項( Reaction Kinetics term ) 6 二、 涂林方程式的線性穩定分析 8 三、 初始值對模選擇的影響 18 第三章 瓢蟲花紋模擬 20 一、 模型與線性穩定分析 21 二、 數值模擬 22 三、 模擬的結果 32 (一) 五斑廣盾瓢蟲 32 (二) 七星瓢蟲 34 (三) 杜虹十星瓢蟲 34 (四) 縱條黃瓢蟲 36 (五) 細紋裸瓢蟲 37 第四章 豹紋的模擬 40 一、 模型與線性穩定分析 43 二、 兩階段涂林模型 50 三、 δ對數值模擬的影響 55 四、 數值模擬與結果 58 第五章 二維傅立葉轉換分析 64 第六章 結論 70 附錄A 反應擴散方程去因次的推導 71 附錄B 美洲豹、花豹與獵豹 73 附錄C 催化物-抑制物系統與基質消耗系統 78 附錄D 以兩階段涂林模型模擬台灣的雲豹的毛皮圖案 82 附錄E 論文的相關報導摘錄 83 一、 PHYSICS NEW UPDATE, 26.09.2001 83 二、 Sciscape新聞報導, 28.09.2001 84 三、 SCIENCE NEWS, 06.10.2001 86 四、 Nature, online News, 04.08.2006 89 五、 Nature 442, Research Highlights, 10.08.2006 92 六、 知識通訊評論, 16.08.2006 93 七、 Math Digest, 08.2006 95 八、 LiveScience, 08.08.2006 97 九、 SCIENCE&VIE, 10. 2006 99 十、 2007傑出團隊研究成果 100 十一、動手玩碎形 (天下文化出版) 101 參考資料 102 圖表目次 圖 1-1 七星瓢蟲的成長圖 4 圖 1-2 在化學實驗系統所觀察到的涂林結構圖 5 圖 1-3 催化物-抑制物系統典型的示意圖 5 圖 2-1 特徵值與固定點穩定性之對應關係圖(不含k) 11 圖 2-2 特徵值與固定點穩定性之對應關係圖(含k) 13 圖 2-3 Ãk(k2)行列式的函數曲線圖 17 圖 2-4 不同D值的色散關係圖 17 圖 2-5 滿足涂林不穩定的色散關係圖 18 圖 2-6 典型線性分析得到的色散關係圖 19 圖 3-1 白斑褐瓢蟲( Halyzia sanscrita ) 20 圖 3-2 滿足涂林不穩定的色散關係圖 23 圖 3-3 (fu+gv)與σv的函數關係圖 25 圖 3-4 (fu+gv)與κ的函數關係圖 26 圖 3-5 (fugv+fvgu)與κ的函數關係圖 26 圖 3-6 Dv/Du與κ的函數關係圖 27 圖 3-7 點的大小與Du、Dv/Du的關係 28 圖 3-8 點、線與κ的關係 30 圖 3-9 模擬瓢蟲鞘翅的半球面座標 31 圖 3-10 瓢蟲鞘翅的差異 31 圖 3-11 五斑廣盾瓢蟲 33 圖 3-12 七星瓢蟲 34 圖 3-13 杜虹十星瓢蟲 35 圖 3-14白條菌瓢蟲 37 圖 3-15 細紋裸瓢蟲 39 圖 4-1 貓科動物尾巴的圖案 40 圖 4-2 模擬花豹的玫瑰斑紋 41 圖 4-3 模擬花豹的玫瑰斑紋 41 圖 4-4 模擬花豹的玫瑰斑紋 42 圖 4-5 模擬幼小美洲豹的斑紋 42 圖 4-6 點、線與r2的關係 45 圖 4-7 涂林參數空間與色散關係圖 50 圖 4-8 條紋棘蝶魚( Pomacanthus imperator ) 51 圖 4-9 疊波棘蝶魚( Pomacanthus semicirculatus ) 52 圖 4-10 Weigel提出的假設：貓科動物斑紋的演化途徑 53 圖 4-11 Werdelin與Olsson將貓科動物的斑紋歸類成六種花紋 54 圖 4-12 Werdelin與Olsson認為貓科動物的斑紋的演化途徑 54 圖 4-13 不同δ的色散關係圖 55 圖 4-14 δ與線疏密的關係圖 56 圖 4-15 δ與點疏密的關係圖 57 圖 4-16 不同δ的色散關係圖 60 圖 4-17不同D的色散關係圖 60 圖 4-18 模擬美洲豹的過程中，改變參數D與δ時的色散關係 61 圖 4-19 第一階段以涂林模型模擬點的圖案 61 圖 4-20 模擬花豹的第二階的過程 62 圖 4-21 模擬美洲豹的第二階的過程 62 圖 4-22 α-β參數平面 63 圖 5-1 美洲豹四個典型的圖案與對應的二維傅立葉轉換 65 圖 5-2 二維傅立葉頻譜，水平橫切線的強度分佈 68 圖 5-3 二維傅立葉逆轉換 69 圖 B-J 美洲豹成長過程 74 圖 B-L 花豹成長過程 76 圖 C-1 不同反應系統的null clines 79 圖 C-2 固定態附近的行為模式 80 圖 C-3催化物-抑制物的機制與基質消耗的機制 80 圖 C-4模擬瓢蟲鞘翅斑紋的系統，在相空間中的null clines 81 圖 C-5模擬豹紋的系統，在相空間中的null clines 81 圖 D 以兩階段涂林模型模擬台灣的雲豹的毛皮圖案 8