臺灣博碩士論文加值系統

　　銅藻（Sargassum horneri）是一種溫帶性海洋大型褐藻，廣泛分布於中國、韓國和日本沿海等地區，密集生長於海底形成藻林並可生長3至5公尺之長度。過去這一世紀以來陸續有報導指出北台灣海面上有大量的銅藻漂浮進來，但位處副熱帶氣候地區的台灣並沒有在地的野生銅藻，因為台灣夏季水溫高達28℃，是銅藻無法建立其族群的主要原因。儘管台灣環境並不適合其建立族群，但它有生長快速、龐大生物量和具有經濟效益等特性，使其具養殖及研究之價值。

　　本論文研究每年出現於北台灣的銅藻其生殖能力，以及利用室外和室內養殖等方式來了解其生活史與高溫影響之銅藻的生長率和存活率。本實驗確證漂浮至台灣海岸的銅藻能以有性生殖方式來產生後代，而這些後代可在台灣北部的室外池中被養殖，
並完成其一年生之生活史。本結果發現，在細心的人工養殖環境下，可以成功地在台灣北部的戶外養殖銅藻。

　　由於台灣夏季溫度高，需要較多的勞力培養銅藻，因此須設計某些實驗方法加以解決此問題。本實驗研究了兩種可能方法用以減輕夏季高溫對銅藻造成的影響，其一為以低劑量輻射誘導銅藻之高溫耐受性；其二為利用長期保種的方法，將銅藻胚胎能在夏季被儲存，並在秋季將其恢復於戶外養殖。

　　輻射激效是一種低劑量的游離輻射，可對生物系統產生刺激作用。本論文研究低劑量60Coγ輻射對銅藻的激效作用，發現在實驗室培養中，經過低劑量γ輻射的銅藻，
具有增強生長和耐高溫雙重作用。但是，同樣經過低劑量γ輻射處理的銅藻幼胚，於室外十噸池中培養，並未觀察到這些效果。然而，在室外栽培經過低輻射照射的銅藻其多醣含量分析，顯示經輻射處理過的銅藻比未輻射者有更高的總醣含量，但對多醣的相對組成成分並沒有變化。

　　本研究利用兩種方式來長期保存銅藻的胚胎：低溫黑暗貯藏法，以及包埋在褐藻膠珠之固定化法。實驗結果發現，於實驗室條件下養殖，在低溫黑暗中儲存胚胎可以存活至少6個月而沒有負影響。然而，附有銅藻胚胎的棉繩先儲存在低溫黑暗環境中6個月後，再轉移到室外池培養，卻無法存活。另外，無經過低溫黑暗儲存的銅藻胚胎固定化之生長情形，無論是室外池或實驗室內皆會生長，但低溫黑暗中長期儲存者均不存活。

　　本論文認為，用大量勞力的養殖情況下，台灣是可以進行全年的銅藻養殖。然而應用低劑量γ輻射的生物技術，以提高物種對溫度的耐受性與多醣含量，和長期儲存胚胎避開夏天之酷熱等技術，這樣應可大大減少在台灣或在亞熱帶環境中養殖銅藻所必須付出的大量勞力，可以較經濟性地養殖銅藻。

Sargassum horneri is a temperate marine brown macroalga that typically grows up to 3-5 m long in dense underwater forests with a widespread distribution along the coasts of China, Korea, and Japan. Although floating masses of S. horneri have been reported in northern Taiwan throughout the past century, there are no persistent populations of this species in subtropical Taiwan. High summertime seawater temperatures in Taiwan, which regularly exceed 28˚C, are likely a significant factor in preventing S. horneri from establishing persistent local populations. Despite the inability of this species to establish persistent local populations, it is of particular interest for cultivation in Taiwan for its rapid growth rate, large biomass, and economic utility.

The reproductive viability of the floating S. horneri that arrives in northern Taiwan each year was examined. Laboratory culture, or in vitro, experiments were conducted on S. horneri embryos to understand the life cycle of the species and the effects of high-temperature on its growth and survival. These experiments provided conclusive evidence that the floating S. horneri arriving on the shores of Taiwan are capable of sexual reproduction and producing viable offspring, but they grow optimally in water temperatures between 18 to 24 °C. S. horneri derived from the floating mass of the species was also cultivated in outdoor seawater tanks, or in situ conditions, in northern Taiwan through a full year and the completion of a reproductive life cycle. This outdoor cultivation experiment demonstrated that S. horneri can be cultivated outdoors in northern Taiwan when it is carefully manually maintained.

Experiments were also devised to study methods that allow for less labor intensive cultivation of S. horneri in Taiwan despite the high local summertime temperatures. Two potential strategies for mitigating the effects of high summertime temperatures on the local cultivation of S. horneri were investigated, the induction of high-temperature tolerance in the alga by treatment with low doses of gamma radiation, as well as the utilization of long-term preservation methods that allow embryos of this species to be placed in storage during the summer months and returned to outdoor cultivation in the fall.

Radiation hormesis is a phenomenon in which low doses of ionizing radiation can have stimulatory effects on biological systems. We examined the hormesis effects of low-dose 60Co gamma radiation on S. horneri and found that in laboratory in vitro culture, low-dose gamma radiation has the dual effects of enhancing growth and high-temperature tolerance in the treated algae. However, these effects were not observed when the same low-dose gamma ray treatment was applied to S. horneri embryos that were cultivated in situ in outdoor tanks. Analysis of the polysaccharide content of the outdoor cultivated and irradiated S. horneri revealed that the radiation treated algae had significantly higher total polysaccharide content than the unirradiated controls, but the relative composition of the component polysaccharides was unchanged.

Two preservation methods for the long-term storage of S. horneri embryos were examined: storage in low-temperature and darkness, and immobilization in Ca-alginate gel. These experiments demonstrated that in laboratory in vitro conditions, embryos preserved in low-temperature and darkness could survive at least 6 months in storage with no negative effects. However, S. horneri embryos attached to cotton ropes and stored in low-temperature and darkness were not viable when transferred to outdoor tank culture after 6 months of storage. S. horneri embryos immobilized in Ca-alginate and immediately cultivated in vitro or in situ without undergoing a low-temperature storage period had no significant differences in growth compared to controls, but embryos immobilized in Ca-alginate and stored at low-temperatures and darkness were not viable in either in vitro or in situ culture.

This dissertation shows that cultivation of S. horneri is possible in Taiwan when labor intensive maintenance is employed. However, the application of additional biotechnologies such as low-dose gamma radiation to enhance the temperature tolerance and polysaccharide content of the species, and long-term storage of embryos to avoid the high summertime temperatures can greatly reduce the labor intensive means necessary to cultivate the species and allow for its economic cultivation in Taiwan or other subtropical environments.

Acknowledgments……………………………………………………………………………i
Chinese Abstract…………………………………………………………………………ii
English Abstract…………………………………………………………………………iii
Table of Contents………………………………………………………………………v
List of Tables………………………………………………………………………………vii
List of Figures……………………………………………………………………………viii

Chapter 1: Introduction………………………………………………………1
Background ………………………………………………………………………………………2
Objective and Hypotheses …………………………………………………5
Summary of Findings ………………………………………………………………6
Synthesis …………………………………………………………………………………………8

Chapter 2: An Assessment of the Reproductive Viability and
Cultivation Potential of the Invasive Brown Alga
Sargassum horneri in Northern Taiwan …………………10
Abstract ……………………………………………………………………………………………11
Introduction …………………………………………………………………………………12
Materials and Methods …………………………………………………………13
Results ………………………………………………………………………………………………18
Discussion ………………………………………………………………………………………21
Figures and Tables …………………………………………………………………23

Chapter 3: The Hormesis Effects of Low-Dose 60Co Gamma
Irradiation on High Temperature Tolerance in
Cultivated Sargassum horneri (Fucales, Phaeophyceae)……………………………………………………………………………………………………………………31
Abstract ……………………………………………………………………………………………32
Introduction …………………………………………………………………………………33
Materials and Methods …………………………………………………………35
Results ………………………………………………………………………………………………39
Discussion ………………………………………………………………………………………42
Figures and Tables …………………………………………………………………46

Chapter 4: Germling Preservation and Tissue Culture Strategies for the Mass Cultivation of the Temperate Brown Alga Sargassum horneri in Subtropical
Taiwan …………………………………………………………………………………………………57
Abstract ……………………………………………………………………………………………58
Introduction …………………………………………………………………………………59
Materials and Methods …………………………………………………………61
Results ………………………………………………………………………………………………65
Discussion ………………………………………………………………………………………70
Figures and Tables …………………………………………………………………74

References ………………………………………………………………………………………87

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