臺灣博碩士論文加值系統

草莓 (Fragaria x ananassa Duchessne) ，全臺種植面積達 550 公頃，於 2017 年 年產值近 10 億新台幣。草莓植株因台灣夏季高溫而無法越夏，農民需在隔年以走 莖苗重新進行定植。炭疽病 是 草莓 苗 期之 重大 病 害，造成 草 莓苗 的 大量 損 失，健 康 種苗 的 缺乏 成為 草 莓產 業 之嚴 重問 題。目前 農 民主 要仰 賴 殺菌 劑防 治 炭疽 病，然而 長年 的 殺菌 劑 使用 已導 致 炭疽 病 產生 抗藥 性，因此 需要 新 的防 治 手段 減少 殺 菌劑 之 使用 以及 維 持草 莓 苗之 健康。已有多篇 研究指不同光波長對植物病原有生長、繁殖抑制效果，且具有提升植物對病害抗性 之能力。因此本研究透過 200 PPFD 之不同光波長 (白光、紅光、2/3 紅光+1/3 藍 光、1/3 紅光+2/3 藍光、藍光) 照射在 PDA (Potato dextrose agar) 皿培之 Colletotrichum siamense、C. siamense 接種之草莓葉片及草莓苗、待克利上，尋找最 具有炭疽病抑制效果及草莓生長促進效果之光波長，並觀察光波長是否可以和農 藥同時使用，期望能作為草莓溫室育苗之物理防治策略。結果中發現與白光相比時， 1/3 紅光+2/3 藍光混和波長可抑制 C. siamense 14 %之菌絲生長、5%之產孢量以及 11%之孢子發芽率。同時 1/3 紅光+2/3 藍光波長也具有促進草莓防禦基因 Chitinase2 基因 1.3 倍之表現量之特性。1/3 紅光+2/3 藍光波長在實驗中也展現植株生長促 進之效果，與白光相比時其促進植株 1.1 倍葉片數、2.4 倍之走莖數及 1.3 倍鮮重 之生長。而所有波長皆不影響待克利之有效性及造成藥害。因 1/3 紅光+2/3 藍光具 有抑制 C. siamense 生長、促進草莓防禦基因表現以及促進草莓苗生長之特性，且 不會影響待克利之有效性，因此本研究提倡農民育草莓苗時給予 1/3 紅光+2/3 藍 光混和波長照射，以達到抑制病害並同時促進草莓苗生長之目標。

Strawberry is an important economic crop in Taiwan, the total planting area has reached 550 hectares, and the annual output value is nearly 1 billion NTD. However, due to the high temperature and humidity in Taiwan, strawberry cannot be planted continuously. Instead, strawberry farmers replant with strawberry runners annually. Anthracnose is one of the most severe diseases in the strawberry nursery industry, leading to a great loss on strawberry seedling. The lack of healthy seedlings has become a huge obstacle in the strawberry industry in Taiwan. Currently, farmers mainly rely on fungicides to prevent anthracnose, as a consequence, the massive use of fungicides has given rise to the anthracnose drug resistance. As a result, it is an emerging need of new methods for controlling this disease, in order to reduce the use of fungicides. Numerous studies have shown different light wavelengths capable of suppressing both plant pathogen growth and plant diseases. Here, in this study, we illuminated different light wavelengths (white light, red light, 2/3red+1/3blue, 1/3red+2/3blue, blue light) at 200 PPFD on Colletotrichum siamense on PDA (Potato dextrose agar) , inoculated strawberry leaves and seedlings, and co-treatment with difenoconazole, to obtain the optimal wavelength for controlling anthracnose disease, to promote strawberry seedlings growth, and to implement with difenoconazole, respectively. Our results showed that, in III compared to white light, 1/3red+2/3blue could inhibit 14% C. siamese mycelial growth, 5% of the sporulation and 11% of spore germination. In addition to C.siamense growth inhibition, 1/3red+2/3blue light could also augment the strawberry defense gene expressions, such as Chitinase-2 gene. The 1/3red+2/3blue light was also found to increase leave number, runner number, and overall fresh weight of seedlings than white light does. Furthermore, all light wavelengths would not impair difenoconazole’s effectiveness or enhance phytotoxicity. In conclusion, our study indicates that 1/3red+2/3blue light shows the characteristics of the growth inhibition of C. siamense, defense genes upregulation, and the growth of strawberry. Therefore, we recommend 1/3red+2/3blue light condition during the nursery period can be applied for suppressing anthracnose disease and promoting strawberry seedling growth.

摘要 I
Abstract II
目錄 IV
表目錄 VI
圖目錄 VII
第1章 前言 1
第2章 前人研究 3
2.1 草莓炭疽病原種類及分類 3
2.2 草莓炭疽病之病徵及傳播 4
2.3 炭疽病之防治 5
2.4 光波長對真菌之影響 6
2.5 光波長對病害之影響 8
2.6 光波長促進植物生長 9
第3章 材料與方法 11
3.1 試驗植物及接種源 11
3.2 光源 11
3.3 接種源製備 12
3.4 菌絲生長實驗 12
3.5 孢子萌芽實驗 12
3.6 模擬溫室應用情況下炭疽病抑制實驗 13
3.7 炭疽病菌侵入感染實驗 13
3.8 炭疽病嚴重度實驗 13
3.9 炭疽病子代致病性實驗 14
3.10 草莓防禦基因表現實驗 14
3.11 草莓炭疽病抗性實驗 14
3.12 草莓苗生長實驗 15
3.13 草莓苗炭疽病抑制實驗 15
3.14 待克利有效性實驗 16
3.15 待克利藥害實驗 16
3.16 DNA萃取 16
3.17 RNA萃取 17
3.18 引子設計及Real-time PCR 17
3.19 統計與分析 18
第4章 結果 19
4.1 藍、紫2光波長抑制Colletotrichum siamense菌絲生長、產孢及孢子發芽 19
4.2 模擬溫室應用情況下藍光波長抑制炭疽病 21
4.3 藍光波長影響炭疽病病害進程 21
4.4 藍、紫2光波長促進草莓苗防禦反應 23
4.5 光波長無促進草莓苗對炭疽病抗性 24
4.6 紫1、紫2光波長促進草莓苗生長 24
4.7 藍光波長抑制草莓苗之炭疽病 25
4.8 光波長不影響待克利之有效性及造成藥害 26
4.9 總結 26
第5章 討論 28
5.1 光波長防治之重要性 28
5.2 光波長抑制微生物生長 28
5.3 光促進植物防禦反應 29
5.4 光促進植物生長 30
5.5 光波長作為草莓病害防治手段之可行性 31
第6章 參考文獻 33
第7章 表 37
第8章 圖 42

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