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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張雅君 | zh_TW |
| dc.contributor.advisor | Ya-Chun Chang | en |
| dc.contributor.author | 陳臻 | zh_TW |
| dc.contributor.author | Chen Chen | en |
| dc.date.accessioned | 2021-07-11T15:20:25Z | - |
| dc.date.available | 2024-03-05 | - |
| dc.date.copyright | 2019-03-12 | - |
| dc.date.issued | 2019 | - |
| dc.date.submitted | 2002-01-01 | - |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78802 | - |
| dc.description.abstract | 紅龍果 (pitaya) 屬於仙人掌科 (Cactaceae)，臺灣主要栽培品種為三角柱屬 (Hylocereus) 的白肉 (H. undatus) 以及紅肉 (H. polyrhizus & H. costaricensis) 品系。由於紅龍果栽培管理容易、營養價值高以及新品種之育成，為臺灣近年來興起的果樹之一。目前已知感染紅龍果的病毒皆為Potexvirus屬：仙人掌X病毒 (Cactus virus X, CVX)、紅龍果X病毒 (Pitaya virus X, PiVX) 以及蟹爪蘭X病毒 (Zygocactus virus X, ZyVX)。本研究於2017至2018年檢測臺灣田間紅龍果枝條，發現所有樣本均受CVX感染，而PiVX與ZyVX的感染率分別為62.7%和55.2%，且複合感染的比例為82.1%，顯示紅龍果病毒已普遍存在於臺灣田間。為建立適合紅龍果白肉品種和紅肉品種富貴紅的組織培養系統，將田間罹病紅龍果枝條扦繁殖以獲得新生枝條，其莖段培養於3 mg/L 6-benzylaminopurine (BAP) 或玉米素 (zeatin) 搭配0.6 mg/L 1-naphthaleneacetic acid (NAA) 之MS培養基，2週後長出新芽，再經3週後新芽可長至2~5公分，成功建立大量繁殖無菌苗之技術。在莖頂組織培養去除病毒部分，將罹病無菌苗莖頂培養於含有1.5 mg/L BAP與0.15 mg/L NAA的培養基，其回復生長率為87.7~100%，且成功從受病毒單獨感染的材料中獲得紅龍果白肉品種無病毒苗；而複合感染的材料僅有部分病毒被去除，尚未能獲得紅肉品種無病毒苗。另外，本研究證實CVX與ZyVX之熱不活化溫度可達80~85°C，而PiVX為70~75°C。以24 GHz微波處理1.5秒後，紅龍果無菌苗可快速升溫至80~90°C，其莖頂組織回復培養後存活率為40%；由於24 GHz微波處理之植物組織可以在數秒內達到病毒熱不活化溫度以上，可望成為一種新穎的植物去病毒方法，目前已利用此方法獲得紅龍果白肉品種無病毒苗，但其去除病毒之效果仍待進一步提升。此外，本研究經改變試驗藥劑配方與處理時間，以及改良回復生長之培養條件，已建立適合紅龍果無菌苗的玻璃化法 (vitrification) 冷凍治療 (cryotherapy) 處理流程，回復生長率達23%，而去除病毒成功率仍待後續檢測。本研究期望能更進一步優化24 GHz微波處理及確認玻璃化法冷凍治療之去病毒效率，以獲得紅肉品種富貴紅無病毒苗，並期望利用所建立之紅龍果組織培養技術，應用於無病毒苗之大量繁殖，提供健康種苗供田間使用，並協助紅龍果病毒病害之研究。 | zh_TW |
| dc.description.abstract | Pitaya, also known as dragon fruit, is a climbing succulent plant in the family Cactaceae, and major cultivars in Taiwan are white-fleshed (Hylocereus undatus) and red-fleshed (H. polyrhizus & H. costaricensis) pitayas. Since pitaya is easy to grow and full of nutrition, and has new bred cultivars, it becomes important fruit crop in Taiwan in recent years. Three potexviruses, Cactus virus X (CVX), Pitaya virus X (PiVX), and Zygocactus virus X (ZyVX), have been reported to infect pitaya. During the period of 2017 and 2018, our field investigation results showed that all pitaya stem samples were infected by CVX, and the infection rates of PiVX and ZyVX were 62.7% and 55.2% respectively. Besides, the mixed infection rate of pitaya samples was 82.1%. This result indicated that pitaya viruses are widespread in Taiwan orchards. In this study, we aimed to establish a tissue culture system suitable for the white-fleshed and the red-fleshed ‘Fu Gui Hong’ pitayas. To achieve the goal, virus-infected new stems were obtained from cutting propagation, and pitaya stem segments were cultured on MS medium containing 3 mg/L 6-benzylaminopurine (BAP) or zeatin and 0.6 mg/L 1-naphthaleneacetic acid (NAA). Consequently, new shoots were successfully induced in 2 weeks and grew to 2~5 cm long after another 3 weeks, and thus the aseptic plantlets were later used for virus elimination experiments. In terms of virus elimination through shoot tip culture, virus-infected shoot tips were cultured on MS medium containing 1.5 mg/L BAP and 0.15 mg/L NAA, and the recovery rate of shoot tip was 87.7~100%. We successfully obtained virus-free white-fleshed pitayas from singly-infected plant materials but failed in mix-infected and red-fleshed ones. In addition, we determined the thermal inactivation point (TIP) of CVX and ZyVX to be 80~85°C and PiVX to be 70~75°C. By treating with 24 GHz microwave for 1.5 seconds, the temperature of aseptic pitaya plantlets increased up to 80~90°C, and the survival rate of the treated shoot tips was 40%. The rapid temperature elevation to reach the TIP of viruses under 24 GHz microwave treatment has the potential to become a new method for plant virus elimination. So far, we have obtained virus-free white-fleshed pitaya by 24 GHz microwave treatment but the successful rate was low. By modifying the reaction solutions, processing time and recovery condition, a vitrification cryotherapy method suitable for pitaya aseptic plantlets was successfully developed. The current recovery rate of cryotherapy-treated shoot tips was 23%, but the effect of virus elimination has not yet done. For further studies, the conditions in 24 GHz microwave treatment needs to be optimized and the virus elimination rate of vitrification cryotherapy has to be confirmed. All these information will be useful for obtaining virus-free red-fleshed pitaya ‘Fu Gui Hong’ efficiently. Moreover, our tissue culture method can be applied to mass propagation of virus-free pitaya plantlets which can be cultivated in the field and used for the basic studies of pitaya virus diseases. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T15:20:25Z (GMT). No. of bitstreams: 1 ntu-108-R05633004-1.pdf: 4118457 bytes, checksum: 8072410afa50ae16081a75b3e01fe4dc (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | Abstract VI
壹、前言 1 一、紅龍果 1 二、紅龍果常見病害 2 三、紅龍果病毒病害之研究 3 四、植物組織培養與大量增殖 4 五、植物去除病毒方法 7 六、微波處理 11 七、研究目的 12 貳、材料與方法 13 一、組織培養試驗材料 13 二、表面消毒 14 三、紅龍果增殖系統與莖頂組織培養 14 四、莖頂組織培養方式去除病毒之試驗 19 五、微波處理搭配莖頂組織培養方式去除病毒之試驗 19 六、仙人掌X病毒、紅龍果X病毒以及蟹爪蘭X病毒之熱不活化溫度 (thermal inactivation point) 試驗 20 七、玻璃化法 (vitrification) 之冷凍治療 (cryotherapy) 搭配莖頂組織培養方式去除病毒之試驗 22 八、紅龍果病毒檢測方式 26 參、結果 28 一、田間紅龍果枝條病毒之檢測結果 28 二、田間紅龍果枝條扦插繁殖之結果 29 三、紅龍果白肉品種與紅肉品種富貴紅之組織培養以及增殖系統之建立 30 四、莖頂組織培養方式去除病毒之試驗結果 37 五、24 GHz 微波處理搭配莖頂組織培養方式去除病毒之試驗結果 38 六、玻璃化法 (Vitrification) 冷凍治療 (Cryotherapy) 搭配莖頂組織培養方式去除病毒之試驗結果 40 肆、討論 42 一、田間紅龍果枝條之病毒檢測 42 二、田間紅龍果枝條扦插繁殖 43 三、紅龍果白肉品種與紅肉品種富貴紅之組織培養以及增殖系統之建立 43 四、莖頂組織培養方式去除病毒之試驗 49 五、24 GHz微波處理搭配莖頂組織培養去除病毒之試驗 51 六、玻璃化 (Vitrification) 冷凍治療 (Cryotherapy) 搭配莖頂組織培養方式去除病毒之試驗 53 伍、參考文獻 56 陸、附表 66 柒、附圖 80 捌、附錄 107 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 紅龍果 | zh_TW |
| dc.subject | 紅龍果病毒病害 | zh_TW |
| dc.subject | 組織培養 | zh_TW |
| dc.subject | 去除病毒 | zh_TW |
| dc.subject | 莖頂組織培養 | zh_TW |
| dc.subject | 24 GHz微波 | zh_TW |
| dc.subject | 玻璃化法冷凍治療 | zh_TW |
| dc.subject | 24 GHz microwave | en |
| dc.subject | pitaya | en |
| dc.subject | pitaya virus disease | en |
| dc.subject | tissue culture | en |
| dc.subject | virus elimination | en |
| dc.subject | shoot tip culture | en |
| dc.subject | vitrification cryotherapy | en |
| dc.title | 紅龍果組織培養系統及去病毒技術之研發 | zh_TW |
| dc.title | Development of tissue culture system and virus elimination methods for pitaya | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 107-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 王淑珍;楊佐琦 | zh_TW |
| dc.contributor.oralexamcommittee | Shu-Jen Wang;Tso-Chi Yang | en |
| dc.subject.keyword | 紅龍果,紅龍果病毒病害,組織培養,去除病毒,莖頂組織培養,24 GHz微波,玻璃化法冷凍治療, | zh_TW |
| dc.subject.keyword | pitaya,pitaya virus disease,tissue culture,virus elimination,shoot tip culture,24 GHz microwave,vitrification cryotherapy, | en |
| dc.relation.page | 111 | - |
| dc.identifier.doi | 10.6342/NTU201900626 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2019-03-07 | - |
| dc.contributor.author-college | 生物資源暨農學院 | - |
| dc.contributor.author-dept | 植物病理與微生物學系 | - |
| dc.date.embargo-lift | 2029-03-07 | - |
| 顯示於系所單位： | 植物病理與微生物學系 | |
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