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  1. NTU Theses and Dissertations Repository
  2. 生物資源暨農學院
  3. 植物醫學碩士學位學程
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/74388
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor洪挺軒
dc.contributor.authorTsung-Han Leeen
dc.contributor.author李宗翰zh_TW
dc.date.accessioned2021-06-17T08:33:08Z-
dc.date.available2020-08-16
dc.date.copyright2019-08-16
dc.date.issued2019
dc.date.submitted2019-08-09
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/74388-
dc.description.abstract由韌皮部侷限細菌所引起的柑橘黃龍病 (Citrus huanglongbing, HLB) 是影響柑橘生產的重要病害,危害世界各大柑橘產區,而年產量高達50萬公噸以上的臺灣亦受嚴重波及。根據2017年田間調查的結果顯示臺灣約有31.9%之罹病率,且各品種皆會染病,造成果實的品質及產量大打折扣。目前已知病害的感染途徑主要藉由嫁接及媒介昆蟲進行傳播,而自1973年起積極推廣健康柑橘種苗概念之下,柑橘園中的亞洲柑橘木蝨 (Diaphorina citri Kuwayama) 似乎成為病害擴散的一大隱憂,對於媒介木蝨傳病生態學的相關研究,則可提供黃龍病防治策略的重要參考。本研究藉由分析不同生長期的柑橘品種其新梢帶菌的情形,以推估黃龍病菌自病徵明顯的老熟組織移動至幼嫩葉部所需的時間,結果發現柳橙在新梢生長一週後即可測得黃龍病菌,往後隨週數的進展菌量呈微幅增長;椪柑及桶柑則在兩週過後病原菌便開始快速增殖,到第四週時菌量達所有品種中的最高,而黃龍病菌在白金柚中的移動最為緩慢,至三、四週後才可在新梢中檢測到少量存在。病原菌移動至新梢後,因木蝨對嫩葉處具有強烈的偏好,進而提供受吸引前來的木蝨獲菌的機會,文中亦針對可能影響木蝨獲菌的因子進行探討,在調查不同齡期若蟲的獲菌情形中,發現自二齡起木蝨即具獲菌的能力,又以四齡及五齡若蟲的帶菌量最高,五齡若蟲的帶菌量約與初獲菌的二齡若蟲相差近130倍;取食病株的時間長短亦會造成獲菌差異,在14天時初有30%的帶菌蟲比例,到28天時則升高至80%,說明獲菌情形可能隨時間的延長而提升;而不同的柑橘品種及其罹病情形也是影響木蝨獲菌的可能因子,以罹病三個月後的柳橙最早有木蝨獲菌的記錄,椪柑及文旦亦在四、五個月時相繼出現木蝨獲菌的情形,而任一品種皆發生在完整病徵出現以前。進入木蝨體內的黃龍病菌隨開放系統進行循環,以持續取食月橘屬 (Murraya spp.) 植株的試驗中觀察病原菌於媒介昆蟲體內的增殖效率,而長至四週後仍能檢測到帶菌情形,證實病原於蟲體內的保存效力,一旦經有效獲菌後,則可能終生成為潛在感染媒介,在田間造成黃龍病傳播的威脅。zh_TW
dc.description.abstractCitrus huanglongbing (HLB) is a destructive disease among citrus industry caused by a phloem-limited bacteria (Candidatus Liberibacter asiaticus, CLas), and it happens in citrus production areas worldwide. With over than 500,000 tons of annual production, Taiwan also suffers from this serious disease. According to the HLB field survey in 2017, there was 31.9% infection in Taiwan. Because all citrus cultivars are susceptible to HLB, it caused great damage on fruit quality and production. As known, the spread of HLB in the field is mainly due to the grafting of infectious scions and insect vector transmission. However, with the help of citrus healthy budwood program which was firstly promoted in 1973, vector psyllids (Diaphorina citri Kuwayama) seems to play an important role in spreading the disease. Researches on transmission ecology of HLB by the psyllid may contribute to the good solution for HLB management. By analyzing new shoot infection status in different weeks, we attempted to find out the movement of CLas from symptomatic old tissues to young leaves among different cultivars, and the results showed that movement in Liucheng cultivar requiring the least time. CLas reached new shoots in only 1 week, and the copy numbers increased slightly. Same movement tendency could be observed in Mandarins (Ponkan and Tankan cultivars), CLas copy numbers rapidly increased after 2 weeks and reach amount of 10,000 copies in week 4, which is also the highest amount in the study. The movement of CLas was much slower in Oroblanco, high ratio of CLas-infected new shoots could only be observed after 3 weeks. Because of the high preference of psyllids to immature foliar tissues, CLas moving to new shoots may offer a good chance for pathogen acquisition. In this study, we also investigated the factors that might influence CLas acquisition. Among different nymphal stages, CLas was firstly detected in the 2nd nymphal stage, and got the highest average copy numbers per psyllid in the 4th and 5th nymphal stages, which had about 130 times in difference. Also, acquisition access period on HLB-infected plant seemed to be another influence factors, percentages of CLas-infected psyllid raised from 30% to 80% when prolonging the period from 14 to 28 days. Moreover, other factors like plant infection status among different cultivars could lead to variation in CLas acquisition. After CLas inoculation by grafting, pathogen acquisition by D. citri firstly occurred after 3 months on infectious Liucheng, 4 and 5 months on Ponkan and Wentan, respectively; while they all happened during asymptomatic phase of infection. As CLas acquired by the psyllids, they followed the open system and circulated in insect bodies. Continuous feeding tests on Murraya spp. made a good way to observe CLas proliferation in D. citri. The results showed that CLas could be still detected until 4 weeks, and it proved the ability of CLas retention in insect vectors. Indicating that once D. citri acquired CLas via feeding, they could probably become a persistent infectious vector, and post the threats to HLB spreading in the fields.en
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dc.description.tableofcontents口試委員審定書 I
致謝 II
中文摘要 III
英文摘要 IV
表目錄 VIII
圖目錄 IX
壹、 前言 1
貳、 前人研究回顧 4
一、 柑橘黃龍病的發現及歷史 4
二、 柑橘黃龍病的病原學研究 5
三、 臺灣柑橘黃龍病的病菌系統 7
四、 柑橘黃龍病的發病生態及危害 9
五、 媒介木蝨的生物學背景 11
六、 黃龍病菌於寄主柑橘及媒介木蝨間的交互關係 12
七、 黃龍病菌的相關偵測技術 15
參、 材料與方法 18
一、 試驗植物之取得 18
二、 罹病植株之準備及來源 18
三、 試驗柑橘木蝨之取得 18
四、 核酸萃取之方法 19
(一) 植物總核酸萃取 19
(二) 木蝨核酸萃取 20
五、 PCR 檢測方法 20
六、 膠體電泳分析 21
七、 Real-time PCR檢測方法 21
(一) 引子對及探針之選擇 21
(二) 反應之條件與過程 21
(三) 絕對定量之標準曲線 (Standard curve) 建立 22
八、 黃龍病菌於不同柑橘品種寄主體內之移動 22
九、 影響木蝨獲菌之因子探討 23
(一) 若蟲齡期對於木蝨獲菌的影響 23
(二) 取食時間長短對於木蝨獲菌的影響 24
(三) 不同罹病時期植株對於木蝨獲菌的影響 24
十、 黃龍病菌於媒介木蝨體內之消長 25
肆、 結果 26
一、 黃龍病菌於不同柑橘品種寄主體內之移動 26
二、 影響木蝨獲菌因子之探討 28
(一) 若蟲齡期對於木蝨獲菌的影響 28
(二) 取食時間長短對於木蝨獲菌的影響 29
(三) 不同罹病時期植株對於木蝨獲菌的影響 29
三、 黃龍病菌於媒介木蝨體內之消長 31
伍、 討論 32
一、 黃龍病菌的移動於柑橘品種間存在之差異 32
二、 影響木蝨獲菌之因子探討 35
(一) 若蟲齡期與獲菌 36
(二) 病株取食時間的長短與獲菌 37
(三) 植株不同罹病狀態與獲菌 38
三、 黃龍病菌於媒介木蝨體內的消長情形 42
陸、 結語 45
柒、 參考文獻 46
捌、 表 56
玖、 圖 64
dc.language.isozh-TW
dc.title黃龍病菌於柑橘寄主體內之移動與影響亞洲柑橘木蝨 (半翅目:扁木蝨科) 獲菌因子之探討zh_TW
dc.titleStudies on the movement of Candidatus Liberibacter asiaticus in citrus hosts and the factors which influence pathogen acquisition for psyllids, Diaphorina citri Kuwayama (Hemiptera:Liviidae)en
dc.typeThesis
dc.date.schoolyear107-2
dc.description.degree碩士
dc.contributor.coadvisor蕭旭峰
dc.contributor.oralexamcommittee蔡志偉,陳文華
dc.subject.keyword柑橘黃龍病,黃龍病菌,柑橘木蝨,獲菌,媒介昆蟲,zh_TW
dc.subject.keywordCitrus huanglongbing (HLB),Candidatus Liberibacter asiaticus,Diaphorina citri Kuwayama,acquisition,insect vectors,en
dc.relation.page79
dc.identifier.doi10.6342/NTU201902913
dc.rights.note有償授權
dc.date.accepted2019-08-12
dc.contributor.author-college生物資源暨農學院zh_TW
dc.contributor.author-dept植物醫學碩士學位學程zh_TW
顯示於系所單位:植物醫學碩士學位學程

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