臺灣博碩士論文加值系統

番茄黃化捲葉病係由粉蝨傳播的雙生病毒所引起，乃世界上番茄生產之重要限制因子。本研究旨在瞭解台灣地區番茄黃化捲葉病對番茄生長之影響及其可能之病因，以作為田間流行病研究及病害防治的參考。本研究首先以目視病徵法於台灣10個重要番茄栽培縣，進行黃化捲葉病田間罹病率調查，隨後逢機採集罹病株之葉片以酵素連結抗體法(Enzyme-linked Immunosorbent Assay, ELISA)做Geminivirus病毒檢測，再配合電子顯微鏡，觀察病毒顆粒及罹病植株細胞內病變情形。調查結果顯示台灣地區番茄約有17%罹染番茄黃化捲葉病。其中以高雄縣之34%最嚴重，南投縣之2%最輕微；分析結果顯示不同期作及不同栽培地區對田間目視罹病率、室內ELISA檢測及實際罹患番茄黃化捲葉病等結果，都有極顯著的差異影響。電子顯微鏡觀察，可以在罹病番茄葉片組織內發現雙生病毒顆粒；而在罹病番茄葉片的韌皮細胞中可以觀察到細胞核核仁異常腫大，而且內含病變的纖維環(fibrillar ring)、顆粒內含體(granular inclusion)、以及有病毒顆粒聚集等現象。網室內，不同生長期接種病毒對番茄在病徵初次出現之日數之影響，因品種不同差異頗大；接種病毒日數多寡對大部分番茄品種在病徵初次出現之日數影響不大；不同生長期接種病毒對番茄罹病率之影響，因品種不同略有差異；接種病毒日數多寡對番茄罹病率完全沒有影響；病毒在罹染病毒植株內之量有消長現象，其情形在不同番茄品種上皆呈先高而後遞減，最後再遞增；田間不同品種罹染病毒對病徵出現之平均日數，並無顯著影響，而不同生育期罹染病毒對病徵出現之平均日數卻有極顯著影響；相同田間定置植期，感染病毒與否對各品種之始花期均無顯著影響，而不同田間定置期對各品種之始花期則均有極顯著影響；聖女品種番茄之株高在移植後44,51,58天等生育期受病毒感染與否之影響極顯著，但移植後66天受病毒感染與否之影響不顯著；農友301品種番茄之株高在移植後51天受病毒感染與否之影響顯著，但移植後44,58,66天受病毒感染與否之影響不顯著；西施、亞蔬33號及亞蔬6號三個品種番茄之株高在移植後44,51,58,66天受病毒感染與否之影響均不顯著。

Tomato yellow leave curl disease, a disease caused by whitefly-transmitted geminivirus, is a serious production constraint of tomato worldwide. The objective of this research is to realize the influence of tomato yellow leaf curl disease on tomato growth and the possible viral agent causing this disease in Taiwan. Results of this study could be used as reference for epidemic study and disease control in the field. Field inspection was conducted first to estimate the rate of infection of the TYLC disease in the field. The areas inspected included ten of the major tomato-growing counties in Taiwan. Then leaves were collected from infected tomato plants in the field, and subjected to ELISA (Enzyme-Linked Immunosorbent assay) assay in the laboratory for serological detection of the existence of geminivirus. Finally, TEM was used to observe geminivirus particles and the changes in the cells of infected plants to identify viral agent in those TYLC infected plants. The survey indicates that nearly 17% of tomato crop infected by TYLCV-like disease in Taiwan. Kaohsiung Hsien was the worst, having 34% infection. Nantou Hsien had the least injected plants (2%). The analysis shows that both different crop season and different area are significantly affect of the result of the survey in the field, as well as the ELISA assay and real TYLCV-like infection in the field. TEM observation, showed has geminivirus particles occurred in the leaf tissue, expressing in cytopathological changes in phloem cells of tomato leaf infected by TYLCV-like. The changes include nucleolus hypertrophy, segregation of nucleolar material into granular and fibrillar ring, and accumulations of viral particles. In the greenhouse, among the tomato varieties tested, the influence of virus inoculation stage on the days of first symptom appeared were significntly different however there was no difference in the days of the first symptom appeared among the varieties irrespective of the duration of inoculation. The influence of virus inoculation stage on the rate of virus infection has some different, and there is no different whether increase inoculation days. Fluctuation of virus content in the infected tomato plants is high first then decreasing and jumping again finally. In the field, the influence of virus inoculation stage on the days of first symptom appeared was significantly different, but was not different among tomato varieties. The influence of virus infection on days to flowering in the same date move to field no different, but different date move to field has significantly different. On Santa variety, the influence of virus infection on plant height was significantly different in 44, 51and 58 days after the transplant, but was not significantly different in 66 days after the transplant. On Farmers 301 variety, the influence of virus infection on plant height was sinificantly different in 51 days after the transplant, but was not significantly different in 44, 58 and 66 days after the transplant. On the three tomato varieties namely FB、AVRDC6 and AVRDC33 there are no significant difference on plant height influenced by virus infection in 44,51,58 and 66 days after transplant.
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目錄
中文摘要……………………………………………………….…Ⅰ
英文摘要………………………………………………….………Ⅲ
誌謝……………………………………………………….……....Ⅴ
表目錄…………………………………………………….………Ⅸ
圖目錄……………………………………………………………ⅩⅠ
壹、 前言………………………………………………………….1
貳、 材料與方法………………………………………………….5
一、田間病害發生調查…………………………………………5
二、病毒顆粒及番茄罹病細胞之電子顯微鏡觀察……………6
三、番茄不同生育期接種病毒及不同接種病毒日數對病徵初次出現之日數及罹病率之響……………………...………7
四、罹病番茄植株內之病毒消長情形…………………………8
五、不同品種及不同生育期番茄罹染病毒對植株生長之影響…………………………………………………………...8
參、 結果………………………………………………………...10
一、田間病害發生調查………………………………………..10
二、病毒顆粒及番茄罹病細胞之電子顯微鏡觀察…………..10
三、番茄不同生育期接種病毒及不同接種病毒日數對病徵初次出現之日數及罹病率之影響………………………….11
四、罹病番茄植株內之病毒消長情形……………………….12
五、不同品種及不同生育期番茄罹染病毒對植株生長之影
響……………………………………………………...…..15
肆、 討論………………………………………………...………19
伍、 結論與建議…………………………………...……………27
陸、 參考文獻………………………………………………...…28
柒、 作者簡介………………………………………………...…71表目錄
表一、台灣十個主要番茄栽培縣及其栽培面積……………….36
表二、田間調查番茄疑似感染TYLCV之調查範圍及田間目視罹病率、ELISA 檢測及估算實際田間罹病率…………37
表三、不同期作及地區對田間調查番茄疑似感染TYLCV之田間目視罹病率、ELISA檢測及田間實際罹病率之影響..38
表四、不同期作番茄在不同地區之田間目視罹病率的差異…..39
表五、不同期作番茄在不同地區之ELISA檢測罹病率間的
差異……………………………………………………….40
表六、不同期作番茄在不同地區之田間實際罹病率的差異….41
表七、病毒接種時期及接種日數對六個番茄品種之病徵初次出現所需日數之影響…………………………………….…42
表八、番茄六品種在不同接種病毒時期及不同接種病毒日數下之平均初次病徵出現日數……………………………….43
表九、病毒接種時期及接種日數對六個番茄品種之感病率之影響……………………………………………………….....44
表十、不同番茄品種在不同病毒接種時期及不同接種日數下之病毒感染率………………………………………….……45
表十一、不同品種及不同田間定置日對病徵出現所需日數的影響………………………………………………….……48
表十二、五個不同品種番茄在不同田間定置日下所需平均病徵出現日數………………………………………….…....48
表十三、感染病毒與否及不同定置期對不同番茄品種之始花期之影響………………………………………………….49
表十四、各番茄品種在不同田間定置期及感染病毒與否之平均始花日數……………………………………………….49
表十五、感染病毒與否及不同田間定置期對聖女品種番茄株高之影響………………………………………………….50
表十六、不同田間定置期聖女品種番茄感染病毒與否下之平均株高………………………………………………….…50
表十七、感染病毒與否及不同田間定置期對農友301品種番茄株高之影響………………………………………...…..51
表十八、不同田間定置期農友301番茄品種感染病毒與否之平均株高……………………………………………….…52
表十九、感染病毒與否及不同田間定置期對西施品種番茄株高之影響………………………………………………….53
表二十、不同田間定置期西施番茄品種感染病毒與否之平均株高……………………………………………………….53
表二十一、感染病毒與否及不同田間定置期對亞蔬33號品種番茄株高之影響……………………….……………54
表二十二、不同田間定置期亞蔬33號番茄品種感染病毒與否之平均株高……………………………………...…..55
表二十三、感染病毒與否及不同田間定置期對亞蔬6號品種番茄株高之影響……………………………….………56
表二十四、不同田間定置期亞蔬6號番茄品種感染病毒與否之平均株高……………………………………….……56
圖目錄
圖一、田間番茄黃化捲葉病發生情形……………………....57
圖二、田間番茄植株受番茄黃化捲葉病為害之病徵…………60
圖三、罹染番茄黃化捲葉病之番茄病葉粗汁液，以負染色電顯檢查可觀察到雙生病毒顆粒…………………………….62
圖四、罹染番茄黃化捲葉病之番茄病葉組織，經超薄切片電子顯微鏡觀察之韌皮細胞病變…………………………….63
圖五、應用酵素連結抗體法檢測黃化捲葉病於亞蔬六號番茄植株體內病毒消長情形…………………………………….65
圖六、應用酵素連結抗體法檢測黃化捲葉病於聖女番茄植株體內病毒消長情形………………………………………….67
圖七、應用酵素連結抗體法檢測黃化捲葉病於亞蔬33號番茄植株體內病毒消長情形………………………………….69

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