劉麗飛臺灣大學：農藝學研究所周書旭Chou, Sue-ShieSue-ShieChou2007-11-282018-07-112007-11-282018-07-112004http://ntur.lib.ntu.edu.tw//handle/246246/59118中文摘要 近年來常利用基因改造作物解決人類糧食缺乏的問題，其中Bt轉殖作物對病蟲害防治有很好的效果。但是轉殖作物所產生的Bt蛋白對特定害蟲具有高毒性及致死性，為了維持生物多樣性及避免害蟲有害生物小種產生，許多學者均傾向改造植物本身既有的防禦系統來增強對蟲害的耐性或抗性。本試驗將馬鈴薯的蛋白質酶抑制素基因（proteinase inhibitor II，PIN 2 )轉殖於水稻中，並使用PIN 2本身的創傷誘導性啟動子表現此防禦性蛋白，藉此減少能量消耗過多及表達蛋白大量累積時所造成的負面影響，希求此轉殖水稻能透過改變害蟲取食習性及干擾害蟲消化能力，來提高對嚼食性昆蟲的抵禦能力。 本試驗使用台農67號及台稉9號水稻為材料，以粒子槍法及農桿菌法成功轉殖出四種PIN 2的轉殖水稻，並進行田間及倉儲的害蟲防治研究。結果顯示以農桿菌轉殖法得到的轉殖水稻在T0代出現較少白苗、不稔等不良性狀。在田間蟲害防治的研究上，使用二化螟蟲為檢定材料，結果顯示PIN轉殖水稻平均能降低1/4的受害比例，且受害率與PIN活性呈現負相關（相關係數= -0.34），但蟲與蛹的體重有微幅上昇的狀況，需要對於二化螟蟲的取食行為及消化生理的深入討論才能釐清。而利用T4代轉殖糙米對米象進行倉儲害蟲的抗蟲力試驗，經處理三個月以後，結果顯示PIN 2轉殖糙米能有效降低米象1/2至3/4的繁殖速率，且米象繁殖速率亦與糙米PIN活性呈現明顯負相關（相關係數 = -0.66）。綜合以上結果證實PIN轉殖水稻不論在田間或是倉儲害蟲的防治上都有顯著效果，值得進一步的應用與研究。In recent years , the development of genetically engineered crops , especially in Bt transgenic crops has been used to resolve the food deficiency problem in the world . Although the expression of Bt toxin in transgenic crops help in prevention of pests, the extreme toxicity and lethality of Bt against specific target organisms leads to the occurance of resistance to pests and disruption of ecological balance. In order to maintain the biodiversity in the environment , many scientists tend to modify the defense mechanism of plant itself to improve the pest ability. In this study, transgenic rice plants with potato proteinase inhibitor 2 (PIN 2) gene have been produced. Also to reduce the negative effect due to large amount of protein accumulation and bioenergy consumption, the endogenous wounding-inducible PIN 2 promoter was used. By this way, we expected the PIN 2 transgenic rice plants were able to raise the defense ability against behavior insect through the inference of the digestion and changing the chewing habit of insects. To produce PIN 2 transgenic rice plants , two different rice cultivars TNG 67 ( cv. Tainung 67 ) and TK 9 (cv. Taikeng 9 ) were used as starting materials and various PIN 2 gene constructs were delivered into rice by particle bombardment and agrobacterium transformation methods. Followed that, researchs on the field trails and storage test of pest control were conducted. Agrobacterium transformation produced T0 transgenic rices showed less negative traits in sterility and etiolation. The field bioassay for insect resistance of rice stem borer ( Chilo suppressalis ) showed that the introduction of potato PIN 2 gene could help transgenic rices to reduce 1/4 damage of tillings , and the reduction damage of tillings was negatively correlated with PIN activity ( CORR.= -0.34 ). Nevertheless, the larval body weight and size had slightly increased in this assay. This remain to be solved by future invesgation of the feeding behavior and digestion physiology of rice stem borer. On the other hand, the resistance bioassay against rice weevil (Sitophilus ozyzae ) with T4 transgenic TNG 67 seeds showed that after 3 months treatment, the transgenic brown grain could significantly reduced the expansion of rice weevil population by 1/2 to 3/4 folds. Thus PIN activity of brown grain had also shown a markedly negative correlationship with weevil generation ( CORR. = -0.66 ). In conclusion, this research demonstrates the practical usefulness of PIN 2 transgenic plants in resistance to pest and provided an opportunity for future field application and experimental study.圖表目錄．．．．．．．．．．．．．．．．I 縮寫對照表．．．．．．．．．．．．．．．II 中文摘要．．．．．．．．．．．．．．．．IV 英文搞要．．．．．．．．．．．．．．．．V-VI 前言．．．．．．．．．．．．．．．．．．1-2 文獻回顧．．．．．．．．．．．．．．．．3-17 試驗材料．．．．．．．．．．．．．．．．18-23 試驗方法．．．．．．．．．．．．．．．．24-39 結果．．．．．．．．．．．．．．．．．．40-68 討論．．．．．．．．．．．．．．．．．．69-77 參考文獻．．．．．．．．．．．．．．．．78-86 附錄．．．．．．．．．．．．．．．．．．87-886149504 bytesapplication/pdfen-US馬鈴薯蛋白酶二化螟蟲米象抗蟲性抑制素轉殖水稻insect resistancerice weevilpotatotransgenic ricerice stem borderproteinase inhibitor[SDGs]SDG7thesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/59118/1/ntu-93-R89621101-1.pdf