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標題: | 水稻側根形成之研究:甲基茉莉酸鹽與氯化鈷之效應 Studies on lateral root formation in rice: the effects of methyl jasmonate and cobalt chloride |
作者: | Yun-Yen Hsu 許雲雁 |
指導教授: | 高景輝(Ching-Huei Kao) |
關鍵字: | 鈣,氯化鈷,血紅素氧化酵素,側根,甲基茉莉酸鹽,一氧化氮,水稻, calcium,cobalt chloride,heme oxygenase,lateral root,methyl jasmonate,nitric oxide,Oryza sativa, |
出版年 : | 2012 |
學位: | 碩士 |
摘要: | 本論文係利用水稻台中在來一號 (Oryza sativa L. cv. Taichung Native 1, TN1)探討甲基茉莉酸鹽 [methyl jasmonate (MJ)]與氯化鈷 (CoCl2)對水稻側根形成之影響,同時探討一氧化氮(NO)、heme oxygenase (HO)與Ca2+於其中所扮演的角色。
MJ處理會增加水稻側根的數目與NO之形成,同時處理MJ以及NO的清除劑carboxy-2-phenyl-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO)後,會降低水稻側根數目與NO之形成。利用nitrate reductase (NR)抑制劑tungstate (Tu)可完全抑制MJ誘導水稻側根與NO之形成,而使用nitric oxide synthase (NOS)抑制劑NG-nitro-L-arginine methyl ester hydrochloride (L-NAME)則無法完全抑制MJ誘導側根與NO形成,顯示MJ促進NO形成是與NR的活性相關。經NO螢光染色結果,可清楚的看到MJ誘導NO產生與側根根原基形成的位置一致。由此推測MJ所引起的側根發育可能是藉由NO作為傳遞訊息分子。 MJ處理會提升血紅素氧化酵素 (heme oxygenase, HO; EC1.14.99.3)的活性。從時間軸來看,HO活性的增加早於側根出現之前,暗示MJ可能是藉由影響HO活性上升,進而誘導側根形成。處理HO抑制劑zinc protoporphyrinIX (ZnPPIX)以及CO清除劑hemoglobin (Hb)會顯著降低MJ所增加HO活性及側根的數目。外加HO的另一個產物biliverdin (BV)後發現,BV不但促進水稻側根數目,亦使得HO的活性增加。此外,我們亦發現了Ca2+會影響側根之發育,處理Ca2+的螯合物、通道抑制劑及攜鈣素 (camodulin)的拮抗劑均會抑制側根之形成。 CoCl2處理亦可增加水稻側根數目,並提升HO的活性。從時間軸來看,HO活性的增加早於側根形成,暗示CoCl2可能是藉由影響HO活性上升,進而誘導側根形成。處理HO抑制劑ZnPPIX以及一氧化碳清除劑Hb會顯著降低CoCl2所增加HO活性及側根的數目。處理CoCl2不會增加H2O2含量,同時處理過氧化氫 (H2O2)清除劑ascorbic acid (AsA) 亦不影響CoCl2誘導的HO活性及側根數目,指出CoCl2所造成之影響與活化氧族 (reactive oxygen species)無關。 In this thesis, the role of nitric oxide (NO), heme oxygenase (HO) and Ca2+ in regulating methyl jasmonate (MJ)-induced lateral roots (LR) formation in rice cultivar TN1 (Oryza sativa L. cv. Taichung Native 1) was examined. Also included in this thesis is the possible mechanism of CoCl2-promoted LR formation in rice. Application of MJ to rice seedlings promoted LR formation and NO production. Treatment with MJ and NO specific scavenger carboxy-2-phenyl-4, 4, 5, 5- tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) reduced MJ-induced LR formation and NO production. NO was detected by the specific fluorescence probe, 4-amino-5- ethylamino-2’, 7’-difluoroflorescein diacetate (DAF-FM DA). NR inhibitor, sodium tungstates (Tu), completely inhibited MJ-induced LR formation and NO fluorescence. However, nitric oxide synthase (NOS) inhibitor, NG-nitro-L-arginine methyl ester hydrochloride (L-NAME) slightly reduced MJ-induced LR formation and NO generation, suggesting that NR activity is involved in MJ-increased NO production in rice roots. Moreover, the fluorescence of the longitudinal section of roots showed that NO production induced by MJ was localized in the LR primodia. It suggests that NO is a signal molecule involved in the regulation of MJ-induced LR formation. MJ was able to increased rice seedlings HO activity. The time-course analyses of HO activity and LR formation clearly indicated that MJ-induced HO activity occurred before LR formation. Treatment with zinc protoporphyrinIX (ZnPPIX), an HO inhibitor, and hemoglobin (Hb), a CO scavenger, blocked MJ-induced LR formation and HO activity. Moreover, BV, another product of HO, also triggered LR development and HO activity. Pretreatment of Ca2+ chelators, channel blockers and camodulin antagonists inhibited MJ-promoted LR formation, but had no effect on MJ-induced HO activity. Rice seedlings treated with CoCl2 also could promoted LR formation and HO activity. The time-course analyses of HO activity and LR formation clearly indicated that CoCl2-induced HO activity occurred before LR formation. ZnPPIX and Hb were able to reduced HO activity and the number of LR. CoCl2 (20 μM) had no effect on H2O2 production. Moreover, ascorbic acid (AsA) had no effect on CoCl2-induced HO activity and the LR number. It appears that reactive oxygen species (ROS) is not involved in the CoCl2-induced LR formation. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65770 |
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顯示於系所單位: | 農藝學系 |
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