臺灣博碩士論文加值系統

本論文係以水稻台中在來一號 (Oryza sativa L. cv. Taichung Native 1) 為材料，探討鹽分逆境下，Ca2+、nitric oxide (NO) 及auxin 對水稻黃化幼苗主根生長及冠根形成之影響。
高鹽逆境 (150 mM NaCl) 會抑制水稻冠根之形成，而處理氯化鈣 (CaCl2)、一氧化氮釋放劑 sodium nitroprusside (SNP) 及植物生長素indole -3- acetic acid (IAA) 可顯著減緩高鹽逆境對冠根形成的抑制，進一步利用石蠟切片發現冠根原基發育與上述結果一致。而處理一氧化氮清除劑2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl -3-oxide (cPTIO) 結果則抑制CaCl2的作用，顯示高鹽逆境下NO參與Ca2+ 調控冠根發育之機制。利用 nitrate reductase (NR) 抑制劑sodium tungstate處理後可完全抑制高鹽逆境下Ca2+ 恢復冠根發育的作用，但 nitric oxide synthase (NOS) 抑制劑Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME) 處理則無顯著影響，結果顯示高鹽逆境下 Ca2+ 主要經由NR途徑生成NO。以auxin 極性運移influx蛋白抑制劑1-Naphthoxyacetic acid (1-NOA) 及efflux蛋白抑制劑N-1-Naphthylphthalamidic acid (NPA) 處理後，Ca2+ 及NO之作用僅受NPA抑制，推測IAA極性運移也參與Ca2+ 及NO恢復水稻幼苗於高鹽逆境下冠根形成之作用。且高鹽逆境下CaCl2 恢復冠根之形成不受guanylate cyclase (GC) 抑制劑 LY83583 及 calmodulin 拮抗劑 chlorpromazine hydrochloride (CPZ) 影響。
低鹽逆境 (25 mM NaCl) 可誘導水稻冠根之形成，但可被1-NOA、NPA、cPTIO 及L-NAME處理所抑制，且 IAA 能明顯恢復 cPTIO 之抑制效果。CPZ、cyclic ADP ribose (cADPR) 合成抑制劑 NA、Ca2+ 螯合劑 EGTA、Ca2+ 原生質膜通道抑制劑 LaCl3 與 ruthenium red (RR) 處理皆不影響低鹽逆境誘導之冠根形成，但受LY83583、inositol 1,4,5-trisphosphate (IP3) 循環抑制劑 LiCl 所抑制。推測低鹽逆境下可能是經IP3 調控細胞質 Ca2+ 濃度，影響 NO、IAA 及cGMP 進而誘導冠根之形成。
處理CaCl2 可明顯減緩高鹽逆境抑制之主根生長，且CaCl2 之作用可被CPZ所抑制，顯示高鹽逆境下 CaCl2 可能經由 calmodulin 減緩主根生長受抑制之情形。

In this thesis, rice (Oryza sativa L. cv. Taichung Native 1) seedlings were used to investigate the role of Ca2+, nitric oxide (NO) and auxin in the regulation of primary root (PR) growth and crown root (CR) emergence in salt stress.
In high salt stress (150 mM NaCl), the CR of rice seedlings was reduced. Application of calcium chloride (CaCl2), sodium nitroprusside (SNP; a NO donor) and indole-3-acetic acid (IAA) to rice seedlings induced CR emergence in high salt stress. Further research on the CR primordia by paraffin section, the result is consistent with above. The effect is specific for NO because the NO scavenger 2-(4-carboxyphenyl)- 4,4,5,5-tetra-methylimidazoline-1-oxyl-3-oxide (cPTIO) blocked the action of CaCl2. These results suggest the involvement of NO in regulating Ca2+ induced CR emergence in high salt stress. Nitrate reductase (NR) inhibitor sodium tungstate inhibited Ca2+ induced CR emergence in high salt stress. In contrast, nitric oxide synthase (NOS) inhibitor Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME) was not significant. It means that NO generation that occurs in response to Ca2+ might primarily involve NR activity in high salt stress. Treatment of seedlings with auxin polar transport influx protein inhibitors 1-Naphthoxyacetic acid (1-NOA) and efflux protein inhibitors N-1-Naphthylphthalamidic acid (NPA), the result indicate that only NPA reduced to response in both Ca2+ and NO. Moreover, guanylate cyclase (GC) inhibitor LY83583 and calmodulin antagonists (CPZ) had no effect on Ca2+ induced CR emergence in high salt stress.
On the other hand, low salt stress (25 mM NaCl) increased the rice seedlings CR emergence, which could be blocked by 1-NOA, NPA, cPTIO and L-NAME. Moreover, application of rice seedlings with IAA resulted in a significant increase in cPTIO treatment. Supplemental CPZ, cADPR synthesis inhibitor (NA), Ca2+ chelator (EGTA), plasma membrane Ca2+-channels inhibitors (LaCl3, RR) had no effect on increased of CR emergence by low salt stress. However, low salt stress increased CR emergence could be reduced by LY83583 and IP3 cycling inhibitor (LiCl). Therefore, our results demonstrated that NO, IAA and cGMP are involved in Ca2+ induced the rice CR emergence in low salt stress. Moreover, cytosolic levels of Ca2+ maybe regulated by IP3.
Treatment of CaCl2 to rice seedlings also induced PR growth in high salt stress, which could be reduced by CPZ. These results indicate that calmodulin are involved in Ca2+ induced the PR growth in high salt stress.

中文摘要………………………………………………………..i
英文摘要………………………………………………………iii
目錄…………………………………………………………….v
圖表目錄………………………………………………………vi
縮寫字對照表………………………………………………..viii
前言…………………………………………………………….1
前人研究……………………………………………………….3
材料與方法…………………………………………………...17
結果…………………………………………………………...24
討論…………………………………………………………...57
未來研究方向………………………………………………...66
參考文獻……………………………………………………...67

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