單胞藻 (Chlamydomonas reinhardtii) 可利用 CO2 作為自營 (Sueoka high
salt medium + 5% CO2) 培養或利用 acetate 作為混營 (Tris-acetate-phosphate) 培
養。本研究目的為釐清 HSM + 5% CO2 自營培養及 TAP 混營培養之
methionine sulfoxide reductase (MSR) 基因表現之差異及對高光 (1,000 μE m-2 s-1)
之反應，並了解光合電子傳遞鏈參與高光調控基因表現之角色。高光抑制自營培
養和混營培養單胞藻之 PSII 活性 (Fv/Fm 及 Fv'/Fm')，抑制程度相同。高光促進
自營培養之 CrMSR isoform 基因表現數目較混營培養多，兩者受高光促進之
isoform 種類不同，高光促進自營培養之 CrMSRA1、CrMSRA2、CrMSRA3、
CrMSRA5 、CrMSRB1.2 和 CrMSRB2.1 基因表現而抑制 CrMSRA4 和
CrMSRB2.2 基因表現， 高光促進混營培養之 CrMSRA3 、CrMSRA5 和
CrMSRB2.1 基因表現而抑制 CrMSRA1、CrMSRA4 和 CrMSRB2.2 基因表現。
利用光合電子傳遞鏈抑制劑證實光合電子傳遞鏈為高光影響自營培養和混營培
養之 CrMSR isoform 基因表現的因子，但是兩者受電子傳遞鏈之不同位置調
控，尤其是 CrMSRA 基因表現。自營培養的 CrMSRA 表現多受 QA 還原態 (-)
調控，混營培養則多由 PQ 還原態 (-) 調控，自營培養 CrMSRB 表現受光合電
子傳遞鏈影響大，主要由 QA.(-) 和 Cytb6f (-) 調控，混營培養 CrMSRB 表現受
光合電子傳遞鏈影響較小，僅 CrMSRB2.2 受 Cytb6f (-) 調控。高光不誘導自營
培養與混營培養的 H2O2 產生，推測 CrMSR isoform 基因表現與 H2O2 無關。
本研究證實自營培養和混營培養 CrMSR 基因表現受高光調控情形不同，並討論
碳源利用方式與 CrMSR 基因表現調控之關係。

Chlamydomonas reinhardtii can utilize CO2 for autotrophic growth (HSM plus
5% CO2) or acetate for mixotrophic growth (TAP). This study was to elucidate the
differential regulation of methionine sulfoxide reductase (MSR) gene expression
between HSM plus 5% CO2 and TAP cultured cells, and also to determine the
difference of gene expression in response to high light (1,000 μE m-2 s-1). The role of
photosynthetic electron transport (PET) in the regulation of MSR gene expression was
also examined by the use of PET inhibitors. High light inhibited PSII activity (Fv/Fm
and Fv'/Fm') of HSM plus 5% CO2 and TAP cultured cells., while the responses of
CrMSR gene expression in mixotrophically grown cells were different from
autotrophically grown cells, High light increased the expression of CrMSRA1,
CrMSRA2, CrMSRA3, CrMSRA5, CrMSRB1.2, and CrMSRB2.1, but inhibited the
expression of CrMSRA4 and CrMSRB2.2 in autotrophically grown cells. The
expression of CrMSRA3, CrMSRA5, and CrMSRB2.1 in mixotrophically grown cells
was increased by high light but that of CrMSRA1, CrMSRA4, and CrMSRB2.2 was
inhbited. The number of MSR isoform that was up-regulated by high light was greater
in autotrophically grown cell than that in mixotrophically grown cells. Using the PET
inhibitors (3-(3,4-dichlorophenyl)-1,1- dimethylurea (DCMU) and
2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB)), most of the CrMSRA
expression was regulated by reduced QA for autotrophically grown cells while
reduced PQ was the main site for mixotrophically grown cells by high light. The
expression of CrMSRB in autotrophically grown cells was mainy modulated by QA (-)
or Cytb6f (-), while that was not affected by PET, except a role of Cytb6f (-) on the
high light-induced CrMSRB2.2 expression. We fouind that CrMSRB gene expression
in autotrophically grown cells was highly affected by PET but not for micotrophically
grtown cells. The present result that H2O2 did not accumulate in autotrophically and
mixotrophically grown cells suggests that H2O2 may be not involved in the regulation
of high light regulation of CrMSR gene expression. The present study shows that the
mRNA expression of CrMSR isoforms in Chlamydomonas was diffrerentially
regulated between autotrophically and mixttrophically grown cells. The relationship
between the utilization of different C source and CrMSR gene expression will be
discussed.

目錄
論文審定書 ..................................................................................................................... i
誌謝 ............................................................................................................................... ii
中文摘要 .......................................................................................................................iii
英文摘要 ....................................................................................................................... iv
縮寫對照表 ................................................................................................................... vi
目錄 .............................................................................................................................viii
圖目次 ........................................................................................................................... ix
表目次 ........................................................................................................................... xi
附錄目次 ...................................................................................................................... xii
壹、 緒論 ...................................................................................................................... 1
貳、 論文研究目的及實驗架構 ................................................................................ 15
參、 材料與方法 ........................................................................................................ 17
肆、 結果 .................................................................................................................... 27
伍、 討論 .................................................................................................................... 36
陸、 參考文獻 ............................................................................................................ 43
柒、 附錄 .................................................................................................................... 74

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