SWEET醣轉運蛋白不僅調控細胞間或細胞內醣類分配以維持植物生長，其活性也與病原菌感染繁殖所需醣份來源息息相有關。已知AtSWEET2為一根部液胞膜上的葡萄糖專一轉運蛋白，會受Pythium感染誘導以回收分泌至根圈之葡萄糖，進而限制病原菌繁殖，為重要的抗病機制。然而，AtSWEET2表現的訊息調控至目前未有任何研究。從酵母單雜合分析，我們分離出12個可能調控AtSWEET2啟動子的轉錄因子，利用阿拉伯芥原生質同時表現由AtSWEET2啟動子活化的冷光報導基因與轉錄因子分析其在活體中的調控能力，結果發現ANAC046轉錄因子的表達則可誘導AtSWEET2啟動子2倍的表達活性，而AHL26、AHL27、AHL29與TCP17轉錄因子的過表達會顯著抑制AtSWEET2啟動子的表達活性，達50％以上，其中AHL29抑制調控最強，因此聚焦分析。當共同表達1.4、1、0.5 kb 不同長度的AtSWEET2啟動子與AHL29時，發現雖然較短的啟動子顯著降低AtSWEET2的表現，然而AHL29的抑制調控的趨勢仍相同，顯示其結合位點應是位於0.5 kb之內，此與PlantPAN的分析在-1400 bp至-1000 bp之間及-500 bp內具有AHL的結合區。當分析AHL29兩個重要的保守區域突變對調控的影響，發現AT-hook motif (ahl29-6) 突變無法抑制AtSWEET2表達，而PPC/DUF domain (AHL29-ΔG)突變則不影響其抑制調控。為了進一步確定AHL29在植物生長的調控，利用qRT–PCR分析突變株中AtSWEET2基因表現，發現在持續活化的AHL29-D (Gain of function) 葉片中AtSWEET2表現量降低約60%，與瞬時表達分析結果雷同。然而在失去DNA結合力的ahl29-6 (Loss of function) 葉片中AtSWEET2表現卻也同樣降低約40%，推測ahl29-6 點突變可能不影響AHL29結合在AtSWEET2啟動子。
進一步利用原生質體瞬時表現探討AHL29對AtSWEET2表現的調控是否會受到微生物訊息分子及荷爾蒙的影響。結果顯示處理0.1 μM細菌的鞭毛蛋白Flagellin22會促進AHL2抑制AtSWEET2表現，10 μM離層酸處理也有相同的促進抑制的趨勢。而30 μM茉莉酸甲酯與25 μM或50 μM水楊酸則減弱AHL29抑制AtSWEET2表現。此外我們也發現AHL29的調控與葡萄糖訊息有關。而另一與AHL29相同基因家族，在AT-hook motif中胺基酸相似性為66 %，在PPC/DUF domain中胺基酸相似性為68 %的AHL26，也會抑制調控AtSWEET2的表現程度，表現結果推測其在AtSWEET2啟動子的結合位點約在1.4 kb與1.0 kb間，未來將建立AHL26基因專一RNAi以及AHL保守序列 RNAi片段 (AT-hook motif與PPC/DUF296 domain) 進行功能分析。針對許多轉錄因子常會功能互補以致無法分離基因功能，我們也建立利用阻遏蛋白抑制基因家族表現的同型合子CRES-T轉殖株，雖然有檢測到阻遏蛋白的表現，然而AtSWEET2基因表現未有顯著差異。總言之，本研究發現AHL29負向調控AtSWEET2基因表現，且可能參與微生物感染訊息的途徑，未來需重複實驗驗証，也會利用突變株進行功能分析，以解析AtSWEET2的分子調控。

In this study, we performed transient assay to indentify that AHL26, AHL27, AHL29 and TCP may function as a repressor, and ANAC046 may function as an activator to regulate AtSWEET2 expression where AHL29 showed the strongest effect and was further studied. The promoter delection analysis further indicated that the binding sites of AHL26 and AHL29 on AtSWEET2 promoter may be located between 1.4 kb to 1.0 kb upstream of the start site. Moreover, mutation in AT-hook motif (ahl29-6) resulted in reduced AHL29 binding on the AtSWEET2 promoter. Consistently, qRT-PCR analysis showed that expression of AtSWEET2 was significantly inhibited by 60% in leaves of AHL29 gain-of-function mutant plants. Furthermore, treatments of 0.1 μM Flagellin22 and 10 μM ABA will promote the repression of AHL29 on AtSWEET2 expression. On the other hands, treatments 30 μM methyl jasmonate and 25 μM salicylic acid reduced AHL29 repression. In summary, our studies indicated that AHL29 negativly regulate SWEET2 function and may be involved in signaling pathway of microorganism interaction in Arabidopsis roots.

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