【摘要】：甜菜堿醛脫氫酶(betaine aldehyde dehydrogenase,BADH)是甜菜堿(glycine betaine,GB)合成過程中的關(guān)鍵酶。為檢測(cè)BADH基因在木薯(Manihot esculenta)不同組織中的表達(dá)差異,探究干旱與NaCl脅迫下其表達(dá)譜變化規(guī)律,本研究利用生物信息學(xué)方法分析和鑒定木薯BADH基因,定位MeBADHs在染色體上的位置以及確定5'端上游序列順式作用元件;采用qRT-PCR測(cè)定BADH基因在木薯品種SC5根、塊根、莖和葉中的表達(dá)差異,同時(shí)檢測(cè)在PEG-6000模擬脫水脅迫、NaCl脅迫及自然干旱脅迫下MeBADHs基因的轉(zhuǎn)錄水平變化,采用相關(guān)試劑盒測(cè)定木薯中GB含量的變化。結(jié)果顯示,MeBADHs兩個(gè)基因在各組織之中均有表達(dá),在塊根中表達(dá)量顯著高于根(P0.05)。PEG-6000模擬干旱條件下,與0 h相比,1~2 h時(shí)MeBADH1和MeBADH2在葉、莖、根以及塊根中的表達(dá)量變化不大;NaCl處理?xiàng)l件下,葉中MeBADH1在8 h表達(dá)量極顯著高于0 h(P0.01),而MeBADH2則在不同處理時(shí)間下的表達(dá)量水平相差不大,在莖和根中,MeBADHs在處理發(fā)生1 h時(shí)表達(dá)量水平極顯著高于0 h(P0.01);自然缺水干旱條件下,莖和根中MeBADH1和MeBADH2分別在6和18 d時(shí)的表達(dá)量極顯著高于0 d(P0.01)。PEG-6000處理下,GB含量出現(xiàn)了波動(dòng)并呈現(xiàn)略增趨勢(shì);NaCl處理?xiàng)l件下GB含量在前期表達(dá)較穩(wěn)定,8 h時(shí)劇烈響應(yīng),即出現(xiàn)下調(diào)趨勢(shì),24 h時(shí)GB含量顯著低于0 h(P0.05)。在自然干旱脅迫下,GB含量逐漸呈現(xiàn)上調(diào)趨勢(shì),并在9和15 d達(dá)到高峰,顯著高于0 d(P0.05)。研究結(jié)果為進(jìn)一步研究木薯分子抗旱機(jī)制及新品種的選育提供了理論依據(jù)。
[Abstract]:Betaine aldehyde dehydrogenase (betaine aldehyde dehydrogenase,BADH) is a key enzyme in the synthesis of betaine (glycine betaine,GB). In order to detect the difference of BADH gene expression in different tissues of cassava (Manihot esculenta) and to explore the variation of BADH gene expression profile under drought and NaCl stress, the BADH gene of cassava was analyzed and identified by bioinformatics. Locate the position of MeBADHs on chromosome and determine the cis acting element of 5'terminal upstream sequence; QRT-PCR was used to detect the difference of expression of BADH gene in SC5 roots, tubers, stems and leaves of cassava cultivars. The transcription level of MeBADHs gene was also detected under PEG-6000 simulated dehydration stress, NaCl stress and natural drought stress. The change of GB content in cassava was determined by correlation kit. The results showed that two MeBADHs genes were expressed in all tissues, and the expression level in tuber root was significantly higher than that in root (P0.05). Under PEG-6000 simulated drought, compared with 0 h, MeBADH1 and MeBADH2 were in leaves and stems at 1h and 2 h, respectively. The expression level of root and tuber root showed little change. Under the condition of NaCl treatment, the expression of MeBADH1 in leaves at 8 h was significantly higher than that in 0 h (P0.01), while the expression level of MeBADH2 in stem and root was not different under different treatment time. The expression level of MeBADHs at 1 h was significantly higher than that at 0 h (P0.01). The expression of MeBADH1 and MeBADH2 in stem and root were significantly higher than that in 0 d (P0.01) at 6 d and 18 d, respectively. Under the condition of natural water shortage and drought, the GB content fluctuated and increased slightly. Under the condition of NaCl treatment, the expression of GB was stable in the early stage, and the response was intense at 8 h, which showed a downward trend, and the GB content at 24 h was significantly lower than that at 0 h (P0.05). Under natural drought stress, the content of GB increased gradually, and reached its peak at 9 and 15 days, which was significantly higher than that of 0 days (P0.05). The results provide theoretical basis for further study on molecular drought resistance mechanism of cassava and breeding of new varieties.
【作者單位】： 海南大學(xué)農(nóng)學(xué)院;中國(guó)熱帶農(nóng)業(yè)科學(xué)院熱帶生物技術(shù)研究所/農(nóng)業(yè)部熱帶作物生物學(xué)與遺傳資源利用重點(diǎn)實(shí)驗(yàn)室;中國(guó)環(huán)境科學(xué)研究院;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(No.31201603) 2016農(nóng)業(yè)部熱作技術(shù)試驗(yàn)示范項(xiàng)目(No.15162130106232022) 海南省自然科學(xué)基金項(xiàng)目(No.312051) 國(guó)家高技術(shù)研究發(fā)展計(jì)劃(863計(jì)劃)子課題(No.2012AA101204-2) 海南省重大科技項(xiàng)目子課題(No.ZDZX2013023-1) 海南省研究生創(chuàng)新課題(S03) 環(huán)境保護(hù)部生物多樣性保護(hù)專項(xiàng)(No.2096001006)
【分類號(hào)】：S533

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