本文選題：水稻 + Os��； 參考：《中國(guó)農(nóng)業(yè)科學(xué)》2017年12期

【摘要】：【目的】農(nóng)作物對(duì)逆境脅迫的耐受能力與產(chǎn)量息息相關(guān),是作物育種要考慮的重要因素。文中對(duì)水稻順式還原酮加雙氧酶基因Os ARD1進(jìn)行研究,分析其表達(dá)模式,明確其在水稻應(yīng)對(duì)非生物脅迫中的功能,為水稻耐旱品種的分子設(shè)計(jì)及育種提供參考依據(jù)�！痉椒ā刻崛〔煌M織器官的總RNA,利用RT-PCR方法分析Os ARD1表達(dá)的組織特異性。利用不同的非生物脅迫處理14 d大小的野生型(中花11)植株,在不同時(shí)間點(diǎn)提取總RNA,利用RT-PCR方法分析Os ARD1表達(dá)的受誘導(dǎo)情況。通過農(nóng)桿菌遺傳轉(zhuǎn)化法轉(zhuǎn)化水稻愈傷組織,經(jīng)過一系列分子檢測(cè)后獲得穩(wěn)定遺傳的T1代Os ARD1的過量表達(dá)轉(zhuǎn)基因植株,以轉(zhuǎn)入空載體的野生型植株作為對(duì)照。將在營(yíng)養(yǎng)液中正常培養(yǎng)的12 d大小的野生型和過表達(dá)幼苗移出營(yíng)養(yǎng)液進(jìn)行缺水處理并進(jìn)行恢復(fù)試驗(yàn)。將催芽后的野生型和過表達(dá)轉(zhuǎn)基因植株種子種在含有5%PEG6000的agar培養(yǎng)基中進(jìn)行滲透脅迫處理,以不含PEG6000的agar培養(yǎng)基作為對(duì)照,觀察二者的表型�！窘Y(jié)果】組織特異性表達(dá)分析表明Os ARD1主要在根及成熟的組織中表達(dá),尤其在衰老的組織中有較高表達(dá)。非生物脅迫處理表明Os ARD1的表達(dá)明顯受機(jī)械損傷、高鹽和滲透脅迫的誘導(dǎo)。獲得6個(gè)獨(dú)立株系的可穩(wěn)定遺傳的Os ARD1過量表達(dá)轉(zhuǎn)基因植株。對(duì)過量表達(dá)轉(zhuǎn)基因植株及空載體野生型對(duì)照進(jìn)行干旱脅迫處理,缺水處理5 h后,野生型植株葉片卷曲皺縮成針狀表現(xiàn)出嚴(yán)重的缺水癥狀,但此時(shí)過表達(dá)轉(zhuǎn)基因植株葉片仍處于舒展?fàn)顟B(tài);缺水處理8 h后開始復(fù)水培養(yǎng)3 d,野生型植株的存活率僅為10%,而過表達(dá)植株存活率為80%,遠(yuǎn)遠(yuǎn)高于野生型,說明過量表達(dá)Os ARD1提高了水稻對(duì)缺水的耐受能力。用PEG滲透脅迫模擬干旱脅迫處理6 d后發(fā)現(xiàn),不含PEG6000對(duì)照組中野生型和過表達(dá)植株的幼苗生長(zhǎng)情況沒有明顯的差別;在PEG處理組中,野生型幼苗根的生長(zhǎng)受到嚴(yán)重抑制,而過表達(dá)植株幼苗根的生長(zhǎng)受到抑制較小,根長(zhǎng)明顯長(zhǎng)于野生型對(duì)照植株,說明過量表達(dá)Os ARD1增強(qiáng)了水稻耐受干旱脅迫的能力�！窘Y(jié)論】Os ARD1主要在水稻根及成熟的組織中表達(dá),并且受機(jī)械損傷、高鹽和滲透脅迫的誘導(dǎo)。過量表達(dá)Os ARD1提高了水稻抗旱性能。
[Abstract]:Objective: the tolerance of crops to stress is closely related to yield and is an important factor to be considered in crop breeding. In this paper, the gene Os ARD1 of cis-reducing ketone and dioxygenase in rice was studied, its expression pattern was analyzed, and the function of Os ARD1 in response to abiotic stress in rice was clarified. [methods] Total RNAs of different tissues and organs were extracted, and tissue specificity of Os ARD1 expression was analyzed by RT-PCR method. Wild type (Zhonghua 11) plants were treated with different abiotic stress for 14 days and total RNAs were extracted at different time points. The induction of Os ARD1 expression was analyzed by RT-PCR method. Transgenic rice callus was transformed by Agrobacterium tumefaciens genetic transformation. After a series of molecular tests, transgenic plants of stable T1 generation Os ARD1 were obtained, and wild type plants with empty vector were used as control. The wild type and over-expressed seedlings which were cultured in nutrient solution for 12 days were removed from the nutrient solution for water deficiency treatment and the recovery test was carried out. The seeds of wild type and overexpression transgenic plants were treated with osmotic stress in agar medium containing 5%PEG6000, and the agar medium without PEG6000 was used as control. [results] tissue specific expression analysis showed that Os ARD1 was mainly expressed in roots and mature tissues, especially in aging tissues. Under abiotic stress, the expression of Os ARD1 was induced by mechanical damage, high salt and osmotic stress. Stable and hereditary Os ARD1 transgenic plants of 6 independent lines were obtained. Transgenic plants and wild type control were treated with drought stress. After 5 hours of water stress treatment, leaf curls of wild type plants showed severe dehydration symptoms. The survival rate of wild type plants was only 10, while that of over-expressed plants was 80%, which was much higher than that of wild type plants. The results showed that the over-expression of Os ARD1 increased the tolerance of rice to water shortage. After simulated drought stress with PEG osmotic stress for 6 days, it was found that there was no significant difference between the growth of wild type seedlings and over-expressed plants in the control group without PEG6000, while in the PEG treatment group, the growth of wild type seedling roots was seriously inhibited. However, the growth of overexpressed seedlings was inhibited slightly, and the root length was significantly longer than that of wild type control plants. [conclusion] Os ARD1 was mainly expressed in roots and mature tissues of rice, and was induced by mechanical damage, high salt and osmotic stress. Overexpression of Os ARD1 improved drought resistance of rice.
【作者單位】： 天津師范大學(xué)生命科學(xué)學(xué)院/天津市動(dòng)植物抗性重點(diǎn)實(shí)驗(yàn)室;
【基金】：天津市自然科學(xué)基金重點(diǎn)項(xiàng)目(16JCZDJC33400) 天津市中青年骨干教師創(chuàng)新培養(yǎng)計(jì)劃(ZX110GG017) 天津師范大學(xué)博士基金(52XB1612,52XB1611)
【分類號(hào)】：S511

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