發(fā)布時間：2018-11-28 11:49

【摘要】：超氧化物歧化酶(SOD,superoxide dismutase)是植物中一種主要的抗氧化酶,在植物應(yīng)對逆境脅迫及抗衰老中起重要作用。本研究從基因芯片數(shù)據(jù)中篩選獲得小麥Cu/Zn-SOD基因的EST序列,通過序列比對后拼接得到小麥Cu/Zn-SOD的候選基因,利用PCR技術(shù)在小麥光溫敏雄性不育材料BS366中克隆并獲得該基因。通過對Cu/Zn-SOD基因序列進行生物信息學(xué)分析,結(jié)果表明,該基因擁有連續(xù)且完整的開放閱讀框,長495bp,編碼164個氨基酸。氨基酸序列分析發(fā)現(xiàn),該蛋白具有保守的Cu/Zn-SOD功能結(jié)構(gòu)域與典型的Cu/Zn-SOD三維結(jié)構(gòu),且定位于細胞質(zhì)中。通過同源進化分析表明,該蛋白與二穗短柄草(Brachypodium distachyon(L.)Beauv.)和大麥(Hordeum vulgare L.)的Cu/Zn-SOD蛋白親緣關(guān)系較近,相似度分別為89%和94%。利用實時熒光定量PCR技術(shù)對其在小麥不同組織的表達特異性及不同逆境脅迫下的表達模式進行分析,結(jié)果表明,該基因在根、莖、葉、雌蕊、雄蕊、穎殼中均有表達,屬于組成型表達,且在小麥的地上部含葉綠體的組織中含量較高;同時受多種脅迫誘導(dǎo),可能參與了多種脅迫誘導(dǎo)調(diào)控途徑。通過對該基因在不同育性環(huán)境中BS366育性轉(zhuǎn)換期花藥中的表達模式分析,發(fā)現(xiàn)可育環(huán)境下,在小孢子母細胞時期和減數(shù)分裂期的表達量分別約為對照的8倍與16倍;而不育環(huán)境下,該基因表達水平無明顯變化。因而推測,小麥Cu/Zn-SOD基因可能參與了光溫敏雄性不育系BS366的育性調(diào)控。本研究為深入研究Cu/Zn-SOD基因在小麥中的作用機理奠定了重要基礎(chǔ)。
[Abstract]:Superoxide dismutase (SOD,superoxide dismutase) is a major antioxidant enzyme in plants, which plays an important role in coping with stress and anti-senescence. In this study, the EST sequence of wheat Cu/Zn-SOD gene was screened from gene chip data, and the candidate gene of wheat Cu/Zn-SOD was obtained by sequence alignment. The PCR technique was used to clone and obtain the gene from wheat photo-thermosensitive male sterile line BS366. The results of bioinformatics analysis of Cu/Zn-SOD gene showed that the gene had a continuous and complete open reading frame with a length of 495bpand encoding 164 amino acids. Amino acid sequence analysis showed that the protein had a conserved Cu/Zn-SOD functional domain and a typical Cu/Zn-SOD three-dimensional structure, and was located in the cytoplasm. The result of homologous evolution analysis showed that the protein was similar to (Brachypodium distachyon (L.) Beauv.). Barley (Hordeum vulgare L.) The similarity of Cu/Zn-SOD protein was 89% and 94%, respectively. The expression specificity of the gene in different wheat tissues and its expression patterns under different stresses were analyzed by real-time fluorescence quantitative PCR. The results showed that the gene was expressed in roots, stems, leaves, pistil, stamens and glumes. It belongs to the constitutive expression, and the content of chloroplast is higher in the tissue with chloroplast in the aboveground part of wheat. At the same time, it may be involved in the regulation of multiple stress induced by multiple stresses. By analyzing the expression pattern of the gene in the anthers of BS366 fertility transition in different fertility environments, it was found that in fertile environment, the expression of the gene in microspore mother cell stage and meiosis stage was about 8 times and 16 times as much as that of the control, respectively. However, there was no significant change in the expression level of the gene in infertile environment. It is suggested that the Cu/Zn-SOD gene may be involved in the fertility regulation of the photo-temperature sensitive male sterile line BS366. This study laid an important foundation for further study on the mechanism of Cu/Zn-SOD gene in wheat.
【作者單位】： 北京農(nóng)學(xué)院植物科學(xué)技術(shù)學(xué)院;北京市農(nóng)林科學(xué)院雜交小麥工程技術(shù)研究中心;雜交小麥分子遺傳北京市重點實驗室;首都師范大學(xué)生命科學(xué)學(xué)院;
【基金】：國家科技支撐計劃項目(2013BAD04B01,2014BAD01B09) 國家自然科學(xué)基金(31371699) 北京市自然科學(xué)基金項目(5131001) 北京市農(nóng)林科學(xué)院科技創(chuàng)新能力建設(shè)專項(KJCX20140405);北京市農(nóng)林科學(xué)院青年基金(QNJJ201427) 北京市科技計劃項目(Z141100002314018)
【分類號】：S512.1

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