本文選題：扁蓿豆 + MrSKIP基因　； 參考：《內(nèi)蒙古大學》2017年碩士論文

【摘要】：本研究用含20%PEG 6000的MS液體培養(yǎng)基模擬生理脫水脅迫處理扁蓿豆幼苗,以上述脅迫處理6 h的扁蓿豆幼苗葉片總RNA為模板進行反轉(zhuǎn)錄,運用RACE技術(shù)和RT-PCR方法,克隆得到扁蓿豆抗逆相關(guān)RNA剪切因子MrSKIP基因cDNA全長。序列分析結(jié)果顯示,MrSKIP基因cDNA全長為2280bp,含有一個1833bp的完整開放閱讀框,序列推測編碼610個氨基酸含一條S-N-W-K-N的多肽,分子量為68.86145 KDa,等電點為8.95,生物信息學分析表明MrSKIP含有該家族的典型結(jié)構(gòu)域SNW_SKIP。進一步克隆得到了扁蓿豆基因組的MrSKIP基因,序列分析表明,該基因無內(nèi)含子。利用tasiRNA技術(shù)和發(fā)根農(nóng)桿菌轉(zhuǎn)化法,在截形苜蓿108R根部干擾MrSKIP同源基因的表達,對轉(zhuǎn)化外源基因的截型苜蓿根系進行干旱、高鹽和低溫脅迫處理,鑒定MrSKIP基因的功能,結(jié)果顯示脅迫處理下干擾MrSKIP同源基因的的轉(zhuǎn)基因根系樣品,其根長和根鮮重均顯著小于對照。研究結(jié)果初步表明MrSKIP基因表達量的降低影響植物對干旱、低溫和高鹽脅迫的耐受性,特別對干旱脅迫耐受性的降低尤為顯著。
[Abstract]:In this study, a MS liquid medium containing 20 and PEG 6000 was used to simulate physiological dehydration stress to treat almond bean seedlings. The leaf total RNA of almond bean seedlings treated with above stress for 6 h was used as template for reverse transcription. Race technique and RT-PCR method were used. The full-length cDNA of rskIP gene was obtained by cloning. Sequence analysis showed that the full length cDNA of Mr skp gene was 2280bp. it contained a complete open reading frame of 1833bp. The sequence was deduced to encode 610 amino acids containing a polypeptide of S-N-W-K-N. The molecular weight is 68.86145 KDaand the isoelectric point is 8.95. The bioinformatics analysis shows that MrskIP contains the typical domain SNWSKIPs of this family. The MRSKIP gene was further cloned and sequenced, which showed that the gene had no intron. Using tasiRNA technique and Agrobacterium rhizogenes transformation method, the expression of MRSKIP homologous gene was interfered at the root of truncated alfalfa (108R). The root system of truncated alfalfa transformed with exogenous gene was treated with drought, high salt and low temperature stress to identify the function of MRSKIP gene. The results showed that the root length and fresh weight of the transgenic root samples which interfered with the MRSKIP homologous gene under stress were significantly lower than that of the control. The results showed that the decreased expression of MrskIP gene affected the tolerance of plants to drought, low temperature and high salt stress, especially to drought stress.
【學位授予單位】：內(nèi)蒙古大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：Q943.2

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