本文選題：獨(dú)行菜 + GRAS轉(zhuǎn)錄因子　； 參考：《分子植物育種》2017年09期

【摘要】：以耐受低溫萌發(fā)停滯期的種子為實驗組,以低溫萌發(fā)停滯期的種子為對照組,對獨(dú)行菜(Lepidium apetalum Willd.)種子轉(zhuǎn)錄組進(jìn)行了高通量測序�；跍y序數(shù)據(jù)庫資源,本研究對獨(dú)行菜GRAS類轉(zhuǎn)錄因子進(jìn)行了分析,除去冗余,發(fā)現(xiàn)獨(dú)行菜種子共有GRAS家族270個成員。其中138個成員相對于低溫萌發(fā)停滯組,在耐受低溫萌發(fā)的獨(dú)行菜種子中上調(diào)表達(dá),128個下調(diào)表達(dá),4個表達(dá)水平無差異。進(jìn)一步選擇了一個極顯著上調(diào)表達(dá)的GRAS基因片段,通過Blastn同源序列比對分析表明,該基因片段與擬南芥GRAS基因家族SCL18全長基因一致性最高,達(dá)到96%,故本研究將其命名為La SCL18(Gen Bank登錄號KY466159)。結(jié)合同源序列克隆法、采用RT-PCR技術(shù)克隆獲得該GRAS基因c DNA序列,全長1 633 bp,編碼一個完整的開放閱讀框。生物信息學(xué)分析表明,該c DNA序列編碼435個氨基酸、具有GRAS轉(zhuǎn)錄因子家族特有的保守結(jié)構(gòu)域、親水性較高、是一個水溶性較好的球狀蛋白。編碼蛋白分子質(zhì)量139.883 k D、理論等電點(diǎn)為4.97、不具有跨膜區(qū)和DNA結(jié)合位點(diǎn),表明其在調(diào)控相關(guān)基因轉(zhuǎn)錄時可能不直接與DNA序列結(jié)合,而是與其它轉(zhuǎn)錄因子相互作用后發(fā)揮功能。實時熒光定量PCR分析表明,獨(dú)行菜幼苗在低溫脅迫處理后La SCL18基因表達(dá)量迅速顯著升高,初步推測La SCL18基因表達(dá)與獨(dú)行菜幼苗耐受低溫有關(guān)。
[Abstract]:The seeds of Lepidium apetalum Willd.were used as experimental group and control group respectively. High throughput sequencing of seed transcriptome was carried out. Based on sequencing database resources, GRAS transcription factors were analyzed in this study. After removing redundancy, 270 members of GRAS family were found. Compared with the cold germinating arrest group, 138 members up-regulated the expression in the seeds of the cold germinating alone, 128 down-regulated expression, and 4 expression levels had no difference. A highly up-regulated GRAS gene fragment was further selected. The results of Blastn homologous sequence alignment analysis showed that the gene fragment had the highest consistency with the full-length SCL18 gene of Arabidopsis GRAS gene family. This study named La SCL18(Gen Bank login number KY466159. Combined with homologous sequence cloning, the GRAS gene c DNA sequence was cloned by RT-PCR technique. The cDNA sequence was 1 633 BP, encoding a complete open reading frame. Bioinformatics analysis showed that the c DNA sequence encodes 435 amino acids, has the conserved domain of GRAS transcription factor family, has high hydrophilicity, and is a better water-soluble globular protein. With a molecular weight of 139.883 KD and a theoretical isoelectric point of 4.97, the encoded protein does not have a transmembrane region or a DNA binding site, indicating that it may not directly bind to the DNA sequence in regulating the transcription of the related genes, but function after interacting with other transcription factors. The results of real-time fluorescence quantitative PCR analysis showed that La SCL18 gene expression increased rapidly after low temperature stress. It was preliminarily assumed that La SCL18 gene expression was related to tolerance to low temperature.
【作者單位】： 新疆特殊環(huán)境物種保護(hù)與調(diào)控生物學(xué)實驗室;北京師范大學(xué)生命科學(xué)學(xué)院抗性基因資源與分子發(fā)育北京市重點(diǎn)實驗室;
【基金】：國家自然科學(xué)基金(3166010371) 新疆維吾爾自治區(qū)高等學(xué)�？蒲杏媱濏椖�(XJEDU2014I027) 新疆維吾爾自治區(qū)大學(xué)生科研創(chuàng)新項目(201610762095) 新疆師范大學(xué)校級研究生科研創(chuàng)新項目(XSY201602003)共同資助
【分類號】：Q943.2

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