本文關(guān)鍵詞：大豆逆境脅迫誘導(dǎo)的miRNA鑒定及功能分析　出處：《中國農(nóng)業(yè)科學(xué)院》2015年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 大豆 miRNA 非生物脅迫 HD-Zip III 遺傳多樣性 遺傳標(biāo)記

【摘要】：micro RNA是一類的非編碼的單鏈小分子RNA,普遍存在于植物基因組中并參與轉(zhuǎn)錄后基因調(diào)控。mi RNA已被證明參與了植物抵抗環(huán)境脅迫(如干旱、高鹽,低溫和高溫等)的過程。近年來,對mi RNA在各作物逆境脅迫中的作用進(jìn)行了大量研究。在環(huán)境脅迫下,許多mi RNA已經(jīng)在擬南芥和水稻中證明存在表達(dá)差異,并且其中的一部分更是鑒定了功能,而在大豆中卻少有mi RNA涉及這方面的研究。雖然mi RNA僅代表轉(zhuǎn)錄本相對豐度的,但在不同物種、不同組織和不同環(huán)境脅迫條件下mi RNA的表達(dá)水平差異卻很大。本研究中我們通過Northern印記雜交分析證明了如gma-mi R166a、gma-mi R168a、gma-mi R398a和gma-mi R1508a等mi RNA會被干旱、高鹽、低溫脅迫誘導(dǎo)表達(dá)。表明這些mi RNA可能是大豆幼苗逆境脅迫的應(yīng)答mi RNA。因?yàn)榇蠖沟纳锒鄻有允重S富,因此我們深入的研究了各種脅迫條件下不同基因型大豆幼苗中g(shù)ma-mi R166a的表達(dá)。mi R166是一個即存在于單子葉植物又存在于雙子葉植物中的保守mi RNA,相關(guān)研究表明它們主要和植物的發(fā)育相關(guān),目前還沒有其與非生物脅迫相關(guān)的研究結(jié)果。本研究采用5'RACE方法從15個與脅迫相關(guān)的預(yù)測靶基因中鑒定了gma-mi R166切割Glyma09g02750和Glyma07g01950兩個基因,其中Glyma09g02750(命名為Gm PHB)屬于HD-Zip III轉(zhuǎn)錄因子家族,與擬南芥PHABULOSA(At2g34710)基因有78%的相似度。實(shí)時定量PCR結(jié)果表明,各脅迫條件下Gm PHB與gma-mi R166a在不同脅迫時間的表達(dá)量呈顯著負(fù)相關(guān)關(guān)系,這為進(jìn)一步研究mi R166在大豆逆境脅迫中的作用提供了科學(xué)依據(jù)。此外,在擬南芥中過表達(dá)gma-MIR166a,即抑制了At PHB的表達(dá),又增強(qiáng)了擬南芥的耐鹽能力,這意味著gma-MIR166a可能在大豆耐鹽能力方面起到了積極的調(diào)節(jié)作用。近年來,隨著公共基因組數(shù)據(jù)庫的使用,分子標(biāo)記已經(jīng)開始從基于PCR的勞動密集型標(biāo)記向基于基因分型和基因功能的標(biāo)記轉(zhuǎn)變,還常被RNA信息相結(jié)合。然而,目前為止基于mi RNA的分子標(biāo)記還僅在水稻、煙草和棉花中開發(fā)出來。本研究,通過對1669份來自中國、韓國、日本和俄羅斯的栽培大豆和一年生野生大豆中的一個特殊的具有遺傳變異的單克隆位點(diǎn)的分析,開發(fā)了第一個大豆功能mi RNA分子標(biāo)記,gma-mi R1511-In Del。結(jié)果表明,gma-mi R1511位點(diǎn)在栽培大豆中都有存在,而在一年生野生大豆中卻存在3種等位變異�；谠摻Y(jié)果,我們認(rèn)為In Del標(biāo)記可用于遺傳變異的評估。gma-mi R1511-In Del標(biāo)記數(shù)據(jù)和地理信息相結(jié)合進(jìn)行分析發(fā)現(xiàn),來自黃河流域的一年生野生大豆具有較高的遺傳多樣性,這為一年生野生大豆的基因多樣性和大豆的起源學(xué)說提供了更多的分子證據(jù)。另外,gma-mi R1511和其靶基因Gm RPL4a在脅迫處理下的表達(dá)也存在負(fù)相關(guān)關(guān)系,這表明gma-mi R1511可能具有調(diào)節(jié)大豆發(fā)育的功能。gma-mi R1511及其靶基因目前還未在其它豆科植物中發(fā)現(xiàn),因此,gma-mi R1511-In Del是可用于研究大豆遺傳多樣性、種質(zhì)的基因分型和進(jìn)化的大豆特有的分子標(biāo)記。這種通過mi RNA序列定位其前體區(qū)域序列來開發(fā)分子標(biāo)記的方法亦是首次報(bào)道。
[Abstract]:Micro RNA is a small single stranded non encoding RNA class, commonly exist in plant genome and involved in regulating gene transcription of.Mi RNA have been shown to be involved in plant resistance to environmental stress (such as drought, high salt, low temperature and high temperature) of the process. In recent years, done a lot of research in the MI RNA as a matter of stress in the role. Under the environmental stress, many mi RNA have proved that there were differential expression in Arabidopsis and rice, and a part of them is more functional identification in soybean and little mi RNA involved in the research. Although the MI RNA represents the relative abundance of transcripts. But in different species, different organizations and different environmental stress difference in MI expression under the condition of RNA is great. In this study we analyzed Northern blot proved that such as gma-mi R166a, gma-mi R168a, gma-mi R398a and gma-mi R1508a mi RNA Will be drought, high salt, low temperature stress induced expression. Suggest that these mi RNA may be the soybean seedling stress response of MI RNA. because soybean biological diversity is very rich, so we studied the different genotypes of Soybean Seedlings under the stress of gma-mi in the expression of R166a.Mi R166 is a mi that is conservative RNA in monocotyledonous plants also exist in dicotyledonous plants, related research shows that they mainly related to plant development, yet with the abiotic stress related research results. This research uses the 5'RACE method to predict the target gene from 15 stress-related identified gma-mi R166 cut Glyma09g02750 and Glyma07g01950 two the Glyma09g02750 gene (named Gm PHB) belongs to HD-Zip III family of transcription factors, and Arabidopsis PHABULOSA (At2g34710) 78% similarity gene. The results of real-time PCR Show that the stress condition Gm PHB and gma-mi R166a showed a significant negative correlation in the expression of different stress time, which provides a scientific basis for further research of MI R166 in soybean stress effect. Moreover, overexpression of gma-MIR166a in Arabidopsis, which inhibited the expression of At PHB, and enhanced the Arabidopsis salt tolerance, which means that gma-MIR166a may be the salt tolerance in soybean has played a positive role. In recent years, with the use of public genome database, molecular marker has begun to change from the marker gene classification and gene function based on labor intensive marker based on PCR, RNA is often combined with information. However, now mi RNA based on molecular markers also developed only in rice, tobacco and cotton. In this study, based on the 1669 from China, South Korea, Japan and Russia and cultivated soybean Analysis of Annual Wild Soybean in a special genetic variation with monoclonal sites, developed the first soybean Mi molecular markers RNA, gma-mi and R1511-In Del. results showed that gma-mi R1511 loci exist in cultivated soybean, but there are 3 alleles in wild soybean. Based on the results. We think that In Del markers can be used for the evaluation of.Gma-mi R1511-In genetic variation of Del marker data and geographic information combination analysis, genetic diversity from the Yellow River basin annual wild soybean has the high performance, the annual wild soybean genetic diversity and soybean origin theory provides a more molecular evidence. In addition, there are also a negative correlation between the expression of gma-mi R1511 and its target gene Gm RPL4a in stress treatments, suggesting that gma-mi R1511 may regulate the function of soybean growth.G Ma-mi R1511 and its target gene has not yet been found in other leguminous plants, therefore, gma-mi R1511-In Del can be used for research on soybean genetic diversity, molecular marker gene specific soybean germplasm classification and evolution. This method of locating the precursor region sequences by Mi RNA sequence to the development of molecular markers is also reported for the first time.

【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：Q943.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 ;Detection of genetically modified ingredients in seed samples from Heilongjiang soybean field[J];Chinese Science Bulletin;2005年10期

2 ;Studies on the overexpression of the soybean GmNHX1 in Lotus corniculatus: The reduced Na~+ level is the basis of the increased salt tolerance[J];Chinese Science Bulletin;2006年11期

3 ;Cloning and expressional characterization of soybean GmLls1 gene[J];Chinese Science Bulletin;2006年10期

4 ;Cloning and expression of putative ethylene receptor genes in soybean plant[J];Progress in Natural Science;2007年10期

5 ;Isolation and functional analysis of transcription factor GmWRKY57B from soybean[J];Chinese Science Bulletin;2008年22期

6 ;MiR1511 co-regulates with miR1511* to cleave the GmRPL4a gene in soybean[J];Chinese Science Bulletin;2012年Z2期

7 ;Construction of a soybean linkage map using an F_2 hybrid population from a cultivated variety and a semi-wild soybean[J];Chinese Science Bulletin;1997年12期

8 ;Characterization of root architecture in an applied core collection for phosphorus efficiency of soybean germplasm[J];Chinese Science Bulletin;2004年15期

9 郭倍奇,車延武,楊福愉;FLUIDITY OF MITOCHONDRIAL H~+-ATPASECONTAINING LIPOSOMES RECONSTITUTED IN THE PRESENCE OF Mg~(2+)[J];A Monthly Journal of Science;1982年11期

10 ;AN ANALYSIS ON THE STRUCTURE TYPES AND BENEFITS OFSLOPELAND AGROFORESTRY SYSTEMS-Taking the Three Gorge Reservoir Area As An Example[J];The Journal of Chinese Geography;1995年04期

相關(guān)會議論文 前10條

1 ;Genetic analysis of salt tolerance gene in wild soybean ND36-87[A];第十三屆全國植物基因組學(xué)大會論文集[C];2012年

2 ;Effects of Glyphosate on target-sites and non-target sites of herbicide in soybean apical bud[A];第十三屆全國植物基因組學(xué)大會論文集[C];2012年

3 XUE Jin-Ai;MAO Xue;YANG Zhi-Rong;JIA Xiao-Yun;WU Yong-Mei;GAO Xiu-Qing;HUANG Chun-Guo;ZHANG Li;WANG Ji-Ping;LI Run-Zhi;;Silencing of soybean Acyl-ACP thioesterase B (GmFatB) increases palmitoleic acid accumulation in soybean oil[A];中國遺傳學(xué)會第九次全國會員代表大會暨學(xué)術(shù)研討會論文摘要匯編（2009-2013）[C];2013年

4 Xiaoqing Liu;Zhaolong XU;Ling Xu;Yihong Huang;Xiaolan He;Jinxin Yi;Dayong Zhang;;Cloning and expression analysis of GmNAC8 in soybean[A];第十四屆全國植物基因組學(xué)大會摘要[C];2013年

5 ;Differential responses of the diazotrophic community to aluminum-tolerant and -sensitive soybean genotypes in acidic soil[A];江蘇省遺傳學(xué)會第八屆會員代表大會暨學(xué)術(shù)研討會論文集[C];2010年

6 ;Genetic map construction and oil contents QTL localization analysis in soybean[A];第十一屆全國植物基因組學(xué)大會摘要集[C];2010年

7 ;Identification of QTL underlying individual and total Vitamin E contents in soybean seeds under multiple environments[A];第十一屆全國植物基因組學(xué)大會摘要集[C];2010年

8 ;Isolation and analysis of FB gene of soybean[A];第十一屆全國植物基因組學(xué)大會摘要集[C];2010年

9 ;Computational identification of disease resistance gene analogs from soybean EST and their in silico mapping[A];第十一屆全國植物基因組學(xué)大會摘要集[C];2010年

10 ;Rhizosphere isoflavones levels and their relation to the soil microbial community structure in a long-term mono-cropping soybean field[A];中國第五屆植物化感作用學(xué)術(shù)研討會論文摘要集[C];2011年

相關(guān)博士學(xué)位論文 前2條

1 Nang Myint Phyu Sin Htwe;大豆逆境脅迫誘導(dǎo)的miRNA鑒定及功能分析[D];中國農(nóng)業(yè)科學(xué)院;2015年

2 Imran Haider Shamsi;大豆鎘毒害耐性的基因型差異和鎘與鋁、鈉和鉀的互作效應(yīng)[D];浙江大學(xué);2006年

，

本文編號：1428332