本文選題：青楊異源三倍體群體　切入點：DNA甲基化　出處：《北京林業(yè)大學(xué)》2016年博士論文

【摘要】：基因的表達(dá)受表觀遺傳修飾和轉(zhuǎn)錄后調(diào)控因子的調(diào)控,開展青楊異源三倍體群體形成過程中表觀遺傳變異和轉(zhuǎn)錄后調(diào)控因子的變異研究,可以從不同層次探討青楊三倍體生長優(yōu)勢形成的分子機(jī)制。本研究以來源于不同類型2n雌配子[FDR型(First division restitution)、SDR型(Second division restitution)、PMR型(Post-meiotic restitution)]的全同胞青楊異源三倍體群體(triploid-F, triploid-S, triploid-P)和其同親本的雜種二倍體群體(diploid-F1),及其母本‘哲引3號楊’[(P. pseudo-simonii)× (P. nigra L. var. italica)]、父本‘北京楊’(P. ×beijingensis)為研究材料,采用群體取樣的策略,利用甲基化敏感擴(kuò)增多態(tài)性(Methylationsensitive amplification polymorphism, MSAP)標(biāo)記和小RNA高通量測序技術(shù),從群體水平分析了青楊異源三倍體群體中全基因組DNA甲基化變異和miRNA表達(dá)變異,綜合評價了雜交和多倍化對DNA甲基化變異和niRNA表達(dá)變異的影響,初步探討了表觀遺傳修飾和轉(zhuǎn)錄后調(diào)控因子在青楊異源三倍體速生優(yōu)質(zhì)等性狀形成過程中的作用。主要研究結(jié)果如下：(1)雜交和多倍化可導(dǎo)致青楊異源三倍體群體DNA甲基化水平和模式發(fā)生變異。3個異源三倍體群體的總甲基化水平都低于其親本和二倍體群體,其中,triploid-F群體的總甲基化水平為19.88%,顯著低于triploid-S群體和triploid-P群體(21.77%和21.66%)；而diploid-F1群體的總甲基化水平為23.93%,顯著高于親本和異源三倍體群體,且4個子代群體中的DNA甲基化均以全甲基化類型為主。此外,絕大多數(shù)檢測位點的DNA甲基化模式可以由親本穩(wěn)定的遺傳給子代群體,但一些位點也發(fā)生了變異,變異類型以超甲基化為主。其中,triploid-F群體發(fā)生超甲基化模式變異的頻率最低,為2.04%,而diploid-F1群體發(fā)生超甲基化模式變異的頻率達(dá)到4.97%,顯著高于3個異源三倍體群體,但3個異源三倍體群體發(fā)生去甲基化的頻率均高于二倍體。由于青楊異源三倍體群體發(fā)生超甲基化的的頻率低于二倍體群體,且去甲基化頻率高于二倍體群體,導(dǎo)致其整體甲基化水平顯著低于二倍體群體,進(jìn)而引起某些基因表達(dá)水平的相對升高,推測這可能是青楊異源三倍體生長性狀優(yōu)于雜種二倍體的原因之一。(2)親本‘哲引3號楊’和‘北京楊‘之間存在差異表達(dá)的miRNA,主要以母本中表達(dá)上調(diào)為主,其中29個miRNA在母本中特異表達(dá)；青楊異源三倍體群體和雜種二倍體群體中miRNA的表達(dá)具有明顯的偏好性,即miRNA表達(dá)在子代與父本間的差異遠(yuǎn)大于與母本間的差異。而且,各子代群體中還檢測到親本表達(dá)水平顯性的miRNA,母本顯性的niRNA數(shù)量遠(yuǎn)大于父本顯性的miRNA數(shù)量；母本顯性的miRNA主要與植物的生長發(fā)育過程相關(guān),而父本顯性的miRNA主要與植物脅迫響應(yīng)和基因組穩(wěn)定性有關(guān)。三個青楊異源三倍體群體與雜種二倍體群體相比,所有檢測到的miRNA(2273個)的表達(dá)水平在不同倍性群體間均無顯著差異；三個來源于不同誘導(dǎo)途徑的異源三倍體群體間的niRNA表達(dá)也無顯著差異。本研究結(jié)果表明,青楊異源三倍體群體miRNA的表達(dá)無劑量效應(yīng),導(dǎo)致miRNA對靶基因的負(fù)調(diào)控作用相對減弱,是青楊異源三倍體生長優(yōu)勢形成的重要分子機(jī)制之一。(3) miRNA在高生長突出的青楊異源三倍體和雜種二倍體植株中的表達(dá)與父本間的差異均顯著大于與母本間的差異,表現(xiàn)出明顯的偏好性,且子代樣本中miRNA的表達(dá)變異主要以上調(diào)為主。四組高生長突出的子代樣本中表現(xiàn)出母本顯性的miRNA也顯著多于父本顯性miRNA,母本顯性的niRNA主要參與木質(zhì)素合成和分解代謝、多細(xì)胞器官發(fā)育等與植物生長發(fā)育相關(guān)的生物學(xué)過程,而父本顯性的miRNA主要與植物營養(yǎng)生長向生殖生長的轉(zhuǎn)變,花器官的發(fā)育、抵抗外界刺激等過程相關(guān)。需要特別指出的是,在3個異源三倍體高生長突出的植株中,還檢測到一些表達(dá)被顯著抑制的miRNA,主要與植物次生細(xì)胞壁的合成和積累,細(xì)胞內(nèi)大分子的代謝活動等相關(guān),推測部分niRNA的非加性表達(dá)也可能與異源三倍體突出的生長優(yōu)勢有關(guān)。此外,三組高生長突出的青楊異源三倍體植株與雜種二倍體植株間miRNA的表達(dá)也均無顯著差異,該結(jié)果進(jìn)一步驗證了青楊異源三倍體中miRNA的表達(dá)無劑量效應(yīng)的特點。
[Abstract]:Gene expression by regulating factor table epigenetic modification and post transcriptional regulation factors, variation of the formation process of developing poplar allotriploid group epigenetic variation and post transcriptional mechanisms from different levels of poplar triploid growth advantage can be formed. This study originated from different types of 2n female gametes ([FDR First division restitution (Second), SDR division restitution), PMR (Post-meiotic restitution)] full sib poplar allotriploid (triploid-F, triploid-S group, triploid-P) and its parent hybrid diploid group (diploid-F1), and its female parent "Zhe Yang [3 '(P. * (P. nigra pseudo-simonii) L. var. italica)], the male" Beijing Yang "(P. * beijingensis) as research materials, using the group sampling method, by using the methylation sensitive amplified polymorphism (Methylationsensiti Ve amplification polymorphism, MSAP) markers and small RNA high-throughput sequencing technology, from the population level analysis of variation of whole genome DNA methylation and miRNA expression of poplar allotriploid group, comprehensive evaluation of the effects of variation of hybridization and polyploidy on the expression variation of DNA methylation and niRNA, discussed the epigenetic modification and transcription after the regulatory factors in the fast-growing poplar allotriploid quality traits in the process of formation. The main results are as follows: (1) hybridization and polyploidization can lead to the total methylation level of poplar allotriploid group DNA methylation level and pattern variation of.3 allotriploid population were lower than that of their parents and diploid populations, among them, the total methylation level of triploid-F group was 19.88%, significantly lower than that of triploid-S group and triploid-P group (21.77% and 21.66%); and group diploid-F1 total methylation The level is 23.93%, significantly higher than that of their parents and allotriploid groups, and 4 sub populations in DNA methylation with full methylation types. In addition, the DNA methylation pattern of majority of testing sites may be stable by genetic parents to offspring groups, but some sites with the variation of variation type hypermethylation. Among them, triploid-F group has the lowest frequency, hypermethylation pattern variation is 2.04%, while the diploid-F1 group showed hypermethylation pattern mutation rate reached 4.97%, significantly higher than 3 allotriploid groups, but the 3 groups of allotriploid demethylation frequency were higher than diploid. Due to hypermethylation the group of poplar allotriploid frequency lower than the diploid group, and the methylation frequency is higher than the diploid group, resulting in the overall methylation level was significantly lower than the diploid group, and Due to some relative increase in gene expression, speculated that this may be one of the reasons for poplar growth traits than allotriploid hybrids were diploid. (2) the parents' 3 Zhe Yang Yang "and" Beijing "the differences between the expression of miRNA, mainly in the female as the main expression, one of the 29 miRNA in the female specific expression; the expression of miRNA and diploid hybrid poplar allotriploids groups in the group have a clear preference, namely the expression of miRNA in the difference between offsprings and far greater than the differences between parents and offspring. Moreover, the group also detected parental dominant expression level of miRNA, the number of miRNA niRNA number of female dominance is far greater than the male dominant; female dominant miRNA mainly in plant growth and development process, and the male dominant miRNA and plant stress response and genome stability. Three poplar Compared with groups of allotriploid hybrids were diploid populations, all detected miRNA (2273) the expression level had no significant difference in different ploidy groups; three from different ways of inducing allotriploid groups niRNA expression was no significant difference. The research results show that no dose effect expression of heterologous poplar the triploid group miRNA, resulting in relatively weak negative regulatory effect of miRNA on target gene, is one of the important molecular mechanism of poplar allotriploids growth advantage formation. (3) differential expression of paternal miRNA in high growth prominent poplar allotriploid and hybrid diploid plants in between were significantly greater than the differences between the performance and the female parent. A clear preference, and the expression variation of miRNA progeny in the sample above the main tone based. Four groups of high growth of outstanding progeny samples showed maternal dominant miRNA also significantly With more than the male dominant miRNA, maternal dominant niRNA mainly involved in lignin biosynthesis and catabolism, multicellular organ development and biological processes related to plant growth and development, while the male dominant miRNA main vegetative growth to reproductive growth, floral organ development, resistance to external stimulation in particular process. In 3, the high growth of allotriploid prominent plants, also detected some expression was significantly inhibited by miRNA, and the main plant secondary cell wall synthesis and accumulation of macromolecules within the cell metabolism and other related activities, the growth advantage of non additive expression that niRNA may also be allotriploid prominent. In addition, the expression of three groups of high growth prominent poplar allotriploid plants and hybrid diploid plants of miRNA also showed no significant difference, the results further validate the poplar The expression of miRNA in the heterologous triploid is characterized by no dose effect.

【學(xué)位授予單位】：北京林業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S792.113

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 Mustafaev A.S.;楊宗萬;;異源三倍體桑葉的葉質(zhì)對桑蠶生物學(xué)指數(shù)的效果[J];廣東蠶絲通訊;1978年02期

2 康向陽,朱之悌,張志毅;毛白楊異源三倍體形態(tài)和減數(shù)分裂觀察[J];北京林業(yè)大學(xué)學(xué)報;1999年01期

3 覃欽博;戴婧;劉少軍;劉筠;;異源三倍體鯽魴的遺傳組成和生殖特性觀察[J];水產(chǎn)學(xué)報;2014年03期

4 龔建國 ,李德梅 ,張有全 ,曹建久 ,劉坤;異源三倍體鯽的引進(jìn)養(yǎng)殖試驗[J];齊魯漁業(yè);2005年10期

5 劉劍;王昭萍;孔靜;于瑞海;王有廷;王廷軍;;香港巨牡蠣♀×太平洋牡蠣♂異源三倍體的初步研究[J];海洋湖沼通報;2012年01期

6 Mustafaev A.S.;楊宗萬;;異源三倍體桑葉的葉質(zhì)的養(yǎng)蠶技術(shù)指數(shù)效果[J];廣東蠶絲通訊;1978年02期

7 楊金水;鄒高治;葛扣麟;葉鳴明;;異源三倍體水稻體細(xì)胞培養(yǎng)與染色體加倍研究初報(摘要)[J];上海農(nóng)業(yè)科技;1983年01期

8 朱藍(lán)菲;桂建芳;梁紹昌;蔣一s，

本文編號：1676473