本文選題：小麥　切入點：ERF轉(zhuǎn)錄因子　出處：《中國農(nóng)業(yè)科學院》2015年碩士論文　論文類型：學位論文

【摘要】：小麥作為全世界廣泛種植的禾本科作物,具有悠久的種植栽培歷史,是世界上最主要的糧食作物之一。人口的持續(xù)增長加大了對糧食需求的壓力。在當前全球可耕地面積不斷減少的情況下,只有提高小麥的產(chǎn)量和質(zhì)量,才能滿足日益增長的人口需求。然而,近幾十年來極端氣候時有發(fā)生,對小麥穩(wěn)產(chǎn)提出了更高要求。因此,迫切需要發(fā)掘小麥種質(zhì)資源中蘊藏的優(yōu)異的基因,培育高產(chǎn)穩(wěn)產(chǎn)的小麥新品種。TaERF5是一個ERF類轉(zhuǎn)錄因子家族成員基因。前期工作已經(jīng)對該基因CDS區(qū)和5’UTR區(qū)進行了分析,發(fā)現(xiàn)該基因非常保守。因此,本研究擬克隆Ta ERF5基因的3’UTR區(qū)的序列,通過單倍型鑒定和關聯(lián)分析研究其與小麥農(nóng)藝性狀的關系;對Ta ERF5-A、Ta ERF5-B、Ta ERF5-D基因進行組織特異性表達分析,明確同源基因的功能;將Ta ERF5-B基因轉(zhuǎn)入日本特早熟粳稻品種Kitaake中和小麥品種Fielder進行功能驗證。主要研究結果如下:1.Ta ERF5的單倍型分類。獲得Ta ERF5 3’UTR序列。在二倍體、缺四體和親本材料擴增與測序分析,發(fā)現(xiàn)在Ta ERF5-A的3’UTR區(qū)存在1個SNP,該SNP可將檢測的18份六倍體小麥材料分成2種單倍型;在Ta ERF5-B的3’UTR與5’UTR區(qū)共檢測到3個SNP和1個In Del,利用這些SNP可將Ta ERF5-B分成4種單倍型。Ta ERF5-D 3’UTR在六倍體中沒有多態(tài)性。依據(jù)Ta ERF-A、Ta ERF-B基因3’UTR區(qū)的多態(tài)性位點開發(fā)了3個AS-PCR分子標記,并對自然群體(包括野生材料、地方品種和現(xiàn)代育成品種)和導入系材料進行基因分型。2.TaERF5不同單倍型的功能分析。TaERF5單倍型與重要農(nóng)藝性狀的關聯(lián)分析發(fā)現(xiàn),TaERF5-A與小麥拔節(jié)期、抽穗期和成熟期顯著相關,其中單倍型Hapl 1在小麥野生群體中是優(yōu)勢單倍型;而Hapl 2在小麥馴化后頻率提高,推測與農(nóng)藝性狀相關。Ta ERF5-B與小麥株高、穗粒數(shù)、千粒重和產(chǎn)量性狀極顯著相關,其中Hapl 2表現(xiàn)為植株較矮、穗粒數(shù)和千粒重高、產(chǎn)量高的特點,偃展1號導入系群體證明Hapl 2與高產(chǎn)顯著相關,證明該單倍型為優(yōu)異單倍型。3.TaERF5的轉(zhuǎn)錄表達分析。TaERF5-A、TaERF5-B、TaERF5-D三個同源基因在中國春小麥幼苗的根、莖節(jié)、莖、葉鞘、旗葉、幼穗及偃展1號和復壯30小麥材料的穗發(fā)育不同時期表達分析發(fā)現(xiàn),Ta ERF5-A在根中表達最高;Ta ERF5-B在旗葉和幼穗中的表達量最高;Ta ERF5-D在葉鞘中的表達量最高�，F(xiàn)代育成品種偃展1號中,Ta ERF5-B基因在穗發(fā)育過程中起著主導的作用,在穗發(fā)育的中期(9.5cm)表達量達到峰值,隨后表達量下降;Ta ERF5-D對偃展1號穗發(fā)育的影響小于Ta ERF5-B;復壯30是我國上世紀50年代育成品種,Ta ERF5-D表達量最高。Ta ERF5同源基因的組織表達特征還有待深入研究。
[Abstract]:Wheat is widely planted in the world and has a long history of cultivation. Is one of the most important food crops in the world. The continued growth of population has increased the pressure on the demand for food. In the current situation of decreasing arable land in the world, only to improve the yield and quality of wheat, In order to meet the growing population needs. However, extreme weather has occurred from time to time in recent decades, placing higher demands on stable wheat production. Therefore, there is an urgent need to explore the excellent genes contained in wheat germplasm resources, TaERF5, a new wheat variety with high and stable yield, is a member of the transcription factor family of ERF. The previous work has analyzed the CDS and 5UTR regions of the gene and found that the gene is very conserved. In this study, we cloned the sequence of 3 UTR region of Ta ERF5 gene, studied its relationship with wheat agronomic traits by haplotype identification and association analysis, and analyzed the tissue specific expression of Ta ERF5-Agna Ta ERF5-BnTa ERF5-D gene to clarify the function of homologous gene. Ta ERF5-B gene was transferred into Kitaake and wheat cultivar Fielder for functional verification. The main results were as follows: 1. Haplotype classification of Ta ERF5. Ta ERF5 3 UTR sequence was obtained. Amplification and sequencing of diploid, tetralogy and parents were performed. It was found that there was one SNPs in the 3UTR region of Ta ERF5-A, which could be divided into two haplotypes by SNP. Three SNP and one in Delin were detected in the 3 and 5 ERF5-B regions of Ta ERF5-B. Using these SNP, Ta ERF5-B could be divided into four haplotypes. Ta ERF5-D 3UTR had no polymorphism in hexaploid. Three AS-PCR molecular markers were developed based on the polymorphism sites of Ta ERF-An Ta ERF-B region. The functional analysis of different haplotypes of TaERF5 and important agronomic characters showed that TaERF5-A was associated with wheat at jointing stage, and the natural population (including wild materials, local varieties and modern bred varieties) and imported lines were genotyped. 2. The functional analysis of different haplotypes of TaERF5 haplotypes and important agronomic traits showed that TaERF5-A was associated with wheat at jointing stage. There was a significant correlation between heading date and maturity stage, among which haplotype Hapl 1 was the dominant haplotype in wild population, while Hapl 2 frequency increased after acclimation. It was inferred that Hapl 2 was related to agronomic characters, such as plant height and grain number per spike. There was a significant correlation between 1000-grain weight and yield traits, in which Hapl _ 2 was characterized by short plant, high grain number per spike and 1000-grain weight, and high yield. It was proved that Hapl _ 2 was significantly correlated with high yield in Yanchang No. 1 import line population. It is proved that the haplotype is an excellent haplotype. 3. The transcriptional expression of TaERF5. TaERF5-B (TaERF5-B) TaERF5-D homologous genes in the roots, stem nodes, stems, leaf sheaths and flag leaves of spring wheat seedlings in China. The analysis of the expression of Ta ERF5-A at different stages of ear development in young panicle and Yanxian 1 and Fuzhuang 30 wheat materials showed that the highest expression of Ta ERF5-B in root was found in flag leaf and young ear. The expression of Ta ERF5-D in leaf sheath was the highest in modern breeding. Ta ERF5-B gene plays a leading role in panicle development. At the middle stage of panicle development, the expression reached a peak value. The effect of Ta ERF5-D on panicle development was lower than that of Ta ERF5-B, and the tissue expression characteristics of the highest ERF5-D expression of Ta ERF5-D in -50s in China need to be further studied.
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：Q943.2;S512.1

【參考文獻】

相關期刊論文 前2條

1 劉強,張貴友,陳受宜;植物轉(zhuǎn)錄因子的結構與調(diào)控作用[J];科學通報;2000年14期

2 ;Functions and Application of the AP2/ERF Transcription Factor Family in Crop Improvement[J];Journal of Integrative Plant Biology;2011年07期

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本文編號：1606576