小麦TaERF5基因多样性与功能分析

发布时间：2018-03-13 13:26

本文选题：小麦　切入点：ERF转录因子　出处：《中国农业科学院》2015年硕士论文　论文类型：学位论文

【摘要】：小麦作为全世界广泛种植的禾本科作物,具有悠久的种植栽培历史,是世界上最主要的粮食作物之一。人口的持续增长加大了对粮食需求的压力。在当前全球可耕地面积不断减少的情况下,只有提高小麦的产量和质量,才能满足日益增长的人口需求。然而,近几十年来极端气候时有发生,对小麦稳产提出了更高要求。因此,迫切需要发掘小麦种质资源中蕴藏的优异的基因,培育高产稳产的小麦新品种。TaERF5是一个ERF类转录因子家族成员基因。前期工作已经对该基因CDS区和5’UTR区进行了分析,发现该基因非常保守。因此,本研究拟克隆Ta ERF5基因的3’UTR区的序列,通过单倍型鉴定和关联分析研究其与小麦农艺性状的关系;对Ta ERF5-A、Ta ERF5-B、Ta ERF5-D基因进行组织特异性表达分析,明确同源基因的功能;将Ta ERF5-B基因转入日本特早熟粳稻品种Kitaake中和小麦品种Fielder进行功能验证。主要研究结果如下:1.Ta ERF5的单倍型分类。获得Ta ERF5 3’UTR序列。在二倍体、缺四体和亲本材料扩增与测序分析,发现在Ta ERF5-A的3’UTR区存在1个SNP,该SNP可将检测的18份六倍体小麦材料分成2种单倍型;在Ta ERF5-B的3’UTR与5’UTR区共检测到3个SNP和1个In Del,利用这些SNP可将Ta ERF5-B分成4种单倍型。Ta ERF5-D 3’UTR在六倍体中没有多态性。依据Ta ERF-A、Ta ERF-B基因3’UTR区的多态性位点开发了3个AS-PCR分子标记,并对自然群体(包括野生材料、地方品种和现代育成品种)和导入系材料进行基因分型。2.TaERF5不同单倍型的功能分析。TaERF5单倍型与重要农艺性状的关联分析发现,TaERF5-A与小麦拔节期、抽穗期和成熟期显著相关,其中单倍型Hapl 1在小麦野生群体中是优势单倍型;而Hapl 2在小麦驯化后频率提高,推测与农艺性状相关。Ta ERF5-B与小麦株高、穗粒数、千粒重和产量性状极显著相关,其中Hapl 2表现为植株较矮、穗粒数和千粒重高、产量高的特点,偃展1号导入系群体证明Hapl 2与高产显著相关,证明该单倍型为优异单倍型。3.TaERF5的转录表达分析。TaERF5-A、TaERF5-B、TaERF5-D三个同源基因在中国春小麦幼苗的根、茎节、茎、叶鞘、旗叶、幼穗及偃展1号和复壮30小麦材料的穗发育不同时期表达分析发现,Ta ERF5-A在根中表达最高;Ta ERF5-B在旗叶和幼穗中的表达量最高;Ta ERF5-D在叶鞘中的表达量最高。现代育成品种偃展1号中,Ta ERF5-B基因在穗发育过程中起着主导的作用,在穗发育的中期(9.5cm)表达量达到峰值,随后表达量下降;Ta ERF5-D对偃展1号穗发育的影响小于Ta ERF5-B;复壮30是我国上世纪50年代育成品种,Ta ERF5-D表达量最高。Ta ERF5同源基因的组织表达特征还有待深入研究。
[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.
【学位授予单位】：中国农业科学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：Q943.2;S512.1

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