普通小麦千粒重相关基因TaPTF1和Tabas1的克隆与表达分析

发布时间：2018-07-12 18:53

本文选题：小麦 + 千粒重　；参考：《安徽农业大学》2016年博士论文

【摘要】：小麦是我国主要的粮食作物之一,随着人口的增长、耕地面积的减少,高产且稳产是我国小麦育种的重要目标。千粒重是构成产量的三要素之一,灌浆时期旗叶叶绿素含量、籽粒可溶性糖含量以及逆境胁迫等因素都影响千粒重的形成。鉴定千粒重相关基因,开发功能标记有助于加快高产小麦新品种的培育进程。PTF1是一种具有bHLH结构域的转录因子。水稻中,转OsPTF1基因的植株在低磷水平下单穗重比野生型高20%左右。BAS1是一种过氧化物还原蛋白。水稻中,NTRC-BAS1通路是一种用于保护叶绿体免遭氧化损伤的氧化还原系统。上述两种基因在水稻籽粒的形成过程中均起着重要作用。但PTF1基因和bas1基因在小麦籽粒的形成过程中的作用还不清楚。本研究通过电子克隆以及比较基因组学方法,克隆了小麦TaPTF1、Tabas1-B1,以及TaGW-B1基因部分片段,利用不同群体材料分析其对千粒重表型的贡献,探索其调控籽粒形成以及粒重的分子机制,并分析不同等位基因型在我国小麦种质资源以及国外引进材料中的分布情况。主要结果如下:1、从京411与红芒春21中克隆了TaPTF1基因,该基因包含8个外显子和7个内含子,编码480个氨基酸(~51.29kDa);利用一套中国春缺体-四体材料,将其定位于第七同源群,分别命名为TaPTF1-A1、TaPTF1-B1和TaPTF1-D1;TaPTF1-A1与TaPTF1-D1基因在千粒重差异显著的品种间无序列差异,而TaPTF1-B1基因在千粒重差异显著的品种第7个内含子位置存在16个碱基的插入/缺失,并开发了一个InDel标记Ta7B。2、TaPTF1-B1基因共存在两种等位基因类型,即TaPTF1-B1a和TaPTF1-B1b;携带TaPTF1-B1a基因型的小麦品种的千粒重显著高于携带TaPTF1-B1b基因型的小麦品种的千粒重(P0.01),为优异等位类型。3、我国主要麦区中,除西南冬麦区(38.0%)和东北春麦区(50.0%)外,TaPTF1-B1a等位类型的分布频率均高于TaPTF1-B1b等位类型。在所引进的国外材料中,除挪威(40.0%)、德国(25.0%)、匈牙利(25.0%)和法国(8.9%)外,TaPTF1-B1a等位类型的分布频率也均高于TaPTF1-B1b等位类型。4、TaPTF1-B1基因与京411/红芒春21群体中控制千粒重的QTL共同定位于7BL染色体区段(Xgwm400-Xgwm573),可解释千粒重表型变异的23.9%-39.1%。5、在灌浆中期的茎、旗叶以及花后不同发育时期的籽粒中,TaPTF1-B1a等位基因的表达水平显著低于TaPTF1-B1b等位基因的表达水平,说明TaPTF1基因负向调控小麦籽粒发育。6、从小麦品种京411中克隆了千粒重相关基因Tabas1,该基因包括7个外显子和6个内含子,编码262个氨基酸(~28.24 kDa);千粒重差异显著的品种间,Tabas1基因在第2个内含子区域存在1-bp的In/Del突变,并开发了一个InDel标记TaS1。利用中国春缺体-四体材料,将Tabas1基因定位于2B染色体,命名为Tabas1-B1基因。7、Tabas1-B1基因共存在两种等位基因类型,即Tabas1-B1a和Tabas1-B1b;携带TaPTF1-B1a等位类型的小麦品种的千粒重显著高于携带TaPTF1-B1b等位类型的小麦品种的千粒重(P0.01);在我国小麦品种资源中,Tabas1-B1a等位类型的分布频率略高于Tabas1-B1b等位类型,说明Tabas1-B1a等位基因类型在育种过程中并没有被充分选择和利用。8、Tabas1-B1基因与京411/红芒春21群体中控制千粒重和旗叶叶绿素含量的QTL共同定位于SSR标记Xbarc167与Xcfa2278之间,分别解释千粒重表型变异的9.5%-15.5%,以及旗叶叶绿素含量表型变异的9.0%-19.2%。9、在不同发育时期的籽粒中(除花后10天外),Tabas1-B1a等位基因的表达水平明显高于Tabas1-B1b等位基因的表达水平,对千粒重起正调控作用。10、从小麦品种红芒春21中克隆得到千粒重相关的未知功能基因的片段(暂命名为TaGW);千粒重差异显著的品种间,该基因片段在外显子部位存在5个碱基的插入/缺失,前者与高千粒重显著相关(P0.01),命名为TaGW-B1a;后者与低千粒重显著相关(P0.01),命名为TaGW-B1b。利用中国春缺体-四体材料,将TaGW基因片段定位于3B染色体。TaGW-B1基因片段与京411/红芒春21群体中控制千粒重的QTL共同定位于SSR标记Xbarc68与Xwmc418之间,可解释千粒重表型变异的34.2%-58.2%;TaGW-B1a等位类型在我国小麦品种资源以及国外引进材料中的分布频率均较低。
[Abstract]:Wheat is one of the main grain crops in China. With the increase of population, the reduction of cultivated land area and high yield and stable yield are the important targets of wheat breeding in China. 1000 grain weight is one of the three factors that constitute the yield. .PTF1 is a transcriptional factor with bHLH domain. In rice, the plant of the transgenic OsPTF1 gene is about 20%.BAS1 higher than the wild type under the low phosphorus level in rice. The NTRC-BAS1 pathway is one of the NTRC-BAS1 pathway in rice. These two genes play an important role in the formation of rice grain, but the role of PTF1 and bas1 genes in the formation of wheat grain is not clear. This study cloned TaPTF1, T of Wheat by electronic cloning and comparative genomics methods. Abas1-B1, and part of TaGW-B1 gene fragment, use different population materials to analyze its contribution to 1000 grain weight phenotype, explore its molecular mechanism to regulate grain formation and grain weight, and analyze the distribution of different alleles in Wheat Germplasm Resources and foreign imported materials in China. The main results are as follows: 1, from Beijing 411 and red mans spring The TaPTF1 gene was cloned in 21, which contains 8 exons and 7 introns and 480 amino acids (~51.29kDa). Using a set of Chinese Spring deficient body four body materials, the gene is located in seventh homologous groups, named TaPTF1-A1, TaPTF1-B1 and TaPTF1-D1, and TaPTF1-A1 and TaPTF1-D1 genes are disordered among the varieties with significant difference in 1000 grain weight. The TaPTF1-B1 gene has 16 bases insertion / deletion in seventh intron sites with significant difference in 1000 grain weight, and a InDel marker Ta7B.2 is developed. There are two alleles in the TaPTF1-B1 gene, namely, TaPTF1-B1a and TaPTF1-B1b, and the 1000 grain weight of the wheat varieties carrying the TaPTF1-B1a genotype is significantly higher than that of the TaPTF1. The 1000 grain weight (P0.01) of the -B1b genotypes of wheat is an excellent allele.3. In the main wheat areas of our country, the distribution frequency of the TaPTF1-B1a allele is higher than the TaPTF1-B1b allele except in the southwest Winter Wheat Region (38%) and the Northeast Spring Wheat Region (50%). In the foreign materials introduced, except Norway (40%), Germany (25%), Hungary (25%), In France (8.9%), the distribution frequency of the TaPTF1-B1a allele is also higher than that of the TaPTF1-B1b allele type.4. The TaPTF1-B1 gene localizes in the 7BL chromosome section (Xgwm400-Xgwm573) in the 7BL chromosome section (Xgwm400-Xgwm573) with the QTL that controls 1000 grain weight in the 21 population of the Beijing 411/ red Mans, which can explain the 23.9%-39.1%.5 of the 1000 grain weight phenotypic variation. The expression level of TaPTF1-B1a allele was significantly lower than that of TaPTF1-B1b allele, indicating that the TaPTF1 gene negatively regulates the development of.6 in wheat grain, and the 1000 grain weight related gene Tabas1 was cloned from the wheat variety Jing 411, which includes 7 exons and 6 introns and encoded 262 amino acids (~28.2). 4 kDa); among the varieties with significant difference in 1000 grain weight, the Tabas1 gene has the In/Del mutation in the 1-BP in the second intron regions, and a InDel marker TaS1. is developed to locate the Tabas1 gene in the 2B chromosome, named Tabas1-B1 gene.7, and the Tabas1-B1 gene has two alleles, that is Tabas1-B1a. The 1000 grain weight of wheat varieties carrying TaPTF1-B1a allele was significantly higher than that of wheat varieties carrying TaPTF1-B1b allele (P0.01), and the distribution frequency of Tabas1-B1a alleles was slightly higher than that of Tabas1-B1b alleles in China's wheat germplasm resources, indicating that the Tabas1-B1a allele types were in the breeding process. .8, Tabas1-B1 and QTL, which control the 1000 grain weight and the chlorophyll content of flag leaf in the 21 population of the Beijing 411/ red mans spring, were located between the SSR markers Xbarc167 and Xcfa2278, explaining the 9.5%-15.5% of the 1000 grain weight phenotypic variation and the phenotypic variation of the flag leaf chlorophyll content, and at different developmental stages. In grain (except 10 days after flower), the expression level of Tabas1-B1a allele is obviously higher than that of Tabas1-B1b allele, and it plays a positive role in controlling 1000 grain weight,.10. The fragment of 1000 grain weight related unknown function gene is cloned from red mans spring 21 of wheat variety (temporary name is named TaGW). There were 5 bases insertion / deletion in exons, the former was significantly correlated with high 1000 grain weight (P0.01), named TaGW-B1a, and the latter was significantly correlated with low 1000 grain weight (P0.01), and named TaGW-B1b. using Chinese Spring deficient body four body material, the TaGW gene fragment was located in the 3B chromosome.TaGW-B1 gene fragment and the Beijing 411/ red awn spring 21 population. QTL, which controls 1000 grain weight, localizes between the SSR marker Xbarc68 and Xwmc418, which can explain the 34.2%-58.2% of the phenotypic variation of 1000 grain weight; the TaGW-B1a allele types are low in the distribution frequency of wheat variety resources and imported materials in our country.
【学位授予单位】：安徽农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S512.1

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