白菜型油菜PHT1基因家族的克隆及功能分析
发布时间：2019-04-28 21:34
【摘要】：磷是高等植物生长发育所必需的第二大营养元素。因土壤中的磷大部分以有机态或矿物盐的形式存在,而不能被植物所吸收利用,使得磷元素成为植物生长发育的主要限制因素之一。油菜是一种磷敏感的油料作物,低磷环境是制约油菜增产的主要限制元素之一。PHT1(Phosphate Transporter I,PHT1)是一类H+/Pi协同转运的跨膜蛋白,位于细胞质膜,是植物从外界环境获取磷的一类关键的磷转运载体蛋白。尽管PHT1在国内外诸多植物中有所报道,但在油菜研究甚少。因此,开展BrPHT1家族的工作对解析油菜响应低磷的分子机制有着重要的意义。本研究确定了白菜型油菜PHT1基因家族的30个成员,并分析了该家族成员的结构特征、染色体分布、基因的复制方式及表达模式等。20个基因家族成员存在共线性关系,有3个成员由转座复制得到,14个成员存在串联复制,8个成员既进行了染色体的大片段复制又进行了串联复制,在白菜基因组中WGD事件及串联复制是PHT1基因家族成员扩张的方式。本研究克隆得到了22个BrPHT1基因的CDS及14个BrPHT1基因的启动子,并将BrPHT1基因和启动子分别构建在经改造的植物表达载体pHZM13和启动子活性验证载体pHZM37上。通过互补酵母突变体,得知BrPT21、BrPT22为高亲和磷转运载体蛋白基因。利用RT-qPCR的方法分析了白菜型油菜PHT1基因的时空表达规律及在短期、长期磷胁迫下的表达模式,BrPT07和BrPT21在油菜根中高效表达,对磷吸收具有重要的作用;BrPT01、16和24在生殖器官中的表达量高,对维持生殖器官的磷水平有着重要作用。本研究结果对进一步了解油菜磷转运蛋白的进化及功能提供了重要参考,对提高油菜耐低磷能力的改良有着重要的意义。
[Abstract]:Phosphorus is the second most important nutrient element for the growth and development of higher plants. Because most of the phosphorus in the soil exists in the form of organic or mineral salts and can not be absorbed by plants, phosphorus has become one of the main limiting factors of plant growth and development. Rape (Brassica napus L.) is a kind of phosphorus sensitive oil crop, and low phosphorus environment is one of the main limiting factors to increase yield of rape. PHT1 (Phosphate Transporter I, PHT1) is a kind of H / Pi co-transport transmembrane proteins located in cytoplasmic membrane. It is a kind of key phosphorus transport vector proteins for plants to obtain phosphorus from the external environment. Although PHT1 has been reported in many plants at home and abroad, little research has been done on rape. Therefore, it is important to carry out the work of BrPHT1 family to analyze the molecular mechanism of response to low phosphorus in oilseed rape (Brassica napus L.). In this study, 30 members of the PHT1 gene family in Brassica napus were identified, and their structural characteristics, chromosome distribution, gene replication and expression patterns were analyzed. 20 members of the gene family had a collinear relationship. Three members were obtained by transposable replication, 14 members were duplicated in tandem, and 8 members were duplicated in tandem as well as large fragments of chromosomes. WGD events and tandem replication in Chinese cabbage genome are the ways of expanding the members of the PHT1 gene family. In this study, 22 BrPHT1 gene CDS and 14 BrPHT1 gene promoters were cloned, and the BrPHT1 gene and promoter were constructed on the modified plant expression vector pHZM13 and promoter activity verification vector pHZM37, respectively. Through complementary yeast mutants, BrPT21,BrPT22 was found to be a highly compatible phosphorus transport vector protein gene. The spatio-temporal expression pattern of PHT1 gene in Brassica napus L. and its expression pattern under short-term and long-term phosphorus stress were analyzed by RT-qPCR. BrPT07 and BrPT21 were highly expressed in rape roots, which played an important role in phosphorus uptake. The high expression of BrPT01,16 and 24 in reproductive organs plays an important role in maintaining the phosphorus level of reproductive organs. The results provide an important reference for further understanding the evolution and function of phosphorus transporter in rape, and it is of great significance to improve the tolerance to low phosphorus in Brassica napus.
【学位授予单位】：牡丹江师范学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2;S565.4
本文编号：2467987
[Abstract]:Phosphorus is the second most important nutrient element for the growth and development of higher plants. Because most of the phosphorus in the soil exists in the form of organic or mineral salts and can not be absorbed by plants, phosphorus has become one of the main limiting factors of plant growth and development. Rape (Brassica napus L.) is a kind of phosphorus sensitive oil crop, and low phosphorus environment is one of the main limiting factors to increase yield of rape. PHT1 (Phosphate Transporter I, PHT1) is a kind of H / Pi co-transport transmembrane proteins located in cytoplasmic membrane. It is a kind of key phosphorus transport vector proteins for plants to obtain phosphorus from the external environment. Although PHT1 has been reported in many plants at home and abroad, little research has been done on rape. Therefore, it is important to carry out the work of BrPHT1 family to analyze the molecular mechanism of response to low phosphorus in oilseed rape (Brassica napus L.). In this study, 30 members of the PHT1 gene family in Brassica napus were identified, and their structural characteristics, chromosome distribution, gene replication and expression patterns were analyzed. 20 members of the gene family had a collinear relationship. Three members were obtained by transposable replication, 14 members were duplicated in tandem, and 8 members were duplicated in tandem as well as large fragments of chromosomes. WGD events and tandem replication in Chinese cabbage genome are the ways of expanding the members of the PHT1 gene family. In this study, 22 BrPHT1 gene CDS and 14 BrPHT1 gene promoters were cloned, and the BrPHT1 gene and promoter were constructed on the modified plant expression vector pHZM13 and promoter activity verification vector pHZM37, respectively. Through complementary yeast mutants, BrPT21,BrPT22 was found to be a highly compatible phosphorus transport vector protein gene. The spatio-temporal expression pattern of PHT1 gene in Brassica napus L. and its expression pattern under short-term and long-term phosphorus stress were analyzed by RT-qPCR. BrPT07 and BrPT21 were highly expressed in rape roots, which played an important role in phosphorus uptake. The high expression of BrPT01,16 and 24 in reproductive organs plays an important role in maintaining the phosphorus level of reproductive organs. The results provide an important reference for further understanding the evolution and function of phosphorus transporter in rape, and it is of great significance to improve the tolerance to low phosphorus in Brassica napus.
【学位授予单位】：牡丹江师范学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2;S565.4
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1 张倩;白菜型油菜PHT1基因家族的克隆及功能分析[D];牡丹江师范学院;2016年
，本文编号：2467987
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