【摘要】：磷是高等植物生長發(fā)育所必需的第二大營養(yǎng)元素。因土壤中的磷大部分以有機(jī)態(tài)或礦物鹽的形式存在,而不能被植物所吸收利用,使得磷元素成為植物生長發(fā)育的主要限制因素之一。油菜是一種磷敏感的油料作物,低磷環(huán)境是制約油菜增產(chǎn)的主要限制元素之一。PHT1(Phosphate Transporter I,PHT1)是一類H+/Pi協(xié)同轉(zhuǎn)運(yùn)的跨膜蛋白,位于細(xì)胞質(zhì)膜,是植物從外界環(huán)境獲取磷的一類關(guān)鍵的磷轉(zhuǎn)運(yùn)載體蛋白。盡管PHT1在國內(nèi)外諸多植物中有所報道,但在油菜研究甚少。因此,開展BrPHT1家族的工作對解析油菜響應(yīng)低磷的分子機(jī)制有著重要的意義。本研究確定了白菜型油菜PHT1基因家族的30個成員,并分析了該家族成員的結(jié)構(gòu)特征、染色體分布、基因的復(fù)制方式及表達(dá)模式等。20個基因家族成員存在共線性關(guān)系,有3個成員由轉(zhuǎn)座復(fù)制得到,14個成員存在串聯(lián)復(fù)制,8個成員既進(jìn)行了染色體的大片段復(fù)制又進(jìn)行了串聯(lián)復(fù)制,在白菜基因組中WGD事件及串聯(lián)復(fù)制是PHT1基因家族成員擴(kuò)張的方式。本研究克隆得到了22個BrPHT1基因的CDS及14個BrPHT1基因的啟動子,并將BrPHT1基因和啟動子分別構(gòu)建在經(jīng)改造的植物表達(dá)載體pHZM13和啟動子活性驗(yàn)證載體pHZM37上。通過互補(bǔ)酵母突變體,得知BrPT21、BrPT22為高親和磷轉(zhuǎn)運(yùn)載體蛋白基因。利用RT-qPCR的方法分析了白菜型油菜PHT1基因的時空表達(dá)規(guī)律及在短期、長期磷脅迫下的表達(dá)模式,BrPT07和BrPT21在油菜根中高效表達(dá),對磷吸收具有重要的作用;BrPT01、16和24在生殖器官中的表達(dá)量高,對維持生殖器官的磷水平有著重要作用。本研究結(jié)果對進(jìn)一步了解油菜磷轉(zhuǎn)運(yùn)蛋白的進(jìn)化及功能提供了重要參考,對提高油菜耐低磷能力的改良有著重要的意義。
[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.
【學(xué)位授予單位】：牡丹江師范學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：Q943.2;S565.4

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1 張倩;白菜型油菜PHT1基因家族的克隆及功能分析[D];牡丹江師范學(xué)院;2016年

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本文編號：2467987