大麥抗條紋病基因定位及7H短臂SSR引物開發(fā)檢測
發(fā)布時間：2018-09-08 15:37
【摘要】：大麥(Hordeum vulgare L.)作為最主要的谷物類作物之一,在遺傳上的優(yōu)勢和經(jīng)濟(jì)的重要性被當(dāng)作禾本科的模式作物進(jìn)行研究。近年來,由于啤酒工業(yè)的大力發(fā)展,啤酒大麥的重要性愈加明顯。在啤酒大麥的發(fā)展和生產(chǎn)過程中,大麥條紋病爆發(fā)較為嚴(yán)重,并逐年增加,對啤酒大麥的質(zhì)量和產(chǎn)量產(chǎn)生嚴(yán)重的影響。由于國內(nèi)大麥抗條紋病種質(zhì)資源稀缺,急需培育抗病新品種。本研究利用100份大麥親本材料進(jìn)行抗性鑒定,篩選免疫品種與高感品種進(jìn)行雜交,利用BSA分析法對抗條紋病基因進(jìn)行定位,后期在定位到的7H短臂開發(fā)新的SSR標(biāo)記,并對這些標(biāo)記進(jìn)行多態(tài)性分析。本研究中將新發(fā)現(xiàn)的抗大麥條紋病基因命名為Rdg3,該基因是國內(nèi)首次定位的大麥抗條紋病基因,也是世界上第三個定位的抗大麥條紋病基因。該成果對培育抗條紋病大麥新品種具有重要作用,并對降低農(nóng)藥使用、減輕環(huán)境污染具有重要意義。主要結(jié)果總結(jié)如下:1、利用QWC菌株對100份大麥材料進(jìn)行接菌,通過抗性鑒定,篩選出免疫材料1份,高抗材料10份,22份材料表現(xiàn)為抗病,感病材料和高感材料分別37份和30份;其中免疫品種為甘啤2號,高感品種Alexis感病最為嚴(yán)重,發(fā)病率為88.24%。2、利用免疫品種甘啤2號和高感品種Alexis同時進(jìn)行正反交,構(gòu)建雜交組合,F1抗性鑒定發(fā)病率為0;對F2群體的276個單株進(jìn)行接菌,統(tǒng)計發(fā)病后抗病單株197株,感病單株79株,經(jīng)卡方檢測抗/感比符合3:1的分離比例,確定抗病基因?qū)Ω胁轱@性,且該抗大麥條紋病基因是由單基因控制的。3、利用大麥染色體1-7H的525對引物在兩親本間進(jìn)行多態(tài)性篩選,共篩選103對有多態(tài)的引物(其中7H多態(tài)性引物11對);利用BSA分析法發(fā)現(xiàn)位于7H短臂的標(biāo)記Bmag206、Bmag7和scssr07970與目標(biāo)基因Rdg3連鎖,因此將抗條紋病基因定位到染色體7H短臂上。在7H短臂開發(fā)SSR標(biāo)記42對,其中在甘啤2號和Alexis中表現(xiàn)多態(tài)的共6對,利用該6對引物和7H有多態(tài)的11對SSR標(biāo)記掃描群體,繪制遺傳連鎖圖譜,發(fā)現(xiàn)目標(biāo)基因位于Bmag206和Bmag7中間,且距這兩個標(biāo)記之間的遺傳距離分別是1.78cM和2.86cM。4、選取大麥親本材料48份,利用新開發(fā)的42對SSR引物進(jìn)行多態(tài)性掃描。其中有多態(tài)的引物共29對。29對引物檢測出的等位變異為125個,變幅在2-9之間,平均每個標(biāo)記4.3個。標(biāo)記Bmg24和Bmg25檢測到等位變異最多,為9個;等位變異頻率的變幅為0.2979-0.9130;標(biāo)記Bmg1的等位變異頻率最低,為0.2979。29對引物PIC變幅介于0.1462-0.8234之間,平均值為0.5132。標(biāo)記Bmg25的PIC值最高,為0.8234,標(biāo)記Bmg43的值最低,為0.1462。多樣性的平均值為0.5596,雜合度平均值為2.98%。
[Abstract]:Barley (Hordeum vulgare L.) As one of the most important cereal crops, genetic superiority and economic importance are studied as model crops of Gramineae. In recent years, with the development of beer industry, the importance of malting barley has become more and more obvious. In the course of development and production of malting barley, the outbreak of barley stripe disease is more serious and increases year by year, which has a serious effect on the quality and yield of beer barley. Because germplasm resources of barley resistant to stripe disease are scarce in China, it is urgent to develop new resistant varieties. In this study, 100 barley parents were used to identify the resistance, to select the immune varieties and to cross with the highly susceptible varieties, to use BSA analysis to localize the gene against stripe disease, and to develop a new SSR marker in the located 7H short arm at the later stage. The polymorphism of these markers was analyzed. In this study, the new gene of resistance to barley stripe disease was named Rdg3, which is the first gene of barley resistance to stripe disease located in China, and the third gene of resistance to barley stripe disease in the world. It is important for breeding new barley varieties resistant to stripe disease, reducing pesticide use and reducing environmental pollution. The main results were summarized as follows: 1 strain of QWC was used to inoculate 100 barley materials. Through identification of resistance, 1 strain of immune material was screened out, and 22 materials of 10 high resistant materials showed resistance to disease, 37 and 30 of susceptible materials and highly susceptible materials, respectively. Gan Beer 2 was the immunized variety, and the highly susceptible variety Alexis was the most serious, the incidence rate was 88.24.2. The positive and negative cross was carried out by using the immunized variety Gan Beer 2 and the highly susceptible variety Alexis at the same time. The incidence of F _ 1 resistance identification in F _ 2 population was 0, 197 resistant plants and 79 susceptible plants were collected from 276 F _ 2 population, and the ratio of resistance to disease was in accordance with the 3:1 isolation ratio by chi-square test. It was determined that the resistance gene was dominant to the susceptible disease, and the resistance gene was controlled by single gene. 525pairs of primers from chromosome 1-7H of barley were used to select polymorphism between the two parents. A total of 103 pairs of polymorphic primers (including 11 pairs of 7H polymorphic primers) were screened, and the marker Bmag206,Bmag7 and scssr07970 located on the 7H short arm were identified by BSA analysis to be linked to the target gene Rdg3, so the gene resistant to stripe disease was located on the chromosome 7H short arm. 42 pairs of SSR markers were developed in 7H short arm, of which 6 pairs were polymorphic in Gan Beer 2 and Alexis. Using the 6 pairs of primers and 11 pairs of SSR markers with 7H polymorphism, the genetic linkage map was drawn, and the target gene was found to be located between Bmag206 and Bmag7. The genetic distance between the two markers was 1.78cM and 2.86cM.4. 48 barley parents were selected, and 42 pairs of SSR primers were used to carry out polymorphic scanning. The alleles of 29 pairs of polymorphic primers were 125, ranging from 2 to 9, with an average of 4.3 markers per marker. The highest allelic variation was detected by marker Bmg24 and Bmg25, and the frequency of allelic variation was 0.2979-0.9130, while that of marker Bmg1 was the lowest, ranging from 0.1462-0.8234 to 0.2979.29, with an average of 0.5132. The PIC value of marked Bmg25 was the highest (0.8234), and the value of marked Bmg43 was lowest (0.1462). The average diversity and heterozygosity were 0.5596 and 2.98 respectively.
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S435.123
本文編號：2230945
[Abstract]:Barley (Hordeum vulgare L.) As one of the most important cereal crops, genetic superiority and economic importance are studied as model crops of Gramineae. In recent years, with the development of beer industry, the importance of malting barley has become more and more obvious. In the course of development and production of malting barley, the outbreak of barley stripe disease is more serious and increases year by year, which has a serious effect on the quality and yield of beer barley. Because germplasm resources of barley resistant to stripe disease are scarce in China, it is urgent to develop new resistant varieties. In this study, 100 barley parents were used to identify the resistance, to select the immune varieties and to cross with the highly susceptible varieties, to use BSA analysis to localize the gene against stripe disease, and to develop a new SSR marker in the located 7H short arm at the later stage. The polymorphism of these markers was analyzed. In this study, the new gene of resistance to barley stripe disease was named Rdg3, which is the first gene of barley resistance to stripe disease located in China, and the third gene of resistance to barley stripe disease in the world. It is important for breeding new barley varieties resistant to stripe disease, reducing pesticide use and reducing environmental pollution. The main results were summarized as follows: 1 strain of QWC was used to inoculate 100 barley materials. Through identification of resistance, 1 strain of immune material was screened out, and 22 materials of 10 high resistant materials showed resistance to disease, 37 and 30 of susceptible materials and highly susceptible materials, respectively. Gan Beer 2 was the immunized variety, and the highly susceptible variety Alexis was the most serious, the incidence rate was 88.24.2. The positive and negative cross was carried out by using the immunized variety Gan Beer 2 and the highly susceptible variety Alexis at the same time. The incidence of F _ 1 resistance identification in F _ 2 population was 0, 197 resistant plants and 79 susceptible plants were collected from 276 F _ 2 population, and the ratio of resistance to disease was in accordance with the 3:1 isolation ratio by chi-square test. It was determined that the resistance gene was dominant to the susceptible disease, and the resistance gene was controlled by single gene. 525pairs of primers from chromosome 1-7H of barley were used to select polymorphism between the two parents. A total of 103 pairs of polymorphic primers (including 11 pairs of 7H polymorphic primers) were screened, and the marker Bmag206,Bmag7 and scssr07970 located on the 7H short arm were identified by BSA analysis to be linked to the target gene Rdg3, so the gene resistant to stripe disease was located on the chromosome 7H short arm. 42 pairs of SSR markers were developed in 7H short arm, of which 6 pairs were polymorphic in Gan Beer 2 and Alexis. Using the 6 pairs of primers and 11 pairs of SSR markers with 7H polymorphism, the genetic linkage map was drawn, and the target gene was found to be located between Bmag206 and Bmag7. The genetic distance between the two markers was 1.78cM and 2.86cM.4. 48 barley parents were selected, and 42 pairs of SSR primers were used to carry out polymorphic scanning. The alleles of 29 pairs of polymorphic primers were 125, ranging from 2 to 9, with an average of 4.3 markers per marker. The highest allelic variation was detected by marker Bmg24 and Bmg25, and the frequency of allelic variation was 0.2979-0.9130, while that of marker Bmg1 was the lowest, ranging from 0.1462-0.8234 to 0.2979.29, with an average of 0.5132. The PIC value of marked Bmg25 was the highest (0.8234), and the value of marked Bmg43 was lowest (0.1462). The average diversity and heterozygosity were 0.5596 and 2.98 respectively.
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S435.123
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