分子標(biāo)記輔助選擇Pi9與Pigm基因培育抗稻瘟病秈稻新品系
發(fā)布時(shí)間：2018-11-09 18:00
【摘要】：水稻是全球半數(shù)以上人口的主要口糧,特別是在一些東南亞國(guó)家。稻瘟病是水稻生產(chǎn)中最嚴(yán)重的病害之一,普遍分布于各個(gè)水稻產(chǎn)區(qū),每年因此造成大量糧食損失。本論文研究分別以含稻瘟病廣譜抗性基因Pi9的水稻品系75-1-127和含稻瘟病廣譜抗性基因Pigm的水稻品種谷梅4號(hào)作供體親本,以35份秈稻品種或品系作為受體親本及輪回親本,設(shè)計(jì)并利用Pi9基因內(nèi)功能標(biāo)記和Pigm基因緊密連鎖標(biāo)記開展分子標(biāo)記輔助選擇(Molecular Marker-assisted Selection, MAS)育種實(shí)踐,定向改良受體親本的稻瘟病抗性,獲得了如下研究結(jié)果：1.根據(jù)已克隆的廣譜持久抗瘟基因Pi9的基因序列信息設(shè)計(jì)基因內(nèi)特異功能標(biāo)記Clon2-1,該標(biāo)記為共顯性DNA標(biāo)記,用它在35份水稻受體親本和Pi9基因供體親本75-1-127進(jìn)行基因型分析,結(jié)果表明在雙親間多態(tài)率為97.1%。2.根據(jù)廣譜持久抗瘟基因Pigm的基因精細(xì)定位信息設(shè)計(jì)了緊密連鎖DNA標(biāo)記T9E3,基因型分析結(jié)果表明該標(biāo)記為共顯性標(biāo)記,在供體親本谷梅4號(hào)與35份受體親本間的多態(tài)率為100%。3.田間病圃抗性鑒定表明,除Pi9基因供體75-1-127與Pigm基因供體谷梅4號(hào)外,35份秈稻受體親本的稻瘟病抗性均不強(qiáng)、抗譜不廣,都表現(xiàn)不同程度的感病；室內(nèi)接種抗性鑒定與抗菌譜分析結(jié)果表明,75-1-127與谷梅4號(hào)對(duì)17份來自不同國(guó)家或地區(qū)的稻瘟菌小種(菌株)的抗性頻率分別達(dá)到88.2%與94.1%,供試的25份受體親本的抗性頻率在16.7%-82.3%之間,大部分受體親本的稻瘟病抗性急需改良。4.對(duì)回交群體中隨機(jī)取樣個(gè)體的基因型分析和田間病圃對(duì)應(yīng)表型鑒定結(jié)果,證明分子標(biāo)記Clon2-1和T9E3對(duì)稻瘟病抗性表型選擇效率均可達(dá)到100%,說明可以用這兩個(gè)標(biāo)記有效開展MAS育種實(shí)踐。5.通過分子標(biāo)記輔助選擇和長(zhǎng)沙三亞兩地連續(xù)回交育種,定向改良35份秈稻受體親本的稻瘟病抗性,獲得了11個(gè)受體親本的48份含Pi9基因(位點(diǎn))的高抗稻瘟病水稻新品系,并用其中4個(gè)抗病純系配制了4個(gè)雜交種,同時(shí)獲得了其他24份受體親本的不同世代回交群體；獲得了10個(gè)受體親本的含pigm基因(位點(diǎn))的BC6F1高代回交群體,以及25個(gè)受體親木的不同世代回交群體。這些工作為進(jìn)一步培育廣譜高抗稻瘟病水稻新品種(組合)奠定了基礎(chǔ)。
[Abstract]:Rice is the staple food for more than half of the world's population, especially in some Southeast Asian countries. Rice blast is one of the most serious diseases in rice production. In this paper, rice lines 75-1-127 containing broad-spectrum resistance gene Pi9 and rice variety Gumei 4 containing Pigm gene were used as donor parents, respectively. 35 indica rice varieties or lines were used as recipient parents and recurrent parents to design and utilize Pi9 gene internal function marker and Pigm gene close linkage marker to carry out molecular marker-assisted selection (Molecular Marker-assisted Selection, MAS) breeding practice). The rice blast resistance of directed modified receptor parents was studied as follows: 1. According to the gene sequence information of the cloned broad-spectrum persistent blast resistance gene Pi9, a special gene marker Clon2-1, was designed. The marker was codominant DNA marker. The genotypes of 35 rice recipient parents and 75-1-127 donor parents of Pi9 gene were analyzed. The results showed that the polymorphism rate between parents was 97.1.2. A tightly linked DNA marker T9E3 was designed based on the gene fine mapping information of broad-spectrum persistent blast resistance gene Pigm. The results of genotype analysis showed that the marker was codominant. The polymorphic rate between donor parent Gu Mei 4 and 35 recipient parents was 100. 3. The identification of field disease resistance showed that, except for Pi9 gene donor 75-1-127 and Pigm gene donor Gamei 4, 35 indica rice receptor parents had low resistance to rice blast, and their resistance spectrum was not wide, and all of them were susceptible to different degrees of disease. The results of indoor inoculation resistance identification and antimicrobial spectrum analysis showed that the resistance frequency of 75-1-127 and Gumei No. 4 to 17 strains (strains) of Magnaporthe grisea from different countries or regions were 88.2% and 94.1%, respectively. The resistance frequency of 25 recipient parents ranged from 16.7- 82.3%, and the resistance of most recipient parents to rice blast was in urgent need of improvement. 4. Genotypic analysis of randomly sampled individuals in backcross population and identification of phenotypic characteristics in field disease beds showed that the selection efficiency of molecular markers Clon2-1 and T9E3 for blast resistance could reach 100%. These two markers can be used to develop MAS breeding practice effectively. 5. 5. By means of molecular marker-assisted selection and continuous backcross breeding in Sanya, Changsha, 35 indica rice receptor parents were directed to improve their rice blast resistance. 48 new rice lines with high resistance to rice blast with Pi9 gene (locus) were obtained from 11 recipient parents. Four hybrids were prepared with 4 resistant pure lines, and the backcross populations of other 24 recipient parents in different generations were obtained. The high generation backcross populations of BC6F1 containing pigm gene (loci) of 10 recipient parents and 25 backcross populations of different generations of recipient parents were obtained. These work have laid a foundation for further breeding new rice varieties (combinations) with wide spectrum and high blast resistance.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S435.111.41
[Abstract]:Rice is the staple food for more than half of the world's population, especially in some Southeast Asian countries. Rice blast is one of the most serious diseases in rice production. In this paper, rice lines 75-1-127 containing broad-spectrum resistance gene Pi9 and rice variety Gumei 4 containing Pigm gene were used as donor parents, respectively. 35 indica rice varieties or lines were used as recipient parents and recurrent parents to design and utilize Pi9 gene internal function marker and Pigm gene close linkage marker to carry out molecular marker-assisted selection (Molecular Marker-assisted Selection, MAS) breeding practice). The rice blast resistance of directed modified receptor parents was studied as follows: 1. According to the gene sequence information of the cloned broad-spectrum persistent blast resistance gene Pi9, a special gene marker Clon2-1, was designed. The marker was codominant DNA marker. The genotypes of 35 rice recipient parents and 75-1-127 donor parents of Pi9 gene were analyzed. The results showed that the polymorphism rate between parents was 97.1.2. A tightly linked DNA marker T9E3 was designed based on the gene fine mapping information of broad-spectrum persistent blast resistance gene Pigm. The results of genotype analysis showed that the marker was codominant. The polymorphic rate between donor parent Gu Mei 4 and 35 recipient parents was 100. 3. The identification of field disease resistance showed that, except for Pi9 gene donor 75-1-127 and Pigm gene donor Gamei 4, 35 indica rice receptor parents had low resistance to rice blast, and their resistance spectrum was not wide, and all of them were susceptible to different degrees of disease. The results of indoor inoculation resistance identification and antimicrobial spectrum analysis showed that the resistance frequency of 75-1-127 and Gumei No. 4 to 17 strains (strains) of Magnaporthe grisea from different countries or regions were 88.2% and 94.1%, respectively. The resistance frequency of 25 recipient parents ranged from 16.7- 82.3%, and the resistance of most recipient parents to rice blast was in urgent need of improvement. 4. Genotypic analysis of randomly sampled individuals in backcross population and identification of phenotypic characteristics in field disease beds showed that the selection efficiency of molecular markers Clon2-1 and T9E3 for blast resistance could reach 100%. These two markers can be used to develop MAS breeding practice effectively. 5. 5. By means of molecular marker-assisted selection and continuous backcross breeding in Sanya, Changsha, 35 indica rice receptor parents were directed to improve their rice blast resistance. 48 new rice lines with high resistance to rice blast with Pi9 gene (locus) were obtained from 11 recipient parents. Four hybrids were prepared with 4 resistant pure lines, and the backcross populations of other 24 recipient parents in different generations were obtained. The high generation backcross populations of BC6F1 containing pigm gene (loci) of 10 recipient parents and 25 backcross populations of different generations of recipient parents were obtained. These work have laid a foundation for further breeding new rice varieties (combinations) with wide spectrum and high blast resistance.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S435.111.41
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